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Effect of exogenous progesterone on pregnancy rates after surgical embryo transfer in mares.

A completely randomized experimental design was used to investigate the effect of supplemental progesterone on pregnancy rates of recipient mares. Every other recipient mare received daily 200 mg progesterone in oil beginning the day of surgical embryo transfer and lasting until either Day 120 of pregnancy or until pregnancy failure was confirmed by ultrasound. Progesterone supplementation did not affect pregnancy rate (P > 0.05). Overall, embryos that did not result in pregnancy were of greater mean diameter than embryos that resulted in pregnancy (P < 0.05). Pregnancy rates tended (P < 0.1) to be greater in recipients that were detected to be ovulating the same day or prior to that of the donor and that had been supplemented with progesterone (75 %) as opposed to untreated control mares of the same synchrony group (40 %). Progesterone supplementation did not affect the incidence of embryonic loss; however, there was a slightly higher loss of pregnancies between Day 15 and 30 in treated versus untreated recipients. There was no effect (P > 0.05) of treatment on pregnancy rate for embryos recovered from fertile versus subfertile donor mares. However, overall, there tended (P < 0.1) to be fewer pregnancies with embryos recovered from subfertile (50 %) as compared to fertile donors (75 %). It was concluded that supplemental progesterone at the dosage and frequency described was not beneficial in improving pregnancy rates in cyclic recipient mares after surgical embryo transfer.

Journal Article↗

The promoter of a metallothionein-like gene from the tropical tree Casuarina glauca is active in both annual dicotyledonous and monocotyledonous plants.

A chimeric gene consisting of the beta-glucuronidase (gusA) reporter gene under the control of the metallothionein-like promoter cgMT1 from the tropical tree Casuarina glauca was introduced into Nicotiana tabacum via Agrobacterium tumefaciens and into Oryza sativa by particle bombardment. The strongest histochemical staining for GUS activity was observed in the root system of the transgenic plants, and especially in lateral roots. In contrast, a relatively low level of reporter gene expression was seen in the aerial tissues and GUS staining was located mainly in the plant vascular system. The average ratio of GUS activity between root and leaf was found to be 13:1 in tobacco and 1.5:1 in rice. The pattern of cgMT1 promoter activity in floral organs was found to be different in tobacco and rice. High levels of gusA gene expression were detected in the ovules, pollen grains and tapetum, whereas in rice PcgMT1 directs expression to the vascular system of the floral organs. These results suggest that PcgMT1 is potentially useful in molecular breeding to express genes of interest whose products are preferentially needed in roots.

Cotyledon↗

The purification of luteinizing-hormone-releasing factor with some observations on its properties.

Bullock median-eminence tissue was used as a source of luteinizing-hormone-releasing factor for small-scale experiments to explore methods for its isolation. The presence of luteinizing-hormone-releasing factor was detected by the ovulation response in rabbits after intrapituitary infusion of the extract. Gel filtration was found to be suitable for the purification of these extracts. The releasing factor appeared to be a basic peptide of molecular weight in the range 1200-2500. On a larger scale, an extract of hypothalamic tissue from sheep was used to establish a multi-stage isolation procedure that resulted in a 200000-fold purification of luteinizing-hormone-releasing factor. After the initial extraction the isolation process consisted of: (1) two cycles of gel filtration; (2) anion-exchange chromatography; (3) gel filtration in a partially organic medium; (4) thin-layer chromatography on cellulose. Stage (3) separated two zones of activity each containing peptides. One of these was purified further by stage (4) to give a preparation that was active at a dose of 6mug. of peptide/animal, although activity diminished seriously during storage. This preparation contained only five or six components, but the small amounts of peptides obtained at this stage of purity were insufficient for full characterization.

Animals↗

[Reactions to stress during the normal menstrual cycle].

In the present study, we questioned, whether the variability of physiological parameters within a menstrual cycle are connected with psychological variables like mood, state of health and positive/negative daily occurrences. We examined 20 women with normal cycles. The definition of a normal cycle was: 28 (+/- 5) days of length, standard weight according to Broca, 3-5 days menstrual bleeding, no oral contraceptive during the last 6 months, no IUP, inconspicuous hormones (no raised prolactin or testosterone levels), gynecologically unremarkable findings. Gynecological and psychological data were gathered daily during one menstrual cycle: hormone levels, vaginal ultrasound for detecting follicle growth, ovulation, corpus luteum and endometrial development. The psychological data on mood, state of health and positive/negative occurrences were recorded in parallel in the form of a diary, in which new entries were made daily. The results showed, that only 6 out of the 20 women had a normal menstrual cycle as defined above. Women with a normal (physiological) cycle, report in their diary: less negative bodily tendencies, more positive everyday situations, but the same amount of negative situations as women with noticeable cycles. Both the normal cycle group and the conspicuous cycle group report more negative body states and more negative moods in the premenstruum than the middle phase. The implications of these results are discussed.

