Lipopolysaccharide and cytokines modulate leukotriene (LT)B4 and LTC4 production by porcine endometrial endothelial cells

Uterine inflammatory response is mediated by inflammatory mediators including eicosanoids and cytokines produced by immune and endometrial cells. Interactions between lipopolysaccharide (LPS) and cytokines, and leukotrienes (LTs) in endothelium, important for the host defence during the inflammation, are unknown. We studied the effect of LPS, tumour necrosis factor (TNF)‐α, interleukin (IL)‐1β, IL‐4 and IL‐10 on 5‐lipooxygenase (5‐LO), LTA₄ hydrolase (LTAH) and LTC₄ synthase (LTCS) mRNA and protein expression, LTB₄ and LTC₄ release from porcine endometrial endothelial cells, and cell viability. For 24 hr, cells were exposed to LPS (10 or 100 ng/ml of medium) and cytokines (each 1 or 10 ng/ml). 5‐LO mRNA/protein expression augmented after incubation with larger doses of LPS, TNF‐α, IL‐4 and IL‐10 and smaller dose of IL‐1β. Larger dose of TNF‐α, smaller doses of LPS and IL‐1β and both doses of IL‐10 increased LTAH mRNA/protein expression. LTAH protein content was up‐regulated by larger dose of LPS, but it was reduced in response to both doses of IL‐4. LTCS mRNA expression was elevated by larger doses of LPS, IL‐4 and IL‐10 or both doses of TNF‐α and IL‐1β. LTCS protein level increased after treatment with both doses of IL‐1β, IL‐4 and IL‐10, smaller dose of LPS and larger dose of TNF‐α. Both doses of LPS and larger doses of TNF‐α and IL‐10 increased LTB₄ release. LPS, IL‐1β and IL‐10 at smaller doses, or TNF‐α and IL‐4 at larger doses stimulated LTC₄ release. Smaller doses of TNF‐α and IL‐1β or both doses of IL‐4 enhanced the cell viability. This work provides new insight on the participation of LPS, TNF‐α, IL‐1β, IL‐4 and IL‐10 in LTB₄ and LTC₄ production/release from porcine endometrial endothelial cells, and the effect of above factors on these cells viability. The used cellular model gives the possibility to further establish the interactions between inflammatory mediators.     

Secondary sexual traits and semen characteristic of chicken germline chimeras

Birds obtained by embryo engineering are used to study embryo development and to produce transgenic birds. As this method of producing birds still generate strong emotions of the public opinion head ornaments, testes and semen characteristics of sex chimera roosters were examined to check whether they differ from chickens obtained by non‐manipulated methods. Measurements of head ornaments, testes and semen were correlated with each other. Semen quality factor (SQF) was calculated, as well as the level of fluctuating asymmetry (FA) of bilateral traits (wattles and testes). Positive correlation was found for comb width and wattle length and comb thickness and sperm concentration. Semen characteristics and FA did not exceed the level encounter in other chicken lines. Results obtained indicate that germline chimeras are similar in appearance of secondary sexual traits, and semen and testes characteristics to chickens produced in non‐manipulated way.     

