Investigations into the effects of cyclical rhythm and hormonal contraception on serum fat-mobilizing activity, glycerol, cholesterol and blood glucose.

The effects were investigated of cyclical rhythm and hormonal contraception on serum fat-mobilizing activity, glycerol, cholesterol and whole blood glucose during 2 menstrual cycles in a group of normally menstruating young women and a second group of young women using hormonal contraception. A control group of normal young men was also investigated. There was no evidence of any change in mean level of any of the parameters measured, among the follicular, ovulatory and luteal phases. No cyclical pattern was discernable in the male subjects. The mean value for serum cholesterol concentration in women using hormonal contraception was higher than the value for the untreated human female group. The overall mean value for serum glycerol concentration in the women was significantly (0.01 > P > 0.001) higher than the mean value obtaining in the men

Changes in the ejaculate of the male tammar wallaby (Macropus eugenii) parallel seasonal reproductive activity in the female

The reproductive tract of the male tammar wallaby increases in size and secretory activity when females are in oestrus, perhaps in response to a release of female pheromones (CATLING and SUTHERLAND 1980; INNS 1982; MCCONNELL et al. 1984). This study investigated the effect of changes in male reproductive structures on semen quality throughout the year in this highly promiscuous species. Body size, semen quality and reproductive structures were measured monthly in adult males from January to November. Although there was no significant difference in the weight of males sampled throughout the year, heavier males had heavier testes and epididymides and produced larger ejaculates. The main breeding season in late January, when all females were in synchronous oestrus, coincided with the heaviest testis, epididymis, prostate, Cowper’s glands, crus penis and urethral bulb. Sperm motility, semen volume and coagulation of the ejaculate were greatest at this time. In July, during an almost total absence of mating in the wild, all structures regressed, resulting in significantly reduced sperm number and motility in small volumes of semen that did not coagulate. During the subsidiary breeding season in October/November, when only pubertal females undergo their first oestrus, all male reproductive structures, except the prostate, returned to maximum weights and sperm number and motility were high. In contrast, prostate weight, semen volume and coagulation of the ejaculate reached intermediate levels. Semen volume was positively correlated with prostate weight while the proportion of motile sperm was positively correlated with epididymal weight. Sperm concentration was negatively correlated with semen volume, and sperm numbers remained low when coagulation of ejaculates was greatest. These results show that semen quality is influenced by the change of androgen-dependent reproductive structures in the male tammar wallaby. The male reproductive tract is apparently primed to produce high quality semen when females are in oestrus. Semen quality may therefore improve in response to the number of available oestrous females, which may in turn provide an advantage in sperm competition

Distribution of spermatozoa and copulatory plug in relation to the time of mating and ovulation in the female tammar wallaby (Macropus eugenii)

In the monovular macropodid marsupial, the tammar wallaby (Macropus eugenii), the cervices are the primary selective barrier to spermatozoa, resulting in differential transport to the non-gravid uterus where a sperm reservoir is established (Tyndale-Biscoe CH and Rodger JC 1978 J. Reprod. Fertil. 52, 37–43). However, due to limited sample size, the dynamics of sperm transport could not be thoroughly examined. In this study, the distribution of spermatozoa, the size of the copulatory plug in the reproductive tract at various times after mating, and the timing of ovulation were characterized in 28 naturally mated female tammars. After the first postpartum (p.p.) mating, adult females were isolated and their reproductive tracts dissected at 0.5, 6, 18, 36, and 40 h post-coitum (p.c.). Each tract was ligated into 13 major anatomical sections, and spermatozoa and eggs were recovered by flushing. Mating occurred 21.7 ± 2.5 h p.p. (mean ± SEM; n = 20) in these animals that were checked frequently and lasted 7.8 ± 0.7 min (n = 15). Within 0.5 h after a single mating (n = 5) the tract contained 2.6 ± 1.0 × 107 spermatozoa and 21.6 ± 8.8 g of copulatory plug, 96 and 70% of which was lost within 6 h p.c., respectively. Spermatozoa reached the uterus, isthmus, and ampulla of the oviduct ipsilateral to the developing follicle within 0.5, 6, and 18 h p.c. respectively, and a uterine population of 2.6 ± 1.2 × 104 spermatozoa (n = 24) was maintained for over 40 h (ANOVA, P > 0.05). Sperm numbers were reduced at the cervix (up to 57-fold) and utero-tubule junction (8-fold), and only 1 in ∼7600 ejaculated spermatozoa (3.4 ± 0.9 × 103; n = 14) reached the oviduct on the side of ovulation. Although sperm numbers were reduced in the gravid uterus (n = 24), differential transport of spermatozoa was not observed (ANOVA, P > 0.05). Ovulation and recovery of sperm-covered eggs from the isthmus of the oviduct occurred 36–41 h p.c. (49–72 h p.p.) (n = 8). Like many eutherian mammals, but in contrast to polyovular dasyurid and didelphid marsupials, the tammar ejaculates large numbers of spermatozoa, but transport is relatively inefficient and sperm storage in the tract before ovulation is limited

