Total Oocytes Research Articles

Effective method for emergency fertility preservation: random-start controlled ovarian stimulation

To determine whether random-start controlled ovarian stimulation (COS), in which a patient is stimulated on presentation regardless of her menstrual-cycle phase, has outcomes similar to conventional early follicular phase-start COS for fertility preservation in cancer patients. Retrospective cohort study. Academic medical center. Women recently diagnosed with cancer and in preparation for gonadotoxic therapy. Random- versus conventional-start COS. number of mature oocytes retrieved; secondary outcomes: pattern of follicular development, oocyte yield, and fertilization rate. The number of total and mature oocytes retrieved, oocyte maturity rate, mature oocyte yield, and fertilization rates were similar in random- (n = 35) and conventional-start (n = 93) COS cycles. No superiority was noted when comparing COS started in the late follicular (n = 13) or luteal phase (n = 22). The addition of letrozole, in the case of estrogen-sensitive cancers, did not adversely affect COS outcomes or oocyte maturity and competence in either random- or conventional-start protocols. Random-start COS is as effective as conventional-start COS in fertility preservation. This protocol would minimize delays and allow more patients to undergo fertility preservation and still proceed with cancer treatment within 2-3 weeks.

Aspects of reproduction of striped venus Chamelea gallina in the Gulf of Cádiz (SW Spain): Implications for fishery management

This study analyzes the main reproductive aspects of a natural population of Chamelea gallina on the SW Spanish coast (Gulf of Cadiz): gametogenic cycle, size at first maturity, size at sexual differentiation and partial fecundity. Methodology involved the use of standard histological techniques, image analysis and the assessment of the variation of gonadal growth.Monitoring throughout one year (May 2010–April 2011) identified 5 gametogenic developmental stages and evidenced the existence of a long reproductive period between March and September. In May 70% of individual were in stage 4, reproduction period (sub-stages 4A: maturity and 4B: partial emission), whereas in June most individuals were partially emitting gametes (4B). An important peak of gamete emission took place between June and July, after that recovery stages were detected (sub-stage 4C). In September most of individuals were in stage 5 (spent). It was not possible to define a single spawning event, but signs of partial spawning of greater or lesser intensity were identified throughout the whole reproduction period. This period was followed by a short period of sexual rest (stage 1; October), before beginning a new gametogenic cycle and of energy storage processes in November. Most of the individuals remained in a pre-active stage (stage 2: initiation of gametogenesis) until February, although advanced gametogenesis stages (stage 3) were also observed in January. The size (shell length) at first maturity was estimated for males (8.41mm), females (10.29mm) and the whole population (9.34mm) using standard histological techniques.For the range of shell lengths studied (20–30mm), between 40% and 60% of the visceral mass of C. gallina was devoted to reproduction. Gonadal volume was dependent on shell length and was between 37.25 and 205.95mm3. Gonadal volume and oocyte volume fraction were the most decisive factors in estimating fecundity. C. gallina is a multiple partial spawner and partial fecundity values were estimated and comprised between 76,835 and 797,424 oocytes per female. Partial fecundity was linearly related with both shell length and live weight. Since the oocyte output highly varies as shell length increases, reducing the minimum legal catch size (25mm) or overfishing legal size classes (>25mm) could diminish the total oocyte output and, consequently, affect the recruitment and the size structure of the population. Therefore, fishery management decisions must be taken carefully.

Serum Estradiol Level as a Predictor of Ovarian Response and Pregnancy Outcome During Controlled Ovarian Hyperstimulation in Women from Gaza Strip

