Presynchronization with gonadotropin-releasing hormone increases the proportion of Bos indicus-influenced females ovulating at initiation of synchronization but fails to improve synchronized new follicular wave emergence or fixed-time artificial insemination conception rates using intravaginal progesterone, gonadotropin-releasing hormone, and prostaglandin F2alpha1.

The objectives were to evaluate the effect of presynchronization with GnRH on pregnancy rates (Exp. 1) and ovarian events (Exp. 2) in Bos indicus-influenced females synchronized with CO-Synch + controlled internal drug release (CIDR) as described below. In Exp. 1, a total of 135 Brahman x Hereford (F(1)) females were assigned randomly after stratification to 1) Presynch; presynchronization followed by CO-Synch + CIDR, or 2) No Presynch; CO-Synch + CIDR without presynchronization. On d -7, cattle received 100 microg of GnRH or 2 mL of intramuscular (i.m.) saline. On d 0, all cattle received a CIDR and i.m. injection of GnRH (GnRH-1), followed by CIDR removal and i.m. injection of 25 mg of PGF(2alpha) (PGF) on d 7, and i.m. injection of GnRH (GnRH-2) and timed AI (TAI) 66 h after CIDR removal (d 10). An additional 77 cows not involved in the Presynch comparison were also treated with CO-Synch + CIDR. Pregnancy rates for Presynch (37.3 +/- 6%) and No Presynch (48.5 +/- 6.1%) did not differ (P = 0.6). Pregnancy rate for all CO-Synch + CIDR-treated cattle combined was 41.9 +/- 6.1 (n = 145). In Exp. 2, we examined ovarian events in 98 Brahman x Hereford (F(1)) cows assigned randomly to Presynch or No Presynch. Ovulatory response to GnRH-1 was greater (P < 0.01) in the Presynch (58 +/- 7.1%) than in the No Presynch (27.1 +/- 6.5%) group. However, emergence of a synchronized new follicular wave after GnRH-1 and ovulation rate after GnRH-2 did not differ between groups (P = 0.4). Presynchronization increased the proportion of females ovulating after GnRH-1, but this did not increase synchrony of new follicular wave emergence or ovulation after GnRH-2, and did not improve TAI pregnancy rates.

Evaluation of vaginal electrical resistance as an indicator of follicular maturity and suitability for timed artificial insemination in beef cows subjected to a synchronization of ovulation protocol.

Objectives were to test the hypothesis that vaginal electrical resistance (VER) could be used to identify cows without a large (<10mm) follicle at timed-AI (TAI) following a synchronization of ovulation protocol and thus serve as a prospective decision aid for determining cows that should not be inseminated. Brahman x Hereford (F1) females (n=233) were synchronized with the CO-Synch+CIDR protocol that consisted of a controlled internal drug release (CIDR) insert and i.m. injection of GnRH (GnRH-1; 100 microg) on day 0, removal of CIDR and i.m. injection of prostaglandin F2alpha (PGF; 25mg) on day 7, and i.m. injection of GnRH (GnRH-2, 100 microg) and TAI 66h after CIDR removal (day 10). Vaginal electrical resistance was determined with a commercially available device (Ovascan; Animark Inc., Aurora, CO) at days 0, 7, and 10. Transrectal ultrasonography was used on day 10 to assess ovarian morphology at TAI in all cattle and in a subset of females (n=98) on days 0 and 7. Mean (+/-S.E.M.) age, body condition score (BCS), BW and days postpartum were 7.2+/-0.3 years, 5.2+/-0.1, 538+/-5.3kg, and 77+/-1.1 days, respectively. Mean VER (Omega) was greatest (101.4+/-0.8) on day 0 and declined (P<0.01) to 95.2+/-0.8 and 82.0+/-0.8 Omega, respectively, on days 7 and 10. Mean diameter of the largest follicle and VER values in females conceiving after TAI differed (P=0.05) from those that did not conceive. Mean VER on days 7 and 10 and VER difference (VER on day 10 minus VER on day 7) did not differ between females with a small (<10mm) or large (>or=10mm) follicle at TAI. Timed-AI pregnancy rate was greater (P<0.01) for females with large follicles (43%) than those with small follicles (22%). Vaginal electrical resistance difference values, categorized as negative (<0 Omega) or neutral/positive (>or=0 Omega), did not differ between females that conceived to TAI compared with those that did not. We conclude that VER measurements, as used in the present study, are not adequately sensitive to differentiate between females with and without a large follicle and thus are unable to serve as a prospective decision aid for determining suitability for TAI after synchronization.

Hormonal patterns in normal and hyperleptinemic mares in response to three common feeding-housing regimens.

