Simple, rapid and semiquantitative enzyme-linked immunoassay for detection of the preovulatory LH excretion in the urine.
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Twenty-two healthy women participated in a study to determine whether roxithromycin (a new macrolide antibiotic agent) obtunds the activity of a triphasic oral contraceptive. The duration of the study was four menstrual cycles. Medication was given as follows: (1) cycle 1, no medication to demonstrate ovulation; (2) cycle 2, triphasic oral contraceptive daily to suppress ovulation; (3) cycle 3, triphasic oral contraceptive daily plus roxithromycin, 150 mg b.i.d.; and (4) cycle 4, triphasic oral contraceptive daily plus rifampin, 300 mg daily. Sonography of the ovaries was performed on day 13, and serum progesterone was measured on day 21 of each cycle. Elevated progesterone indicated ovulation. The presence of a maturing follicle supported this finding. All volunteers ovulated in the first cycle and no volunteers ovulated in the second and third cycles. However, 11 women ovulated when rifampin and the triphasic oral contraceptive were given concomitantly. The findings suggest there is no reason to believe that roxithromycin interferes with the efficacy of oral contraceptives.
The efficiency of superovulating mares with an enriched fraction of equine follicle-stimulating hormone (feFSH) and an equine pituitary extract (EPE) with similar FSH content but differing in the LH amount was compared. Mares were randomly assigned to an feFSH (n = 5) or EPE (n = 5) treatment. The experimental period was of 2 successive estrous cycles, with the first cycle as the control. At Days 6 and 7 of the estrous cycle, the mares received 250 micrograms i.m. cloprostenol. The treatments consisted of daily injections of 25 mg feFSH or EPE beginning on Day 6 post ovulation. Mares were inseminated every other day until the last ovulation was detected. When the mares in the control and treatment cycles developed at least 1 or 2 > or = 35-mm follicle, respectively, the treatment was interrupted, and a single injection of EPE (25 mg, i.v.) was administered to induce ovulation(s). Nonsurgical embryo recovery was performed 6 or 7 d after ovulation in both control and treatment cycles. The number of ovulations per mare was not significantly different (P > 0.05) between feFSH and EPE groups, but both were higher (P < 0.05) than that of the control cycle. The number of recovered embryos per ovulation was similar (P > 0.05) for control, feFSH and EPE groups. The high amount of LH presented in EPE did not affect the superovulatory response of the mares. Superovulatory treatments increased the ovulation rate of mares but did not affect the embryo recovery rate per ovulation.
One hundred and twenty-two patients who attended the author's Gynaecological Clinic at The Logas University Teaching Hospital complaining of infertility, amenorrhoea or irregular menstruation were each studied during a minimum of three menstrual cycles. Cytological Curves based on weekly karyopyknotic indices of their vaginal epithelial cells were plotted throughout the duration of the study. Basal body temperatures and endometrial biopsies were also obtained from all of the patients. In the absence of vaginitis, cytological studies were found to be useful in determining the period of maximal fertility in infertile women and in detecting abnormal oestrogen excretion in women with functional menstrual abnormalities.
Application of time series analysis to a database containing serial pregnanediol data from 113 complete ovulatory menstrual cycles contributed by 83 women of proven fertility and 68 cycles for which pregnanediol values were available over the ovulatory period, detected the first statistically significant risk in pregnanediol excretion for all cycles for which a baseline was available (n = 170). However, even at the 99% confidence level, for 22% of cycles a rise was observed before the presumed day of ovulation. Therefore, a threshold value for pregnanediol was sought from the database as a better marker for the end of fertility. A value of 1.4 mg per 24 h was not reached before day 2 after the pre-ovulatory estrogen peak day for 96% of the cycles. In the remaining 4% of cycles it was reached one day after the total estrogen peak day. The validity of this threshold was confirmed in extensive studies using the Ovarian Monitor where the equivalent is 6.3 mumol per 24 h of pregnanediol glucuronide and measurements are performed on timed urine specimens with a minimum collection time of three hours. These studies were as follows: 1) a World Health Organization study on the use of the Ovarian Monitor as a fertility self test in the home (108 cycles), 2) a multicenter study on returning fertility during breast feeding conducted by Family Health International (73 women), and 3) the general application of the Ovarian Monitor for pregnancy achievement and avoidance during the past ten years (over 250,000 PdG assays performed in ten countries). With rare exceptions, the use of these threshold values is applicable for all women provided correction is made for urine volume.
