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Follicular changes throughout the oestrous phase have been poorly documented in queens because of the location and the small size of ovaries. We investigated follicular development in queens treated with a combination of equine chorionic gonadotropin (eCG) and human chorionic gonadotropin (hCG) and evaluated the effects of vaginal stimulation by a tomcat on ovulation induction. A hormonal treatment was administered using a simple crossover design. Four queens were administered 150 IU of eCG (day 1) and 250 IU of hCG on day 5 and 6. Half of the queens were mated with a vasectomised tomcat for 3 days after hCG injection. Ultrasound imaging of the ovaries clamped at a subcutaneous site was performed once a day from day 1 to 7, and on day 13, and the serum concentrations of oestradiol and progesterone were examined on day 1, 5, 7 and 13. The mean number of follicles gradually increased with the eCG treatment and decreased after hCG injection. The ovulation rate of follicles was significantly higher in the vaginal stimulation group (70.0%) than in the control group (42.6%). During the hormonal treatments, the serum concentration of oestradiol and progesterone did not differ between the two groups. Ultrasound imaging of the ovaries clamped at a subcutaneous site showed that eCG and hCG treatment promoted the follicular growth and corpus luteum formation, respectively. The combination of hCG injection with vaginal stimulation by a vasectomised tomcat enhanced the ovulation rate of follicles., Ultrasound imaging of the ovaries clamped at a subcutaneous site showed that the combination of hCG injection with vaginal stimulation by a vasectomised male cat enhanced the ovulation rate of follicles.

Ovarian follicular dynamics is not well known in dogs. Imaging of ovaries is technically difficult; however, ovaries clamped at a subcutaneous site can more easily be monitored using ultrasound imaging. This study investigated the follicular development of canine ovaries stimulated by hormone treatment using ultrasound imaging of the ovaries clamped at a subcutaneous site. Oestrus was induced using subcutaneous administration of 500 IU equine chorionic gonadotropin (eCG) and 1000 IU human chorionic gonadotropin (hCG) (eCG/hCG). Five bitches were given 1000 IU hCG 11 days after eCG/hCG administration. Examinations with ovarian ultrasonography using a 7.5‐MHz sector transducer, vaginal cytology, and assays of serum oestrogen and progesterone were performed daily until 20 days after eCG/hCG administration. Serosanguineous vaginal discharges and vaginal cytology of two of the bitches were observed. New follicular growth (>1.0 mm in diameter) was observed in all bitches from 2 to 8 days after eCG/hCG administration. The mean diameter of follicles and maximum numbers of follicles per ovary ranged from 2.8 to 5.5 mm and 4 to 16, respectively. The elevation in oestrogen concentrations after eCG/hCG administration was observed in all bitches, and elevation in progesterone concentration (>2 ng mL−1) was observed in three bitches. However, no follicles ovulated until 9 days after hCG administration. In conclusion, although the number of examined bitches were limited, follicular growth in ovaries clamped at a subcutaneous site can be monitored using ultrasound imaging. Ovarian ultrasonography showed that eCG/hCG administration induced new follicular growth and hCG administration induced increases in oestrogen concentrations but not ovulation by hCG administration.

Contents The current applications for cat cloning include production of models for the study of human and animal diseases. This study was conducted to investigate the optimal fusion protocol on in vitro development of transgenic cloned cat embryos by comparing duration of electric pulse. Cat fibroblast cells containing a human artificial chromosome (HAC) vector were used as genetically modified nuclear donor cells. Couplets were fused and activated simultaneously with a single DC pulse of 3.0 kV/cm for either 30 or 60 μs. Low rates of fusion and embryo development to the blastocyst stage were observed in the reconstructed HAC-transchromosomic embryos, when the duration of fusion was prolonged to 60 μs. In contrast, the prolongation of electric pulse duration improved the embryo development and quality in the reconstructed control embryos without HAC vector. Our results suggested that the optimal parameters of electric pulses for fusion in cat somatic cell nuclear transfer vary among the types used for donor cells.