Adult↗

Cloning and localization of two multigene receptor families in goldfish olfactory epithelium.

Goldfish reproduction is coordinated by pheromones that are released by ovulating females and detected by males. Two highly potent pheromones, a dihydroxyprogesterone and a prostaglandin, previously have been identified, and their effects on goldfish behavior have been studied in depth. We have cloned goldfish olfactory epithelium cDNAs belonging to two multigene G-protein coupled receptor families as a step toward elucidating the molecular basis of pheromone recognition. One gene family (GFA) consists of homologs of putative odorant receptors (approximately 320 residues) found in the olfactory epithelium of other fish and mammals. The other family (GFB) consists of homologs of putative pheromone receptors found in the vomeronasal organ (VNO) of mammals and also in the nose of pufferfish. GFB receptors (approximately 840 residues) are akin to the V2R family of VNO receptors, which possess a large extracellular N-terminal domain and are homologs of calcium-sensing and metabotropic glutamate receptors. In situ hybridization showed that the two families of goldfish receptors are differentially expressed in the olfactory epithelium. GFB mRNA is abundant in rather compact cells whose nuclei are near the apical surface. In contrast, GFA mRNA is found in elongated cells whose nuclei are positioned deeper in the epithelium. Our findings support the hypothesis that the separate olfactory organ and VNO of terrestrial vertebrates arose in evolution by the segregation of distinct classes of neurons that were differentially positioned in the olfactory epithelium of a precursor aquatic vertebrate.

Amino Acid Sequence↗

Value and limitations of vaginal ultrasonography--a review.

Vaginal ultrasonography has become an indispensable method in infertility and early pregnancy investigations, in spite of its comparatively recent introduction. Initially this technique was introduced as an alternative for abdominal ultrasonography in in-vitro fertilization treatment for monitoring follicle growth and harvesting oocytes. In due course, the technique was used in many other fields of gynaecology and obstetrics, e.g. ovulation induction, early detection of (ectopic) pregnancy, oncology and placenta localization. The value and limitations of vaginal ultrasonography are discussed as well as patients' acceptance.

Female↗

Post-partum anovulation in nursing mothers.

An analysis of breastfeeding and ovulation data from 72 nursing mothers is presented. A proportional hazards model of the data strengthens the hypothesis that a pattern of short, frequent bouts of nursing defines a risk-set of women less susceptible to post-partum ovulation than those who choose to nurse on a schedule of relatively lengthy and infrequent bouts. In contrast, no effect of maternal weight-for-height on the month-specific rate of post-partum ovulation could be detected in the study population comprised of American mothers trained in self-observation of fertility signs.

Anovulation↗

Steady-state concentrations of messenger ribonucleic acid encoding cytochrome P450 side-chain cleavage and 3 beta-hydroxysteroid dehydrogenase/delta 5,delta 4 isomerase in ovine corpora lutea during the estrous cycle.

The objective of these experiments was to determine the pattern of mRNA expression for cytochrome P450 side-chain cleavage (P450scc) and 3 beta-hydroxysteroid dehydrogenase/delta 5,delta 4 isomerase (3 beta-HSD) during luteinization of the follicle and in ovine luteal tissue on Days 3, 6, 9, 12, and 15 of the estrous cycle. Mean concentration of mRNA for P450scc was not different in follicles collected 4 or 24 h after the onset of estrus but increased (p < 0.05) 3-fold by 48 h (corpus hemorrhagicum). With the methods used, mRNA for 3 beta-HSD was not detected until after ovulation and formation of the corpus hemorrhagicum (48 h after onset of estrus). In luteal tissue, mean concentration of mRNA for P450scc increased from Days 3 to 9 (p < 0.05) and had decreased (p < 0.05) by Day 15. Mean concentration of mRNA for P450scc was higher (p < 0.05) in small luteal cells on Day 9 than on Day 15, with values on Days 6 and 12 being intermediate. In large luteal cells, mean concentrations of P450scc mRNA increased (p < 0.05) between Days 6 and 12 and then decreased (p < 0.05) on Day 15. Mean concentration of mRNA for 3 beta-HSD was not different (p = 0.33) in luteal tissue on any day examined. In small luteal cells, mean concentrations of mRNA for 3 beta-HSD decreased between Days 6 and 15 (p < 0.05) while in large luteal cells, mean concentrations decreased (p < 0.05) between Days 12 and 15.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Ectopic expression of carpel-specific MADS box genes from lily and lisianthus causes similar homeotic conversion of sepal and petal in Arabidopsis.