Background: Currently, controlled ovarian hyperstimulation (COH) is monitored by serum estradiol (E2) levels which are believed to primarily detect functional activity of follicles. Objective: To evaluate estradiol level as a predictor of ovarian response and pregnancy outcome during COH in women from Gaza Strip. Methods: This prospective cohort study consisted of 75 women attending in vitro fertilization (IVF) at Al-Basma Fertility Center in Gaza City. Blood withdrawal for E2 hormone measurement was performed in all the patients and the number of oocytes and embryos were recorded for each female and the occurrence of pregnancy was followed for three months. Obtained data were computer analyzed using SPSS statistical package version 18. Results: The mean age of the study population was 29.2±5.9 years. Questionnaire interview showed that the cause of infertility was mostly referred to husbands. More than half of women seeking IVF had no children and had repeated IVF. The mean level of E2 showed the highest value of 2194.4 (pg/ml) at age group 26-35 years. However, the difference in E2 levels among the age groups was not significant (F=0.940 and P=0.395). When related to the number of oocytes retrieved, E2 level showed general increase with increase ovarian response, recording values of 1642.7, 1665.1, 2156.8 and 1798.7 pg/ml with <4, 4-8, 9-16 and >16 oocytes, respectively, but this change was not significant (F=0.219 and P=0.883). The mean level of E2 showed its maximum value of 2143.6 pg/ml in positive pregnancy. However the difference in E2 levels among the different categories of IVF outcome was not significant (F=0.423 and P=0.656). The numbers of total and mature oocytes, and embryo were significantly increased with increased levels of E2, showing a good response at E2 level=1000-2000 pg/ml. The number of mature oocytes showed positive correlations with E2 and number of embryo (r= 0.159, P= 0.177 and r= 0.890, P=0.000, respectively) and negative correlation with age (r=-0.276, P=.017). Conclusion: Estradiol level 1000-2000 pg/ml at mean age of 27.8±4.9 years, could be a predictor of ovarian response and pregnancy outcome during COH. Consequently, for women to undergo a more likely successful IFV program, it is recommended to optimize E2 level at 1000-2000 pg/ml.DOI: http://dx.doi.org/10.3329/jom.v14i1.14537 J MEDICINE 2013; 14 : 52-56

CYP19 gene variants affect the assisted reproduction outcome of women with polycystic ovary syndrome

Objective: Cytochrome P450 aromatase catalyzes the irreversible transformation of androgens into estrogens. The association of CYP19(TTTA)n polymorphism with the hormonal profile and the assisted reproduction outcome of women with polycystic ovary syndrome (PCOS) was explored.Methods: One hundred and thirty-two women with PCOS and 200 with male-factor infertility, as controls, participated in the current study. The CYP19(TTTA)n polymorphism was genotyped, while the hormonal profile was determined at the third day of the menstrual cycle. During oocyte retrieval, the follicular size, the follicle and oocyte numbers were recorded.Results: Genotype analysis revealed 6 CYP19(TTTA)n alleles with 7–12 repeats. In PCOS women, the CYP19(TTTA)7 allele presence was associated with lower serum E2 levels at the third day of the menstrual cycle (p < 0.009), lower large follicle (p < 0.02) and total oocyte numbers (p = 0.006), but with significantly higher pregnancy rates after assisted reproduction (p < 0.004).Conclusions: Potential associations of the CYP19(TTTA)7 allele with ovarian response to standard gonadotrophin stimulation and with assisted reproduction outcome were found in PCOS women, probably due to androgen/estrogen ratio alterations.

Influence of heparin or the presence of cumulus cells during fertilization on the in vitro production of goat embryos

Considerable research has been focused on in vitro production (IVP) of goat embryos to improve its efficiency. In Experiment 1, the effect of the cumulus cells by comparing slaughterhouse-oocytes denuded on purpose (DOP) prior to IVF to intact COC, and the effect of heparin during IVF were assessed. In Experiment 2, oocytes that were already denuded at collection (DOC), DOP and intact COC were studied. Three treatments used oocytes denuded at collection: DOC oocytes were cultured alone for both IVM and IVF; DOC and COC were cultured together for both IVM and IVF or DOC were IVM alone and then mixed with COC for IVF. In other treatments, COC were allocated to four IVF treatments: Intact COC; COC were denuded prior to IVF; COC were denuded and IVF with added cumulus cells; COC were denuded and IVF mixed with intact COC giving two sub-treatments: Denuded oocytes that were IVF with COC; and COC that were IVF with denuded oocytes. After fertilization, all presumptive zygotes were cultured for 8 days. In Experiment 1, the yield of blastocysts as a proportion of total oocytes was greater (P<0.05) for COC that were IVF in the presence of heparin (54%) than without heparin (42%) or oocytes already denuded at collection that were IVF with or without heparin (41%; 38%; respectively). In Experiment 2, the developmental potential of oocytes denuded at collection was reduced (cleavage and blastocyst rates calculated from total oocytes: 34%; 11%, respectively) as compared to COC (77%; 59%, P<0.05). However, when equal numbers of both were mixed at the start of IVM, the rates were not significantly different to COC alone (68%; 45%), but when both were mixed equally only for IVF, the rates were reduced (57%; 40%, P<0.05). Denuded oocytes co-cultured with cumulus cells were not significantly different to intact COC (76%; 55%). The effect of adding COC during IVF to oocytes denuded after IVM was similar to adding cumulus cells to the same type of oocytes. In conclusion, both the use of heparin and the association of oocytes with cumulus cells, either detached or in intimate contact, during IVM and/or IVF significantly improve IVP of goat embryos. Furthermore, some oocytes that are already denuded at collection will develop satisfactorily to blastocysts when matured and fertilized with intact COC.