The influence of graft site on the survival of canine follicles and oocytes after autografting was investigated. Hemi-ovaries were autografted to three locations (quadriceps femoris muscle fascia, kidney capsule, and gastrosplenic ligament), and grafted ovaries were recovered (under anesthesia) 28 to 31 d after transplantation. The grafted hemi-ovaries were bisected: one-quarter ovary was used for histological assessment and another quarter for evaluation of oocyte viability. As controls, the remaining fresh hemi-ovaries were used to assess the viability of follicles and oocytes in non-transplanted ovaries. Most follicles in the histological sections of the grafts were classified as primordial or primary follicles. Antral follicles were not observed in the grafts, irrespective of the graft site. The percentages of viable follicles in the sections from control ovaries, and the ovaries grafted to the kidney capsule, the quadriceps femoris muscle fascia, and the gastrosplenic ligament were 17.4, 22.9, 18.3, and 32.4%, respectively. A total of 12 oocytes was recovered from the 15 hemi-ovaries grafted in five bitches, of which five (41.7%) oocytes from the ovaries grafted to the quadriceps femoris muscle fascia and the kidney capsule were cultured for assessment of meiotic competence. Three oocytes were viable but remained in the germinal vesicle stage after 72 h of maturation culture. The quadriceps femoris muscle fascia might be useful for grafting like the kidney capsule, but improvement of follicle survival and meiotic competence of oocytes in the grafts is necessary.

This study reports about follicular development on the surface of canine ovarian tissue after autografting under the fascia of the thoracolumbar muscle and about meiotic resumption of follicle-derived oocyte after maturation culture. After ovarian excision from a bitch, each ovary of the pairs was cut approximately into half. The hemi-ovaries were transplanted into the bitch of origin at three different body sites (under the fascia of the quadriceps femoris muscle and the thoracolumbar muscle, and in the deltoid muscle in the scapular region). All grafted ovaries were recovered from the bitch at 35 days post-transplantation. A visible antral follicle was observed on the surface of the ovary grafted under the thoracolumbar fascia. Histological examination revealed viable follicles at different stages of development irrespective of graft site. Most granulosa cells in the follicles at different stages of development expressed proliferating cell nuclear antigen (PCNA). A total of three oocytes were collected from an ovary grafted under the fascia of the thoracolumbar muscle, wherein an oocyte reached metaphase I after maturation culture. This is the first report to demonstrate follicular development and meiotic resumption of oocytes recovered from autografted canine ovarian tissues.

This study reports about follicular development on the surface of canine ovarian tissue after autografting under the fascia of the thoracolumbar muscle and about meiotic resumption of follicle-derived oocyte after maturation culture. After ovarian excision from a bitch, each ovary of the pairs was cut approximately into half. The hemi-ovaries were transplanted into the bitch of origin at three different body sites (under the fascia of the quadriceps femoris muscle and the thoracolumbar muscle, and in the deltoid muscle in the scapular region). All grafted ovaries were recovered from the bitch at 35 days post-transplantation. A visible antral follicle was observed on the surface of the ovary grafted under the thoracolumbar fascia. Histological examination revealed viable follicles at different stages of development irrespective of graft site. Most granulosa cells in the follicles at different stages of development expressed proliferating cell nuclear antigen (PCNA). A total of three oocytes were collected from an ovary grafted under the fascia of the thoracolumbar muscle, wherein an oocyte reached metaphase I after maturation culture. This is the first report to demonstrate follicular development and meiotic resumption of oocytes recovered from autografted canine ovarian tissues.

The compression-mode isoscalar giant monopole resonance (ISGMR) has been studied in the Sn, Cd and Pb isotopes using inelastic scattering of 400 MeV {alpha}-particles at extreme forward angles, including 0 deg. We have obtained completely 'background-free' inelastic-scattering spectra for the Sn, Cd, and Pb isotopes for a wide angular and excitation-energy range. The various giant resonances excited with different transferred angular momenta were extracted by a multipole-decomposition analysis (MDA). It was found that the centroid energies of the ISGMR in Sn isotopes are significantly lower than the theoretical predictions. The K{sub {tau}} in the empirical expression for the nuclear incompressibility has been determined to be K{sub {tau}} = -550{+-}100 MeV for the Sn isotopes. The extracted value for the Cd isotopes is K{sub {tau}} = -480{+-}100 MeV. These numbers are consistent with values of K{sub {tau}} = -370{+-}120 MeV obtained from an analysis of the isotopic transport ratios in medium-energy heavy-ion reactions, K{sub {tau}} -500{sub -100}{sup +120} MeV obtained from constraints placed by neutron-skin data from anti-protonic atoms across the mass table, and K{sub {tau}} = -500{+-}50 MeV obtained from theoretical calculations using different Skyrme interactions and relativistic mean field (RMF) Lagrangians. Stringent constraints on interactions employed in nuclear structuremore » calculations are obtained on the basis of the experimentally determined values for K{sub {infinity}} and K{sub {tau}}. These parameters constrain as well the equation of state (EOS) of nuclear matter. However, a significant discrepancy still remains. The ISGMR positions in Sn and Cd isotopes are systematically lower than the predictions obtained on basis of determined from the ISGMR in {sup 208}Pb. This raises the question ''Why are Sn and Cd nuclei so soft?''. For a clue to solve the problem, we discuss the preliminary results on the exact positions of the ISGMR in {sup 204,206,208}Pb.« less

