Effect of constant infusion of oxytocin on luteal lifespan and oxytocin-induced release of prostaglandin F2α in heifers

Two experiments were conducted to determine whether constant infusion of oxytocin would prolong the luteal phase and inhibit uterine prostaglandin F2 alpha (PGF2 alpha) secretion in heifers. In Experiment 1, twelve heifers, treated with saline (SAL) or oxytocin (OXY) via jugular cannulae infusions (INF) or osmotic minipumps (OMP), were allotted at estrus into four treatment groups (n = 3). Treatments were: SAL-INF, SAL-OMP, OXY-INF and OXY-OMP. Physiological saline or oxytocin was given from Days 10 to 23 (Day 0 = estrus) of the estrous cycle. Method of treatment (jugular cannula infusion or osmotic minipump) had no effect (P greater than 0.05) on estrous cycle length or pattern of secretion of progesterone; therefore, data were pooled. Estrous cycle lengths were extended (P less than 0.01) for heifers which received oxytocin (25.3 +/- 0.4 d) compared to saline (20.5 +/- 0.4 d). Luteolysis did not occur in oxytocin-treated heifers until after treatment ceased. Experiment 2 was designed and conducted identically to Experiment 1 with the addition of a "challenge" injection of oxytocin (100 IU oxytocin, i.v.) given on Day 16 of the estrous cycle. Treatment of heifers with oxytocin extended (P less than 0.05) estrous cycle length by an average of 3 d compared to heifers treated with saline. The "challenge" injection induced (P less than 0.05) secretion of PGF2 alpha (as measured by the stable PGF2 alpha metabolite, 15-keto-13,14-dihydro-PGF2 alpha) in saline-treated but not oxytocin-treated heifers. In both Experiment 1 and 2, serum concentrations of FSH were elevated (P less than 0.05) in oxytocin-treated heifers. No increase was observed for LH or prolactin. The rise in estradiol-17 beta at luteolysis was not affected (P greater than 0.10) by treatment. In summary, constant infusion of oxytocin extended luteal lifespan, prolonged secretion of progesterone, and inhibited oxytocin-induced secretion of PGF2 alpha. Constant infusion of oxytocin did not affect serum concentrations of estradiol-17 beta, LH or prolactin; however, serum concentrations of FSH were elevated during the oxytocin treatment period.     

Changes in the C-reactive protein and 13,14-dihydro-15-keto-prostaglandin F2α concentrations of uterine lavage samples after the administration of povidone-iodine in cows

Changes in the C-reactive protein (CRP) and 13,14-dihydro-15-keto-prostaglandin F_2α (PGFM) concentrations of uterine lavage fluid were examined in cows given an intrauterine povidone-iodine (PI) infusion. The mean polymorphonuclear leukocyte (PMN) ratios (the ratio of PMN to total cells) and CRP concentration of uterine lavage fluid on the day after the treatment were significantly (P<0.05) greater in the PI infusion group (PMN: 53.0 ± 32.7%, CRP: 50.2 ± 32.3 ng/mL) than in the non-treatment control group (PMN: 7.9 ± 21.9%, CRP: 17.2 ± 5.9 ng/mL), whereas there was no significant difference in the mean PGFM concentration between the two groups. The present findings suggest that the uterine CRP level is a useful biomarker of local uterine inflammation in cows.     

Plasma concentrations of 13,14-dihydro-15-ketoprostaglandin F2 alpha in mares during uterine involution.