We previously reported that a rise in plasma leptin concentrations followed the rise in insulin and glucose in meal-fed horses, whereas horses maintained on pasture had little fluctuations in hormonal patterns. We have also described a hyperleptinemic-hyperinsulinemic condition that occurs in about 30% of our light horse mares of high body condition maintained on pasture. The present experiment was designed to 1) study the effect of 3 common feeding-housing regimens on leptin and other metabolic hormones in mares and 2) determine whether the hyperleptinemic condition interacted with these regimens. Six light horse mares with high body condition (average score = 7) were assigned to 2 simultaneous 3 x 3 Latin squares, 1 with normal mares (leptin = 0.1 to 6 ng/mL) and 1 with mares displaying hyperleptinemia (>10 ng/mL). Three feeding-housing regimens were compared: ad libitum pasture, ad libitum native grass hay in an outdoor paddock, and single morning feedings of a pelleted concentrate and hay at 0700 in a barn. Five days of acclimation to the feeding regimens were followed by a 36-h period of hourly blood collection to characterize the hormonal characteristics. Leptin concentrations were elevated (P < 0.001) in mares predetermined to be hyperleptinemic compared with normal mares, regardless of the feeding regimen. Leptin was greatest (P < 0.01) in mares on pasture and least in mares fed hay. Variations over time (P < 0.01) were present for all hormones and metabolites studied. Glucose and insulin concentrations were greatest (P < 0.01) in mares on pasture, with meal-fed mares exhibiting an immediate rise in plasma concentrations of both after feeding. Mares on hay had low and constant concentrations of glucose, insulin, and leptin, with no apparent fluctuations. Cortisol, prolactin, and IGF-I did not differ with leptin status, whereas GH differed due to feeding-housing regimen (P < 0.02); there was also an interaction of leptin status and feeding-housing regimen for GH concentrations (P = 0.094). It was concluded that 1) estimates of hormonal secretion in horses based on frequent sampling, depending upon the hormone in question, can be profoundly affected by the feeding-housing regimens, and 2) the hyperleptinemic condition persists under differing conditions of feeding-housing.

Skills acquired on virtual reality laparoscopic simulators transfer into the operating room in a blinded, randomised, controlled trial.

Virtual reality surgical simulators have proven value in the acquisition and assessment of laparoscopic skills. In this study, we investigated skill transfer from a virtual reality laparoscopic simulator into the operating room, using a blinded, randomised, controlled trial design. Surgical trainees using the LapSim System performed significantly better at their first real-world attempt at a laparoscopic task than their colleagues who had not received similar training, as measured independently by a number of expert surgical observers using four criteria.

Ovarian, hormonal, and reproductive events associated with synchronization of ovulation and timed appointment breeding of Bos indicus-influenced cattle using intravaginal progesterone, gonadotropin-releasing hormone, and prostaglandin F2alpha.

The objectives of this study were to 1) compare cumulative pregnancy rates in a traditional management (TM) scheme with those using a synchronization of ovulation protocol (CO-Synch + CIDR) for timed AI (TAI) in Bos indicus-influenced cattle; 2) evaluate ovarian and hormonal events associated with CO-Synch + CIDR and CO-Synch without CIDR; and 3) determine estrual and ovulatory distributions in cattle synchronized with Select-Synch + CIDR. The CO-Synch + CIDR regimen included insertion of a controlled internal drug-releasing device (CIDR) and an injection of GnRH (GnRH-1) on d 0, removal of the CIDR and injection of PGF2alpha (PGF) on d 7, and injection of GnRH (GnRH-2) and TAI 48 h later. For Exp. 1, predominantly Brahman x Hereford (F1) and Brangus females (n = 335) were stratified by BCS, parity, and day postpartum (parous females) before random assignment to CO-Synch + CIDR or TM. To maximize the number of observations related to TAI conception rate (n = 266), an additional 96 females in which TM controls were not available for comparison also received CO-Synch + CIDR. Conception rates to TAI averaged 39 +/- 3% and were not affected by location, year, parity, AI sire, or AI technician. Cumulative pregnancy rates were greater (P < 0.05) at 30 and 60 d of the breeding season in CO-Synch + CIDR (74.1 and 95.9%) compared with TM (61.8 and 89.7%). In Exp. 2, postpartum Brahman x Hereford (F1) cows (n = 100) were stratified as in Exp. 1 and divided into 4 replicates of 25. Within each replicate, approximately one-half (12 to 13) received CO-Synch + CIDR, and the other half received CO-Synch only (no CIDR). No differences were observed between treatments, and the data were pooled. Percentages of cows ovulating to GnRH-1, developing a synchronized follicular wave, exhibiting luteal regression to PGF, and ovulating to GnRH-2 were 40 +/- 5, 60 +/- 5, 93 +/- 2, and 72 +/- 4%, respectively. In Exp. 3, primiparous Brahman x Hereford, (F1) heifers (n = 32) and pluriparous cows (n = 18) received the Select Synch + CIDR synchronization regimen (no GnRH-2 or TAI). Mean intervals from CIDR removal to estrus and ovulation, and from estrus to ovulation were 70 +/- 2.9, 99 +/- 2.8, and 29 +/- 2.2 h, respectively. These results indicate that the relatively low TAI conception rate observed with CO-Synch + CIDR in these studies was attributable primarily to failure of 40% of the cattle to develop a synchronized follicular wave after GnRH-1 and also to inappropriate timing of TAI/GnRH-2.