A Stockpile of Ova in the Grass Frog Rana temporaria is Established Once for the Life Span. Do Ovaries in Amphibians and in Mammals Follow the Same Evolutionary Strategy?

Most anuran amphibians produce high numbers of eggs during several consecutive breeding seasons. The question is still open whether oocytes are formed anew as a result of oogonial proliferation after each spawning or the definitive pool of oocytes is established during the juvenile period and is sufficient for the whole reproductive life span of a female. Our quantitative studies show that primary oogonia in adult female frogs can proliferate, but they fail to differentiate further and do not enter meiosis, and thereby there is no supplementation of new generations of oocytes after each spawning. Ovaries of one-year-old grass frogs contain (median) 53,447 diplotene oocytes, in two-years-old frogs this number decreased to 33,583 and eventually reached 25,679 in virgin mature females. More than 50% decrease in the total oocyte number was accompanied by massive degeneration (atresia) of oocytes. The final number of oocytes in a female forms a stock for 11-12 breeding seasons and exceeds the number of eggs produced during the potential reproductive life span of this species. The phylogenetic context of oocyte recruitment modes in the major clades of vertebrates is discussed in respect to their ability to replenish the stock (a renewable stock in ovaries named "open" vs. a non-renewable stock in ovaries named "closed").

Efeitos da suplementação com fontes de ácidos graxos em doadoras da raça Holandesa durante o pré e pós-parto sobre o retorno à ciclicidade e produção in vitro de embriões

The supplementation of dairy cattle with sources of polyunsaturated fatty acids (PUFA) can be use to increase the energy level of the diet in addition to having positive effects on reproductive functions of important tissues including the hypothalamus, pituitary, ovaries and uterus. The aims of this study were to evaluate the reproductive conditions of the postpartum, number of follicles, corpus luteum (CL) presence, concentration of progesterone (P4), aspirated oocytes, amount of viable oocytes and in vitro production of embryos (IVPE) of the Holstein multiparous donors supplemented with rich diet in protected PUFA (especially linoleic acid - n- 6) and non-protected (especially linolenic acid - n-3) during pre and post partum. The diets had been given for pre-partum during 30 d and post partum 60 d. The donors were divided into three groups: Control (n=6), Megalac-E® (n=5; supplemented with protected fat source 100 g/donor/ day in pre-partum and 300 g/donor/day in postpartum) and linseed (n=5; supplemented with fat source unprotected containing 1 kg/donor/day pre-partum and 1.5 kg/donor/day in postpartum). The animals were submitted to ovum pick-up (OPU) on days 30, 45 and 60 d postpartum. The recovered oocytes were selected and the viable ones were submitted to IVPE procedures. The data were analyzed by the method of least squares variance using the GLM protocol. The differences between averages were compared by Tukey test with 5% significance. There was no detectable effect of treatment, aspirations of postpartum days and interactions on variables: CL presence, concentration of P4, amount of viable oocytes, viable oocytes rate, IVPE and embryos production rate. However, was observed in the group supplemented with linseed more follicles and total oocytes than Megalac-E® and Control group. Supplementation with PUFA didn't increase the number of viable oocytes and IVPE.

Comparison of microdose GnRH agonist protocol with GnRH antagonist / letrazole protocol in patients with poor ovarian response