Two MADS box genes, Lily MADS Box Gene 2 (LMADS2) and Eustoma grandiflorum MADS Box Gene 1 (EgMADS1), with an extensive similarity to the petunia (Petunia hybrida) FLORAL BINDING PROTEIN 7/11 and Arabidopsis AGL11, were characterized from the lily (Lilium longiflorum) and lisianthus (Eustoma grandiflorum). The expression of LMADS2 and EgMADS1 mRNA was restricted to the carpel and was absent in the other flower organs or vegetative leaves. LMADS2 mRNA was detected mainly in ovules and weakly in style tissues of the carpel, whereas EgMADS1 mRNA was only expressed in the ovules. Transgenic Arabidopsis plants ectopically expressing LMADS2 or EgMADS1 showed similar novel phenotypes resembling 35S::AGAMOUS plants by significantly reducing plant size, flowering early, and losing inflorescence indeterminacy. Ectopic expression of these two genes also generated similar ap2-like flowers by inducing homeotic conversion of the sepals into carpel-like structures in which stigmatic papillae and ovules were observed. In addition, the petals were converted into stamen-like structures in the second whorl of 35S::LMADS2 and 35S::EgMADS1 transgenic Arabidopsis. Our data indicated that LMADS2 and EgMADS1 are putative D functional MADS box genes in lily and lisianthus with a function similar to C functional genes once ectopically expressed in Arabidopsis.

Amino Acid Sequence↗

Arabidopsis RGL1 encodes a negative regulator of gibberellin responses.

In Arabidopsis, the DELLA subfamily of GRAS regulatory genes consists of GAI, RGA, RGA-LIKE1 (RGL1), RGL2, and RGL3. GAI and RGA are known to be negative regulators of gibberellin (GA) responses. We found that RGL1 is a similar repressor of GA responses, as revealed by RGL1 gain-of-function and loss-of-function phenotypes. Repression of GA responses in Arabidopsis was conferred by a dominant 35S-rgl1 transgene carrying a DELLA domain deletion analogous to the GA-insensitive gai-1 mutation. As in GA-deficient Arabidopsis, the transgenic plants were dark green dwarfs with underdeveloped trichomes and flowers. Expression levels of GA4, a feedback-regulated GA biosynthetic gene, were increased correspondingly. Conversely, a loss-of-function rgl1 line had reduced GA4 expression and exhibited GA-independent activation of seed germination, leaf expansion, flowering, stem elongation, and floral development, as detected by resistance to the GA biosynthesis inhibitor paclobutrazol. RGL1 plays a greater role in seed germination than do GAI and RGA. The expression profile of RGL1 differed from those of the four other DELLA homologs. RGL1 message levels were predominant in flowers, with transcripts detected in developing ovules and anthers. As with RGA, green fluorescent protein (GFP)-tagged RGL1 protein was localized to the nucleus, but unlike GFP-RGA, there was no degradation after GA treatment. These findings indicate that RGL1 is a partially redundant, but distinct, negative regulator of GA responses and suggest that all DELLA subfamily members might possess separate as well as overlapping roles in GA signaling.

Amino Acid Sequence↗

Offspring sex-ratio and reproductive performance in heterogametic females of the South American field mouse Akodon azarae. Reproduction in heterogametic Akodon azarae females.

We have compared the breeding performance of homogametic (XX) and heterogametic (XY*) females of the South American sigmodontine rodent Akodon azarae under laboratory conditions. XY* females showed an enhanced reproductive performance when compared with normal, XX, females. The XY* females had a longer reproductive lifespan. They started to reproduce early, had more frequent litters, and stopped reproduction later than XX females. Their progeny showed a biased 1:2 male:female sex ratio which may be explained by the early loss of YY* zygotes after fertilization. However, litter size at birth was similar both in XY* and XX females, and no difference in ovulation rate was detected between them. This indicates that an "automatic" rather than an "evolved" reproductive compensation mechanism may be acting in heterogametic females. A separate study has shown that self-synapsis of both the X and Y* chromosomes takes place during meiosis, allowing the oocytes to escape from functional deterioration. It is suggested that self-synapsis and "automatic" reproductive compensation account for the preservation of fertility in heterogametic females in spite of the heteromorphic sex chromosomes and the early embryo loss they experience. However, these mechanisms do not account for the enhancement of reproductive lifespan. The possibility that an intrauterine position phenomenon is acting in A. azarae is discussed.