The strength distributions of the giant monopole resonance (GMR) have been measured in the even-A Sn isotopes (A = 112-124) with inelastic scattering of 400-MeV alpha particles in the angular range 0 degrees-8.5 degrees. We find that the experimentally observed GMR energies of the Sn isotopes are lower than the values predicted by theoretical calculations that reproduce the GMR energies in Pb-208 and Zr-90 very well. From the GMR data, a value of K-tau = -550 +/- 100 MeV is obtained for the asymmetry term in the nuclear incompressibility.

The strength distributions of the giant monopole resonance (GMR) have been measured in the even-A Sn isotopes (A=112--124) with inelastic scattering of 400-MeV $\alpha$ particles in the angular range $0^\circ$--$8.5^\circ$. We find that the experimentally-observed GMR energies of the Sn isotopes are lower than the values predicted by theoretical calculations that reproduce the GMR energies in $^{208}$Pb and $^{90}$Zr very well. From the GMR data, a value of $K_{\tau} = -550 \pm 100$ MeV is obtained for the asymmetry-term in the nuclear incompressibility., Comment: Submitted to Physical Review Letters. 10 pages; 4 figures

Follicle-stimulating hormone (FSH) alone can induce oestrus in bitches, but few reports describe oestrous induction by FSH because pregnant mare serum gonadotrophin (PMSG) has been more successful than FSH for oestrus induction. Real-time ultrasonography can show canine ovarian follicle development, but no method can determine or predict ovulation accurately. Moreover, the ovary location and size complicate imaging. Using ultrasonography, we investigated FSH treatment stimulation of canine ovary follicles, with clamping of the ovaries at a subcutaneous site. Bilateral malacotomy of four 5-year-old Beagle bitches (mean weight 10.3 ± 2.0 kg) with normal oestrous cycles was done using a ventral flank abdominal approach with routine techniques and materials. Each ovary that maintained blood circulation from the suspensory ligament was clamped at a subcutaneous site through muscles of the abdomen. After about six months of bilateral malacotomy, four bitches at the anestrous (two bitches) and diestrous (two bitches) stages of the oestrous cycle were given 0.5 Armour units of FSH twice daily for 5 days. Examinations with ovarian ultrasonography with 7.5 MHz sector transducer, vaginal cytology, and serum concentrations of progesterone and oestradiol were performed daily from the day before the start of FSH treatment through 7 days after FSH treatment. After 15 days of ovarian examination, each bitch received the same FSH treatment twice continually at 15-day intervals. No serosanguineous vaginal discharge was observed during the ovarian examination. The concentrations of progesterone (

Follicular growth in bitches is usually detected indirectly through behaviour observation, vaginal smears, and hormonal assay in blood. Although real-time ultrasonography can reveal the development of canine ovarian follicles, no method has been established to determine or predict ovulation accurately. Moreover, the location and small size of the ovaries make imaging technically difficult. This study was conducted to investigate follicular waves of canine ovaries stimulated by hormone treatment, in which ovaries had been clamped at a subcutaneous site. Bilateral malacotomy of 3 bitches (4 years of age) at the anestrous (2 bitches) and proestrous (1 bitch) stages of the oestrous cycle was performed using a ventral flank abdominal approach with routine techniques and materials. Each ovary that maintained blood circulation from the suspensory ligament was clamped at a subcutaneous site through muscles of the abdomen. Oestrus was induced using subcutaneous administration of 500 IU of eCG and 1000 IU of hCG (eCG/hCG). Each bitch was given 1000 IU of hCG at 11 days after eCG/hCG administration. Examinations with ovarian ultrasonography using a 7.5-MHz sector transducer, vaginal cytology, and serum progesterone assay were performed daily until 20 days after eCG/hCG administration, and every 10 days thereafter from 20 days to 60 days. Serosanguineous vaginal discharges and vaginal cytology of 2 of the bitches were observed. Follicular growth (>1.1 mm in diameter) was observed in all bitches after eCG/hCG administration. The appearance of new follicular growth was observed on 2 days, 6 days, and 8 days after eCG/hCG administration. The mean diameter of follicles reached 4.3 to 5.5 mm, and the maximum numbers of follicles in bitches were 11 to 16. However, all follicles regressed, irrespective of hCG administration. Elevation in progesterone levels (>2 ng mL–1) after eCG/hCG administration was observed from 2 days to 12 days after eCG/hCG administration. No correlation was found between follicular development, progesterone profiles, and vaginal smear characteristics. Follicular growth clamped at the subcutaneous site can be monitored easily using ultrasound without an experienced operator. Moreover, ultrasonography proved that hormonal stimulation can induce follicular growth, but the day of appearance of new follicles varied.