Leptin secretion in horses: effects of dexamethasone, gender, and testosterone.

Five experiments were performed to evaluate the effects of dexamethasone (DEX), gender, and testosterone on plasma leptin concentrations in horses. In experiment 1, plasma leptin, insulin, glucose, and IGF-1 concentrations were increased (P < 0.01) in stallions following five daily injections of DEX (125 microg/kg BW). In experiment 2, leptin concentrations increased (P < 0.01) in mares, geldings, and stallions following a single injection of DEX, and the response was greater (P < 0.01) in mares and geldings than in stallions. The gender effect was confounded by differences in body condition scores and diet; however, based on stepwise regression analysis, both BCS and gender were significant sources of variation in the best fit model for pre-DEX leptin concentrations (R(2) = 0.65) and for maximum leptin response to DEX (R(2) = 0.75). In experiment 3, in which mares and stallions were pair-matched based on age and body condition and fed similar diets, mares again had higher (P < 0.01) leptin concentrations than stallions after DEX treatment as used in experiment 2. In experiment 4, there was no difference (P > 0.1) in plasma leptin response in mares following four single-injection doses of DEX from 15.6 to 125 microg/kg BW. In experiment 5, treatment of mares with testosterone propionate every other day for 5 days did not alter (P > 0.1) plasma leptin concentrations or the leptin response to DEX. In conclusion, multiple injections of DEX increase leptin concentrations in stallions, as does a single injection in mares (as low as 15.6 microg/kg BW), geldings and stallions. The greater leptin levels observed in mares and geldings relative to stallions were due partially to their greater body condition and partially to the presence of hyperleptinemic individuals; however, even after accounting for body condition and diet, mares still had greater leptin concentrations than stallions after DEX administration. Elevation of testosterone levels in mares for approximately 10 days did not alter leptin concentrations in mares.

Effect of dexamethasone, feeding time, and insulin infusion on leptin concentrations in stallions.

Three experiments tested the hypotheses that daily cortisol rhythm, feeding time, and/or insulin infusion affect(s) leptin secretion in stallions. Ten mature stallions received ad libitum hay and water and were fed a grain concentrate once daily at 0700. In Exp. 1, stallions received either a single injection of dexamethasone (125 microg/kg BW i.m.; n = 5) or vehicle (controls; n = 5) at 0700 on d -1. Starting 24 h later, blood samples were collected every 2 h for 36 h via jugular venipuncture. Cortisol in control stallions varied (P < 0.01) with time, with a morning peak and evening nadir; dexamethasone suppressed (P < 0.01) cortisol concentrations. Leptin and insulin were greater (P < 0.01) in the treated stallions, as was the insulin response to feeding (P < 0.01). Leptin in control stallions varied (P < 0.01) in a diurnal pattern, peaking approximately 10 h after onset of eating. This pattern of leptin secretion was similar, although of greater magnitude (P < 0.01), in treated stallions. In Exp. 2, five stallions were fed the concentrate portion of their diet daily at 0700 and five were switched to feeding at 1900. After 14 d on these regimens, blood samples were collected every 4 h for 48 h and then twice daily for 5 d. Cortisol varied diurnally (P = 0.02) and was not altered (P = 0.21) by feeding time. Insulin and leptin increased (P < 0.01) after feeding, and the peaks in insulin and leptin were shifted 12 h by feeding at 1900. In Exp. 3, six stallions were used in two 3 x 3 Latin square experiments. Treatments were 1) normal daily meal at 0700; 2) no feed for 24 h; and 3) no feed and a bolus injection of insulin (0.4 mIU/kg BW i.v.) followed by infusion of insulin (1.2 mIU.kg BW(-1).min(-1)) for 180 min, which was gradually decreased to 0 by 240 min; sufficient glucose was infused to maintain euglycemia. Plasma insulin increased (P < 0.01) in stallions when they were meal-fed (to approximately 150 microIU/mL) or infused with insulin and glucose (to approximately 75 microIU/mL), but insulin remained low (10 microIU/mL or less) when they were not fed. The increases in insulin were paralleled by gradual increases (P < 0.01) in leptin concentrations 3 to 4 h later in stallions fed or infused with insulin and glucose. When stallions were not fed, leptin concentrations remained low. These results demonstrate that feeding time, and more specifically the insulin increase associated with a meal, not cortisol rhythm, drives the postprandial increase in plasma leptin concentrations in horses.

Pregnancy in dairy cows after synchronized ovulation regimens with or without presynchronization and progesterone.

