Effect of Deslorelin and/or Human Chorionic Gonadotropin on Inducing Ovulation in Mares During the Transition Period Versus Ovulatory Season

The objective of this study was to compare the rate of ovulation when deslorelin and/or human chorionic gonadotropin (hCG) was administered in mares in both the transition period and the ovulatory season. A total of 200 Paint Horses, Quarter Horses, and crossbred mares were used during the transition season (July to September) and the ovulatory season (October to February) of the southern hemisphere. The animals were divided into four groups. In the control group (n = 72), mares received 1 mL of saline; in deslorelin group (n = 171), 1.5 mg of deslorelin was administered by intramuscular (IM) injection; in hCG group (n = 57), 1,667 IU of hCG was administered IV; and in hCG + deslorelin group (n = 438), 1.5 mg of deslorelin (IM) and 1,667 IU of hCG (IV) were administered. The drugs were administered after follicles ≥35 mm in diameter were identified and grade III uterine edema was observed. At 48 hours after application, ultrasonography was performed to detect ovulation. During the transition period, the ovulation rates were 4.3% (control), 78.6% (deslorelin), 50% (hCG), and 73.3% (hCG + deslorelin). During ovulatory season, the ovulation rates were 16.4% (control), 68.8% (deslorelin), 60% (hCG), and 73% (hCG + deslorelin). There was no significant difference (P > .05) in the ovulation rate between the groups or the periods, except that the control group was lower than all others. Furthermore, both hCG and deslorelin are viable options for inducing ovulation during the transition period before ovulation season.     

Follicular Fluid Prolactin and the Periovulatory Prolactin Surge in the Mare

Prolactin may play multiple roles in equine reproduction. Prolactin appears to be associated with seasonal reproduction, and fluctuating prolactin levels during the estrous cycle suggest that it may play a role in estrous cyclicity as well. The purpose of this research was to investigate the activity of prolactin during the follicular phase of the estrous cycle. In experiment 1, prolactin concentrations were determined from plasma samples collected at least every other day throughout the estrous cycle. Periovulatory (ovulation ± 1 day) prolactin concentrations were compared with concentrations during early diestrus (days 2−10 postovulation). In experiment 2, prolactin concentrations were measured in follicular fluid collected from 74 follicles of various sizes. Follicles were grouped into small (≤20 mm), medium (21−35 mm), and large (>35 mm) size categories. Prolactin concentrations increased during the periovulatory period in cycling mares. This periovulatory surge was superimposed on baseline prolactin concentrations that varied with season. Prolactin was present in significant quantities in the follicular fluid. Follicular fluid prolactin concentrations were lowest in small follicles and increased in medium and large follicles. Concentrations did not differ between medium and large follicles. Follicular fluid prolactin concentrations were lower in autumnal follicles compared with summer follicles of comparable size. It is possible that the short-term surge in circulating prolactin around ovulation could be linked to the significant levels of prolactin in follicular fluid. Ovulation releases a relatively large volume of fluid into the peritoneum. The prolactin in this fluid could be a contributor to the periovulatory prolactin surge.     

Effect of Follicle Size and Follicle-Stimulating Hormone on Ovulation Induction and Embryo Recovery in the Mare

The timing of ovulation is an important component to many equine breeding strategies. The action of luteinizing hormone on ovulation induction has been recognized; however, potential effects of follicle-stimulating hormone (FSH) have been less defined. Objectives of this study were to determine whether (1) mares could be induced to ovulate follicles ≤30 mm; (2) equine FSH (eFSH) has a positive effect on ovulation induction, and (3) ovulation of small follicles would affect embryo recovery. Light-horse mares (n = 12) between 4 and 10 years of age were assigned to treatments when they had a dominant growing follicle with a mean diameter of 24, 28, or 35 ± 2 mm and endometrial edema. Treatments were (1) H35, human chorionic gonadotropin (hCG) at 35 ± 2 mm; (2) F35, eFSH at 35 ± 2 mm; (3) H28, hCG at 28 ± 2 mm; (4) FH28, eFSH and hCG at 28 ± 2 mm; (5) D28, deslorelin (gonadotropin-releasing hormone [GnRH] analog) at 28 ± 2 mm; (6) FH24/H24, hCG or eFSH and hCG at 24 ± 2 mm. Mares’ reproductive tracts were scanned at 24 ± 2-hour intervals after treatment to detect ovulation. Mares were inseminated, and embryos were collected. Numbers of mares that ovulated within 48 ± 2 hours after treatment were: H35, 8/8 (100%); F35, 8/14 (57%); H28, 7/12 (58%); FH28, 9/12 (75%); D28, 3/7 (43%) and FH/H24, 4/14 (29%). The number of mares that ovulated in 48 ± 2 hours for H35 was not different from that for FH28 but was higher (P < .05) than all other groups. Embryo recovery rates, diameters, developmental stages, and morphology scores were not different for mares ovulating 48 hours or less versus more than 48 hours after treatment or among treatment groups. Results of this study demonstrate that follicles ≤30 mm can be induced to ovulate with no effect on embryo recovery or quality, as assessed by stereomicroscopy.     

