Leukotrienes Affect Secretory Function of Ovarian Cells In Vitro*

The aim of this study was to determine which cells are the source of production and target for leukotriene (LTs) action within the bovine ovary. Luteal (CL, days 14–16 of the oestrous cycle), steroidogenic cells (LSC) and endothelial cells (LEC) of the bovine corpus luteum (CL), and granulosa cells (GC) were isolated enzymatically, cultured in a monolayer and incubated with LTC₄, LTB₄, Azelastine (an antagonist of LTC₄) or Dapsone (an antagonist of LTB₄). Then cells were collected for determination of mRNA expression for LT receptors (LTRs) and 5‐lipoxygenase (5‐LO) by real time RT‐PCR, and media were collected for determination of prostaglandin (PG)E₂, F_2α, progesterone (P4; LSC only), endothelin‐1 (ET‐1; LEC only) and 17‐β oestradiol (E2; GC only). The greatest mRNA expression for LTR‐II and 5‐LO were found in LEC, whereas LTR‐I mRNA expression did not differ among cell types. The level of PGE₂ increased after LTs treatment in each type of ovarian cell, excluding LTC₄ treatment in LEC. The secretion of PGF_2α was also increased by LTs, but decreased after LTB₄ treatment of LSC. In GC cultures, both LTs stimulated E2 secretion; in LEC cultures, LTB₄ stimulated whereas LTC₄ inhibited P4 secretion; in LEC cultures, LTC₄ stimulated but LTB₄ inhibited ET‐1 secretion. The results show that LTs are produced locally and are involved in PGs production/secretion in all examined cells (LSC, LEC and GC) of bovine ovary. Leukotriene treatment modulate secretion of E2, by GC, P4 by LSC and ET‐1 by LEC, which indicates that LTs are involved in regulation of ovarian secretory functions.     

Expression of the Vascular Endothelial Growth Factor (VEGF‐A) and its Receptors in the Umbilical Cord in the Course of Pregnancy in the Pig

The umbilical cord (UC) and the placenta are important organs through which respiratory gases, nutrients, wastes and biologically active substances are exchanged between the maternal and the foetal system. A rapid placental vascularization observed in the second half of pig pregnancy is positively correlated with the mRNA expression of the vascular endothelial growth factor (VEGF). Based on these findings, we hypothesized that VEGF may have a stimulatory effect in the dynamically growing UC. To further understand the role of the VEGF–VEGFR system during UC development, mRNA and protein expression as well as the cellular localization of VEGF‐A, VEGFR‐1 and VEGFR‐2 in UC were examined on days 40, 60, 75 and 90 of pregnancy and after physiological delivery in the pig (day 114 of pregnancy). Real Time RT‐PCR analysis showed an increase in the mRNA levels of VEGF120 and VEGF164 from day 90 of pregnancy. VEGFR‐1 mRNA expression was significantly increased on day 75 of pregnancy. No significant changes in VEGFR‐2 mRNA expression were detected. In turn, western blot analysis revealed an increase in VEGF‐A protein expression on day 40, compared to the later days of pregnancy. A rapid increase in the VEGFR‐1 protein level was noted on day 75 and 90 of gestation. No significant changes in VEGFR‐2 protein expression were detected on any of the analysed days of pregnancy. Immunohistochemical staining enabled detection of VEGF–VEGFR system, in endothelial and tunica media cells of the umbilical vessels and in allantoic duct and amniotic epithelium on all analysed days of pregnancy. Positive reactions for VEGF‐A and VEGFR‐1, but not VEGFR‐2, were also observed in myofibroblasts. In conclusion, this data shows that members of the VEGF–VEGFR system are temporally and spatially well localized for playing key roles during umbilical cord formation and its intensive growth observed after day 75 of pregnancy.     

Response of Thawed Epidi dymal Red Deer Spermatozoa to Increasing Concentrations of Hydrogen Peroxide,and Importance of Individual Male Variability