Two experiments examined pregnancy after synchronized ovulation (Ovsynch) with or without progesterone (P4) administered via controlled internal drug release (CIDR) intravaginal inserts. In experiment 1, 262 lactating cows in one herd were in 3 treatments: Ovsynch (n = 91), Ovsynch + CIDR (n = 91), and control (n = 80). The Ovsynch protocol included injections of GnRH 7 d before and 48 h after an injection of PGF20. Timed artificial insemination (TAI; 57 to 77 d postpartum) was 16 to 20 h after the second GnRH injection. Cows in the Ovsynch + CIDR group also received a CIDR (1.9 g of P4) insert for 7 d starting at first GnRH injection. Control cows received A-I when estrus was detected using an electronic estrus detection system. Based on serum P4, 44.1% of cows were cyclic before Ovsynch. Pregnancy rates at 29 d (59.3 vs. 36.3%) and 57 d (45.1 vs. 19.8%) after TAI and embryo survival (75.9 vs. 54.5%) from 29 to 57 d were greater for Ovsynch + CIDR than for Ovsynch alone. In experiment 2, 630 cows in 2 herds received TAI at 59 to 79 d postpartum after 6 treatments. Estrous cycles were either presynchronized (2 injections of PGF2alpha 14 d apart; n = 318) or not presynchronized (n = 312). Within those groups, Ovsynch was initiated 12 d after second presynchronization PGF2alpha, and used alone (n = 318) or with CIDR inserts for 7 d (1.38 g of P4/insert, n = 124 or 1.9 g of P4/insert, n = 188). Before Ovsynch, 80% of cows were cyclic. Presynchronization increased pregnancy (46.8 vs. 37.5%) at 29 d after TAI, but CIDR inserts had no effect on pregnancy in experiment 2. Overall embryonic survival between 29 and 57 d in experiment 2 was 57.7%. Use of CIDR inserts with Ovsynch improved conception and embryo survival in experiment 1 but not in experiment 2, in part due to differing proportions of cyclic cows at the outset. Presynchronization before Ovsynch enhanced pregnancy rate.

Endocrine responses in mares and geldings with high body condition scores grouped by high vs. low resting leptin concentrations.

Previous observations from this laboratory indicated that horses with high BCS could have resting plasma leptin concentrations ranging from low (1 to 5 ng/mL) to very high (10 to 50 ng/mL). To study the possible interactions of leptin secretion with other endocrine systems, BCS and plasma leptin concentrations were measured on 36 mares and 18 geldings. From mares and geldings that had a mean BCS of at least 7.5, five with the lowest (low leptin) and five with the highest (high leptin) leptin concentrations were selected. Jugular blood samples were collected twice daily for 3 d from the 20 selected horses to determine average resting hormone concentrations. Over the next 12 d, glucose infusion, injection of thyrotropin-releasing hormone (TRH), exercise, and dexamethasone treatment were used to perturb various hormonal systems. By design, horses selected for high leptin had greater (P < 0.0001) leptin concentrations than horses selected for low leptin (14.1 vs. 2.8 +/- 0.92 ng/mL, respectively). In addition, mares had greater (P = 0.008) leptin concentrations than geldings. Horses selected for high leptin had lower (P = 0.027) concentrations of GH but higher (P = 0.0005) concentrations of insulin and thriiodothyronine (T3) than those selected for low leptin. Mares had greater (P = 0.0006) concentrations of cortisol than geldings. There was no difference (P > 0.10) in concentrations of IGF-1, prolactin, or thyroid-stimulating hormone (TSH). Horses selected for high leptin had a greater (P = 0.0365) insulin response to i.v. glucose infusion than horses selected for low leptin. Mares had a greater (P = 0.0006) TSH response and tended (P = 0.088) to have a greater prolactin response to TRH than geldings; the T3 response was greater (P = 0.047) in horses selected for high leptin. The leptin (P = 0.0057), insulin (P < 0.0001), and glucose (P = 0.0063) responses to dexamethasone were greater in horses selected for high leptin than in those selected for low leptin. In addition, mares had a greater (P < 0.0001) glucose response to dexamethasone than geldings. Cortisol concentrations were decreased (P = 0.029) by dexamethasone equally in all groups. In conclusion, differences in insulin, T3, and GH associated with high vs. low leptin concentrations indicate a likely interaction of these systems with leptin secretion in horses and serve as a starting point for future study of the cause-and-effect nature of the interactions.

Thyrotropin releasing hormone interactions with growth hormone secretion in horses.

