Size Of Ovarian Follicles Research Articles

Hypothyroidism Reduces the Size of Ovarian Follicles and Promotes Hypertrophy of Periovarian Fat with Infiltration of Macrophages in Adult Rabbits.

Ovarian failure is related to dyslipidemias and inflammation, as well as to hypertrophy and dysfunction of the visceral adipose tissue (VAT). Although hypothyroidism has been associated with obesity, dyslipidemias, and inflammation in humans and animals, its influence on the characteristics of ovarian follicles in adulthood is scarcely known. Control and hypothyroid rabbits were used to analyze the ovarian follicles, expression of aromatase in the ovary, serum concentration of lipids, leptin, and uric acid, size of adipocytes, and infiltration of macrophages in the periovarian VAT. Hypothyroidism did not affect the percentage of functional or atretic follicles. However, it reduced the size of primary, secondary, and tertiary follicles considered as large and the expression of aromatase in the ovary. This effect was associated with high serum concentrations of total cholesterol and low-density lipoprotein cholesterol (LDL-C). In addition, hypothyroidism induced hypertrophy of adipocytes and a major infiltration of CD68+ macrophages into the periovarian VAT. Our results suggest that the reduced size of ovarian follicles promoted by hypothyroidism could be associated with dyslipidemias, hypertrophy, and inflammation of the periovarian VAT. Present findings may be useful to understand the influence of hypothyroidism in the ovary function in adulthood.

Effects of excess metabolizable protein on ovarian function and circulating amino acids of beef cows: 2. Excessive supply in varying concentrations from corn gluten meal

In the dairy industry, excess dietary CP is consistently correlated with decreased conception rates. However, amount of excess CP effects on reproductive function in beef cattle is largely undefined. The objective of this experiment was to determine the effects of excess metabolizable protein (MP) supplementation from a moderately abundant rumen undegradable protein (RUP) source (corn gluten meal: 62% RUP) on ovarian function and circulating amino acid (AA) concentrations in beef cows consuming low quality forage. Non-pregnant, non-lactating beef cows (n=16) were allocated by age, BW and body condition score (BCS) to 1 of 2 isocaloric supplements designed to maintain BW for 60 days. Cows had ad libitum access to corn stalks and were individually offered a corn gluten meal-based supplement daily at 125% (MP125) or 150% (MP150) of National Research Council (NRC) MP requirements. After a 20-day supplement adaptation period, cows were synchronized for ovulation. After 10 days of synchronization, follicular growth was reset with gonadotropin releasing hormone. Daily thereafter, transrectal ultrasonography was performed to diagram ovarian follicular waves, and blood samples were collected for hormone, metabolite and AA analyses. After 7 days of observation of estrus, corpus luteum (CL) size was determined via ultrasound. Data were analyzed using the MIXED procedures of SAS. No differences (P⩾0.21) in BW and BCS existed throughout the study; however, plasma urea N at ovulation was greater (P=0.04) in MP150. Preovulatory ovarian follicle size at dominance, duration of dominance, size at spontaneous luteolysis, length of proestrus and wavelength were not different (P⩾0.11) between treatments. However, ovulatory follicles were larger (P=0.04) and average antral follicle count was greater (P=0.01) in MP150 than MP125. Estradiol concentration and ratio of estradiol to ovulatory follicle volume were not different due to treatment (P⩾0.25). While CL volume 7 days post-estrus was greater (P<0.01) in MP150 than MP125, circulating progesterone 7 days post-estrus and ratio of progesterone to CL volume were not different (P⩾0.21). Total AA were not different (P⩾0.76) at study initiation or completion; however, as a percent of total AA, branched-chain AA at ovulation were greater (P=0.02) in MP150. In conclusion, supplementation of CP at 150% of NRC MP requirements from a moderately undegradable protein source may enhance growth of the ovulatory follicle and subsequent CL compared with MP supplementation at 125% of NRC MP requirements.

12 OVARIAN STRUCTURES, ESTRUS EXPRESSION, AND PREGNANCY RATE IN BEEF HEIFERS USING ESTRADIOL CYPIONATE OR GnRH AS OVULATION INDUCTORS IN TIMED AI PROTOCOL