Animals↗

Pituitary luteinizing hormone (LH), follicle stimulating hormone (FSH) and prolactin from birth to puberty in female and male rats.

Pituitary content and concentration of LH, FSH and prolactin were measured by radioimmunoassay (RIA) at 2-day intervals from birth to puberty in female and male rats. During the first 2 to 3 weeks of life all hormones were low in pituitary content and concentration in both sexes. They all increased in females during the third and fourth week, but decreased sharply during the days before vaginal opening. During the first ovulatory cycle pituitary content and concentration of LH and prolactin increased again, FSH, however, remained low. In males, pituitary LH, FSH and prolactin content reached peak levels during puberty. Our results show a distinct sexual dimorphism for pituitary FSH. Pituitary LH and prolactin content and concentration patterns show similar tendencies in both sexes with a delay of several days in males. The dramatic changes in female pituitary hormone concentrations just before the first ovulation were not detected in males before the first occurrence of mature spermatozoa in the tubuli of the testes.

Age Factors↗

Human chorionic gonadotropin-induced alterations in ovarian follicular dynamics during the estrous cycle of heifers.

This experiment was designed to characterize hCG (3,000 IU)-induced alterations in ovarian follicular and corpus luteum (CL) dynamics during the estrous cycle (EC) in heifers. Following synchronization of estrus (norgestomet implant for 7 d with injection of PGF2alpha 1 d before implant removal), 13 heifers were treated with either hCG (n = 6) or saline (control group; n = 7) on d 5 of the EC (d 0 = day of estrus). Blood sampling from the jugular vein and ultrasonography of both ovaries were conducted daily until confirmation of ovulation following a detected estrus. Treatment with hCG at d 5 induced formation of an accessory CL in all hCG-treated heifers. Subsequent plasma progesterone (P4) concentrations were higher (P < .01) between d 9 and 17 for the hCG group than for the control group. More (P < .05) hCG-treated heifers had EC with three waves of follicles than control heifers. The second-wave dominant follicle (DF) emerged earlier in hCG than in control heifers (7.3 vs 10.4 d; P < .01). A group x wave interaction (P < .01) was detected for duration of the second and third wave follicles; hCG treatment decreased duration of the second wave (6.3 vs 9.3 d) and increased duration of the third wave (9.2 vs 5.3 d). The hCG-treated heifers had preovulatory follicles that lasted longer (P < .01) on the ovary than those in control heifers. However, plasma estradiol concentrations did not differ (P > .10) between hCG-treated and control heifers during the preovulatory period. A wave x duration interaction (P < .01) was detected for P4 concentrations, between d 13 and 22, for heifers with two vs three spontaneous-wave EC. Higher P4 concentrations during the luteal phase may contribute to a greater rate of follicular turnover or frequency of three-wave cycles, and lower P4 concentrations were associated with two-wave cycles.

Animals↗

Insemination of Holstein heifers at a preset time after estrous cycle synchronization using progesterone and prostaglandin.

Two experiments were conducted to study estrous cycle control regimens that combine progesterone administration via an intravaginal device ( PRID ) with a single injection of prostaglandin F2 alpha (PG). In Exp. I, 242 Holstein heifers were assigned randomly to one of three treatment groups at 14 to 18 mo of age. Treatments were: 1) control, 2) PRID -6 + PG-6 ( PRID in place for 6 d plus PG on the day of PRID removal) and 3) PRID -7 + PG-6 ( PRID in place for 7 d plus PG on the day before PRID removal). Heifers were observed for estrous activity and were inseminated at 8 to 20 h after estrus was detected. Estrus and ovulation were effectively synchronized after both PRID + PG treatments. Ninety-nine percent of the heifers in each group were in estrus within 168 h after PG injection. However, the interval from PG administration to the onset of estrus was longer after PRID -7 + PG-6 (75 +/- 2 h) than after PRID -6 + PG-6 (66 +/- 2 h). A lower variance in the interval from PG treatment to estrus was observed after PRID -7 + PG-6, suggesting that the 24 h delay in PRID withdrawal improved the synchrony of the onset of estrus. Pregnancy rates (72 to 82%) did not vary across treatment groups. Two-hundred seventy-four heifers were assigned to Exp. II. Treatments were 1) control, 2) 2 X PG (two injections of PG at an 11 d interval) and 3) PRID -7 + PG-6.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Embryo reduction before day 11 in mares with twin conceptuses.

