Expression of cyclooxygenase-1 and -2 in naturally occurring squamous cell carcinomas in horses

OBJECTIVE: To assess expression of cyclooxygenase (COX)-1 and -2 in naturally occurring squamous cell carcinomas (SCCs) and the analogous normal tissues in horses. SAMPLE POPULATION: Tissue samples collected from 3 conjunctival, 2 vulvar, 4 preputial, and 5 penile SCCs during surgical excision in 14 horses and from corresponding body regions (conjunctiva [n = 5 horses], vulva [2], prepuce [3], and penis [3]) in 5 horses euthanized for reasons unrelated to neoplasia. PROCEDURES: Tissue samples were snap frozen in liquid nitrogen and stored at -80 degrees C until analysis. Protein was extracted from the frozen tissues, and western blot analyses were performed. Nonneoplastic and abnormal tissues from each body region were run on the same blot, and blots were run in triplicate. Molecular-weight markers and COX-1 and 2 ovine standards (positive control samples) were run concurrently on the gels; negative control samples were not used. RESULTS: All tissues, including the nonneoplastic and SCC tissues, expressed both COX-1 and -2 proteins. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that the expression of COX proteins in both nonneoplastic and SCC-affected tissues in horses is markedly different from that in other species. The reason for the potential benefit of COX-2 inhibitors in horses and other species is unknown. Further research needs to be performed to evaluate the efficacy of COX-2 inhibitors as cancer treatments in horses. Investigation of the mechanisms of tumor development in horses should be performed to increase understanding of this disease and ascertain how the mechanisms differ from those in other animals.     

Identification of infrared absorption spectral characteristics of synovial fluid of horses with osteochondrosis of the tarsocrural joint

OBJECTIVE: To determine the feasibility of the use of Fourier-transform infrared (FTIR) spectroscopy within the midinfrared range to differentiate synovial fluid samples of joints with osteochondrosis from those of control samples. ANIMALS: 33 horses with osteochondrosis of the tarsocrural joint and 31 horses free of tarsocrural joint disease. PROCEDURES: FTIR spectroscopy of synovial fluid was used. Sixty-four synovial fluid samples from the tarsocrural joint were collected. Of these, 33 samples were from horses with radiographic evidence of osteochondrosis of the tarsocrural joint and 31 from control joints. Disease-associated features within infrared spectra of synovial fluid were statistically selected for spectral classification, and the variables identified were used in a classification model. Linear discriminant analysis and leave-one-out cross-validation were used to develop a classifier to identify joints with osteochondrosis. RESULTS: 12 significant subregions were identified that met the selection criteria. The stepwise discriminant procedure resulted in the final selection of 6 optimal regions that most contributed to the discriminatory power of the classification algorithm. Infrared spectra derived from synovial fluid of joints with osteochondrosis were differentiated from the control samples with accuracy of 77% (81% specificity and 73% sensitivity). CONCLUSIONS AND CLINICAL RELEVANCE: The disease-associated characteristics of infrared spectra of synovial fluid from joints with osteochondrosis may be exploited via appropriate feature selection and classification algorithms to differentiate joints with osteochondrosis from those of control joints. Further study with larger sample size including age-, breed-, and sex-matched control horses would further validate the clinical value of infrared spectroscopy for the diagnosis of osteochondrosis in horses.     

Effects of the cyclooxygenase inhibitor meloxicam on recovery of ischemia-injured equine jejunum

OBJECTIVE: To determine the effect of meloxicam and flunixin meglumine on recovery of ischemia-injured equine jejunum. ANIMALS: 18 horses. PROCEDURES: Horses received butorphanol tartrate; were treated IV with saline (0.9% NaCl) solution (SS; 12 mL; n = 6), flunixin meglumine (1.1 mg/kg; 6), or meloxicam (0.6 mg/kg; 6) 1 hour before ischemia was induced for 2 hours in a portion of jejunum; and were allowed to recover for 18 hours. Flunixin and SS treatments were repeated after 12 hours; all 3 treatments were administered immediately prior to euthanasia. Selected clinical variables, postoperative pain scores, and meloxicam pharmacokinetic data were evaluated. After euthanasia, assessment of epithelial barrier function, histologic evaluation, and western blot analysis of ischemia-injured and control jejunal mucosa samples from the 3 groups were performed. RESULTS: Meloxicam- or flunixin-treated horses had improved postoperative pain scores and clinical variables, compared with SS-treated horses. Recovery of transepithelial barrier function in ischemia-injured jejunum was inhibited by flunixin but permitted similarly by meloxicam and SS treatments. Eighteen hours after cessation of ischemia, numbers of neutrophils in ischemia-injured tissue were higher in horses treated with meloxicam or flunixin than SS. Plasma meloxicam concentrations were similar to those reported previously, but clearance was slower. Changes in expression of proteins associated with inflammatory responses to ischemic injury and with different drug treatments occurred, suggesting cyclooxygenase-independent effects. CONCLUSIONS AND CLINICAL RELEVANCE: Although further assessment is needed, these data have suggested that IV administration of meloxicam may be a useful alternative to flunixin meglumine for postoperative treatment of horses with colic.     