Twelve mares were allowed to foal naturally, after which they were monitored to study uterine involution. Starting on day 3 after parturition, the internal genital tract was examined per rectum manually and ultrasonographically every other day for changes in uterine characteristics and ovarian activity. By day 5, gravid and nongravid uterine horns were similar in size, and by day 7, uterine fluid was absent. On day 7 after parturition, endometrial biopsy samples were obtained for histologic evaluation, and uterine swab specimens were obtained for microbiologic culture. Uterine swab specimens from 10 of 12 mares had slight bacterial growth. The uteri of 8 of the 12 mares were histologically involuted by day 7. All mares ovulated 7 to 12 days after parturition. Concentrations of 13,14-dihydro-15-keto-PGF2 alpha (PGFM) were measured in jugular plasma samples obtained daily for 21 days after parturition. Concentrations of PGFM were low by the day after parturition, and there was no significant correlation between uterine involution and PGFM concentrations in these mares. All 12 mares were bred at the first estrus after parturition, and 9 became pregnant.     

Retention of a functional corpus luteum and peripheral concentrations of 13,14-dihydro-15-keto-prostaglandin F2alpha following metestrus administration of Syncro-Mate-B.

This study was conducted to examine the effects of metestrus administration of SyncroMate-B (SMB) on PGF2alpha secretion and corpus luteum (CL) development. In a study replicated over 2 yr, cows were observed for spontaneous estrus in yr 1, and cows received an injection of 25 mg of PGF2alpha and were observed for subsequent estrus in yr 2. At standing estrus (estrus = d 1), cows were randomly allotted to receive either the standard SMB regimen (n = 40) on d 3 of the estrous cycle or no treatment (n = 8). Fifty percent (n = 20) of SMB-treated cows were administered PGF2alpha on d 10 of the estrous cycle 48 h prior to implant removal. Twice-daily blood samples were collected in the morning (AM) and evening (PM) from d 2 AM through d 14 AM of the treated estrous cycle and subsequently analyzed for progesterone (P4) and PGF2alpha metabolite (PGFM). Prior to statistical analysis, SMB- and SMB/PGF2alpha-treated cows were sorted according to P4 concentration at d 10 of the treated estrous cycle to either a CL functional group (P4 > or = 1 ng/mL; n = 20) or a CL nonfunctional group (P4 < 1 ng/mL; n = 17). Following d 10 AM administration of PGF2alpha, functional and nonfunctional groups were further subdivided based on treatment. The groups were as follows: untreated control cows (n = 8); SMB-treated cows retaining a functional CL (SMB-F; n = 8); SMB-treated cows with a nonfunctional CL (SMB-N; n = 11); SMB/PGF2alpha-treated cows retaining a functional CL (SMB/PG-F; n = 12); and SMB/PGF2alpha-treated cows with a nonfunctional CL (SMB/PG-N; n = 6). Of all SMB-treated cows, 54% retained a functional CL through d 10 AM of the treated estrous cycle. Mean serum P4 concentrations increased for cows in all groups until d 7, after which P4 concentrations increased for cows in SMB/PG-F, SMB-F, and control groups and decreased for cows in SMB/PG-N and SMB-N groups. Following PGF2alpha administration on d 10, mean serum P4 concentrations remained < 1 ng/mL for cows in SMB/PG-N and SMB-N groups, decreased to < 1 ng/mL for cows in the SMB/ PG-F group, and remained > 1 ng/mL for cows in SMB-F and control groups. Mean serum PGFM concentrations tended (P = .06) to increase in cows with nonfunctional CL compared with control cows on d 8 AM and were greater (P < .05) in cows with functional CL on d 8 PM through d 9 PM. These results indicate that retention of a functional rather than a nonfunctional CL following metestrus administration of SMB is dependent on a premature release of uterine PGF2alpha.     

Effects of arachidonic acid and oxytocin on equine endometrial PGF2α during normal cycles and pseudopregnancy