Light horse mares, stallions, and geldings were used to 1) extend our observations on the thyrotropin releasing hormone (TRH) inhibition of GH secretion in response to physiologic stimuli and 2) test the hypothesis that stimulation of endogenous TRH would decrease the normal rate of GH secretion. In Exp. 1 and 2, pretreatment of mares with TRH (10 microg/kg BW) decreased (P < 0.001) the GH response to exercise and aspartate infusion. Time analysis in Exp. 3 indicated that the TRH inhibition lasted at least 60 min but was absent by 120 min. Administration of a single injection of TRH to stallions in Exp. 4 increased (P < 0.001) prolactin concentrations as expected but had no effect (P > 0.10) on GH concentrations. Similarly, 11 hourly injections of TRH administered to geldings in Exp. 5 did not alter (P > 0.10) GH concentrations either during the injections or for the next 14 h. In Exp. 5, it was noted that the prolactin and thyroid-stimulating hormone responses to TRH were great (P < 0.001) for the first injection, but subsequent injections had little to no stimulatory effect. Thus, Exp. 6 was designed to determine whether the inhibitory effect of TRH also waned after multiple injections. Geldings pretreated with five hourly injections of TRH had an exercise-induced GH response identical to that of control geldings, indicating that the inhibitory effect was absent after five TRH injections. Retrospective analysis of pooled, selected data from Exp. 4, 5, and 6 indicated that endogenous GH concentrations were in fact lower (P < 0.01) from 45 to 75 min after TRH injection but not thereafter. In Exp. 7, 6-n-propyl-2-thiouracil was fed to stallions to reduce thyroid activity and hence thyroid hormone feedback, potentially increasing endogenous TRH secretion. Treated stallions had decreased (P < 0.01) concentrations of thyroxine and elevated (P < 0.01) concentrations of thyroid-stimulating hormone by d 52 of feeding, but plasma concentrations of GH and prolactin were unaffected (P > 0.10). In contrast, the GH response to aspartate and the prolactin response to sulpiride were greater (P < 0.05) in treated stallions than in controls. In summary, TRH inhibited exercise- and aspartate-induced GH secretion. The duration of the inhibition was at least 1 h but less than 2 h, and it waned with multiple injections. There is likely a TRH inhibition of endogenous GH episodes as well. Reduced thyroid feedback on the hypothalamic-pituitary axis did not alter basal GH and prolactin secretion.

Effects of deslorelin acetate implants in horses: single implants in stallions and steroid-treated geldings and multiple implants in mares.

Three experiments were performed to test the following hypotheses: 1) stallions and/or progesterone-estradiol-treated geldings could serve as models for the effects of a single implant of the GnRH analog, deslorelin acetate, on LH and FSH secretion by mares; and 2) multiple implants of deslorelin acetate could be used as a means of inducing ovarian atrophy in mares for future study of the mechanisms involved in the atrophy observed in some mares after a single implant. In Exp. 1, nine light horse stallions received either a single deslorelin implant (n = 5) or a sham injection (n = 4) on d 0. In Exp. 2, 12 geldings received daily injections of progesterone on d -20 through -4, followed by twice-daily injections of estradiol on d -2 to 0. On the morning of d 0, geldings received either a single deslorelin implant (n = 6) or a sham injection (n = 6). Daily injections of progesterone were resumed on d 2 through 15. In Exp. 1, plasma LH and FSH were elevated (P < 0.05) in the treatment group relative to controls at 4, 8, and 12 h after implant insertion. In the treated stallions, FSH was decreased (P < 0.05) on d 3 to 13, and LH was decreased on d 6 to 13. In Exp. 2, plasma LH and FSH were elevated (P < 0.05) at 4,8, and 12 h after deslorelin implant insertion. Plasma LH was suppressed (P < 0.05) below controls on d 2 to 7, 9, and 11 to 15; plasma FSH was suppressed (P < 0.05) on d 4 to 15. In Exp. 3, 21 mares were used to determine whether multiple doses of deslorelin would cause ovarian atrophy. Mares received one of three treatments: 1) sham injections; 2) three implants on the first day; or 3) one implant per day for 3 d (n = 7 per group). Treatment with multiple implants increased (P < 0.05) the interovulatory interval by 14.8 d and suppressed (P < 0.01) LH and FSH concentrations for approximately 25 d; no mare exhibited ovarian atrophy. In conclusion, after an initial short-term increase in LH and FSH secretion, deslorelin implants caused long-term suppression of both gonadotropins in stallions as well as in geldings treated with progesterone and estradiol to mimic the estrous cycle. It is likely that either of these models may be useful for further study of this suppression in horses. Although multiple implants in mares suppressed gonadotropin secretion longer than a single implant, the lack of ovarian atrophy indicates that the atrophy observed after a single implant in previous experiments was likely due to the susceptibility of individual mares.

Supplemental norgestomet, progesterone, or melengestrol acetate increases pregnancy rates in suckled beef cows after timed inseminations.