Adrenomedullin (AM) and adrenomedullin binding protein (AM-BP) in the bovine mammary gland and milk: Effects of stage of lactation and experimental intramammary E. coli infection

Adrenomedullin (AM) has been characterized as an endogenous tissue survival factor and modulator of many inflammatory processes. Because of the increased susceptibility of the mammary gland to infection during the time surrounding parturition in the cow, we investigated how milk and tissue content of AM and its binding protein (AM-BP) might be affected by the stage of lactation and the udder health status. Milk and mammary biopsy samples were obtained from Holstein cows 21 days prior to and at various times after calving to represent the dry period and early and mid-stages of lactation. Additional cows received an intramammary challenge with Escherichia coli for immunohistochemical characterization of AM and AM-BP. Milk AM concentrations were relatively constant across the stages of lactation while AM-BP increased two-fold (P<0.04) between early and mid-lactation. Milk AM (P<0.04) and AM-BP (P<0.03) increased as somatic cell counts (SCCs) increased within a given stage of lactation. Tissue content of both (AM and AM-BP) were significantly affected by stage of lactation, lowest in the dry period and progressively increasing to peak at mid-lactation as well as increasing in association with higher levels of SCCs. Following E. coli challenge, AM increased in epithelial cells surrounding mammary alveoli presenting high levels of SCCs. The data suggest that AM and AM-BP are cooperatively regulated in the mammary gland during lactation; changes in localized tissue AM and AM-BP content reflect a dynamic regulation of these tissue factors in the bovine mammary gland consistent with their protective effects within inflamed tissue.     

Cyclooxygenase (COX) inhibitors and the intestine

Nonsteroidal anti-inflammatory drugs (NSAIDs) have long been used for the treatment of pain and inflammation because of their inhibitory effects on cyclooxygenase (COX). For almost as long as NSAIDs have been in use, multiple adverse effects have been noted. Assessment of many of these adverse effects have been complicated because of the discovery of multiple splice variants of the cox gene, and a greater array of COX inhibitors, especially the COX-2 selective inhibitors have become available. Some of these adverse effects cannot be readily explained by the effect of these drugs on COX. This has sparked a new field of investigation into the COX-independent effects of the COX inhibitors. The major noncyclooxygenase targets of the COX inhibitors of particular relevance to inflammation and the gastrointestinal tract are phosphatidylinositol 3'-kinase Akt signaling, uncoupling of oxidative phosphorylation, PPARgamma, nuclear factor KB, mitogen activated protein kinases, and heat shock proteins.     

Comparison between the penetration characteristics of methyl bromide and ethanedinitrile through the bark of pine (Pinus radiata D.Don) logs

BACKGROUND

Ethanedinitrile (EDN) is a fumigant being commercialized worldwide as an alternative phytosanitary treatment to methyl bromide (MB) for forest products. The penetration characteristics of MB and EDN were measured through the bark of wooden blocks (100 × 100 × 50 mm) cut from the upper (average bark thickness 5 ± 2 mm) and lower (average bark thickness 25 ± 5 mm) trunk of recently felled pine (Pinus radiata D.Don) trees. Doses of 48 g m⁻³ MB and 50 g m⁻³ EDN were applied to chambers at 10 and 20°C for 10 h.

RESULTS

Penetration of MB was influenced by the interaction between fumigation time and temperature, with concentrations increasing at a higher rate at 20°C compared with at 10°C. After 10 h, an average concentration of 8.05 ± 0.89 g m⁻³ had penetrated the bark of log sections at 20°C, whereas 5.20 ± 0.89 g m⁻³ was measured at 10°C. By contrast, the factors examined in this study did not significantly impact the penetration of EDN. Concentration × time (CT) values for MB under the bark were 35.20 ± 2.30 g h m⁻³ at 10°C and 55.85 ± 9.58 g h m⁻³ at 20°C; whereas for EDN, CT values were 19.50 ± 6.80 g h m⁻³ at 10°C and 19.08 ± 4.10 g h m⁻³ at 20°C.