The aim of this study was to evaluate the effect of presence of corpus luteum (CL) at the onset of timed AI (TAI) protocol, size of ovulatory follicle and oestrus expression on pregnancy rate using E2 cypionate (EC) or GnRH as ovulation inductor in Angus heifers. On day 0, heifers [n = 414; 317 ± 23 kg; 3.08 ± 0.21 body condition score (1 to 5 scale) and 22–24 months of age] received a 1-g progesterone intravaginal device (Primer®, Agener União, Brazil) and 2 mg of oestradiol benzoate, IM, (RIC-BE®, Agener União, Brazil). On day 8, the device was removed, were administered 500 μg of sodium cloprostenol, IM, (Estron®, Agener União, Brazil) and heifers were divided in 2 groups: EC [n = 213; 0.5mg EC, IM, (SincroCP®, Ouro Fino, Brazil) on day 8] and GnRH [n = 201; 25 μg of Licerelin, IM, (GestranPlus®, Agener União, Brazil) at TAI] TAI was performed 50 h after device removal. Heifers were tail painted (Tell Tail®, GEA, New Zealand) to control oestrus expression from device removal to TAI. Heifers were scanned (7.5-MHz linear transducer MediSono® P3V, USA) to determine presence of CL (Day 0), ovulatory follicle diameter (OFD; Day 10) and ovulation (Day 11). Pregnancy was diagnosed 30 days after TAI. Oestrus expression was confirmed when marking ink was removed on day 10. Data were analysed using logistic regression and ANOVA. Oestrus expression was higher (P &lt; 0.05) in EC (93.90%, 200/213) compared to GnRH group (67.66%, 136/201). The OFD differed (P &lt; 0.05) according to presence of CL on Day 0 (10.3 mm) or not (9.6 mm), but was similar in treatment groups. Ovulation rate did not differ in EC (85%, 17/20) and GnRH (95%, 19/20) groups. Pregnancy rates were similar (P &gt; 0.05) between EC (52.58%, 112/213) and GnRH (59.20%, 119/201) groups; however, in heifers that expressed oestrus, rates were higher (P &lt; 0.05) (EC = 56%, 112/200; GnRH = 71.3%, 97/136) than in those did not show (EC = 0%, 0/13; GnRH = 33.8%, 22/65). Presence of CL at onset of TAI protocol was associated with higher pregnancy (P &lt; 0.05) in GnRH group (68%, 68/100) compared with EC (54.46%, 61/112) group, but in heifers without CL, pregnancy were similar for GnRH (50.5%, 51/101) and EC (50.5%, 51/101) groups. Heifers were also classified in 3 groups according OFD: small (≤9.5 mm), medium (9.6–11.5 mm), and large (≥11.6 mm) to verify its effect on oestrus expression and pregnancy. Medium and large OFD were implicated in higher (P &lt; 0.05) oestrus expression [(EC = 100%, 51/51; 96.97%, 32/33) and (GnRH = 83.02%, 44/53; 97.44%, 38/39)] than small OFD [(EC = 85%, 51/60) and (GnRH = 23.8%, 15/63)]. Pregnancy was lower (P &lt; 0.05) in small OFD groups (EC = 28.3%, 17/60; GnRH = 25.4%, 16/63). In medium and large OFD, pregnancy was similar in EC (72.55%, 37/51; 54.55%, 18/33) groups, but differed (P &lt; 0.05) between GnRH large (82.05%, 32/39) and medium (62.26%, 33/53). Higher pregnancy was achieved in heifers with CL in the beginning of TAI protocol treated with GnRH. Size of ovulatory follicle was associated with higher oestrus behaviour and pregnancy rate in EC and GnRH treatments.

Ovarian Dynamics in Free-Ranging Loggerhead Sea Turtles (Caretta caretta)

Vitellogenin (VTG) is an egg yolk-precursor protein that serves as a nutrient source for developing embryos in oviparous vertebrates. The hormonal control of this protein has been studied in a variety of taxa, but details about the dynamics of this protein remain to be elucidated in sea turtle species. To investigate the dynamics of VTG in a multi-clutch species under natural conditions, 38 adult Loggerhead females entrained in the Florida Power and Light St. Lucie Nuclear Plant intake canal in Hutchinson Island, Florida were sampled from May–August of 2014. Blood samples were drawn to measure testosterone, estradiol 17β, and vitellogenin (T, E2, and VTG, respectively) using enzyme-linked immunosorbent assays. Ultrasound imaging of the gonads was used to determine ovarian status and to measure ovarian follicle and oviductal egg size. Results showed that VTG concentration increased from May (8.27 mg mL−1) to June (15.37 mg mL−1) and declined into July and August (9.44 mg mL−1); this decline corresponded wi...

Female reproductive cycle and clutch size of Aspidoscelis costata costata (Squamata: Teiidae) from Tonatico, Estado de México

Female reproductive cycle and clutch size are important life history traits in lizards that can be influenced by biotic and/or abiotic factors. Here, the ovarian cycle and clutch size of a high-elevation population (1500–1600m) of Aspidoscelis costata costata in Central Mexico are described. Females were collected monthly between 2005 and 2007, and their reproductive condition was evaluated based on appearance and size of ovarian follicles and presence/absence of oviductal eggs. The reproductive cycle was markedly seasonal with asynchrony in reproductive condition among females. Vitellogenic and gravid females were recorded from April (end of the dry season) through July (middle of the rainy season). Although this population inhabits a semi-humid warm climate, the timing of female reproduction in A. costata costata is similar to the cycle of temperate-zone species of Aspidoscelis. Females produced a single clutch annually with an average clutch size of 7.7±0.44 eggs (median 8 eggs, range 4–14), which is the largest reported to date within the genus Aspidoscelis.