The effect of excess arachidonic acid or oxytocin on equine endometrial prostaglandin F_2α(PGF_2α) synthesis was measured in vitro under physiologic and pathophysiologic conditions. Endometrial tissues obtained by uterine biopsy at 5, 10, 12, 14, 16 and 20 days post-ovulation from cycling mares, after 0, 2, 4, 6, 8, 10, 12, 14 or 16 days of progesterone (P₄) in ovariectomized mares and at 30 days postovulation in mares undergoing spontaneously prolonged corpus luteum (SPCL) activity were incubated in vitro with and without added arachidonic acid or oxytocin. Endometrial PGF_2α content and synthetic capacities were determined by radioimmunoassay. PGF_2α production increased significantly at Days 12–16. Arachidonic acid did not alter this effect. Oxytocin stimulated additional PGF_2α production on Days 5, 16, and 20. SPCL tissues had minimal PGF_2α production which was increased significantly by arachidonic acid but not oxytocin. PGF_2α synthesis in ovariectomized P₄ treated mares was minimal and did not vary with length of progesterone exposure or addition of arachidonic acid. These results suggest that a) oxytocin may play a role in luteolysis in the equine, b) although arachidonic acid appears not to be limiting to PGF₂α production under normal physiological conditions, its absence may play a role in pathophysiological conditions, c) factors in addition to progesterone and arachidonic acid are required to initiate PGF_2α synthesis in the mare.     

Effect of altrenogest and Lutalyse on parturition control, plasma progesterone, unconjugated estrogen and 13,14-dihydro-15-keto-prostaglandin F2 alpha in sows

To investigate control of parturition time, 154 sows farrowing 220 litters at three locations were treated with altrenogest and Lutalyse (PG). The four treatment groups were: 1) no treatment (control group); 2) an im injection of 15 mg of PG at 1000 on d 111, 112 or 113 of gestation (d 0 = first day of estrus and gestation); 3) altrenogest (20 mg X sow-1 X d-1) fed twice daily for 4 d starting on d 109, 110 or 111; and 4) altrenogest and an injection of PG at 1000 on the day after the last feeding of altrenogest. Control sows at the University of Delaware (UD), University of Maryland (UM) and USDA, Beltsville Agricultural Research Center (BARC) had mean gestation lengths of 113.5, 114.2 and 115.7 d and live pigs/litter were 10.5, 11.0 and 7.4, respectively. Altrenogest started by d 110 prevented unscheduled early farrowing and increased (P less than .01) gestation length by 1.7 and 1.1 d, respectively, at UD and UM, but had not effect at BARC. The time from PG to parturition was 24.3, 22.6 and 34.4 h, respectively, at UD, UM and BARC. More sows at UD and UM farrowed between 0700 and 1700 on the expected day of parturition after injection of PG (59.3%) than with no PG (20.7%; P less than .05). The high incidence of small litters (less than six pigs) from sows inseminated with frozen semen at BARC resulted in negative correlations of live pigs/litter with gestation length (r = -.533, P = .0001) and with time from PG injection to birth of first pig (r = -.425, P = .017); these correlations were not significant at UD and UM where only natural service was used.(ABSTRACT TRUNCATED AT 250 WORDS)     

Changes in plasma estrogen, luteinizing hormone, follicle-stimulating hormone and 13,14-dihydro-15-keto-prostaglandin F2 alpha during blockade of luteolysis in pigs after human chorionic gonadotropin treatment

An injection of human chorionic gonadotropin (HCG) or estrogen on d 12 of the estrous cycle delays luteolysis in the pig. In an experiment to determine if HCG stimulated estrogen secretion, 21 cyclic pigs received one of five different amounts of HCG-(A) 0, (B) 125, (C) 250, (D) 500 or (E) 1,000 IU-as a single, im injection in 2 ml of distilled water on d 12 of the estrous cycle. Blood was collected from the jugular vein immediately before HCG injection and once daily thereafter until d 20 of the estrous cycle. Plasma progesterone, estrogen (unconjugated) and 13,14-dihydro-15-keto-prostaglandin F2 alpha (PGFM) were quantified for pigs in all groups; luteinizing hormone (LH) and follicle-stimulating hormone (FSH) were quantified for pigs in groups A and E. The HCG injection exerted a dose-related increase on the mean interestrus interval (groups A, B, C, D and E were 20.5, 20.2, 22.5, 31.0 and 61.4 d, respectively) and on the delay of luteolysis as measured by mean plasma progesterone on d 16 (A, B and C vs D and E, respectively, 1.9, 1.2 and 10.4 vs 34.1 and 47.1 ng/ml; P less than .05). The HCG injection caused a transitory increase in plasma estrogen from d 12 (5 to 10 pg/ml before treatment) to d 15 (35.5 pg/ml, group D) and to d 16 (90.2 pg/ml, group E) before it decreased to preinjection levels on d 17 (group D) and 18 (group E).(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of GnRH in combination with PGF2α on the dynamics of follicular and luteal cells in post-pubertal Holstein heifers