Endocannabinoids as endometrial inflammatory markers in lactating Holstein cows

The objective of this study was to consider endocannabinoid system as inflammatory markers in bovine endometrium to better understand the role of this system in regulating many of the functions that are related to inflammatory condition. At day 26 post‐partum, fourteen cows were divided into two groups depending on the inflammatory condition: 1‐ subclinical endometritis (n = 7, with purulent or mucopurulent uterine discharge detectable in the vagina) and 2‐ healthy (n = 7, No (muco)) purulent discharge. Blood samples were collected at 26 and 30 days relative to calving to determine plasma tumour necrosis factor (TNF) and lipopolysaccharide‐binding protein (LBP) concentrations; moreover, uterine biopsy was carried out on day 26 post‐partum to measure mRNA abundance of TNF, interleukin‐1B (IL1B), interleukin‐6 (IL‐6), C‐X‐C motif chemokine ligand 8 (CXCL8), endocannabinoid receptor (CNR2), N‐acyl phosphatidylethanolamine phospholipase D (NAPEPLD), fatty acid amide hydrolase (FAAH), N‐acylethanolamine acid amidase (NAAA) and monoglyceride lipase (MGLL) by real‐time PCR. Results showed mean plasma concentrations of TNF and LBP were lower in healthy cows compared to subclinical endometritis cows (p < .05). Relative mRNA expression for NAAA and FAAH was decreased (p < .05), and relative mRNA expression for CNR2 and NAPEPLD increased in cows with subclinical endometritis compared to healthy cows. In conclusion, relative mRNA expression of TNF, IL1B and CXCL8 and plasma concentration of LBP increased during inflammatory condition along with decreased endocannabinoids hydrolyzing enzyme (NAAA and FAAH), increased enzymes that synthesize endocannabinoids (NAPEPLD) and relative gene expression of the endocannabinoid receptor; together, these contribute to increased endocannabinoids levels during inflammation. Overall, we provide evidence that endocannabinoid system is altered in endometrium tissue during inflammation through increased mRNA expression of CNR2 and synthesis enzyme and decreased mRNA expression of hydrolyzing enzymes interfere with pro‐cytokine production and signalling, which may interfere with the onset and progression of inflammation.     

Idiopathic eosinophilic colitis lesions of the equine small (descending) colon

Five horses with a primary surgical lesion of the small (descending) colon were diagnosed with eosinophilic colitis based on visual and histopathological examination. These were evident as visibly striking, hyperaemic, focal lesions of the small colon, with serosal petechiation, oedema and marked thickening of the intestinal wall at the site. Areas of focal necrosis were also evident. The gross appearance of the lesions were considered to be sufficiently severe in all cases to merit resection, due to concerns about intestinal necrosis and septic peritonitis. An inability to fully exteriorise the affected portion of intestine to perform a resection and anastomosis necessitated intraoperative euthanasia of one horse. A total of three horses survived to hospital discharge. Eosinophilic colitis lesions are a rare cause of severe small colon disease, but should be considered in cases with similar visual characteristics.     

Successful small intestinal resection and anastomosis in a late term broodmare with colic via a standing left flank laparotomy

A late term broodmare presented with low‐grade intestinal colic. Clinical findings were consistent with a small intestinal lesion requiring surgical intervention. The risks of general anaesthesia to the fetus, combined with clinical findings in an otherwise quiet natured horse influenced the decision to choose an alternative standing left flank approach first. A small intestinal resection and anastomosis was performed successfully and the mare delivered a healthy foal without complications a month later. Standing colic surgery might be a preferred alternative to general anaesthesia in certain circumstances as discussed in this case report.     

Accessory nictitating membrane in a Lipizzaner colt

A 2‐year‐old Lipizzaner colt presented for removal of a mass over the left eye. The colt had no blepharospasm or significant ocular discharge, but the cornea underlying the mass was mildly oedematous. The mass protruded from the conjunctiva at the dorsotemporal aspect of the globe and was covered in normal conjunctiva. It had a gross appearance similar to the cartilaginous flap of a nictitating membrane, but in an aberrant location. It was moveable to cover the dorsal aspect of the cornea when the globe was retropulsed. Excision of the mass under general anaesthesia was elected. Histopathology supported a diagnosis of an accessory nictitating membrane, consisting of a cartilaginous band surrounded by glandular epithelium. The colt recovered from surgery without complication and no problems were reported by the owner at 8.5 months post‐operatively. To the author's knowledge, there have not been previous reports of accessory or ectopic nictitating membranes in equine or other species.     

Intra‐abdominal cystic lymphangiomatosis in a Thoroughbred foal

Cystic lymphangiomas are rare malformations of the lymphatics that result in the formation of a cystic mass. Where multiple cysts are seen, the condition is termed cystic lymphangiomatosis. This case describes the diagnosis and unique management of cystic lymphangiomatosis in a 10‐day‐old Thoroughbred foal. Ultrasonography, histopathology and laparoscopy were essential for diagnosis and appreciation of the extent of disease. Ultimately, the cystic lymphangiomatosis was so extensive in this foal that complete surgical excision was considered impossible and the presence of adhesions within the abdomen indicated a very poor long‐term prognosis; the owners elected for euthanasia at age 14 weeks. Although rare, lymphangioma and lymphangiomatosis should be considered as a differential diagnosis for an intra‐abdominal mass in a young horse.