Relationship between follicular volume and oocyte competence, blastocyst development and live-birth rate: optimal follicle size for oocyte retrieval.

OBJECTIVE: To analyze oocyte competence in gonadotropin-releasing hormone agonist (GnRHa) stimulation cycles with regard to maturity, fertilization and blastocyst rate, as well as clinical outcome (pregnancy and live-birth rate), in relation to follicular volume, measured by three-dimensional transvaginal sonography (3D-TVS), and follicular fluid composition. METHODS: This was a prospective single-center study conducted between June 2012 and June 2014, including 118 ovum pick-ups with subsequent embryo transfer. Ovarian stimulation was performed using the GnRHa long protocol. Of 1493 follicles aspirated individually, follicular volume was evaluated successfully in 1236 using automated 3D-TVS during oocyte retrieval. Oocyte maturity and blastocyst development were tracked according to follicular volume. Intrafollicular concentrations of estradiol, testosterone, progesterone, luteinizing hormone, follicle-stimulating hormone and granulocyte-colony stimulating factor were quantified by immunoassay. Clinical outcome, in terms of implantation rate, (clinical) pregnancy rate, miscarriage and live-birth rate (LBR), was evaluated. RESULTS: Follicles were categorized, according to their volume, into three arbitrary groups, which included 196 small (8-12 mm/0.3-0.9 mL), 772 medium (13-23 mm/1-6 mL) and 268 large (≥ 24 mm/> 6 mL) follicles. Although oocyte recovery rate was significantly lower in small follicles compared with medium and large ones (63.8% vs 76.6% and 81.3%, respectively; P < 0.001), similar fertilization rates (85.1% vs 75.3% and 81.4%, respectively) and blastocyst rates (40.5% vs 40.6% and 37.2%, respectively) per mature metaphase II oocyte were observed. A trend towards higher LBR after transfer of blastocysts derived from small (< 1 mL) follicles compared with medium (1-6 mL) or large (> 6 mL) follicles (54.5% vs 42.0%, and 41.7%, respectively) was observed. No predictive value of follicular fluid biomarkers was identified. CONCLUSIONS: Our data indicate that the optimal follicular volume for a high yield of good quality blastocysts with good potential to lead to a live birth is 13-23 mm/1-6 mL. However, oocytes derived from small follicles (8-12 mm/0.3-0.9 mL) still have the capacity for normal development and subsequent delivery of healthy children, suggesting that aspiration of these follicles should be encouraged as this would increase the total number of blastocysts retrieved per stimulation. Copyright © 2017 ISUOG. Published by John Wiley & Sons Ltd.

Higher sensitivity of Adamts12-deficient mice to tumor growth and angiogenesis.

ADAMTS (a disintegrin and metalloproteinase domain with thrombospondin motifs) constitute a family of endopeptidases related to matrix metalloproteinases. These proteases have been largely implicated in tissue remodeling and angiogenesis associated with physiological and pathological processes. To elucidate the in vivo functions of ADAMTS-12, we have generated a knockout mouse strain (Adamts12(-/-)) in which Adamts12 gene was deleted. The mutant mice had normal gestations and no apparent defects in growth, life span and fertility. By applying three different in vivo models of angiogenesis (malignant keratinocyte transplantation, Matrigel plug and aortic ring assays) to Adamts12(-/-) mice, we provide evidence for a protective effect of this host enzyme toward angiogenesis and cancer progression. In the absence of Adamts-12, both the angiogenic response and tumor invasion into host tissue were increased. Complementing results were obtained by using medium conditioned by cells overexpressing human ADAMTS-12, which inhibited vessel outgrowth in the aortic ring assay. This angioinhibitory effect of ADAMTS-12 was independent of its enzymatic activity as a mutated inactive form of the enzyme was similarly efficient in inhibiting endothelial cell sprouting in the aortic ring assay than the wild-type form. Altogether, our results show that ADAMTS-12 displays antiangiogenic properties and protect the host toward tumor progression.

Regulation of progesterone during pregnancy in the cat: studies on the roles of corpora lutea, placenta and prolactin secretion.