To investigate the effects of prostaglandin (PGF 2alpha) plus GnRH at different stages of the luteal phase 13 ewes received PGF 2alpha on Day 9 of the synchronized cycle, followed 36 h later by GnRH. This control regimen resulted in ovulation and normal corpus luteum (CL) function. In the next cycle, the ewes were treated simultaneously with PGF 2alpha and GnRH either on Day 4 (early, n = 7) or Day 9 (late, n = 6). Ovarian activity was monitored daily by ultrasonography, and blood samples were obtained to monitor hormonal patterns. Size of the largest follicle present when GnRH was administered was similar in all groups, but the preceding growth rate was greatest for the early group. In the 36 h after injection of PGF 2alpha, serum progesterone (P4) had declined to basal levels in the control cycles when GnRH was administered, but P4 concentrations were higher in the early group and were highest in the late group when the GnRH was administered with PGF 2alpha. The LH surges induced by GnRH were highest in the control cycles, and were lower in the 2 treated groups. In the early group, 6 of 7 ewes demonstrated ovulation within 48 h of GnRH, resulting in the formation of normal CL. In the late group, ovulation was delayed for about 5 d in 4 of 6 ewes, and subsequent luteal function was normal; no ovulation was detected in the other 2 ewes of this group, but the follicles became luteinized, resulting in a normal P4 profile in one and subnormal in the other. These results suggest that follicles present during the early luteal phase are capable of ovulating and forming fully functional CL in response to exogenous GnRH. In contrast, follicles present during the late luteal phase fail to ovulate in response to GnRH while P4 levels are high, even though the LH stimulus is adequate; however, these follicles persist and subsequently ovulate after P4 levels have decreased. Therefore, the endocrine milieu to which a follicle was exposed may be more important than its size in determining its ability to undergo ovulation and development into a normal CL.
Precise data on fertility results following peri- and postovulatory insemination in spontaneously ovulating gilts is lacking. Using transcutaneous sonography every 4 h during estrus as a tool for diagnosis of ovulation, the effects of different time intervals of insemination relative to ovulation were investigated with liquid semen (Experiment 1, n=76 gilts) and frozen semen (Experiment 2, n=80 gilts). In Experiment 3 (n=24 gilts) the number of Day-28 embryos related to the various intervals between insemination and ovulation was determined after the use of liquid semen. Using liquid semen the fertilization rates based on Day-2 to Day-5 embryos and the number of accessory spermatozoa decreased significantly in gilts inseminated with 2 x 10(9) spermatozoa per dosage in intervals of more than 12 h before or more than 4 h after ovulation. In the time interval 4 to 0 h before ovulation, comparable fertilization rates were obtained using frozen semen (88.1%) and liquid semen (92.5%). Fertilization rates and numbers of accessory spermatozoa decreased significantly when gilts were inseminated with frozen semen more than 4 h before or 0 to 4 h after the detection of ovulation. The percentage of Day-28 embryos was significantly higher following preovulatory insemination compared to inseminations 0 to 4 h and 4 to 8 h after ovulation. It is concluded that the optimal time of insemination using liquid semen is 12 to 0 h before ovulation, and 4 to 0 h before ovulation using frozen semen. The results stress the importance of further research on sperm transport and ovulation stimulating mechanisms, as well as studies on the time of ovulation relative to estrus-weaning intervals and estrus duration.
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Legumes can obtain nitrogen from symbiotic nitrogen fixation in root nodules. The glutamine synthetase/glutamate synthase cycle is responsible for the initial nitrogen assimilation. This work reports the analysis of transgenic Lotus japonicus plants with the chimeric gene containing the alfalfa cytosolic glutamine synthetase (GS1) (EC 6.3.1.2) gene controlled by the Sesbania rostrata leghemoglobin gene promoter (Srglb3p). Surprisingly, all of the transgenic primary transformants analysed were sterile. Two transformants designated GS39 and GS44 were further analysed. GS in nodules of GS39 and GS44 plants was upregulated, at the level of transcript and protein. The transgenic plants had 2-fold higher nodule GS activity and similar root GS activity compared to control plants. The GS39 and GS44 sterile plants showed morphological alterations in pollen grains and in ovules. An increase in GS transcript abundance and enzyme activity was measured during early and late stages of flower development of GS plants. Flowers of GS plants showed higher glutamine content, resulting in an increased glutamine/glutamate ratio. The GS transcript and protein were detected in ovules. These data indicate that overexpression of GS1 in reproductive organs critically affects their development and might be a reason for sterility of L. japonicus plants.
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Plasma FSH, LH, estradiol (E2) and progesterone (P4) profiles and patterns of follicular growth and regression by ultrasonography were determined after E2 treatment (1 microg/kg) in anestrous ewes. Fifteen ewes were treated with one (group I, n=7) or two (group II, n=4) i.m. injections of E2 with a 24h interval, or two oil injections with a 24h interval (group C, n=4). Blood samples for E2, P4, FSH and LH determinations were collected daily 4 days before the initiation of the treatment (day 0), when bleeding increased to every 2h starting 2h before treatment until 56h after the first injection and from then on every 6h until day 8, and twice per day till the end of the experiment (day 9). During the experimental period (days -4 to 9), transrectal ultrasonic examinations were carried out daily using a 7.5 MHz linear array probe. Number and size of follicles > or =3mm in diameter were recorded. No estrous was detected before, during or after treatment. LH and FSH surges were observed 10-18h after the first E2 injection. The second E2 injection stimulated another release of LH but no surges. E2 inhibited FSH levels before the surge and the second E2 injection induced a longer inhibition. No ovulation was detected by ultrasonography during the experimental period and P4 levels remained low (<0.7 nmol/l) before, during and after the treatment in all ewes. There was an effect of E2 treatment on the diameter of the largest follicle, a decrease could be observed 3 days after the first injection in both ewes of groups I and II. The E2-treated groups had a higher frequency of ewes showing wave emergence on day 3 (day 1.5+/-1,2.4+/-0.4 and 2.5+/-0.5 for control, groups I and II). LH and FSH surges were observed after E2 treatment, but were not able to provoke ovulation neither luteinization. In contrast, the treatment was associated with the regression of the largest follicle and with emergence of a new follicular wave on day 3.