In Exp. 1, 187 lactating beef cows were treated with injections of GnRH 7 d before and 48 h after prostaglandin F2alpha (PGF2alpha; Cosynch) or with Cosynch plus a 7-d treatment with an intravaginal progesterone (P4)-releasing insert (CIDR-B; Cosynch + CIDR). In Exp. 2, 183 lactating beef cows were treated with the Cosynch protocol or with Cosynch plus a 7-d treatment with norgestomet (Cosynch + NORG). In Exp. 1 and 2, blood samples for later P4 analyses were collected on d -17, -7 (first GnRH injection), 0 (PGF2alpha injection), and at timed artificial insemination (TAI; 48 h after PGF2alpha). In Exp. 3, 609 lactating beef cows were treated with the Cosynch + CIDR protocol or were fed 0.5 mg of melengestrol acetate (MGA) per day for 14 d before initiating the Cosynch protocol 12 d after the 14th d of MGA feeding (MGA + Cosynch). Blood samples were collected as in Exp. 1 and 2, plus additional samples on d -33 and -19 before PGF2alpha. In Exp. 4, 360 lactating beef cows were treated with a Cosynch + CIDR protocol, with TAI occurring at either 48 or 60 h after PGF2alpha, while receiving either GnRH or saline to form four treatments. Blood samples were collected as in Exp. 1 and 2. In Exp. 1, addition of P4 reduced the ability of the first GnRH injection to induce ovulation in anestrous cows with low P4 before PGF2alpha but improved (P = 0.06) pregnancy rates (61 vs 66%). In Exp. 2, the addition of NORG mimicked P4 by likewise increasing (P < 0.01) pregnancy rates (31 vs 51%) beyond those after Cosynch. In Exp. 3, the Cosynch + CIDR protocol increased (P < 0.001) pregnancy rates from 46 to 55% compared to the MGA + Cosynch protocol. In Exp. 4, administration of GnRH at TAI improved (P < 0.05) pregnancy outcomes (50 vs 42%), whereas timing of TAI had limited effects. We conclude that a progestin treatment concurrent with the Cosynch protocol improved pregnancy outcomes in all experiments, but pretreatment of cows with MGA was not as effective as the CIDR insert or NORG implants in this Cosynch-TAI model. Most of the improvement in pregnancy rates was associated with the increase in pregnancy rates of anestrous cows, regardless of whether ovulation was successfully induced in response to GnRH 7 d before PGF2alpha. Injection of GnRH at TAI following the Cosynch + CIDR protocol increased pregnancy rates in cycling cows with high P4 before the PGF2alpha injection and in anestrous cows with low P4 before PGF2alpha injection.

Effects of dexamethasone, glucose infusion, adrenocorticotropin, and propylthiouracil on plasma leptin concentrations in horses.

In experiment 1, nine light horse geldings (three 3 x 3 Latin squares) received dexamethasone (DEX; 125 microg/kg BW, i.m.), glucose (0.2 g/kg BW, i.v.), or nothing (control) once per day for 4 days. DEX increased (P < 0.001) glucose, insulin, and leptin concentrations and resulted in a delayed increase (P < 0.001) in IGF-I concentrations. In experiment 2, mares were similarly treated with DEX (n = 6) or vehicle (n = 6). DEX again increased (P < 0.01) glucose, insulin, and leptin concentrations; the delayed elevation in IGF-I concentrations occurred on day 10, 12, and 19, relative to the first day of treatment. In experiment 3, six light horse geldings received either 200 IU of adrenocorticotropin (ACTH) i.m. or vehicle twice daily for 4 days. ACTH increased (P < 0.001) cortisol concentrations. Further, ACTH resulted in increases (P < 0.01) glucose, insulin, and leptin concentrations. In experiment 4, plasma samples from four light horse stallions that were fed 6-n-propyl-2-thiouracil (PTU) at 6 mg/kg BW for 60 days to induce hypothyroidism were compared to samples from control stallions. On day 52, stallions receiving PTU had lower concentrations of thyroxine (P < 0.05) and triiodothyronine (P < 0.01) and higher (P < 0.01) concentrations of TSH. Leptin concentrations were higher (P < 0.01) in PTU-fed stallions from day 10 through 52. In conclusion, circulating concentrations of leptin in horses was increased by administering DEX. Treatment with ACTH increased cortisol and resulted in lesser increases in leptin, glucose, and insulin. In addition, PTU feeding results in lesser increases in leptin concentrations.

Pituitary responsiveness to GnRH in mares following deslorelin acetate implantation to hasten ovulation.

The present experiment characterized the pituitary responsiveness to exogenous GnRH in the first 10 d after ovulation following commercially available deslorelin acetate implantation at the normal dosage for hastening ovulation in mares. Twelve mature, cyclic mares were assessed daily for estrus and three times weekly for ovarian activity starting May 1. Mares achieving a follicle at least 25 mm in diameter or showing signs of estrus were checked daily thereafter for ovarian characteristics. When a follicle >30 mm was detected, mares were administered either a single deslorelin acetate implant or a sham injection and then assessed daily for ovulation. On d 1, 4, 7, and 10 following ovulation, each mare was challenged i.v. with 50 microg GnRH, and blood samples were collected to characterize the LH and FSH responses. The size of the largest follicle on the day of treatment did not differ (P = 0.89) between groups. The number of days from treatment to ovulation was shorter (P < 0.001) by 2.0 d for the treated mares indicating a hastening of ovulation. The size of the largest follicle present on the days of GnRH challenge was larger in the treated mares on d 1 (P = 0.007) but smaller on d 10 (P = 0.02). In addition, the interovulatory interval was longer (P = 0.036) in the treated mares relative to controls by 4.4 d. Concentrations of FSH in plasma of the treated mares were lower (P < 0.05) than control concentrations from d 3 to 12; LH concentrations in the treated mares were lower (P < 0.05) relative to controls on d 0 to 5, d 7, and again on d 20 to 23. Progesterone values were the same (P = 0.99) for both groups from 2 d before ovulation though d 23. There was an interaction of treatment, day, and time of sampling (P < 0.001) for LH and FSH concentrations after injection of GnRH. Both the LH and FSH responses were suppressed (P < 0.009) in the treated mares relative to controls on d 1, 4, and 7; by d 10, the responses of the two groups were equivalent. In conclusion, deslorelin administration in this manner increased the interovulatory interval, consistently suppressed plasma LH and FSH concentrations, and resulted in a complete lack of responsiveness of LH and FSH to GnRH stimulation at the dose used during the first 7 d after the induced ovulation. Together, these results are consistent with a temporary down-regulation of the pituitary gland in response to deslorelin administered in this manner.