The regulation of progesterone secretion and release by the corpus luteum and/or by the placenta in cats during pregnancy has been poorly studied and the relative roles of the corpus luteum and the placenta in the release of progesterone into the plasma is not well defined. It is generally assumed that after 40-45 days of pregnancy, circulating progesterone is mainly produced by the placenta and that after 45-50 days the corpus luteum is no longer necessary to maintain pregnancy, even if remnants of corpora lutea are detected in the ovaries during the following interoestrous period. In this study, the relative roles of the placenta and ovary in progesterone production after 30 days of pregnancy have been analysed in an attempt to define the role of progesterone in pregnant cats. Different experiments involved (1) analysis of the effects of an ovariectomy performed at 30 days of pregnancy, and of megestrol acetate supplementation on plasma progesterone during the second half of pregnancy, (2) analysis of the effects of an ovariectomy performed at 45 days on plasma progesterone production and on pregnancy and (3) analysis of the effects of an antiprolactin agent (cabergoline) at days 30 and 40 of pregnancy on progesterone secretion and pregnancy. These results were all compared with the normal plasma progesterone profiles during (a) pseudopregnancy and (b) pregnancy.(ABSTRACT TRUNCATED AT 250 WORDS)

Comparison of the clinical utility of medetomidine/ketamine and xylazine/ketamine combinations for the ovariectomy of cats.

A controlled trial was conducted to assess suitability of combinations of medetomidine and ketamine for the ovariectomy of cats, to investigate the possible side effects, and to compare medetomidine/ketamine with a combination of xylazine and ketamine. Three hundred and thirty-seven cats were submitted to surgery; 100 were anaesthetised with 80 micrograms/kg medetomidine and 5 mg/kg ketamine, 137 with 80 micrograms/kg medetomidine and 7.5 mg/kg ketamine, and 100 were anaesthetised with 1 mg/kg xylazine and 10 mg/kg ketamine. The combinations were injected intramuscularly in the same syringe. The anaesthesia provided by the medetomidine/ketamine combinations was characterised by good muscle relaxation, good analgesia and minimal side effects. The only difference between the two doses of ketamine was the length of the period of anaesthesia. The advantages of the medetomidine/ketamine combination in comparison with xylazine/ketamine were the need for a lower dose of ketamine, a longer duration of action and better analgesia. Similar side effects were observed with both medetomidine/ketamine and xylazine/ketamine combinations.

Sensitivity of anterior hypothalamic areas to gonadal steroid implantation in androgenized female rats.

A single injection of 1 mg of a complex of testosterone esters on day 5 of life was used to prepare constantly oestrous rats. Such androgenized female rats were then ovariectomized and submitted to stereotaxical implantation of 1 microgram oestradiol benzoate, 5 microgram testosterone isobutyrate or, as a control, 10 microgram cholesterol in the anterior hypothalamic areas. The effects of the steroids on plasma and pituitary FSH and LH were assessed by radioimmunoassay. As reported previously by us in normal female and male rats, the preoptic-suprachiasmatic area (POA) was able to control synthesis and secretion of both gonadotrophins and did not lose its sensitivity to oestradiol and testosterone in andorgenized rats. Evidence for enhanced prolactin secretion in androgenized rats was derived from immunofluorescence studies of the pituitary gland and from histology of the mammary glands. In this respect the condition of the androgenized females was opposite to that of the males. The present work demonstrated that stimulation of prolactin secretion in androgenized female rats resulted from oestrogen action due to permanent oestrus rather than from impairment of hypothalamo-hypophysial relationships. Indeed, prolactin stimulation was suppressed when the androgenized rats were ovariectomized and restored when they were subsequently implanted with oestradiol in the POA.

Effects of anterior hypothalamic implantation of gonadal steroids on serum and pituitary gonadotrophins (follicle-stimulating hormone and luteinizing hormone) in the castrated male rat.

Castrated rats were stereotaxically implanted with 1 microng oestradiol benzoate, 5 microng testosterone isobutyrate or, as a control, 10 microng cholesterol, in the hypothalamus. The effects of the steroids on plasma and pituitary gonadotrophins (FSH and LH) were assessed by radioimmunoassay. Our results indicate that, in the male rat, in addition to the arcuate nucleus-median eminence complex, the preoptic suprachiasmatic area is able to control synthesis and secretion of both gonadotrophins, and that it is sensitive to oestradiol and testosterone.

[Bovine 17-beta-estradiol during estrous cycle]. / Variations du 17 beta aestradiol au cours du cycle aestral chez la Vache

Using a highly specific antiserum, a method of plasma estradiol-17 beta determination by R.I.A. without chromatographic separation, was developed. Increasing on day 18, estradiol-17 beta reached a peak level of 8.6 pg/ml (6.4-12.6) at the onset of estrus. It then sharply decreased, and remained at a low level 1.75 pg/ml with secondary peaks on day 4-5, 12 and sometimes on day 8.

[Radioimmunoassay of bovine serum progesterone]. / Le dosage radio-immunologique de la progestérone plasmatique chez la vache

Radioimmunoassay determination of plasma progesterone in the cow without chromatographic purification was developed. The level ranged from 0.6 ng/ml during estrus to 6.4 ng/ml during the luteal phase. The progesterone level declined rapidly 48 to 72 h before estrus beginning.