Holstein heifers were randomly allotted by weight, age and body condition score to one of three treatments to test the hypothesis that GnRH administration concurrent with PGF_2α injection would advance follicle or corpus luteum (CL) development parallel to an induced luteolysis of the pre-existing CL. Heifers in the control group (n = 14) received two treatments of PGF_2α(25 mg, im) given 10 days apart. Groups 2 (n = 14) and 3 (n = 14) received an additional treatment of GnRH (100 μg, im) after the first and second PGF_2α respectively. Estrus detection began immediately after PGF_2α and continued for 80 h. Blood sampling was initiated 7 days prior to the first PGF_2α (day − 7) and continued on days 0, 7, 10 (prior to the second PGF_2α), 17 and 24. Heifers were artificially inseminated after the second PGF_2α and pregnancy diagnosed at 60 days. There was a trend (P < .10) toward a lower estrus response in group 3 when compared to the other groups. Pregnant heifers in group 2 had lower progesterone (0.44 ± 0.09 vs. 1.72 ± 0.56 ng/ml) a week after the second PGF_2α than the non-pregnant animals in that group (P < .05). Similar results were observed in the control group but only within the responding heifers (0.61 ± 0.08 vs. 0.93 ± 0.03 ng/ml; P < .05). Progesterone in heifers in group 2 remained high on day 0, 7, and 10 (1.48 ± 0.37, 1.23 ± 0.39, 1.96 ± 0.36 ng/ml) in spite of the treatment with PGF_2α. This data suggest that administration of GnRH following PGF_2α alters bovine luteal and/or follicular cell function.     

Twin reduction by PGF2α intraluteal instillation in dairy cows

The objective of this study was to determine whether induced luteolysis of one of the two corpora lutea in twin pregnancies would provoke spontaneous twin reduction. In Experiment 1, 12 post‐partum cows with two corpora lutea in the same ovary were assigned to (three cows per group): Group I, Group II, Group III or Group IV receiving into one of the corpora lutea puncture with no treatment, 0.5 mg dinoprost, 1.5 mg dinoprost and 2.5 mg dinoprost, respectively. One of the two corpora lutea showed clear signs of luteolysis on Day 2 and was practically non‐detectable on Day 7 after treatment in the three cows of the Group IV. In Experiment 2, 11 cows carrying live twins with two corpora lutea on Day 28 of gestation, eight bilateral and three unilateral, received 2.5 mg dinoprost into one of the corpora lutea. Corpus luteum reduction and embryo reduction after treatment were registered in 10 and 9 cows, respectively. In bilateral twin pregnancies, four cows suffering embryo reduction remained pregnant. In unilateral twin pregnancies, membrane detachment resulted in the death of both cotwins. In conclusion, although observations were based on few animals, there seems to be a mechanism that operates locally to transfer ovarian progesterone to the uterus, and also a quantitative relationship between the amount of progesterone secreted and support of conceptuses, resulting in death of one twin embryonic vesicle when one corpus luteum regresses.     