Physiological predictors of ovulation and pregnancy risk in a fixed-time artificial insemination program.

The objective of this study was to determine the relative importance and contribution of several physiological factors as predictors of pregnancy risk in an Ovsynch (GnRH-1 - 7d - PGF2α - 56h - GnRH-2 - 16h - artificial insemination) timed artificial insemination program: (1) age of the corpus luteum (CL; original CL, new CL, or both in response to GnRH-1) and resulting progesterone concentrations as they affected luteolysis, ovulation after GnRH-2, and pregnancy risk; (2) progesterone concentration before GnRH-1 and GnRH-2 on subsequent ovulatory response to GnRH-1 and GnRH-2 as well as pregnancy risk; and (3) a combination of these factors in a multivariable logistic regression model to predict pregnancy risk. Original data from 7 published studies were combined including ovulatory responses to both GnRH injections, blood progesterone concentrations before GnRH-1, before PGF2α, and at 48h after PGF2α, pregnancy per artificial insemination at d 32 and 60 after artificial insemination, and intervening pregnancy loss. Ovulation outcomes were greater at lesser progesterone concentrations after both GnRH injections despite the fact that pregnancy outcomes were greatest when progesterone exceeded 3ng/mL before GnRH-1 and PGF2α, suggesting that greater progesterone concentration before GnRH-1 and incidence of ovulation act via different mechanisms to improve subsequent fertility. Ovulation after GnRH-2 and subsequent pregnancy outcome were positively related to lesser concentrations of progesterone at 48h after PGF2α; however, for maximal pregnancy outcome, progesterone should be <0.5ng/mL. Cows with a CL that also formed a new CL after GnRH-1 had the greatest pregnancy outcome, whereas cows with only a new CL (i.e., anovulatory cows and those at or near estrus without a CL at GnRH-1) had the poorest pregnancy outcome. Pregnancy loss was not affected by CL status during the timed artificial insemination program. Receiver operator curves produced cut points of progesterone concentration that predicted ovulation and pregnancy risk. Selected cut points of progesterone concentration 48h after PGF2α produced true positive risks greater than 90% and false positive risks less than 25%, thus demonstrating the high predictability of ovulation after GnRH-2 and subsequent pregnancy outcome based on progesterone concentration. We conclude that progesterone concentration is highly predictive of ovulation and pregnancy risk in addition to body condition, size of ovulatory follicle, and parity.

Follicular wave emergence in Santa Inês ewes subjected to long-term, progesterone-based estrous synchronization protocols at different times of the year

This study was conducted to document the pattern of antral follicular wave emergence throughout the 14-day, progesterone (P4)-based estrous synchronization protocol in ewes that were maintained in subtropical conditions, during the period of increasing day lengths (ID), decreasing day lengths (DD), and the transitional period (TP). In addition, the influence of ovarian status (i.e., size of ovarian antral follicles and the presence of corpora lutea) at the outset of P4 treatment on ensuing ovarian follicular wave development was examined. Sexually mature Santa Inês ewes (n=70) were subjected to one of the two estrous synchronization protocols in the three seasons. On Day 0, the ewes received an i.m. injection of prostaglandin F2α and an intravaginal P4-releasing device that remained in place for 14days (G-1CIDR) or was replaced on Day 7 (G-2CIDR). Daily ultrasonography of ovaries was conducted from Days 0 to 15. Mean (±SEM) numbers of follicular waves per ewe were 3.7±0.1 and 3.6±0.1 for G-1CIDR and G-2CIDR (P>0.05). The number of emerging follicular waves was greater (P<0.05) during the ID period than during the TP and DD periods (4.0±0.1, 3.4±0.1 and 3.6±0.1, respectively). The presence of medium-sized antral follicles (4.0 to 5.75mm) in the absence of corpora lutea at the time of CIDR insertion tended to advance follicular wave emergence. Although the long-term P4 treatment was not originally designed to synchronize follicular waves, there was a distinctive pattern of follicular wave dynamics during the period of application of CIDRs that was affected mainly by the number of emerging follicular waves and ovarian status at CIDR insertion.

Developmental programming: rescuing disruptions in preovulatory follicle growth and steroidogenesis from prenatal testosterone disruption.