Objective: To compare microdose GnRH agonist (MDL) protocol with GnRH antagonist/letrazole (AL) protocol in patients with poor ovarian response. Design: Randomised clinical trial. Setting: Kocaeli University Assisted Reproductive Centre Patients: Fifty-five patients with the diagnosis of poor ovarian response (POR) were randomized and performed either MDL or AL protocol. Interventions: POR was diagnosed according to ESHRE Bologna criteria in case of presence of 2 out of 3 criteria. In the MDL group (n=27), 40µg/0.2 cc subcutaneous leuprolid acetate was given two times a day starting on the first day of menstrual cycle. On the second day of the cycle, gonadotrophin stimulation was started at 450-600 IU and/or 150 IU human menopausal gonadotrophin (hMG) along with leuprolid. In the AL group (n=28), 5 mg letrazole was given for 5 days starting on the second day of the spontaneous cycle and after the 5th day of the cycle, 450-600 IU and/or 150 IU hMG was given. Main Outcome Measures: To compare the outcomes of MDL protocol with AL protocol in patients with poor ovarian response. Results: Stimulation days, total gonadotrophin dose (IU), peak E2 (pg/ml) levels, and endometrial thickness were found to be statistically significantly lower in the AL group. While no cases of cycle cancellation was detected in the MDL group, statistically significantly higher rates of cycle cancellation (%42.8, p<0.001) was detected in the AL group. No statistically significant differences were detected in total oocyte numbers (3.7±2.9 vs 2.7±2.2), number of transferred embryos (1.4±0.5 vs 1.4±1.9) and pregnancy rates (3 vs 2) in between the groups. Comment: Although AL protocol is not superior to MDL protocol in poor ovarian response patients, it may be used as an alternative protocol.

Follicle flushing in monofollicular in vitro fertilization almost doubles the number of transferable embryos

Follicle flushing has been proved to be ineffective in polyfollicular in vitro fertilization. To analyze the effect of flushing in monofollicular in vitro fertilization we aspirated and then flushed the follicles in 164 cycles. Total oocyte yield/aspiration was 44.5% in the aspirate, 20.7% in the 1st flush, 10.4% in the 2nd flush and 4.3% in the 3rd flush. By flushing, the total oocyte yield increased (p < 0.01) by 80.9%, from 44.5 to 80.5%. The total transfer rate increased (p < 0.01) by 91.0%, from 20.1 to 38.4%. The results indicate that the oocyte yield and the number of transferable embryos can be increased significantly by flushing.

Clomiphene combined with gonadotropins and GnRH antagonist versus conventional controlled ovarian hyperstimulation without clomiphene in women undergoing assisted reproductive techniques: systematic review and meta-analysis

To compare the effectiveness and safety of controlled ovarian hyperstimulation (COH) using clomiphene citrate associated with gonadotropin and GnRH antagonist (CC + Ant) versus conventional COH without clomiphene citrate (Non-CC) for women undergoing assisted reproductive techniques (ART). Systematic review and meta-analysis of randomized controlled-trials comparing CC + Ant versus non-CC. The last search was performed in Apr 13 2012. The following outcomes were retrieved from included trials and compared between CC + Ant versus non-CC: live birth, clinical pregnancy, ovarian hyperstimulation syndrome (OHSS), miscarriage, multiple pregnancy, endometrial thickness, total oocytes retrieved, MII oocytes retrieved, total gonadotropin used, and duration of COH. Seven trials (702 participants) were included. There was no significant difference in live birth (55/182 = 30.2 % vs. 47/181 = 26.0 %, p = 0.26, CC + Ant vs. non-CC, respectively), clinical pregnancy (98/346 = 28.3 % vs. 84/356 = 23.6 %, p = 0.12), miscarriage (6/35 = 17.1 % vs. 7/32 = 21.9 %, p = 0.42), endometrial thickness, and on the number of oocytes retrieved. There was a significant reduction in OHSS (1/216 = 0.5 % vs. 9/217 = 4.1 %, p = 0.01), consumption of gonadotropins, and duration of COH. Compared to non-CC, CC + Ant is likely to reduce the risk of OHSS, medication costs, and the duration of COH without evidence of effect on live birth and clinical pregnancy. More trials are still needed to improve the quality of the evidence. Future studies should evaluate women with PCOS or at increased risk of OHSS, quality of life, satisfaction with the treatment, total cost per live birth, and frequency of congenital anomalies.

Fecundity and Length-Weight Relationship of Two Commercially Important Fish Species in Minna, Niger State

Two commercially important fresh water fish (Clarias gariepinus and Oreochoromis niloticus) were investigated to access their fecundity and length/eight relationship. Morphometric measurements such as Standard length (SL), Total length (TL), Total fish body weight (TBW), Total ovary weight (TOW), oocyte weight and fecundity were carried out on 20 samples of each of these two species sourced from Minna market, Niger State. Fecundity of Clarias gariepinus were moderately higher (1225 – 45640 eggs/ fish of total length 19.50 -14.00cm) than Oreochromis niloticus (369 – 22460eggs/fish of TL 15.50 – 29.50cm). T –test analysis showed significant difference (P&gt;0.05) in length and weight relationship.