The relationship between body condition, leptin, and reproductive and hormonal characteristics of mares during the seasonal anovulatory period.

An experiment was conducted to determine the effects of high vs low body condition scores (BCS) produced by restricted feeding on reproductive characteristics, hormonal secretion, and leptin concentrations in mares during the autumnal transition and winter anovulatory period. Mares with BCS of 6.5 to 8.0 were maintained on pasture and/or grass hay, and starting in September, were full fed or restricted to produce BCS of 7.5 to 8.5 (high) or 3.0 to 3.5 (low) by December. All but one mare with high BCS continued to ovulate or have follicular activity during the winter, whereas mares with low BCS went reproductively quiescent. Plasma leptin concentrations varied widely before the onset of restriction, even though all mares were in good body condition. During the experiment, leptin concentrations gradually decreased (P < 0.0001) over time in both groups, but were higher (P < 0.009) in mares with high vs low BCS after 6 wk of restriction, regardless of initial concentration. No differences (P > 0.1) between groups were detected for plasma concentrations of LH, FSH, TSH, GH, glucose, or insulin in samples collected weekly; in contrast, plasma prolactin concentrations were higher (P < 0.02) in mares with high BCS, but also decreased over time (P < 0.008). Plasma IGF-I concentrations tended (P = 0.1) to be greater in mares with high vs low BCS. The prolactin response to sulpiride injection on January 7 did not differ (P > 0.1) between groups. During 12 h of frequent blood sampling on January 12, LH concentrations were higher (P < 0.0001), whereas GH concentrations (P < 0.0001) and response to secretagogue (EP51389; P < 0.03) were lower in mares with high BCS. On January 19, the LH response to GnRH was higher (P < 0.02) in mares with high BCS; the prolactin response to TRH also was higher (P < 0.01) in mares with high BCS. In conclusion, nutrient restriction resulting in low BCS in mares resulted in a profound seasonal anovulatory period that was accompanied by lower leptin, IGF-I, and prolactin concentrations. All but one mare with high BCS continued to cycle throughout the winter or had significant follicular activity on the ovaries. Although leptin concentrations on average are very low in mares with low BCS and higher in well-fed mares, there is a wide variation in concentrations among well-fed mares, indicating that some other factor(s) may determine leptin concentrations under conditions of high BCS.

A curved edge moderates high pressure generated by a laparoscopic grasper.

BACKGROUND: Very high pressures are generated at the interface between the tip of laparoscopic graspers and tissue. The pressure profile suggests that the high pressure is due to stress concentration at the edge of the jaw. Stress concentration at an edge can be modified by curving the edge. This study sought to determine the effect of a curved edge on pressures generated by the jaw of a laparoscopic grasper. METHODS: Pressure generated at the instrument tissue interface was measured by a thin film pressure transducer. The two instruments that we compared were identical in every respect except the profile of the jaw edge. One was unmodified; the other had a rounded grasping edge. Load and handle pressure were constant throughout. Tissue pressures generated by the grasper were recorded as the angle of load was increased from vertical (0 degrees ) to 135 degrees. RESULTS: Localized pressures at the tip of the unmodified instrument reached a maximum of 1500 kPa. Pressures at the tip of the curved instrument reached a maximum of 920 kPa under identical conditions. CONCLUSION: High pressures generated at the tip of laparoscopic graspers can be reduced by rounding the edge of the jaw.

Resistance forces acting on suture needles.

Understanding the resistance forces encountered by a suture needle during tissue penetration is important for the development of robotic surgical devices and virtual reality surgical simulators. Tensile forces applied to skin and tendon during suturing were measured. Fresh sheep achilles tendons were tensioned with a static load 4.9 N, 9.8 N or 19.6 N and sheepskin with 0.98 N, 2.9 N or 4.9 N static load. A straight 2/0 cutting suture needle in series with a load cell on a materials testing machine penetrated the tissue at 90 degrees with a velocity of 1, 5 or 10mm/s for each tissue tension (n=5). Continuous load versus displacement data was obtained and penetration load and stiffness were noted. The load versus displacement curve for skin during needle penetration demonstrated two characteristic peaks, corresponding to initial penetration and emergence of needle from the undersurface of the tissue. Increasing the tension within the tissue (skin and tendon) increased the amount of force required to penetrate the tissue with a suture needle (p<0.05). Needle displacement rate did not affect the resistance to needle penetration (p<0.05). This study provides a simple model for measuring force-feedback during needle penetration of soft tissues and is a good starting point for future studies of the penetration resistance properties of human tissues.

Stage of cycle, incidence, and timing of ovulation, and pregnancy rates in dairy cattle after three timed breeding protocols.