Female infertility can be categorised into those who fail to ovulate (anovulatory infertility) because of some defect at hypothalamic-pituitary-ovarian axis and those who are ovulatory (ovulatory infertility), but are infertile because of some lesion present in genital tract. The role of vaginal hormonal cytology, endometrial biopsy and endocrinological evaluation in the detection of ovulation and various ovulatory dysfunction was studied in 42 infertile female patients. On the basis of cytological findings, of the 42 patients, 14 were found to be ovulatory, 26 anovulatory (which include 5 cases of atrophic changes) and 2 inconsistent due to inflammatory changes. Endometrial biopsy showed evidence of ovulation in 15, anovulation in 27 cases. Hormonal evaluation indicated some sort of endocrinological disorders in 15 patients, which may underlie anovulatory infertility in these patients, while results were within normal range in the rest 27 patients. Results of vaginal cytology and endometrial biopsy showed correlation in respect to ovulation in 93.33% of the cases.
A computerized pump for continuous pulsatile HMG administration was used with 5 patients having prolonged anovulatory infertility, for 14 consecutive cycles. All of these women had previously failed to respond to various treatment regimens, or repeatedly developed ovarian overstimulation syndrome. In 12 out of 14 cycles, ovulation was detected by hormonal and ultrasonic evaluation, and pregnancy was achieved in 2 out of 5 cases. A mild form of overstimulation was observed in only 3 out of 14 cycles. A summary of the results of the present and similar previously published series demonstrated overall ovulation and pregnancy rates of 75% and 23.5% respectively. We concluded that inducing ovulation by administering pulsatile HMG is an appropriate alternative method of treatment when ovulation and conception fail with conventional regimens.
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Changes in follicular and luteal structures were assessed and concentrations of estradiol and progesterone were measured in 13 Hereford X Angus suckled beef cows during resumption of estrous cycles. Transrectal ultrasonography was used to monitor follicular size, ovulation, and formation and regression of the corpus luteum (CL). The interval from parturition to first postpartum ovulation (FO) was 82 +/- 4.7 d. Serum progesterone remained low before FO. One cow exhibited standing estrus, two cows showed other signs of estrus, and 10 displayed no signs of behavioral estrus preceding FO. All cows exhibited standing estrus before the second postpartum ovulation (SO). All cows had a short luteal phase after FO, with an average interval of 8.5 +/- .2 d between FO and SO. Concentrations of estradiol in serum during the 8 d preceding ovulation were similar before FO and SO. Maximal diameter of the preovulatory follicle was similar before FO and SO. However, the ovulatory follicle was larger in diameter at 2 d (P = .02) and 3 to 8 d (P less than .005) before FO than before SO. The time from detection until ovulation was less (P = .005) for the ovulatory follicle preceding SO than for the follicle associated with FO (8.5 vs 10.2 d, respectively, SE = .4). The second-largest follicle was larger (P less than .005) in diameter during the 8 d preceding the FO than before the SO. The difference in size between the ovulatory follicle and the second-largest follicle on the day before ovulation was greater (P less than .005) preceding SO than preceding FO (8.7 vs 6.6 mm, respectively, SE = .4).(ABSTRACT TRUNCATED AT 250 WORDS)
Induction of ovulation is indicated for anovulatory and amenorrheic women as well as for women who have an inadequate luteal phase. It is also indicated as a strategy for recruiting multiple follicles for women with unexplained infertility and those who are undergoing assisted reproductive technologies. The use of various agents and detection of ovulation are described. This includes a discussion of clomiphene citrate, bromocriptine, human menopausal gonadotropins, urinary follicle stimulating hormone and pulsatile gonadotropin releasing hormone therapy. Regimens, success rates and potential complications of each form of therapy are reviewed. Also discussed is the use of combination therapy and partial ovarian destruction.
A review of the in vitro fertilization and embryo transfer program between July 1979 and December 1980 at The Royal Women's Hospital, Melbourne, is presented. A total of 402 treatment cycles involving 186 patients have produced five pregnancies. Two pregnancies resulted from spontaneous ovulatory cycles and three from stimulated cycles. One live birth of a normal female child is reported. The protocol for both spontaneous and stimulated cycles and the problems that each produces are discussed. It is concluded that the stimulated cycle is to be preferred, because it allows higher rates of ovum recovery and embryo transfer. The fertilization and cleavage rates are the same for both methods. Associated research into in vitro blastocyst development, steroidogenesis in the corona cells, and the detection of other "early pregnancy factors" is presented and discussed. The paper concludes with a short statement on the ethical and moral problems posed by the research.