Time of ovulation and artificial insemination following superovulation in the southern hairy-nosed wombat (Lasiorhinus latifrons)

The southern hairy-nosed (SHN) wombat (Lasiorhinus latifrons) is an ideal model for developing assisted reproductive techniques (ART) which may be used to increase the population size of its critically endangered sister species, the northern hairy-nosed (NHN) wombat (Lasiorhinus krefftii). Previously in SHN wombats, we demonstrated that multiple follicles are recruited and maturation to metaphase II occurs in 37 % of follicular oocytes recovered after treatment with 14. 3 mg porcine FSH (pFSH) every 12 h for 7 days, followed by a single 25 mg pLH injection 12 h later (DRUERY et al. 2003). The aims of this study were (1) to determine the timing of ovulation after superovulation and (2) to attempt artificial insemination (AI) after superovulation to produce live offspring. Six captive female wombats had their ovaries stimulated using the 7-day pFSH/pLH treatment protocol described above, and the timing of ovulation in relation to the pLH injection was determined using repeated laparoscopic examination (3 hourly). Ovulations were first observed 36 h after pLH injection (1 out of 33 mature follicles) and most follicles greater than 5 mm (39/51) had ovulated by 48 h post-pLH. Progesterone levels increased slowly after ovulation, indicating the presence of a luteal phase. This ovulation window was then used to time the delivery of semen by intra-uterine Al (IUAI) in SHN wombat females. Nine females were treated with the same pFSH/pLH protocol and inseminated 34h after the pLH injection with electro-ejaculated sperm (range 1.5 - 3.0 x 10 /uterus). Females were examined using ultrasound and laparoscopy on -day 18 post-IUAI for evidence of pregnancy and monitored for births at hourly intervals for 48 h from the expected time of parturition. Two pouch checks were then made at weekly intervals after the expected time of birth had _passed. At the time of AI these females responded to pFSH by producing multiple ovanan follicles. By day 17 - 18 post-IUAI, laparoscupic observations of enlarged uteri and ultrasonographic images of structures resembling gestational sacs strongly suggested up to five females were pregnant. However, no live births were observed. The exact point at which embryo viability was compromised was not determined. This study has confirmed successful ovulation using the pFSH/pLH protocol and determined the window for ovulation in female SHN wombats to be 36 - 48 h post­pLH injection. Further refinement of the insemination procedure is now required to increase fertilisation rates and improve post-conception survival, development and successful parturition of wombat neonates

Timing of ovulation in the gonadotrophin-stimulated southern hairy-nosed wombat, Lasiorhinus latifrons

The southern hairy-nosed wombat (SHW), Lasiorhinus latifrons, is a model species in which to develop assisted breeding techniques for the endangered northern hairy-nosed wombat, Lasiorhinus krefftii. We recently showed that anoestrus SHW respond to eutherian gonadotrophins by production of multiple ovarian follicles, but ovulation had not occurred at the time of examination 24 h post-LH (Druery GV et al. 2003 Theriogenology 59, 391 abst). This study investigated the timing of ovulation in six anoestrus captive adult female SHW (n = 3 per group) after ovarian superstimulation using porcine FSH (200 mg total, Folltropin-V, Bioniche, Belleville, Ontario, Canada) administered s.c. at 12-h intervals over 7 days. Ovulation was triggered by a single s.c. dose of porcine LH (25 mg Lutropin-V, Bioniche) 12 h after the final FSH injection. Superstimulatory response was determined by laparoscopy immediately after the final FSH injection on Day 7 prior to LH. Group 1 was re-examined at 33, 36, and 39 h post-LH, and Group 2 at 42, 45, and 48 h post-LH, for evidence of ovulations using laparoscopy and transabdominal ultrasonography. Laparoscopy on Day 7 revealed an ovarian follicular response in all six females, which coincided with the highest levels of estradiol. The reproductive tract also responded to the treatment (swollen fimbriae and enlarged, highly vascular uteri). Multiple follicles (range 16–31) up to 11 mm in diameter were observed in five females. One female had ovulated, as determined by the presence of corpora lutea. Transabdominal ultrasonographic imaging was unable to confirm the number of follicles in stimulated ovaries. Ovulation had commenced by 36 h post-LH, with the majority occurring 39–45 h post-LH. Ovulation was recorded as having occurred if a dark red, highly vascular crater on the surface of the newly formed corpus hemorrhagicum was observed. Increased circulating levels of progesterone were confirmed 9 days after the last laparoscopies. These results have important implications for the development of assisted reproductive technologies in the SHW: (1) transabdominal ultrasound imaging is ineffective for determining ovarian activity; (2) laparoscopy is a well-tolerated, repeatable minor surgical procedure that can be used for intrauterine AI in this species in which nonsurgical AI is unlikely to succeed (Paris DBBP et al. 2003 Theriogenology 59, 401 abst); and (3) knowledge of the timing of ovulation will enable insemination of spermatozoa into the uterus prior to ovulation