BackgroundPrenatal testosterone (T) excess from days 30-90 of gestation disrupts gonadotropin surge and ovarian follicular dynamics and induces insulin resistance and functional hyperandrogenism in sheep. T treatment from days 60-90 of gestation produces a milder phenotype, albeit with reduced fecundity. Using this milder phenotype, the aim of this study was to understand the relative postnatal contributions of androgen and insulin in mediating the prenatal T induced disruptions in ovarian follicular dynamics.MethodsFour experimental groups were generated: 1) control (vehicle treatment), 2) prenatal T-treated (100 mg i.m. administration of T propionate twice weekly from days 60-90 of gestation), 3) prenatal T plus postnatal anti-androgen treated (daily oral dose of 15 mg/kg/day of flutamide beginning at 8 weeks of age) and 4) prenatal T and postnatal insulin sensitizer-treated (daily oral dose of 8 mg/day rosiglitazone beginning at 8 weeks of age). Follicular response to a controlled ovarian stimulation protocol was tested during their third breeding season. Main outcome measures included the determination of number and size of ovarian follicles and intrafollicular concentrations of steroids.ResultsAt the end of the controlled ovarian stimulation, the number of follicles approaching ovulatory size (≥6 mm) were ~35 % lower in prenatal T-treated (6.5 ± 1.8) compared to controls (9.8 ± 2.0). Postnatal anti-androgen (10.3 ± 1.9), but not insulin sensitizer (5.0 ± 0.9), treatment prevented this decrease. Preovulatory sized follicles in the T group had lower intrafollicular T, androstenedione, and progesterone compared to that of the control group. Intrafollicular steroid disruption was partially reversed solely by postnatal insulin sensitizer treatment.ConclusionsThese results demonstrate that the final preovulatory follicular growth and intrafollicular steroid milieu is impaired in prenatal T-treated females. The findings are consistent with the lower fertility rate reported earlier in these females. The finding that final follicle growth was fully rescued by postnatal anti-androgen treatment and intrafollicular steroid milieu partially by insulin sensitizer treatment suggest that both androgenic and insulin pathway disruptions contribute to the compromised follicular phenotype of prenatal T-treated females.

The effect of ovulatory follicle size at the time of insemination onpregnancy rate in lactating dairy cows

The aims of this study were to determine the factors that affect follicle size at the time of artificial insemination (AI) and to detect the relationship between ovulatory follicle size at the time of AI and pregnancy rate in dairy cows. A total of 1428 follicle size measurements were obtained from Holstein-Friesian (HF) and Swedish Red (SR) cows that were inseminated with the fixed-timed AI protocols used in this study. Follicle size was affected by breed (P = 0.0001), milk production (P = 0.01), parity (P = 0.05), and season (P = 0.04). Follicle size was greater (P = 0.0001) in the HF (15.55 mm) than the SR cows (14.88 mm). Multiparous cows had larger (P = 0.04) follicles (15.35 mm) than the primiparous cows (15.07 mm). Cows with follicle sizes between 13.5 and 17.5 mm were more likely to be pregnant than cows with other sizes follicles (P < 0.01). Embryonic loss was lower (P < 0.01) in cows with follicle sizes between 13.5 and 16.5 mm. Thus, the follicle size was affected by breed, milk production, parity, and season. Pregnancy and embryonic loss in lactating dairy cows were significantly related to follicle size.

Expression of integrins and GDF9 mRNAs is associated with ovarian follicle size and donor puberty status in pigs

Ovarian folliculogenesis and oogenesis has a significant impact on embryo growth and development in preimplantation stages. Although both processes are widely understood in several species of mammals, including pigs, the factors influencing the proper maturation capability of oocytes, as well as the developmental competence of the surrounding somatic granulosa cells (GCs) and cumulus cells (CCs), are still not entirely known. This study aimed to investigate the expression of growth differentiation factor 9 (GDF9) and integrins (ITGB1, ITGB2, ITGB3 and ITGB4) in porcine oocytes isolated from follicles of various size and donors characterized by different puberty status. The relative abundance of GDF9, ITGB1, ITGB2, ITGB3, and ITGB4 mRNAs in porcine oocytes isolated from medium follicles of cycling sows (MFCS), small follicles of juvenile gilts (SFJG), and small follicles of cycling sows (SFCS) was assessed by an RT-qPCR assay. We found an increased expression of GDF9 in oocytes isolated from the small follicles of juvenile gilts as compared to the other two groups (P&lt;0.001). A significant down-regulation of ITGB1 and ITGB2 oocyte mRNAs collected from medium follicles of cycling sows was observed (P&lt;0.05 and P&lt;0.001, respectively). The ITGB3 mRNA expression was significantly decreased in oocytes isolated from small follicles of juvenile gilts (P&lt;0.001), whereas a lower expression of ITGB4 in oocytes from both medium follicles (cycling sows) and small follicles (juvenile gilts) was observed. In conclusion, GDF9 may be recognized as the main factor regulating follicle growth at early stages of folliculogenesis. The expression of ITGBs is significantly regulated by the puberty status of donor pigs, and different follicular sizes may play a subordinate role in integrin expression during in vivo follicle development in pigs.