Association of CXCR1 polymorphisms with apoptosis,necrosis and concentration of milk neutrophils in early lactating dairy heifers

Associations between polymorphisms in the candidate gene CXCR1, encoding the chemokine (C-X-C motif) receptor 1, and udder health have been identified before. In the present study, associations between the CXCR1 genotype (whole coding region) and apoptosis, necrosis, and concentration of milk polymorphonuclear neutrophilic leukocyte (PMNL) of 292 quarters belonging to 73 early lactating dairy heifers were studied. In uninfected quarters, % milk PMNL apoptosis was higher in c.980GG heifers [least squares means (LSM) 27%] compared to c.980AG heifers (LSM 16%), whereas in infected quarters, % milk PMNL apoptosis was higher in c.642GG heifers (LSM 29%) compared to c.642AG heifers (LSM 18%). Differences in milk PMNL concentration between infected and uninfected quarters were smaller in c.980AG heifers than in c.980GG heifers. An association between the CXCR1 genotype and necrosis of milk PMNL could not be demonstrated. Results indicate that CXCR1 polymorphisms influence viability and concentration of milk PMNL and provide a foundation for future research.     

Duration of estrus induced after GnRH‐PGF2α protocol in dairy heifer

Estrous expressions in dairy cows have been shortened and weakened. Dairy heifers, on the other hand, may not have had such changes in estrous signs as observed in cows, since they have less stresses than cows. The aim of this study was to describe the duration of estrus in a herd of dairy heifers. A total of 56 Holstein Friesian heifers estrus was synchronized using two different hormonal protocols. They were checked for primary and secondary estrous signs with the help of heat detection devices for 48 h at an interval of 4 h starting at 16.00 hour, one day after PGF_2α treatment. Onset and end of standing estrus during 48 h observation period was recorded in 35 of the 44 heifers coming into estrus within 5 days after PGF_2α treatment during the observation period. The duration of standing estrus on the average (±SD) was 9.7 ± 5.3 h. Percentage of heifers with standing estrus longer than 12 h was 40%, and 53% showed standing estrus only for 4–8 h. It is indicated that duration of estrus in dairy heifers has been shortened recently.     

The effect of prostäglandin F2α and oxytocin on porcine myometrium in vitro *

Acht geschlechtsreife Jungsauen wurden ungefähr 24 Stunden nach Eintritt der Brunst geschlachtet und 8 trächtige Jungsauen 30–60 Tage nach der Besamung geschlachtet. Von deren Uteri wurden Uterusstreifen präpariert und im Durchlaufbad eingespannt. Die Motilität der Muskelstreifen war bei den nicht trächtigen Uteri nach 2 Stunden regelmäβig mit 1–2 Kontraktionen pro Minute, während die spontane Motilität des Myometriums bei der Frühträchtigkeit sehr unregelmäβig war und manchmal ganz fehlte. PGF₂α und Oxytocin ergaben immer eine Stimulanz der Motilität in den Muskelstreifen bei den nicht trächtigen als auch bei den trächtigen Jungsauen.     

Retinol, β-carotene and β-tocopherol concentrations in mare and foal plasma and in colostrum

Fat-soluble vitamins transfer poorly across the diffuse epitheliochorial placenta of the mare, so the neonatal foal is dependent on its supply via colostrum. Concentrations of retinol (RT), β-carotene (BC), and β-tocopherol (AT) were assayed in samples of plasma, colostrum and milk from nine mares of mixed light breeding and their foals at parturition and days 1, 2, and 4 postpartum. Samples were analyzed simultaneously for RT, BC, and AT using a new, less time-consuming reverse phase high-pressure liquid chromatography (HPLC) method. Feeds were analyzed, and mean daily intakes calculated. Mare plasma RT increased from day 1 to day 4 (P = .033), and plasma AT declined linearly from day 0 to day 4 (P = .006). Colostrum concentrations of all vitamins increased from parturition to day 1, and then declined rapidly over the 4 days. Foal plasma BC increased from parturition to day 4 (P = .080), and plasma AT increased from parturition to day 2 (P < .001), and 4 (P = .060). These observations suggest that three times the current recommendation of vitamin A is sufficient for pregnant mares, in keeping with previous studies of growth, pregnancy, and lactation. In contrast, the linear decrease in mare plasma AT suggests that 1.2 times the current recommendation of vitamin E may be less than optimal for pregnant mares.