34 PRE-IMPLANTATION DEVELOPMENT OF HORSE SOMATIC CELL NUCLEAR TRANSFER EMBRYOS ORIGINATED FROM METAPHASE I OOCYTES

Because of the limited availability of horse oocytes and their wide maturation window that leads to the existence of a subpopulation of oocytes with significantly longer maturation period, horse cloning would benefit from the possibility of also using metaphase I oocytes (Choi et al. 2009 Cloning Stem Cells). The scope of this work was to compare the developmental ability of cloned horse embryos constructed using oocytes in metaphase I (MI) and II (MII). Oocytes of slaughtered mares were matured for 27 h in DMEM/F12 with 10% FCS, 1 µL mL–1 of insulin-transferrin-selenium (ITS), 1 mM sodium pyruvate, 50 ng mL–1 of long-epidermal growth factor, 100 ng mL–1 of long-insulin-like growth factor-I, and 0.1 IU mL–1 each of FSH and LH, denuded of cumulus and enucleated using a zona-free method (Lagutina et al. 2005 Reproduction). Oocytes with a visible polar body were classified as MII and those with no polar body as potential MI. During enucleation, oocytes having a metaphase plate only were confirmed as MI; oocytes in anaphase and telophase were classified as MII. Adult skin fibroblasts of passages 2 to 10 were cultured in TCM 199/DMEM with 10% FCS and serum starved for 1 to 2 days before NT. Nuclear-transfer embryos were constructed after washing of zona-free oocytes in 400 µg mL–1 of phytohemagglutinin P and attachment of each to a single cell in HEPES-SOF by fusion with 2 direct-current (DC) pulses of 1.2 kV cm–1 applied for 30 µs in fusion medium. One hour after fusion, embryos were activated by 5 µM ionomycin for 4 min, followed by culture in 5 µg mL–1 of cycloheximide and 1 mM DMAP in SOFaa for 3 h. Embryos were cultured in SOFaa in 5% CO2 and 5% O2 at 38.5°C. Half of the medium was renewed on Day 3 and replaced on Day 5 with DMEM/F12 with 5% FCS and 5% serum replacement. Cleavage was assessed 48 h after activation and the rate of blastocyst formation was recorded at Day 8. The data were compared by chi-square test. The development ability of MI embryos assessed by cleavage and blastocyst formation was significantly lower than of MII embryos (Table 1). The obtained MI blastocysts were smaller than their MII counterparts. These data demonstrate that MI oocytes account for 20% of the total oocytes after 27 h of maturation, have a lower developmental competence to form blastocysts after NT, and the blastocysts obtained are of smaller size and likely less viable. Therefore, the use of MI oocytes can only marginally improve the outcome of horse cloning. Table 1.Embryo development after SCNT using oocytes in metaphase I or II

38. Impact of microwave processing on dehydration and structural integrity of germinal vesicles from domestic cat oocytes