In experiment 1, 705 cows were assigned to three treatments: 1) the Ovsynch protocol (a GnRH injection given 7 d before and another 48 h after one PGF2alpha injection); 2) PGF2alpha + Ovsynch (one PGF2alpha injection given 12 d (d -22) before initiating Ovsynch (d -10); and 3) 2xPG12 (two PGF2alpha injections 12 d apart; d -15 and -3, followed 48 h later by a GnRH injection. All cows were inseminated (d 0) 16 to 20 h after the GnRH injection on d -1. Cyclic status was estimated by serum progesterone. More cows were in early diestrus at d -10 for PGF2alpha + Ovsynch (36%) and 2 x PG12 (29%) versus Ovsynch (19%). Multiparous cows receiving PGF2alpha + Ovsynch had greater pregnancy rates via ultrasonography at d 28 after AI (42%) than contemporaries after Ovsynch (28%) or 2xPG12 (27%) but did not differ significantly at palpation 10 to 30 d later (28, 19, and 17%, respectively). Pregnancy of first-parity cows was similar across treatments at 28 d (41%) or at palpation (33%). Pregnancy rates for 128 anestrous cows were lower, regardless of treatment. Overall embryo survival from d 28 until palpation was 72% but was only 44% in 2xPG12 cows that were anestrus through d -10. Experiment 2 included the three treatments above plus controls (one GnRH injection 7 d before PGF2alpha and AI after estrus). Preovulatory follicles were 6 to 11% larger near estrus in controls than on d -1 in cows receiving GnRH. More controls ovulated by 32 h after onset of estrus than were treated cows by 32 h after GnRH, but percentages (79 to 94%) were similar by 40 h. In multiparous cows, PGF2alpha before Ovsynch increased pregnancy rates, whereas the 2xPG12 protocol produced similar pregnancy rates as Ovsynch across parities. Ovulation was effectively induced by 40 h after GnRH.

An alternative AI breeding protocol for dairy cows exposed to elevated ambient temperatures before or after calving or both.

Our objective was to determine if a timed artificial insemination (AI) protocol (Ovsynch) might produce greater pregnancy rates than AI after a synchronized, detected estrus during summer. Lactating Holstein cows (n = 425) were grouped into breeding clusters and then assigned randomly to each of two protocols for AI between 50 and 70 days in milk. All cows were treated with GnRH followed 7 d later by PGF2alpha. Ovsynch cows then were treated with a second injection of GnRH 48 h after PGF2alpha and inseminated 16 to 19 h later. Controls received no further treatment after PGF2alpha and were inseminated after detected estrus. Pregnancy was diagnosed once by transrectal ultrasonography (27 to 30 d after AI) and again by palpation (40 to 50 d). Based on concentrations of progesterone in blood collected before each hormonal injection, only 85.4% of 425 cows were considered to be cycling. Although conception rates were not different between protocols at d 27 to 30, AI submission rates and pregnancy rates were greater after Ovsynch (timed AI) than after detected estrus. A temperature-humidity index > or = 72 was associated with fewer controls detected in estrus with lower conception than for controls detected in estrus when index values were < 72, whereas the reverse was true for cows after the Ovsynch protocol. We concluded that a timed AI protocol increased pregnancy rates at d 27 to 30 because its success was independent of either expression or detection of estrus. However, because of poorer embryonic survival in Ovsynch cows during heat stress only (39.5 vs. 69.2% survival for Ovsynch and control, respectively), pregnancy rates were not different by d 40 to 50 after timed AI.

Grasper trauma during laparoscopic cholecystectomy.

BACKGROUND: The present study characterized the histopathological nature of laparoscopic grasper trauma during laparoscopic cholecystectomy in a prospective, blinded trial in order to establish a model for laparoscopic grasper trauma. The null hypothesis that graspers cause no histologically distinct tissue injury was tested. METHODS: The gall bladders of 19 patients undergoing laparoscopic cholecystectomy were examined. The area of gall bladder that had been grasped by Debakey laparoscopic forceps was excised (sample), along with an area of gall bladder that had not been grasped (control). Paired specimens were examined by a pathologist (blinded) to identify which was 'sample' and which was 'control' and to assess for histological markers of crushed tissue injury. The data were analysed by chi-squared or Fisher's exact tests. RESULTS: The pathologist was able to identify the sample (gripped) specimen in 13 of the 19 cases. In the remaining six cases the pathologist was unable to determine the specimen that had been gripped due to either absence of damage (four cases), or severe inflammation precluding assessment (two cases). The ability of the pathologist to distinguish the sample from the control specimen was significant (chi-squared test, P = 0.003). Of the histological markers of crushed tissue injury, focal thinning of the gall bladder wall and epithelial loss were present in significantly more sample (gripped) specimens than control specimens (chi-squared test, P = 0.0002 and P < 0.0001, respectively). CONCLUSIONS: Laparoscopic graspers cause tissue trauma that can be assessed histologically. The current study presents a relevant, reproducible, ethically acceptable human model for assessing the interaction between laparoscopic graspers and soft tissues.