Introduction

The diffuse epitheliochorial placenta of the mare does not allow fat-soluble vitamins to cross with ease to the fetus,[1] so their status in the neonatal foal is dependent on colostrum. This first milk contains these vitamins, which are necessary for many functions.Vitamin A is required for vision, reproduction, growth, development, and maintenance of epithelial cells, and osteoclast activity in bone formation. In addition, vitamin A deficiencies reduce resistance to disease, can cause neurological degeneration, and congenital defects.[2] Vitamin E, an antioxidant, prevents lipid peroxidation and cell membrane damage. Its deficiency in horses has been associated with a form of muscular dystrophy and impaired immune function. [3]Studies on the mare and foal have described blood concentrations of one or two of these vitamins,[4, 5 and 6] but only one involved all three. [7] In that study, vitamin concentrations were assayed in mare blood and milk, but only once in foal blood at parturition, so data during the periparturant period is incomplete.The objectives of this enumerative study were to determine concentrations of RT, BC, AT in plasma and colostrum, to evaluate colostral transfer of these vitamins, to assess the adequacy of vitamin intakes of mares, and to develop a time-saving single-run HPLC method.

Materials and methods

Animals

Nine healthy aged multiparous mares of mixed light breeding (13.6 ± 1.5 years, 592 ± 20 kg body mass) and their foals were sampled. The institutional animal care and use committee approved the protocol. Mares were kept on meager late-winter mixed grass pasture, with free access to round bale orchard and fescue grass hay, water, and trace mineral/vitamin salt. They were fed 2 kg of concentrate twice daily at 7:00 and 3:00 . The diet met or exceeded current recommendations for pregnant mares at 11 months gestation for energy.[8] Samples of hay and concentrate were submitted for partial proximate analysis to the Virginia Tech Forage Testing Laboratory where standard AOAC [9] methods are used ( Table 1). Samples of feeds were also taken for analysis of RT, BC, and AT ( Table 2).     

Bactrian camel (Camelus bactrianus) integrins αvβ3 and αvβ6 as FMDV receptors: Molecular cloning, sequence analysis and comparison with other species

Integrins are heterodimeric adhesion receptors that participate in a variety of cell–cell and cell–extracellular matrix protein interactions. Many integrins recognize RGD sequences displayed on extracellular matrix proteins and the exposed loops of viral capsid proteins. Four members of the αv integrin family of cellular receptors, αvβ3, αvβ6, αvβ1 and αvβ8, have been identified as receptors for foot-and-mouth disease virus (FMDV) in vitro, and integrins are believed to be the receptors used to target epithelial cells in the infected animals. To analyse the roles of the αv integrins from a susceptible species as viral receptors, we have cloned Bactrian camel αv, β3 and β6 integrin cDNAs and compared them to those of other species. The coding sequences for Bactrian camel integrin αv, β3 and β6 were found to be 3165, 2289 and 2367 nucleotides in length, encoding 1054, 762 and 788 amino acids, respectively. The Bactrian camel αv, β3 and β6 subunits share many structural features with homologues of other species, including the ligand binding domain and cysteine-rich region. Phylogenetic trees and similarity analyses showed the close relationships of integrin genes from Bactrian camels, pigs and cattle, which are each susceptible to FMDV infection, that were distinct from the orders Rodentia, Primates, Perissodactyla, Carnivora, Galliformes and Xenopus. We postulate that host tropism of FMDV may in part be related to the divergence in integrin subunits among different species.     