The ability to compact and inject the cat germinal vesicle (GV) into a recipient cytoplast allows exploration of a new fertility preservation strategy that avoids whole oocyte freezing and could enable drying and storage of GV at supra-zero temperatures. Previous studies demonstrated that (1) 60% of GVs can survive simple air-drying in the presence of trehalose followed by 4 weeks of storage at 4 °C and (2) 10% of reconstructed oocytes with rehydrated GV can achieve nuclear maturation (Graves-Herring et al., 2011). However, air-drying is an uncontrolled process that often results in non-uniform distributions of water within and between samples. Compared to existing techniques, microwave assisted dehydration appears to be a simple and reliable approach to dry biological samples at ambient temperatures (Chakraborty et al., 2008). The objective of the study therefore was to (1) explore the feasibility of drying GV exposed to trehalose using microwave processing and (2) assess the impact on GV structural integrity (chromatin configuration and incidence of DNA damage). Immature cat oocytes from adult ovaries were denuded from the cumulus cells and permeabilized with hemolysin. Groups of five denuded oocytes were exposed to Tris–HCL EDTA (pH 8.0) buffer containing 200 mM trehalose for 10 min and then transferred into 10-μl droplets placed on small pieces of Millipore Isopore Membrane filters (n = 182 total oocytes in 5 replicates). A maximum of eight droplets were arranged on the turntable of a CEM SAM 255 microwave system. Drying was conducted at 20% power for 5, 8, 20, 25 or 30 min. After each time point, samples were weighed immediately to establish the extent of water loss and then were rehydrated before fixation in 4% paraformaldehyde followed by GV chromatin configuration assessment (Hoechst staining) and DNA fragmentation assay (TUNEL). Fresh denuded oocytes (n = 60 total oocytes) were fixed as controls. The average ambient humidity during all measurements was 43.2 ± 1.1%. From time 0 through 30 min of microwave treatment, moisture contents (g water/g dry weight) progressively decreased (0 min: 12.3 ± 0.5; 5 min: 8.3 ± 1.2; 8 min: 8.9 ± 1.2; 20 min: 5.0 ± 0.5; 25 min: 1.9 ± 0.3; 30 min: 1.3 ± 0.4). GV chromatin configuration was not affected by the microwave treatment. The proportion of degenerated oocytes (absence of GV) ranged from 10% to 15% after the different time points and was not different (P > 0.05; Chi-square test) from the fresh controls. Interestingly, pooled proportions of GV with DNA damage following 5–8 min of microwave dehydration were not different (P > 0.05) from fresh controls (14.3% and 10.9%, respectively). However, the incidence of DNA fragmentation was higher (43.5%; P

Protein diets promote the maturation of oocytes and spawning of Piaractus mesopotamicus kept in cages

Summary The objective of this study was to evaluate the effects of balanced diets on the maturation of oocytes and the reproductive performance of P. mesopotamicus in cages. A completely random design with 224 fish in 16 cages measuring 5 m3 was employed for this purpose. The treatments consisted of diets containing 18, 24, 30, and 36% crude protein (CP) provided ad libitum. The external and internal morphological characteristics of the specimens were examined, as well as: the position of the germinal vesicle, the distribution of oocyte diameters, the fertilization and hatching rates, the number of oocytes released, the total number of oocytes, the remaining weight and total weight of the ovaries, the gonadosomatic index, the condition factor (K), and the histology of the oocytes and ovaries post-spawning and during ovarian regression. The diameters of the oocytes collected before the first hormonal application displayed a unimodal distribution for the lowest protein content and a polymodal distribution for the other treatments. A similar situation was seen during spawning. The lowest fertilization and hatching rates were found as a consequence of the treatment with 30% CP (P < 0.05). The greatest hatching rate occurred in the females fed 18% CP. The greatest total oocyte weight was found in the specimens that received between 30 and 36% CP. The lowest K index was found in the females fed 36% CP. In conclusion, a diet containing 18% CP satisfies the reproductive requirements of females adapted to this system.

Predicting successful induction of oocyte maturation after gonadotropin-releasing hormone agonist (GnRHa) trigger

Are there factors predicting the number of total and mature oocytes retrieved after controlled ovarian hyperstimulation (COH) utilizing a gonadotropin-releasing hormone (GnRH) antagonist protocol and a GnRH agonist (GnRHa) to induce oocyte maturation? Peak estradiol (E₂) level, post-trigger LH and progesterone and the magnitude of LH rise are independent predictors of the total number of oocytes and mature oocytes retrieved. Despite multiple follicular development in high responders, oocyte retrieval after a GnRHa trigger in a small subset of patients fails to obtain a substantial number of total oocytes or mature oocytes. A retrospective chart review of all autologous and oocyte donation cycles utilizing a GnRHa antagonist protocol where GnRHa was used for the induction of oocyte maturation between 1 April 2003 and 31 December 2011. A total of 508 autologous and donor IVF/ICSI cycles utilizing a GnRH antagonist protocol for COH and GnRHa for the induction of oocyte maturation at a university-based tertiary fertility center. Peak E₂ on the day of trigger (r = 0.19, P < 0.001), post-trigger LH (r = 0.12, P = 0.009) and progesterone (r = 0.47, P < 001) and LH rise (r = 0.18, P < 0.001) all positively correlated with the number of total and mature oocytes retrieved. The true incidence of empty follicle syndrome was 1.4% (7/508). There was no post-trigger LH or progesterone cut-off value for the prediction of oocyte yield. However, all cases of empty follicle syndrome occurred in patients with post-trigger LH <15 IU/l and P ≤ 3.5 ng/ml. The findings of this study may also be due to chance since it was a retrospective study and not prospectively designed. This is a retrospective chart review and therefore subject to bias. Serum hormone measurements were performed between 8 and 12 h after GnRHa trigger rather than a standardized time period following trigger administration. Therefore, peak levels of LH may have been missed due to the short ascending limb of LH rise lasting approximately 4 h after GnRHa trigger. The results of this study can be generalized to high responders utilizing a GnRH antagonist protocol for COH and a GnRHa for the induction of oocyte maturation. The use of alternative stimulation regimens or medications will limit the ability to generalize the results of this study to other populations. This study was not funded, and there are no conflicts of interest. n/a.