A delay differential equation model of follicle waves in women

ABSTRACTThis article presents a mathematical model for hormonal regulation of the menstrual cycle which predicts the occurrence of follicle waves in normally cycling women. Several follicles of ovulatory size that develop sequentially during one menstrual cycle are referred to as follicle waves. The model consists of 13 nonlinear, delay differential equations with 51 parameters. Model simulations exhibit a unique stable periodic cycle and this menstrual cycle accurately approximates blood levels of ovarian and pituitary hormones found in the biological literature. Numerical experiments illustrate that the number of follicle waves corresponds to the number of rises in pituitary follicle stimulating hormone. Modifications of the model equations result in simulations which predict the possibility of two ovulations at different times during the same menstrual cycle and, hence, the occurrence of dizygotic twins via a phenomenon referred to as superfecundation. Sensitive parameters are identified and bifurcations in model behaviour with respect to parameter changes are discussed. Studying follicle waves may be helpful for improving female fertility and for understanding some aspects of female reproductive ageing.

Factors affecting the size of ovulatory follicles and conception rate in high-yielding dairy cows

Two studies were designed to determine (1) the effects of Heatsynch and Ovsynch protocols versus spontaneous ovulation and (2) the effects of calving problems, clinical uterine infections, and clinical mastitis on the size of the ovulatory follicle, conception rate, and embryonic/fetal (E/F) death in high-yielding dairy cows. In study 1, cows without the history of calving problems, clinical uterine infections, and clinical mastitis were randomly allocated to either an Ovsynch (n = 45) or Heatsynch (n = 39) ovulation synchronization protocol or spontaneous ovulation (n = 43) groups. Blood samples were collected on the day of artificial insemination (AI) to measure progesterone (P4), estradiol-17β, and insulin-like growth factor 1 (IGF-1) and 7 days later to measure P4. Study 2 consisted of cows (n = 351) with or without the history of calving problems, clinical uterine infections, and clinical mastitis which were artificially inseminated after a 55-day voluntary waiting period. Transrectal ultrasonography was performed at the time of AI to measure the ovulatory follicle size and on Days 30 and 68 after AI to diagnose pregnancy in both studies. In study 1, the mean (±standard error of the mean) diameter of the ovulatory follicle was greater (P = 0.0005) and E/F mortality was lower (P = 0.007) for the spontaneous ovulation group compared with Ovsynch and Heatsynch groups. Serum concentration of P4 on Day 7 after AI was correlated with the size of the ovulatory follicle (P = 0.007). Conception rate at Days 30 and 68 was not significantly different between the three experimental groups in study 1. Cows with serum IGF-1 concentrations greater than 55 ng/mL at AI had significantly higher Day 68 conception rate (50% vs. 24%) and lower E/F death (16.6% vs. 40%) compared to cows with serum IGF-1 concentrations lower than 56 ng/mL at AI. The conception rate on Days 30 and 68 for follicles of 10 to 14 mm in diameter (34% and 21.8%) was significantly lower than follicles of 14.1 to 19 mm in diameter (60% and 50%), respectively (P < 0.05). In study 2, the ovulatory follicle in cows with clinical uterine infections was smaller than that in cows without clinical uterine infections (16.4 vs. 17.1 mm; P = 0.04). In conclusion, the size of the ovulatory follicle is affected by ovulation synchronizing protocols and postpartum clinical uterine infections. In addition, cows with higher serum IGF-1 concentrations on the day of AI had higher Day 68 conception rate and lower E/F death.

Comparison of follicular dynamics, superovulatory response, and embryo recovery between estradiol based and conventional superstimulation protocol in buffaloes (Bubalus bubalis).

Aim:To evaluate the follicular dynamics, superovulatory response, and embryo recovery following superstimulatory treatment initiated at estradiol-17β induced follicular wave emergence and its comparison with conventional superstimulatory protocol in buffaloes.Materials and Methods:Six normal cycling pluriparous buffaloes, lactating, 90-180 days post-partum, and weighing between 500 and 660 kg were superstimulated twice with a withdrawal period of 35 days in between two treatments. In superstimulation protocol-1 (estradiol group) buffaloes were administered estradiol-17β (2 mg, i.m.) and eazibreed controlled internal drug release (CIDR) was inserted intravaginally (day=0) at the random stage of the estrous cycle. On the day 4, buffaloes were superstimulated using follicle stimulating hormone (FSH) 400 mg, divided into 10 tapering doses given at 12 hourly intervals. Prostaglandin F2α analogs (PGF2α) was administered at day 7.5 and day 8, and CIDR was removed with the second PGF2α injection. In superstimulation protocol - 2 (conventional group) buffaloes were superstimulated on the 10th day of the estrous cycle with same FSH dose regimen and similar timings for PGF2α injections. In both groups, half of the buffaloes were treated with luteinizing hormone (LH) 25 mg and other half with 100 ug buserelin; gonadotrophin releasing hormone (GnRH) analog at 12 h after the end of FSH treatment. All buffaloes in both protocols were inseminated twice at 12 and 24 h of LH/GnRH treatment. Daily ultrasonography was performed to record the size and number of follicles and superovulatory response.Results:Significantly higher number of small follicles (<8 mm) was present at the time of initiation of superstimulatory treatment in the estradiol group compared to the conventional group (12.5±0.80 vs. 7.3±1.21, respectively, p=0.019), however, the number of ovulatory size follicles (≥8 mm) did not differ significantly between the respective groups (15.5±1.24 vs. 12.2±1.30; p=0.054). Total embryos and transferable embryos recovered were non-significantly higher in the estradiol group compared to the conventional group (5.83±0.86 vs. 4.67±1.16, p=0.328, and 3.67±0.93 vs. 2.67±0.68, p=0.437, respectively). The significant higher proportion of transferable embryos were recovered in buffaloes treated with LH compared to GnRH (73.3% vs. 48.5%; p=0.044).Conclusion:The average number of ovulatory size follicles (>8 mm), corpora lutea, and transferable embryos was higher in buffaloes superstimulated at estradiol-induced follicular wave compared to the conventional protocol: Further the percentage of transferable embryos was significantly higher in buffaloes administered with LH compared to GnRH.