Pharmacokinetics of flunixin and its effect on prostaglandin F2α metabolite concentrations after oral and intravenous administration in heifers

Flunixin meglumine (FM) was administered either orally as granules or intravenously to six heifers in a two period crossover study. Single doses of 2.2 mg/kg body weight were used. Pharmacokinetic variables were calculated using statistical moment methods. The effect exerted by flunixin was measured as changes in the basal plasma concentration of the main metabolite of prostaglandin (PG) F_2α. After oral FM the arithmetic means of pharmacokinetic variables were: MRT = 12.7 h; MAT = 6.3 h; C _max= 0.9 μg/mL; t _max= 3.5 h. The bioavailability was 60% and the mean half-life (harmonic mean) was 6.2 h. Oral administration of FM inhibited as effectively as intravenous administration the prostaglandin biosynthesis. The concentration of the PG metabolite decreased almost as rapidly as after intravenous administration. The duration of the effect was prolonged and the PG metabolite concentration was significantly lower between 10 and 30 h after oral than after intravenous administration. The results indicate that oral dosing of flunixin, in the form of granules, can be an alternative to intravenous administration for therapeutic use in cattle.     

13,14‐Dihydro‐15‐Keto Prostaglandin F2α Release in Response to Oxytocin Challenge Early Post‐Partum in Anoestrous Nelore Cows Submitted to Temporary Calf Removal and Progesterone Priming

The study evaluated, in early post‐partum anoestrous Nelore cows, if the increase in plasma oestradiol (E2) concentrations in the pre‐ovulatory period and/or progesterone priming (P4 priming) preceding ovulation, induced by hormonal treatment, reduces the endogenous release of prostaglandin PGF₂αand prevents premature lysis of the corpus luteum (CL). Nelore cows were subjected to temporary calf removal for 48 h and divided into two groups: GPE/eCG group (n = 10) and GPG/eCG group (n = 10). Animals of the GPE/eCG group were treated with a GnRH agonist. Seven days later, they received 400 IU of eCG, immediately after PGF₂α treatment, and on day 0, 1.0 mg of oestradiol benzoate (EB). Cows of the GPG/eCG group were similarly treated as those of the GPE/eCG group, except that EB was replaced with a second dose of GnRH. All animals were challenged with oxytocin (OT) 9, 12, 15 and 18 days after EB or GnRH administration and blood samples were collected before and 30 min after OT. Irrespective of the treatments, a decline in P4 concentration on day 18 was observed for cows without P4 priming. However, animals exposed to P4 priming, treated with EB maintained high P4 concentrations (8.8 ± 1.2 ng/ml), whereas there was a decline in P4 on day 18 (2.1 ± 1.0 ng/ml) for cows that received GnRH to induce ovulation (p < 0.01). Production of 13,14‐dihydro‐15‐keto prostaglandin F₂α (PGFM) in response to OT increased between days 9 and 18 (p < 0.01), and this increase tended to be more evident in animals not exposed to P4 priming (p < 0.06). In conclusion, the increase in E2 during the pre‐ovulatory period was not effective in inhibiting PGFM release, which was lower in P4‐primed than in non‐primed animals. Treatment with EB promoted the maintenance of elevated P4 concentrations 18 days after ovulation in P4‐primed animals, indicating a possible beneficial effect of hormone protocols containing EB in animals with P4 priming.     

α- and β-adrenoceptors in the sheep urinary bladder

A study was undertaken to determine the presence and distribution of alpha- and beta-adrenoceptors in the sheep bladder body and base. In the bladder body, noradrenaline and isoproterenol induced relaxation which was significantly inhibited by propranolol, pafenolol and butoxamine. In the presence of propranolol (10(-5) M), noradrenaline induced a small contraction, as well as phenylephrine, but B-HT 920 failed to cause any effect on the bladder body. In the bladder base, noradrenaline caused a contraction that was significantly inhibited by prazosin but not by yohimbine. Phenylephrine also induced a contractile response in this structure which was inhibited by prazosin. Isoproterenol caused a relaxation that was significantly inhibited by propranolol and pafenolol but not by butoxamine. Relaxation was mediated by both beta 1 and beta 2-adrenoceptors in the detrusor muscle and by beta 1-adrenoceptors in the bladder base. Alpha 1-adrenoceptors contributed to maintain the detrusor tone and contract the bladder base.     