Incidence of embryo reduction (natural elimination of one member of a twin set) before d 11 was studied by comparing the number of ultrasonically detected conceptuses per ovulation between single and double ovulators. Effect of unilateral (n = 24) vs bilateral (n = 26) double ovulations on the incidence of embryo reduction also was considered. Each of 50 double-ovulating mares was matched with two single ovulators yielding 100 ovulations, or potential embryos, per group. Frequency with which an ovulation resulted in a conceptus was greater for single ovulators (85%, P less than .01) and for bilateral double ovulators (77%, P less than .05) than for unilateral double ovulators (60%). The difference between single and bilateral ovulators was not significant. Expected frequencies for none, one or two conceptuses per mare for double ovulators were calculated using the d-11 pregnancy rate for the single ovulators (85%). For unilateral ovulators, but not for bilateral ovulators, observed frequencies of none, one or two conceptuses were different (P less than .05) from expected (observed and expected, respectively, for no conceptuses, 29% and 2%; one conceptus, 21% and 26%; two conceptuses, 50% and 72%). Results did not indicate the existence of an embryo reduction phenomenon before d 11 in bilateral ovulators; each ovum had the same chance (no significant difference) of developing into a d-11 conceptus as an ovum in single ovulators. In unilateral double ovulators, the lower d-11 pregnancy rate per ovum, compared with bilateral ovulators and single ovulators, was attributable to a greater frequency of mares with no embryonic vesicles rather than to a greater frequency of mares with one vesicle.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

HIV prevention. Assisted reproduction in HIV-discordant couples.

Serodiscordant couples in whom the man is HIV-1-positive and the woman is negative have limited options if they wish to have children safely, because sexual intercourse carries a 1-in-500 risk of transmitting virus in semen to the female partner. Sperm washing is a risk-reduction option in which infected sperm are washed free of virus before insemination into the female partner at the time of ovulation. Absence of detectable HIV is verified before insemination using a polymerase chain reaction nucleic acid-based sequence amplification assay. Pregnancy rate per insemination is 14%, based on a European experience of more than 2000 inseminations; to date, there have been no seroconversions in either mother or child. Washed sperm have also been used in other assisted conception treatments, such as in vitro fertilization. In the United States, the CDC has recommended against insemination of women with semen from men infected with HIV. Current data from programs in Europe would suggest sperm washing to be a safe risk-reduction option for heterosexual couples wishing to bear a child. We suggest that sperm washing should only be carried out in dedicated units using a multidisciplinary approach to ensure that couples receive adequate preconceptual counseling, detailed sexual health and fertility assessment, and careful monitoring of the woman's HIV status during treatment and pregnancy.

Acquired Immunodeficiency Syndrome↗

Reduced pregnancy rates in AID women with unsuspected ovulatory failure.

A review over five years of 232 patients attending an artificial insemination by donor (AID) clinic was undertaken in order to detect abnormalities in ovulation patterns. Ovulation was determined by luteal phase progesterone estimations. Unexpected anovulation was found in 36% of the patients attending despite an apparently normal previous menstrual history and investigations. This group consisted of 22% who were anovulatory from commencement in the programme and 14% who became anovulatory during treatment. The cumulative pregnancy rates at 15 months for these two groups were lower (73% and 55%) than that for the remaining patients who ovulated normally (90%). A proportion (16%) of the latter group were receiving clomiphene therapy for anovulation recognized before entering the programme. The lower pregnancy rate could not be related to other factors influencing conception and occurred in spite of clomiphene therapy. These results should assist in advising patients on their chances of conception, preventing early discouragement but not encouraging long term insemination which may not improve the pregnancy rate.

Anovulation↗

Natural family planning.

Natural family planning (NFP) methods can be effective if taught well and practised by well-motivated couples. Midwives should be able to give couples an understanding of the basic principles of the methods. The observable signs of the fertile cycle are changes in basal body temperature, in the consistency of cervical mucus and in the texture and position of the cervix. Practised properly the ovulation method can have a failure rate of as little as 2.2 pregnancies per 100 woman-years. The ovulation method is a suitable method for mothers who have breastfed to detect the onset of ovulation.

Family Planning Services↗