Prediction of Normal Ovulation by Sonographic Folliculometry Involving Natural Cycles among Women in Ojo, Southwest Nigeria.

Accurate prediction of ovulation is important in the management of female infertility. To determine the sonographic sensitivity of reduction in follicular size and disappearance of ovarian follicle as predictors of imminent ovulation. This was a longitudinal study involving 100 women between the ages of 18 and 35 years. Transvaginal sonography with 6.5 MHz probe frequency was performed with a General Electric (RT 2800) ultrasound machine. Dominant follicles were identified and measured in both the longitudinal and transverse planes and their disappearance was monitored prior to ovulation. Laboratory luteinizing hormone test strips were used to test serum samples collected daily from each patient to confirm the time of ovulation. Pre-ovulation follicular size among the subjects was in the range of 18-36 mm while the mean follicular size was 26.78 ± 4.03 mm. Prior to ovulation, disappearance and reduction in follicular size was noted in 59% and 41% of subjects, respectively. Luteinizing hormone test was also positive and peaked prior to ovulation in 92% of the subjects among whom follicles disappeared in 37% while their size reduced in 55%. There was no statistically significant difference between sonographic and laboratory findings (p > 0.05). Age, height, weight and body mass index do not have significant influence (p > 0.05) on follicular size and ovulation. Sonographic observation of complete disappearance of a dominant follicle and reduction in follicular size of surrogate follicles after follicular rupture appeared to be a reliable predictor of imminent ovulation.

Diameter of the ovulatory follicle at timed artificial insemination as a predictor of pregnancy status in lactating dairy cows subjected to GnRH-based protocols

Ovulation of an early-stage dominant follicle induced by exogenous GnRH reportedly results in a reduced ovulatory follicle size and fertility in beef and dairy cattle. Objectives were to examine factors associated with ovulatory follicle diameter and the relationship between ovulatory follicle diameter and pregnancy per artificial insemination (P/AI) and pregnancy loss in lactating dairy cows subjected to GnRH-based programs for timed artificial insemination (TAI). A total of 1048 ovulations from 1576 breedings of cows that had or had not been presynchronized and subjected to a 5- or a 7-day GnRH-based protocol with or without progesterone supplementation between initial GnRH and PGF2α were analyzed. Ultrasonography was used to determine cyclicity, ovulation, and pregnancy status and to measure diameter of the ovulatory follicle at the time of TAI. Mean (±standard error) diameter of the ovulatory follicle was 16.4 ± 0.1 mm (range, 11–25 mm; n = 1048); the most common diameters were 15 (n = 170), 16 (n = 169), and 17 mm (n = 170). Multiparous cows and those subjected to a 7-day protocol had larger ovulatory follicles (P < 0.05). On average, multiparous cows that did not ovulate after the initial GnRH had the largest ovulatory follicles (17.0 ± 0.1 mm; n = 290), whereas all cows that ovulated after the initial GnRH and were subjected to a 5-day GnRH-based protocol had the smallest ovulatory follicles (15.6 ± 0.2 mm; n = 207). The associations between ovulatory follicle diameter and P/AI at 32 and 60 days after TAI were not significant. However, a significant linear relationship between ovulatory follicle diameter and pregnancy loss between 32 and 60 days after TAI was observed, with an increased (P < 0.05) probability of pregnancy loss in cows with an ovulatory follicle greater than 20 mm in diameter. In summary, the diameter of the ovulatory follicle preceding TAI was affected by interactions between ovulatory response to initial GnRH and parity and between ovulatory response to initial GnRH and the length of ovulation synchronization protocol. Ovulatory follicle diameter was not associated with P/AI at 32 or 60 days, but large ovulatory follicle size was identified as a predictor of pregnancy loss in lactating dairy cows subjected to GnRH-based protocols for TAI.