The Effect of Local Injury to the Endometrium for Implantation and Pregnancy Rates in ICSI –ET Cycles with Implantation Failure: a randomised controlled study

To assess the effect of local injury to the endometrium for implantation, pregnancy and live birth rates in ICSI –ET cycles with recurrent implantation failure. The study was prospectively designed and performed in a university hospital. A group of 100 women, who failed to conceive during one or more cycles of IVF and embryo transfer (ET), was treated with a long protocol for controlled ovarian hyperstimulation. The IVF treatment and ET were preceded by repeated endometrial biopsies, in a randomly selected 50 of a total of 100 women. Age of the women, days of stimulation, total dose of gonadotropins, E2 concentrations and endometrial thickness on hCG day, total MII stage oocytes, the total number of high quality embryos for transfer, the day of transfer and the number of embryos transferred were similar for both groups. Transfer of a similar number of embryos (138 and 141 in the experimental and control patients, respectively) resulted in rates of implantation (34.67 % vs. 30.88 %; p= 0,1384) clinical pregnancy (60 % vs. 34 %; p= 0.009), and live births per ET (44 % vs. 24 %; p= 0.03), which were higher in the experimental group as compared to controls. These results suggest that IVF treatment that is preceded by local injury to the endometrium improved the rates of embryo implantation, clinical pregnancy, and live births in asiste reproductive technique.

Fertility preservation with IVF OR IVM for women with hematological malignancy – experiences and expectations

Women with hematological malignancy are often referred for fertility preservation before commencing potentially gonadotoxic chemotherapy.Treatment is individualized according to age, ovarian reserve, and urgency of chemotherapy. In vitro fertilization (IVF), and more recently, in vitro maturation (IVM), offer the potential to cryopreserve embryos or oocytes.This study compares IVF and IVM techniques for fertility preservation in women with hematological malignancy. Retrospective cohort study,single tertiary center. Women underwent fertility preservation with IVF or IVM for hematological malignancy, during 2003-2011. Age,antral follicle count (AFC), oocytes retrieved (total and MII), MII oocytes after maturation, and number of embryos or oocytes for cryopreservation were compared for each treatment arm. 66 women with hematological malignancy underwent treatment with either IVF (38) or IVM (28) for oocyte and/or embryo cryopreservation. 37 women had Hodgkin's lymphoma (20 IVF, 17 IVM), 19 non-hodgkin's lymphoma (11 IVF/8 IVM), 5 AML(2 IVF/3 IVM), 3 myelodyslastic syndrome (all IVF) and 2 aplastic anemia (both IVF). Age (27.2vs 25.7) and AFC (16.3 vs 20.2) were similar. Women who underwent IVF had significantly more MII oocytes collected compared with those who underwent IVM (8.4 vs. 2.1, P<0.05). However, there was no significant difference in oocytes collected (10.6 vs 11.6), total MII oocytes (8.9 vs 8.4), embryos fertilized (6.7 vs 6.75), or number of embryos (5.9 vs 6.25) or oocytes (8.6 vs 8.25) for cryopreservation. Fertility preservation treatment for women with hematological malignancy should be tailored for ovarian reserve, prognosis, and cancer treatment protocol and timing. Whilst IVM with oocyte or embryo cryopreservation for malignancy is still investigational, it may offer an alternative to IVF for fertility preservation. Larger studies, as well as data regarding pregnancy outcomes, are needed to provide support and evidence for this treatment option.