CONCLUSION

MB can achieve a higher concentration under the bark of log sections during simulated fumigations, but all of the factors examined affected the ability of MB to penetrate the bark of wooden blocks. By comparison, the penetration of EDN through the bark is more consistent than MB under laboratory conditions. © 2022 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.     

Pilot study of the ultrasonographic examination of the intact and transected medial glenohumeral ligament in dogs

Medial glenohumeral ligament injury is commonly reported during medial shoulder joint instability in dogs. Arthroscopy is considered the gold standard procedure, but it is invasive and requires distension of the joint. Ultrasonographic examination of the medial glenohumeral ligament has been studied as a possible, less invasive alternative to arthroscopy however it has not been considered a useful method of assessment due to the interference of the probe with the pectoral muscles. The aims of this prospective analytical randomized pilot study were to develop a standardized ultrasound protocol for visualizing the canine medial glenohumeral ligament and to compare goniometry and ultrasound findings in cadaver dogs with versus without transection of the medial glenohumeral ligament. Nine adult Beagle cadavers (18 shoulders) were used. The first six shoulders were used in a preliminary study to describe an ultrasound technique to identify the medial glenohumeral ligament. Arthroscopy was performed on the remaining 12 shoulders, with six randomly selected medial glenohumeral ligaments from these shoulders, transected during the procedure. Ultrasound examination was performed after each arthroscopic procedure by an ultrasonographer blinded to the patient group. Four medial glenohumeral ligaments (67%) were correctly identified during the preliminary study. Ultrasonographic examination failed to diagnose the transection of all six medial glenohumeral ligaments in the second part of the study. No difference was observed in the ligament thickness between the dogs with and without a transected medial glenohumeral ligament. Dogs with a transected medial glenohumeral ligament had a wider articular space compared to dogs without a transected ligament (P < 0.001), and an articular space wider than 8.2 mm was discriminatory of a transected medial glenohumeral ligament in all the shoulders. In conclusion, the medial glenohumeral ligament could be identified with a medial ultrasonographic approach of the shoulder and a wider articular space can be a sign of a medial shoulder joint instability. Further studies are needed to confirm these preliminary findings in living dogs, with and without shoulder instability.     

Difference in factors explaining growth rate variability in European eel subpopulations: the possible role of habitat carrying capacity

As a key parameter in the management of fish populations, individual growth rate (GR) variations were examined in the European eel (Anguilla anguilla; > 150 mm) using extensive mark–recapture surveys in the lotic habitats of two small rivers of western France: the Frémur, supposed to be saturated, and at the same latitude, the Oir with densities fivefold lower than those of the Frémur. In both systems, generalised linear models were used to test whether spatiotemporal factors such as dominant habitat type or local density affect GR variability. In the presumed unsaturated system, the Oir, GR variability is mainly explained by a set of habitat suitability drivers (density, dominant habitat type). In the Frémur, GRs appear independent of differences in habitat density or productivity. Below saturation, an increase in density will decrease the GR through intraspecific competition. At saturation, intraspecific competition reaches such high levels that regardless of eel density and productivity, the resources available by individual are similar throughout the system. In these circumstances, the effect of density on growth was presumed undetectable. Despite these contrasted results, mean GRs observed in both catchments were closed (~20 mm·year^?1). This is an unexpected result as GR is expected to be higher in unsaturated systems. This similarity could be explained by the difference between the two systems in terms of: (i) sex ratio (the Frémur is dominated by male, whereas the female is dominant in the Oir), (ii) habitat type distribution or (iii) possible interspecific competition (important salmonid populations in the Oir).     

Interactions of insecticides with biological membranes

Many insecticides (e.g. DDT, lindane, pyrethroids) are hydrophobic molecules which bind extensively to biological membranes. Binding of insecticides to phospholipid bilayers frequently shows saturation, with binding increasing with increasing insecticide concentration up to some limit, beyond which no further insecticide is incorporated into the bilayer. For lindane, this limit has been shown to correspond to the concentration at which the aqueous phase becomes saturated. Simple alkanes show similar saturation phenomena, and binding of halogenated alkanes can be followed by fluorescence quenching methods. It is shown that effects of hydrophobic molecules on bilayer fluidity are small. Effects of alkanes and insecticides on the activity of the (Ca²⁺-Mg²⁺)-ATPase purified from sarcoplasmic reticulum follow from direct binding to the ATPase, at sites which are not at the lipid-protein interface of the ATPase.