The efficacy and safety of clomiphene citrate and metformin on ovulation induction in patients suffering from anovulatory infertility

Background: Ovulation disorders are an important cause of infertility. Polycystic ovarian syndrome (PCOS) comprises almost 90% cases of all ovulatory disorders. Clomiphene citrate helps to stimulate ovulation. Recently, use of metformin to treat PCOS due to its insulin sensitizing property is increasing. Hence, this study was undertaken to evaluate efficacy and safety of clomiphene citrate and its combination with metformin on ovulation induction and pregnancy outcome in the treatment of anovulatory infertility. Methods: This study was done in patients of primary infertility due to anovulatory cycles at Civil Hospital, Ahmedabad. Patients were treated with clomiphene citrate in incremental doses from 50 to 150 mg for a period of maximum 6 menstrual cycle. In patients of anovulatory infertility with PCOS, clomiphene citrate was given along with metformin. At every visit, abdominal and pelvic examination, the size of ovarian follicle, endometrial thickness by ultrasonography was recorded. Urine pregnancy test was done once the menstruation was overdue by 9 days. Adverse drug effects, if any were noted. If patient did not conceive, then whole treatment was repeated from the next menstrual cycle. Results: Out of 53 patients, five patients were lost to follow-up. The mean age of the enrolled patients in the study was 26.04±7.78. There were 20 patients (41.67%) with body mass index (BMI)

Vascular endothelial growth factor (VEGF) and placental growth factor (PLGF) directly correlate with ovarian follicle size in women undergoing in vitro fertilization (IVF)

To assess VEGF and PlGF concentrations in FF from women undergoing IVF, stratified according to follicle size, and to correlate these with VEGF and PlGF mRNA expression in corresponding luteinized granulosa cells (GC), as a follow-up of our previous demonstration that these genes are expressed in GC(1). Prospective sample collection for in vitro studies. 18 consenting subjects with normal ovarian function (IVF being undertaken due to male factor) were recruited prior to oocyte retrieval (mean age of the women = 31 years). Only blood-free FF obtained with the first puncture of each ovary was collected, without the use of flushing media to avoid dilution of the FF contents. All specimens were categorized and analyzed as FF derived from small follicles (8-14 mm diameter) or large follicles (>14 mm diameter). VEGF and PlGF concentrations in the FF were measured using validated commercial ELISAs (RayBio®). Levels of FF estradiol (E2) and progesterone (P4) were determined by RIA. VEGF and PlGF mRNA expression was evaluated by real time RT-PCR, using cultured GC derived from the small and large follicles as indicated above. We also evaluated the expression of other relevant ovarian genes. FF VEGF concentrations from large follicles were markedly higher (6788 ± 2863 pg/ml) compared to those in small follicles (1942 ± 879 pg/ml, p = 0.0003). PlGF concentrations in FF from large follicles also was higher (2004 ± 816 pg/ml) than in small follicles (1144 ± 898 pg/ml, p =0.002). By RT-PCR, the expression of VEGF mRNA was ∼2.5 fold greater in GC from large follicles than from small follicles. However, we observed no significant statistical differences in PlGF mRNA expression relative to follicle size. As expected, expression of mRNAs encoding steroidogenic acute regulatory protein (StAR), anti-mullerian hormone (AMH) and aromatase P450 (CYP19) were all increased in cells from large follicles as compared with those from small follicles. VEGF and PlGF protein concentrations in FF are correlated with follicle size; this relationship also applied to E2 concentration and StAR, AMH and CYP19 expression. Moreover, VEGF and PlGF levels were significantly higher in FF than in matched serum. Similar trends were noted for VEGF mRNA, but not PlGF mRNA expression from cultured GC. We conclude that local VEGF and PlGF gene expression are likely to mediate ovarian follicle angiogenesis and may contribute to oocyte development, maturation and selection of dominant follicle.

Comparison of endocrine changes, timing of ovulations, ovarian follicular growth, and efficacy associated with Estradoublesynch and Heatsynch protocols in Murrah buffalo cows (Bubalus bubalis)