Absence of Any Specific Diurnal Pattern in the Time of Ovulation in the Mare

This study investigates a long-held belief that mares tend to ovulate during the night. If this were so, a strategic examination regimen might replace multiple daily examinations when the time of ovulation needs to be known to within a few hours. Over 500 mares were examined at least three times daily to determine the time, to within ±4.25 hours, of 896 ovulations. The time of ovulation was determined as midway between the pre-ovulatory and post-ovulatory examinations. Three broad peaks of incidence of ovulation occurred at 5:00 am, 2:00 pm, and 10:00 pm. The interval between peaks was 9, 8, and 7 hours. during which 34.2% (5:00 am to 2:00 pm), 44.3% (2:00 pm to 10:00 pm), and 21.3% (10:00 pm to 5:00 am) of ovulations occurred. The percentage of ovulations occurring in the period 2:00 pm to 10:00 pm was significantly higher (P < .001) than in the period 10:00 pm to 5:00 am and significantly higher (P < .01) than in the period 5:00 am to 2:00 pm. The data showed that mares ovulated at all times of the day and night, with some increased incidence between 2:00 pm and 10:00 pm. Therefore, no strategic examination regimen was possible.     

Repeatability of Prolactin Responses to a Small Dose of Sulpiride in Estrogen-Primed Geldings in Spring and in Mares During the Estrous Cycle in Summer

Two experiments were conducted to assess the repeatability of prolactin responses to a small dose of sulpiride in estrogen-primed geldings in spring and in mares during the estrous cycle in summer. Six long-term geldings each received a single intramuscular injection of 100 mg of estradiol cypionate on March 31, 2011, and were then challenged with an intravenous injection of dl-sulpiride (5 μg/kg of body weight of the racemic mixture) every other day for a total of 8 days. Jugular blood was collected at 0, 10, 20, 40, and 60 minutes after the injection of sulpiride for prolactin measurement. The experiment was repeated with six mares during the summer (July), except that the number of challenges was extended to 15 over 30 days so that any effect of estrous cycle stage could be assessed. Prolactin responses in geldings during April were robust and were varied in a quadratic manner (P < .003) over the eight sulpiride injections, increasing linearly to a plateau by the fourth injection. Mares also displayed robust prolactin responses to sulpiride injections in July, and there was no effect (P > .1) of day of injection and no effect of stage of estrous cycle (follicular phase, early diestrus, or late diestrus). We concluded that prolactin responses to this dose of sulpiride were sufficiently robust and repeatable for use as a paradigm for studies of the relative competitive efficacy and duration of action of various dopaminergic compounds and their vehicular formulations.     

Long-term Treatment of Insulin-insensitive Mares with Cabergoline: Effects on Prolactin and Melanocyte Stimulating Hormone Responses to Sulpiride and on Indices of Insulin Sensitivity