Oxidative stress represents a challenge during sperm manipulation. We have tested the effect of increasing hydrogen peroxide (H₂O₂) levels on red deer spermatozoa after cryopreservation, and the role of male‐to‐male variation in that response. In a first experiment, eight thawed samples were submitted to 0, 25, 50, 100 and 200 μm H₂O₂ for 2 h at 37°C. Intracellular reactive oxygen species (H₂DCFDA‐CM) increased with H₂O₂ concentration, but we only detected a decrease in sperm function (motility by CASA and chromatin damage by sperm chromatin structure assay) with 200 μm . Lipoperoxidation assessed by the thiobarbituric acid reactive substance (TBARS) method increased slightly with 50 μm H₂O₂ and above. In a second experiment, samples from seven males were submitted to 0 and 200 μm H₂O₂ for 2 h, triplicating the experiment within each male. Males differed at thawing and regarding their response to incubation and H₂O₂ presence. We found that the kinematic parameters reflected male‐to‐male variability, whereas the response of the different males was similar for lipid peroxidation and viability. A multiparametric analysis showed that males grouped differently if samples were assessed after thawing, after incubation without H₂O₂ or after incubation with H₂O₂. Red deer spermatozoa are relatively resilient to H₂O₂ after thawing, but it seems to be a great male‐to‐male variability regarding the response to oxidative stress. The acknowledgement of this individual variability might improve the development of optimized sperm work protocols.     

Use of rotational periosteal flaps in primary repair of sinofacial fractures in seven horses

ABSTRACT

Case history: Medical records were reviewed of horses (n?=?7) undergoing surgery for fracture of one or more facial bones extending into the paranasal sinuses that was repaired primarily within 24 hours of the time of injury using a rotational periosteal flap, between April 2009 and May 2017. A kick from another horse was the cause of the injury of three horses, and one horse was injured when it collided with a tree. The cause of the injury of three horses was unknown.

Clinical findings and treatment: Fractures were of the right maxillary bone in two horses, the left maxillary bone in two horses, the left frontal and left nasal bones in two horses, and the right frontal bones in one horse. The fracture of all but one horse was accompanied by an open wound. The fracture of all seven horses was reduced, stabilised, and covered with a rotational, periosteal flap. Surgery was carried out while standing in six horses, and while anesthetised in one horse. All horses had a deficit in the fractured facial bones after the fracture was reduced. Four horses had complications following surgery, but all horses were reported to have excellent cosmetic outcomes and had retuned to their previous level of activity, as reported by their owners.

Clinical relevance: Covering a primarily repaired sinofacial fracture of a horse with a rotational periosteal flap resulted in good cosmetic outcomes, and may be especially beneficial if the fracture is accompanied by loss of bone.     

Arbuscular Mycorrhizal Fungi Alleviate the Negative Effect of Temperature Stress in Millet Lines with Contrasting Soil Aggregation Potential

Arbuscular mycorrhizal fungi (AMF) establish a mutualistic symbiosis with several plants and play a key role in improving plant growth, tolerance to abiotic and biotic stresses as well as the soil structure. This work aimed at elucidating the AMF temperature stress modulating impact on four pearl millet lines plant growth and soil aggregation. Experimental trials were carried out in both greenhouse and growth chamber to determine the response of the four millet lines to inoculation with two AMF strains (Rhizophagus aggregatus and Funneliformis mosseae) under heat and non-stress conditions. We first investigated the mycorrhizal colonization (MC) and the mycorrhizal growth response (MGR) of millet lines in relation with their soil aggregation potential (root adhering soil/root biomass, MAS/RB) in the greenhouse. Secondly, the four millet lines were grown in two separated growth chambers and subjected to a day/night temperature of 32/28?°C as the control treatment and 37/32?°C as the temperature stress treatment. Plant growth, mycorrhization rate and several physiological, mycorrhizal and soil parameters were measured. Results showed that the mycorrhization rates of millet lines were low and not significantly different. Funneliformis mosseae (31.39%) showed higher root colonization than Rhizophagus aggregatus (22.79%) and control (9.79%). The temperature stress reduced the mycorrhizal colonization rate, shoot and root biomass, and the soil aggregation for all tested lines. L220 and L132 showed more MC rate and MGR than the other lines under control and high-temperature treatment. The MGR was significantly better under temperature stress conditions than in the control. Under the temperature stress conditions, inoculation with R. aggregatus and F. mosseae increased chlorophyll concentration, root dry weight and shoot dry weight as compared to non-inoculated plants. AMF inoculation, particularly with F. mosseae had a positive influence on the tolerance of millet lines to temperature stress. This study demonstrates that AMF play an important role in the response of these four millet lines to temperature stress. AMF is therefore an important component in the adaptation of crops to climatic variations in Sub-Saharan Africa.

     

Ontogenetic movements of juvenile blacktip reef sharks: evidence of dispersal and connectivity between coastal habitats and coral reefs

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