Poor estrus expression and the difficulty encountered in predicting the time of ovulation compromise the reproductive efficiency of Murrah buffalo cows. Synchronization of ovulation and timed artificial insemination are able to precisely control the time of ovulation and thus avoid the need for estrus detection. Recently, the Estradoublesynch protocol (administration of a PGF2α injection 2 days before Heatsynch protocol; GnRH 0, PGF2α 7, estradiol benzoate [EB] 8) was developed that precisely synchronized ovulation twice, i.e., after GnRH and EB injections and resulted in satisfactory pregnancy rates in Murrah buffaloes. The present study was conducted on 104 cycling and 31 anestrus buffaloes to compare (1) the endocrine changes, timing of ovulations, ovarian follicular growth, and efficacy of Estradoublesynch and Heatsynch protocols in cycling and (2) the efficacy of Estradoublesynch and Heatsynch protocols for the improvement of fertility in cycling and anestrus Murrah buffalo cows. Ovulation was confirmed after all GnRH and EB treatments by ultrasonographic examination at 2-hour intervals. Plasma progesterone and total estrogen concentrations were determined in blood samples collected at daily intervals, beginning 2 days before the onset of protocols until the day of second ovulation detection. Ovulatory follicle size was measured by ultrasonography at six time points (first PGF2α administration of Estradoublesynch protocol every 2 days before the onset of Heatsynch protocol, GnRH administration of both protocols, 2 hours before ovulation detection after GnRH administration of both protocols, second PGF2α injection of Estradoublesynch protocol, PGF2α injection of Heatsynch protocol, EB injection of both protocols and, 2 hours before ovulation detection after EB administration of both protocols). Plasma LH, total estrogen, and progesterone concentrations were determined in blood samples collected at 30-minute intervals for 8 hours, beginning GnRH and EB injections, and thereafter at 2-hour intervals until 2 hours after the detection of ovulation. The first ovulatory rate was significantly higher (P < 0.05) in the Estradoublesynch protocol (84.6%) than that in the Heatsynch protocol (36.4%). The first LH peak concentration (74.6 ± 10.4 ng/mL) in the Estradoublesynch protocol was significantly higher (P < 0.05) than that of the Heatsynch protocol (55.3 ± 7.4 ng/mL). In Estradoublesynch protocol, the total estrogen concentration gradually increased from the day of GnRH administration coinciding with LH peak, and then gradually declined to the basal level until the time of ovulation detection. However, in Heatsynch protocol, the gradual increase in total estrogen concentration after GnRH administration was observed only in those buffalo cows, which responded to treatment with ovulation. In both Estradoublesynch and Heatsynch protocols, ovulatory follicle size increased by treatment with GnRH and EB until the detection of ovulation. The pregnancy rate after the Estradoublesynch protocol (60.0%) was significantly higher (P < 0.05) than that achieved after the Heatsynch protocol (32.5%). Satisfactory success rate using the Estradoublesynch protocol was attributed to the higher release of LH after treatment with GnRH, leading to ovulation in most of the animals and hence creating the optimum follicular size at EB injection for ovulation and pregnancy to occur.

Endocrine control of embryonic diapause in the Australian sharpnose shark Rhizoprionodon taylori.

The reproductive cycle of the Australian sharpnose shark, Rhizoprionodon taylori, includes a temporary suspension of development at the commencement of embryogenesis termed embryonic diapause. This study investigated levels of 17β-estradiol (E2), testosterone (T) and progesterone (P4) in plasma samples of mature wild female R. taylori captured throughout the reproductive cycle and correlated them with internal morphological changes. Levels of T were elevated through most of the embryonic diapause period, suggesting a role of this hormone in the maintenance of this condition. Increasing plasma T concentrations from late diapause to early active development were associated with a possible role of androgens in the termination of embryonic diapause. As in other elasmobranchs, a concomitant increase of E2 with ovarian follicle size indicated a direct role of this hormone in regulating vitellogenesis, while a peak in P4 suggested this hormone is associated with preovulation and ovulation. Additionally, significant correlations between photoperiod or water temperature and maximum follicular diameter and hepatosomatic index suggest that these abiotic factors may also play a role triggering and regulating the synchrony and timing of reproductive events.

The Comparison of Ovarian Reserve After Uterine Artery Embolization and Hysterectomy for Uterine Fibroma

Background: The purpose of this study was to compare the ovarian reserve before and after uterine artery embolization and hysterectomy for uterine fibroma treatment. Methods: After obtaining the approval of the Ethics Committee of Kermanshah Medical University, this study was carried out in Imam Reza Hospital. Between March 2011 and May 2013, 66 women who suffered from symptomatic fibroma were studied. Hysterectomy with retained ovaries was done on 33 patients and uterine artery embolization (UAE) was done on the other 33 patients. The levels of hemoglobin (Hb), follicle stimulating hormone (FSH) and ovarian follicle size before and 6 months after the operation was measured. Vaginal bleeding, reintervention, hot flash and cost in both procedures were studied. Results: No significant difference was observed after 6 months between the two groups (P = 0.356). In each group one case with FSH above 10 IU/L was observed. The existence of dominant follicle and the size of it was the same in the two groups (P = 0.390). But the level of Hb was higher in women under embolization 6 months after intervention (P = 0.001). The average level of pain after hysterectomy was higher than embolization. The cost of embolization was 12 times higher than hysterectomy. Conclusion: This study indicated that UAE in young women had no impact on the ovarian reserve and did not increase the level of FSH. Long term studies regarding reproductive assessment and menopausal age in women with history of UEA are necessary. J Clin Gynecol Obstet. 2014;3(2):62-67 doi: http: //dx.doi.o rg/10.14740/jcgo239 w