The main experiment assessed whether the inhibitory effects of the dopamine agonist, cabergoline, on prolactin and α-melanocyte stimulating hormone (MSH) concentrations would persist throughout a longer-term administration (65 days). The possible effect of cabergoline on insulin sensitivity was also studied. Ten mares known to be insulin insensitive were allotted to two groups (treated vs. control). An insulin challenge, a glucose tolerance test, and a sulpiride challenge were administered before treatment. On day 0, treated mares (n = 5) received an injection of 5 mg cabergoline in slow-release vehicle; control mares (n = 5) received an equivalent vehicle injection. Injections were repeated every 10 days for a total of seven injections. Sulpiride challenges were done 1 day before each cabergoline treatment to assess possible refractoriness to the treatment. Behavior and hair coat density were also monitored. Plasma prolactin was suppressed (P < .01) to undetectable levels in mares receiving cabergoline; control mares had robust prolactin responses to each sulpiride injection. There was no indication of refractoriness to cabergoline over time. Plasma MSH concentrations after sulpiride were also suppressed (P < .05) by cabergoline. After treatment, neither the glucose response to insulin nor the insulin response to glucose differed (P > .1) between groups. No behavioral changes were noted because of treatment. Weight of hair samples indicated that cabergoline perturbed (P < .05) winter coat growth. It is concluded that 5 mg of cabergoline in slow-release vehicle administered every 10 days is an effective way of delivering dopaminergic activity to mares that results in no noticeable detrimental effects and no refractoriness to the drug.     

Effect of Repeated Cabergoline Treatment on the Vernal Transition and Hair Shedding of Mares (Year 1) and a Subsequent Comparison of the Effect of Starting Date on Prolactin Suppression (Year 2)

Two studies were conducted to determine efficacy of cabergoline for suppressing prolactin (PRL) and the possible effects on vernal transition in mares. In experiment 1, six mares each received either vehicle or cabergoline (5 mg, intramuscularly) every 10 days for 12 treatments beginning February 4, 2013. Blood samples were drawn regularly, and mares were challenged with sulpiride periodically to assess PRL suppression. Weekly hair samples were obtained to determine shedding. Prolactin was suppressed (P < .05) by cabergoline, but suppression waned in spring. There was no effect (P > .05) of treatment on day of first ovulation, luteinizing hormone, or follicle stimulating hormone. Hair shedding was generally suppressed (P = .05). In 2014 (experiment 2), eight of the same 12 mares were used in a similar experiment to determine if the rise in PRL observed in experiment 1 was due to refractoriness to cabergoline or perhaps another factor. Treatment began on April 6, 2014, corresponding to the increase in PRL in treated mares in experiment 1. Mares were treated with cabergoline or vehicle until June 5. Prolactin was suppressed (P < .05) by cabergoline, and the pattern of apparent escape from suppression was similar to year 1. We conclude that (1) cabergoline at this dose alters hair shedding but does not alter the time of first ovulation in mares and (2) relative to our previous reports of cabergoline treatment in the fall, there is a seasonal effect on the ability of this dose of cabergoline to suppress unstimulated PRL secretion.     

Comparison of Compounded Deslorelin and hCG for Induction of Ovulation in Mares

Ovulation-inducing agents are routinely used in broodmare practice. The objective of this study was to compare the efficacy of two compounded deslorelin products and human chorionic gonadotropin (hCG) in inducing ovulation in a clinical reproduction program. Breeding records of 203 mares administered an ovulation-inducing agent during the 2006 breeding season were reviewed. Estrous cycles were included for comparison if agents were administered when the largest follicle was 35 to 45 mm in diameter and endometrial edema was present. There was no significant difference (P > .05) in interval to ovulation for mares receiving deslorelin (1.9 ± 0.7 days) or hCG (2.0 ± 0.7 days). The percentage of mares that ovulated within 48 hours after treatment was also not significantly different between the agents (90.1% and 88.3%, respectively). In summary, clinical efficacy at inducing a timed ovulation in estrual mares with follicles 35 to 45 mm was similar between compounded deslorelin and hCG.     

Effect of Dose of Cloprostenol on the Interval to Ovulation in the Diestrous Mare: A Retrospective Study

Although the ovulatory effects of prostaglandins are well documented in several domestic species including horses, there has been little attention paid to the use of this ovulatory effect for clinical purposes. Mares often grow large follicles during the luteal phase that may or may not ovulate before progesterone levels decline. Clinical observations of administering prostaglandins in diestrous mares with large follicles suggest that there may be a negative correlation between follicular diameter and interval from treatment to ovulation. The objectives of this study were twofold: to investigate the cloprostenol dose rate effect on interval to ovulation and to confirm the negative correlation between follicular diameter and interval to ovulation. The hypothesis tested was that high doses of cloprostenol given in diestrus to mares with larger follicles would induce ovulation more rapidly than in mares given lower doses or with smaller follicles. To test the hypothesis, a total of 1,234 estrous cycles were induced with different doses of cloprostenol (ranging from 8.75 to 625 μg). All mares had at least one follicle of 28 mm or larger. Dominant follicles were followed by transrectal ultrasound examinations every other day until ovulation was detected. There was a significant effect of dose (P < .000) and follicular diameter (P < .000) on the interval from treatment to ovulation. The shortest mean interval (2.4 days) was observed after administration of 625 μg in mares with follicles 36 mm or larger, whereas the longest (4.9 days) occurred after 8.75 μg in follicles of 28 to 31 mm.     