Identification of Novel Craniofacial Regulatory Domains Located far Upstream of SOX9 and Disrupted in Pierre Robin Sequence.

Mutations in the coding sequence of SOX9 cause campomelic dysplasia (CD), a disorder of skeletal development associated with 46,XY disorders of sex development (DSDs). Translocations, deletions, and duplications within a ∼2 Mb region upstream of SOX9 can recapitulate the CD-DSD phenotype fully or partially, suggesting the existence of an unusually large cis-regulatory control region. Pierre Robin sequence (PRS) is a craniofacial disorder that is frequently an endophenotype of CD and a locus for isolated PRS at ∼1.2-1.5 Mb upstream of SOX9 has been previously reported. The craniofacial regulatory potential within this locus, and within the greater genomic domain surrounding SOX9, remains poorly defined. We report two novel deletions upstream of SOX9 in families with PRS, allowing refinement of the regions harboring candidate craniofacial regulatory elements. In parallel, ChIP-Seq for p300 binding sites in mouse craniofacial tissue led to the identification of several novel craniofacial enhancers at the SOX9 locus, which were validated in transgenic reporter mice and zebrafish. Notably, some of the functionally validated elements fall within the PRS deletions. These studies suggest that multiple noncoding elements contribute to the craniofacial regulation of SOX9 expression, and that their disruption results in PRS

Measurement of the p-pbar -> Wgamma + X cross section at sqrt(s) = 1.96 TeV and WWgamma anomalous coupling limits

The WWgamma triple gauge boson coupling parameters are studied using p-pbar -> l nu gamma + X (l = e,mu) events at sqrt(s) = 1.96 TeV. The data were collected with the DO detector from an integrated luminosity of 162 pb^{-1} delivered by the Fermilab Tevatron Collider. The cross section times branching fraction for p-pbar -> W(gamma) + X -> l nu gamma + X with E_T^{gamma} > 8 GeV and Delta R_{l gamma} > 0.7 is 14.8 +/- 1.6 (stat) +/- 1.0 (syst) +/- 1.0 (lum) pb. The one-dimensional 95% confidence level limits on anomalous couplings are -0.88 < Delta kappa_{gamma} < 0.96 and -0.20 < lambda_{gamma} < 0.20.Comment: Submitted to Phys. Rev. D Rapid Communication

Measurement of the ttbar Production Cross Section in ppbar Collisions at sqrt{s} = 1.96 TeV using Kinematic Characteristics of Lepton + Jets Events

We present a measurement of the top quark pair ttbar production cross section in ppbar collisions at a center-of-mass energy of 1.96 TeV using 230 pb**{-1} of data collected by the DO detector at the Fermilab Tevatron Collider. We select events with one charged lepton (electron or muon), large missing transverse energy, and at least four jets, and extract the ttbar content of the sample based on the kinematic characteristics of the events. For a top quark mass of 175 GeV, we measure sigma(ttbar) = 6.7 {+1.4-1.3} (stat) {+1.6- 1.1} (syst) +/-0.4 (lumi) pb, in good agreement with the standard model prediction.Comment: submitted to Phys.Rev.Let

Measurement of the ttbar Production Cross Section in ppbar Collisions at sqrt(s)=1.96 TeV using Lepton + Jets Events with Lifetime b-tagging

We present a measurement of the top quark pair ($t\bar{t}$) production cross section ($\sigma_{t\bar{t}}$) in $p\bar{p}$ collisions at $\sqrt{s}=1.96$ TeV using 230 pb$^{-1}$ of data collected by the D0 experiment at the Fermilab Tevatron Collider. We select events with one charged lepton (electron or muon), missing transverse energy, and jets in the final state. We employ lifetime-based b-jet identification techniques to further enhance the $t\bar{t}$ purity of the selected sample. For a top quark mass of 175 GeV, we measure $\sigma_{t\bar{t}}=8.6^{+1.6}_{-1.5}(stat.+syst.)\pm 0.6(lumi.)$ pb, in agreement with the standard model expectation.Comment: 7 pages, 2 figures, 3 tables Submitted to Phys.Rev.Let