Anti-mullerian hormone evaluation and initial ovarian response in an apparently good prognosis population treated by intracytoplasmic sperm injection

To evaluate the ability of anti-mullerian hormone (AMH) to predict ovarian response in a selected population treated by intracytoplasmic sperm injection (ICSI). A prognostic study. Data from a total of 100 first attempts were studied. The inclusion criteria included, among others, basal mass index<35, age<38,antral follicle count (AFC, 3-8 in diameter) between 14 and 30, FSH<10 IU/L estimated at the beginning of a non stimulated cycle. A long analog protocol was used, and ovarian stimulation was initiated by 200 international Units of recombinant follicle-stimulating hormone per day. The first estradiol (E2) evaluation was performed on day 4. Treatment was not modified when E2 concentration on day 4 was between 0.5 and 1 mmol/L. The measurement of AMH was determined by the enzyme linked immune-absorbent assay provided by Immunotech Beckman Coulter. AMH concentration remained unknown to the clinical team until the end of the attempt. In this population AMH varied from 8 to 78 pmol/L. AMH and AFC were significantly correlated with E2 on day 4 and with E2 at the end of the stimulation. In a multiple linear regression analysis,only AMH remained significantly correlated with E2 on day 4 (P<0.003). Treatment was modified on day 4 in 62 cases (E2<0.5 nmol/L in 56 cases). In a multiple linear regression, only AMH remained significantly correlated with treatment modification frequency (P<0.003). Cancellation (less than 3 adequate follicles) was decided in 6 cases. Cancellation rate was only correlated with AMH (P<0.004). AMH (P<0.001) and AFC (P<0.03) were significantly correlated with total oocyte number. AMH concentration is significantly correlated with initial ovarian response and cancellation risks independently of AFC, age and basal FSH.

Ratio of euploid embryos following comprehensive chromosomal screening (CCS) in day 5 and day 6 blastocyts

To determine the ratio of euploid embryos obtained from day 5 and day 6 embryos. Prospective cohort study. A total of 778 oocytes were retrieved from 52 patients with intent of trophectoderm biopsy, cryopreservation and analysis with CCS. Embryos were all subjected to intracytoplasmic sperm injection and group cultured with 4 to 5 embryos per microdoplet under oil in a sequential culture system. Day 3 embryos were laser assisted hatched then moved to the blasotcyst segment of the sequential culture system. Blastocysts on day 5 and day 6 the displayed a distinct inner cell mass and multicelluler trophectoderm were subjected to trophectoderm biopsy utilizing a laser sampling technique. All biopsied embryos were cryopreserved using a slow cool methodology. A total of 209 embryos were biopsied in this study. Day 5 resulted in 60 (56%) euploid embryos of the 108 blastocysts biopsied. Alternately, a total of 56 (55%) day 6 embryos were euploid from the 101 blastocysts biopsied (N.S.). There was no significant difference in the total number of blastocysts biopsied between day 5 and day 6 (14% and 13% respectively). Although some preliminary studies have shown that day 5 embryos have a higher rate of euploidy as compared to day 6, these authors have not observed these same results. These data show the inefficiency in the human reproductive potential in that only 116 (15%) euploid embryos were obtained from the 778 total oocytes. Further evaluation may show that CCS is the optiimal way to obtain the best embryo for the overall reduction of multiples with the focus on patient safety.

Fecundity regulation strategy of the yellowfin tuna (Thunnus albacares) in the Western Indian Ocean

The oocyte development process and fecundity regulation (i.e. whether yellowfin tuna shows determinate or indeterminate fecundity) were analyzed to investigate the reproductive strategy of yellowfin tuna, Thunnus albacares. A total of 819 yellowfin ovaries were sampled at sea and at the Seychelles cannery during 2009 and 2010 from purse-seiners operating in the Western Indian Ocean. Histological analysis and automated computer-controlled image analysis software were used to study four main criteria applied for fecundity style determination: (a) oocyte size–frequency distribution, (b) number of cortical alveoli and total vitellogenic oocytes in different ovary maturation phases, (c) differences in mean diameter of tertiary vitellogenic oocytes, and (d) seasonal development of atresia.The results revealed an asynchronous oocyte development and a continuous oocyte size–frequency distribution throughout all ovarian developmental phases over the spawning season. The percentage and number of cortical alveoli and total vitellogenic oocytes remained constant through the spawning season. The mean diameter of tertiary vitellogenic oocytes decreased as spawning progressed. Also, the incidence of atresia was higher at the end of the spawning season as a consequence of over-recruitment of oocytes during this period. These findings revealed that yellowfin tuna exhibit indeterminate fecundity.