Estradiol interactions with dopamine antagonists in mares: Prolactin secretion and reproductive traits

Two experiments studied the effects of pretreatment with estradiol benzoate before treatment with a dopamine antagonist on prolactin secretion and reproductive traits in mares during (1) the seasonal anovulatory period and (2) the normal breeding season. Experiment 1 was performed in winter with 17 mares selected for low follicular activity. Nine mares received estradiol benzoate injections every other day for a total of 10 injections; 8 mares received similar injections of vehicle. Ten days after onset of injections, all mares were placed on daily injections of sulpiride (250 mg) for 35 days or until ovulation. Plasma prolactin concentrations were higher (P < .001) in mares receiving estradiol than in controls for all assessments from days 12 through 36. Plasma luteinizing hormone (LH) concentrations were also increased (P < .05) by estradiol treatment from days 14 to 23. Mean day of first ovulation was 73.6 for control mares and 29.0 for estradiol-treated mares (P = .016). Estradiol treatment greatly enhanced prolactin secretion in response to sulpiride and increased LH secretion in seasonally anovulatory mares, which together hastened the date of first ovulation by an average of 45 days. Experiment 2 was designed to assess the efficacy of a long-acting, single-injection microparticle preparation of another dopamine antagonist, domperidone, for increasing prolactin secretion in cyclic mares in the summer. The experimental design and procedures used in experiment 1 were repeated, except that a single 3-g domperidone-microparticle injection was administered on day 11 rather than 45 days of sulpiride injections. Day 0 was the first day of estrus for each mare. Prolactin concentrations were higher (P < .05) in mares receiving estradiol than in control mares from days 12 through 25 and after a thyrotropin-releasing hormone injection on d 21. Estrous cycle traits (time to ovulation and time of luteal regression) were not affected (P > .1) by treatment. Estradiol enhanced the prolactin response to a single injection of 3 g domperidone in cyclic mares in the summer in a manner similar to the estradiol enhancement of prolactin secretion in response to daily sulpiride injections in anovulatory mares in winter. Thus, the single injection of domperidone could possibly replace the daily sulpiride injections used in experiment 1 to induce ovulation in seasonally anovulatory mares; this needs to be tested in future experiments.     

The Level of Prolactin,Serum Amyloid A,and Selected Biochemical Markers in Mares Before and After Parturition and Foal Heat

The aim of the study was to determine the level of prolactin (PRL), serum amyloid A (SAA), and selected biochemical markers (T-Chol, AST, TP, Mg²⁺, P⁺, and Ca²⁺) in the blood of mares during the perinatal period. The study involved 14 mares of the Polish Coldblood Horse breed, which were in the third trimester of pregnancy. Blood was collected for testing 2 weeks before parturition and then 24 hours after delivery and in the foal heat (9 days) and 9 days after ovulation and breeding. The research revealed significant differences in the level of PRL and SAA before and after delivery. The highest PRL level was found 24 hours after delivery, lowest in foal heat and 9 days after ovulation. Serum amyloid A concentration was within the accepted norms; however, on day 9 after foaling, a significant increase of this protein was observed. All biochemical markers were within physiological limits. However, significant increases in T-Chol, AST, and TP levels was observed 24 hours after the delivery, whereas in foal heat and after ovulation levels of T-Chol and TP significantly decreased and the AST level remained at a similar level. There were no significant changes in electrolyte levels such as Mg²⁺, P⁺, and Ca²⁺. The pregnancy rate in the foal heat was at 43%. Collectively, the results of this study in conjunction with clinical observations demonstrated that when the perinatal period was normal, no disturbances in health related to pregnancy, parturition, lactation, and reproductive status during the postpartum period were found.     