Numerous studies have reported that follicles in ovarian tissue are not only able to survive ovarian grafting but also to grow from the early to advanced stages of follicular development. The site of grafting is very important for tissue survival and follicle development after transplantation. Several grafting techniques have been reported, basically differing only in the location to which the ovarian grafts have been transplanted, such as the bursal cavity, kidney capsule, and subcutaneous sites. The objective of the present study was to determine the viability of canine oocytes from ovaries transplanted in the proximal portion of the body surface. Bilateral ovariectomy was performed using a ventral midline abdominal approach with routine techniques and materials. After excision of ovaries from 10 bitches (4–7 y of age), each ovary of the pairs was cut approximately in half using a scalpel blade. The hemi-ovaries were kept in physiological saline, maintained at 38°C, and then transplanted to the animal of origin in each body site. The ovaries were transplanted under the fascia of the left quadriceps femoris muscle, the left thoracolumbar fascia, and the deltoid muscle in left scapular region. As control, a remaining hemi-ovary was used to examine the morphology and viability of oocytes from fresh ovaries without transplantation. All grafted ovaries were recovered from animals at 28 to 31 days after transplantation under the anesthesia. After collection, each ovary was sliced repeatedly to release oocytes. All oocytes collected from ovaries were examined microscopically for morphological quality and categorized by the classification as previously described (Hewitt et al. 1998 Theriogenology 49, 1083–1101). Grade A oocytes were darkly pigmented and surrounded by one or more layers of cumulus cells, Grade B oocytes were lightly pigmented with incomplete layers of cumulus cells, and Grade C oocytes were pale coloured and irregularly shaped without any cumulus cells. Only Grade A oocytes were cultured in maturation medium for 72 h at 38.5°C in a humidified atmosphere of 5% CO2 in air. After incubation for 72 h, the oocytes were denuded, fixed, and stained by Hoechst 33342 to examine the nuclear maturation. After grafting for 28 to 31 days, a total of 5, 7, and 4 oocytes were recovered, respectively, from hemi-ovaries transplanted into quadriceps femoris fascia, thoracolumbar fascia, and scapular deltoid muscle, of which 1 and 2 oocytes from quadriceps femoris fascia and thoracolumbar fascia were classified into Grade A, respectively. After maturation culture of 3 oocytes, one oocyte reached the metaphase II stage. In the control group, a total number of 183 oocytes were collected from fresh ovaries, and 20.8% of these oocytes were classified into Grade A, of which 6.1% of oocytes reached the metaphase II stage after maturation culture. These results indicate that some oocytes from grafted ovaries can develop and maintain meiotic competence even when the ovaries are transplanted under the fascia in the proximal portion of the body surface.

Human growth hormone (hGH) is a single chain, 22 kd-protein with two intramolecular disulfide bonds. The hGH gene is located on chromosome 17 at band q22-q24 and has four introns separating five coding exons. The expression of hGH is restricted to the pituitary and regulated by GHF-1 which binds to the hGH promoter acting in concert with several other more ubiquitous DNA binding proteins. The secretion of hGH is regulated by GH releasing hormone (GRH) and somatostatin. GRH controls GH synthesis by stimulating transcription of GH mRNA while somatostatin determines the timing and amplitude of GH pulses. Pulsatile GH secretion is influenced by a number of neurogenic, metabolic and hormonal factors.