FC‐33 Expression of IL‐12 receptor β2 gene in atopic dogs

Atopic dermatitis (AD) is very common in dogs, but its pathogenesis is not yet fully understood. It has been suggested that a Th2‐dominant status may be associated with the occurrence of canine AD. IL‐12 is thought to be important for the differentiation of Th1 cells. The IL‐12 receptor β2 (IL‐12Rβ2) gene is considered to play a critical role in signal transduction and is attracting attention as one of the causative genes of AD in humans. The purpose of this study was to investigate the relationship between IL‐12Rβ2 gene expression and canine AD. The canine IL‐12Rβ2 gene was cloned by RT‐PCR and its nucleotide sequences were determined. Canine IL‐12Rβ2 showed 76.8% homology at the amino acid level with human IL‐12Rβ2, and its structural motifs were well conserved. cDNA with a 91 bp deletion including the transmembrane region was also cloned, which consequently produced a frame shift and an early stop codon. The deletion region corresponded to exon 14 of the human IL‐12Rβ2 gene on chromosome 1. The expression of deleted canine IL‐12Rβ2 mRNA in phytohemagglutinin‐stimulated peripheral blood mononuclear cells was examined in seven healthy dogs and 11 AD dogs. Both deleted and intact mRNAs were expressed at constant ratios in healthy and AD dogs. The results indicate that the deletion of the transmembrane region is not associated with the occurrence of AD, and that the expression of the deleted mRNA may be constitutive and produced by alternative splicing. Funding: Self‐funded.     

Plasma concentrations of flunixin in the horse: its relationship to thromboxane B2 production

The effects of the intravenous (i.v.) administration of 1.1 mg/kg of flunixin meglumine on thromboxane B2 (TxB2) concentrations were studied in sedentary and 2-year-old horses in training. The baseline TxB2 serum concentrations generated during clotting were 2.89 +/- 0.81, 2.19 +/- 0.25 and 0.88 +/- 0.12 ng/ml for the 2-year-old Thoroughbreds in training, sedentary horses under 10 and over 10 years old, respectively. There was a significant difference in baseline TxB2 concentrations between older and younger horses (P less than 0.005). Significant reduction in TxB2 production from baseline were noted at 1 (P less than 0.01) and 4 h (P less than 0.01) but not at 8 h after flunixin administration. The percent reduction in serum TxB2 concentration at 1 h after the administration of flunixin was 68.6 +/- 7.3 and 45.2 +/- 6.8 for the training and sedentary horses, respectively; the differences were significant (P less than 0.04). Serum concentrations of TxB2 returned to baseline values by 12-16 h after flunixin administration. The results of this study indicate a difference in the TxB2 concentrations of older vs. younger horses and a difference in the suppression of TxB2 after the administration of flunixin in 2-year-old Thoroughbreds in training compared to sedentary horses. The results of this study suggest that the detection of low concentrations of flunixin in urine 24 h post-administration may not represent pharmacologic effective concentrations of flunixin in plasma.     

Expression of fibronectin and its integrin receptor α5β1 in canine mammary tumours

Fibronectin and its integrin receptor α5β1 were studied by immunohistochemical methods in five normal canine mammary glands, four dysplastic glands and 18 mammary tumours. The aim of the study was to evaluate the possible changes in the α5β1 integrin receptor and its ligand fibronectin in relation to the metastatic capacity of canine mammary neoplasms. The immunostaining of α5β1 was very uniform in the hyperplastic glands but uneven in the mammary tumours. The expression of α5 and β1 was diminished in metastatic tumours but there were some α5-positive cells with pronounced features of malignancy and immaturity. Stromal fibronectin was increased in most cases and cytoplasmic staining of fibronectin was observed in epithelial and myoepithelial cells in mammary neoplasms but not in normal or dysplastic mammary tissue. There was no relationship between the content of α5β1 and the expression of fibronectin in canine mammary tumours.