Inhibitory Effects of Pergolide and Cabergoline Formulations on Daily Plasma Prolactin Concentrations in Geldings and on the Daily Prolactin Responses to a Small Dose of Sulpiride in Mares

Two experiments were conducted to assess the efficacy and duration of action of two dopaminergic compounds, pergolide and cabergoline, on daily prolactin secretion in geldings and on prolactin responses to a small dose of sulpiride over 10 days. In the first experiment, oral administration of 2 mg of pergolide was compared to a single injection of 2 mg of pergolide in a slow-release vehicle and a single injection of 5 mg of cabergoline in slow-release vehicle. Controls received vehicle only. All drug treatments reduced (P < .05) prolactin concentrations relative to that in controls but differed substantially in duration of action (oral pergolide approximately 6 hours or less, injected pergolide 6 to 24 hours, and injected cabergoline at least 6 days). In the second experiment, repeated small doses of sulpiride (2 μg/kg of body weight intravenously) were used to stimulate prolactin release in mares, and the ability of seven daily injections of pergolide (2 mg each) and a single injection of cabergoline (5 mg) in slow-release vehicle to suppress this release were compared. Control mares receiving vehicle injections had robust prolactin responses to the sulpiride injections on all days of injection (days 1, 0, 1, 2, 3, 4, 6, 8, and 10 relative to treatment). Prolactin responses were muted (P < .05) by pergolide and cabergoline treatments on the first day of injection (day 0, 30 min after treatment) and were basically absent on days 1 to 8. The single injection of cabergoline continued to be suppressive through day 10, whereas mares previously treated with pergolide (through day 6) had begun to recover a prolactin response by day 10. We conclude that either daily 2-mg pergolide injections in slow-release vehicle or a single injection of 5 mg of cabergoline in slow-release vehicle is an effective way to apply dopaminergic activity to horses for approximately 7 to 10 days and may have application in the treatment of pituitary pars intermedia dysfunction in affected horses.     

Efficacy of Deslorelin Acetate (SucroMate) on Induction of Ovulation in American Quarter Horse Mares

Equine clinicians rely on ovulation induction agents to provide a timed ovulation in mares for optimal breeding management. Numerous studies have been performed on the efficacy of human chorionic gonadotropin (hCG) to induce ovulation in the mare, but limited clinical data are available for the new deslorelin acetate product SucroMate. This study was designed to evaluate the efficacy of SucroMate (deslorelin) in comparison with hCG to induce ovulation. American Quarter horse mares (n = 256) presented to Colorado State University for breeding management were used in this study. Mares received either deslorelin or hCG when a follicle ≥35 mm was detected by transrectal ultrasound in the presence of uterine edema. Ultrasonographic examinations were subsequently performed once daily until ovulation was detected. Deslorelin was administered to 138 mares during168 estrous cycles, and hCG was given to 118 mares during 136 estrous cycles. Mares administered deslorelin had a similar (P < .05) higher ovulation rate (89.9%) within 48 hours following drug administration than mares administered hCG (82.8%). There are no effects of season or age on ovulation rates in either treatment group. Twenty-one mares administered deslorelin and 11 mares administered hCG were monitored by transrectal ultrasound every 6 hours to detect ovulation as part of a frozen semen management program. Average intervals from deslorelin or hCG administration to ovulation were 41.4 ± 9.4 and 44.4 ± 16.5 hours, respectively. Results of this study indicate that SucroMate is effective at inducing a timed ovulation in the mare.     

The Effect of Manual Removal of Placenta Immediately after Foaling on Subsequent Fertility Parameters in the Mare

Retained placenta is considered to be a common problem in postpartum mares. The incidence varies from 6% to 54% depending on the breed, with higher incidence in heavy draught mares than in light-weight mares. Retained placenta has been linked to lower postpartum oxytocin concentration, impaired uterine involution, and dystocia. The objective of this study was to assess the effect of early manual removal of placenta immediately postpartum on subsequent fertility parameters (development of free intrauterine fluid, inflammatory status of endometrium, and on pregnancy rates) and to compare them with mares with spontaneous expulsion of placenta. A total of 29 mares, mainly Irish draught, were closely monitored during foaling by closed circuit television and allocated to two groups: (1) mares that expelled the placenta spontaneously within 3 hours of foaling; and (2) mares that were cleansed manually immediately after foal delivery. All mares were examined and scanned 5 and 9 days postpartum, and free intrauterine fluid was recorded; endometrial swabs were taken 9 days postpartum for endometrial cytology and culture. None of the fertility parameters analyzed showed statistical difference between groups 1 and 2. Therefore, it can be concluded that early manual removal of placenta has no detrimental effects on subsequent fertility of mares and, therefore, can be recommended when a veterinarian attends a foaling.     

影响母牛排卵的激素因素

本文论述了在牛生产上常用的影响母牛排卵的几种激素因素,对其作用机理、商品制剂,以及在实际生产上的应用进行了探讨和总结。     

超数排卵对荷斯坦牛产奶量的影响

利用来自国家“八五”攻关课题“用ＭＯＥＴ技术选育高产黑白花奶牛的研究”的１３０头中国荷斯坦供体母牛的超数排卵结果和产奶量记录，分析了超数排卵对母牛产奶量的影响。结果表明，总体来说，超数排卵对母牛本胎次泌乳期产奶量、超排当月和下月产奶量，以及下胎次泌乳期产奶量均无显著影响。在不同泌乳月超排对泌乳期产奶量均无显著影响。不同超排次数（单次超排与重复超排）和超排效果的好坏对产奶量的影响无显著差异。超排处理对产奶性能不同（高，中，低）的母牛的影响也无显著差异     

催乳素受体的研究进展

介绍催乳素受体的结构、功能,催乳素受体基因的定位、表达及调节,催乳素受体基因的多态性与生产性状和疾病的关系等。提示催乳素受体基因可作为生产性状及疾病的一个候选基因。     

自制孕马全血对凉山半细毛羊不育母羊的治疗观察

用自制孕马全血对体况较好的不育、适龄凉山半细毛羊母羊进行皮下注射治疗。试验表明：该自制药品对体况较好的凉山半细毛羊不育适龄母羊具有一定疗效。     

The Effect of Cloprostenol on the Incidence of Multiple Ovulation and Anovulatory Hemorrhagic Follicles in Two Mares: A Case Report

Reproductive records of the entire lives of two mares with abnormally high incidence of multiple ovulation and hemorrhagic anovulatory follicles were analyzed retrospectively. Chi-square analysis was used to test statistically the effect of cloprostenol, a prostaglandin F2 analog, on the incidence of multiple ovulations, hemorrhagic anovulatory follicles, and its ultrasonographic appearance. A total of 319 estrous cycles during a 17-year period were analyzed. Cycles induced with cloprostenol were more likely (P < .000) to develop hemorrhagic anovulatory follicles than spontaneous cycles. The incidence of multiple ovulation was higher in induced cycles than in spontaneous cycles in one of the two mares.     

Effect of Administration of Oxytocin During Diestrus on Corpus Luteum Function and Endometrial Oxytocin Receptor Concentration in Cycling Mares

The objectives of this study were to (1) compare the effect of twice versus once daily administration of oxytocin on days 7-14 after ovulation on the duration of corpus luteum (CL) function and (2) determine the effect of oxytocin treatment on endometrial oxytocin receptor concentration in mares. In experiment 1, mares were randomly assigned to three groups on day 7: (1) untreated control group (n = 7), (2) twice daily oxytocin treatment group (n = 7), and (3) once daily oxytocin treatment group (n = 8). Oxytocin-treated mares received 60 U of oxytocin intramuscularly (IM) the respective number of times each day on days 7 through 14. One of seven control mares (14%), five of seven (71%) twice daily oxytocin-treated mares, and five of eight (63%) once daily oxytocin-treated mares had prolonged CL function. There was no significant difference in the proportion of mares with prolonged CL function between the two oxytocin-treated groups, and collectively, oxytocin treatment increased (P < .05) the proportion of mares with prolonged CL function compared with no treatment. In experiment 2, mares were randomly assigned to two groups (n = 5/group): (1) saline-treated control mares, and (2) oxytocin-treated mares. Beginning on day 7, control mares received 3 mL of sterile saline IM twice daily, and oxytocin-treated mares received 60 U of oxytocin IM twice daily through day 14. On day 15, endometrial oxytocin-binding capacity was determined (as a measure of oxytocin receptor concentration), and there was no difference (P > .1) between control and oxytocin-treated mares (1,465.7 ± 108 and 1,382.8 ± 108 fmol/mg protein [mean ± standard error of mean], respectively).