Evaluating the value of day 0 of an ICSI cycle on indicating laboratory outcome.

A number of oocyte characteristics have been associated with fertilization, implantation and live-birth rates, albeit without reaching a consensus. This study aims to delineate possible associations between oocyte characteristics, oocyte behavior during intracytoplasmic sperm injection (ICSI), fertilization potential, and laboratory outcomes. Four-hundred and seventy-seven patients, yielding 3452 oocytes, were enrolled in this prospective observational study from 2015 to 2018. Οoplasm granularity was associated with poor embryo quality and higher probabilities of post-ICSI oocytes and embryos discarded in any developmental stage and never selected for embryo transfer or cryopreservation (p < 0.001). Both sudden or difficult ooplasm aspiration, and high or lack of resistance during ICSI were associated with either a poor Zygote-Score or fertilization failure (p < 0.001). Sudden or difficult ooplasm aspiration and high resistance during ICSI penetration were positively associated with resulting to a post-ICSI oocyte or embryo that would be selected for discard. Evaluation of oocyte characteristics and oocyte behavior during ICSI may provide early information regarding laboratory and cycle outcomes. Particularly, ooplasm granularity, and fragmentation of polar body, along with sudden or difficult ooplasm aspiration and high or lack of resistance during ICSI penetration may hinder the outcome of an ICSI cycle. The associations presented herein may contribute towards development of a grading system or a prediction model. Taking into account information on oocytes and ICSI behavior may effectively assist in enhancing IVF outcome rates.

The use of serum anti-Mullerian hormone (AMH) levels and antral follicle count (AFC) to predict the number of oocytes collected and availability of embryos for cryopreservation in IVF.

AIM: To investigate the predictive value of anti-Mullerian hormone (AMH) and antral follicle count (AFC) on the final number of oocytes retrieved and the availability of embryos for cryopreservation in in vitro fertilization (IVF) cycles. PATIENTS AND METHODS: In this prospective study, one hundred and twenty women in their first IVF treatment were enrolled. The short stimulation agonist protocol was used for controlled ovarian hyperstimulation in all cases. Serum AMH levels were measured during the menstrual cycle preceding treatment. AFC was measured in cycle day 2, just before starting ovarian stimulation. RESULTS: A strong, positive correlation between AMH, AFC and the number of collected oocytes was found. The patients with available and suitable supplementary embryos for cryopreservation had higher levels of AMH and larger numbers of AFC. CONCLUSION: AMH and AFC appear to be valuable markers mainly for ovarian reserve and response to IVF treatment. Serum AMH levels and AFC are significantly associated with the number of retrieved oocytes. Also, a positive correlation with the availability of supernumerary embryos suitable for cryopreservation was observed.

Follicular fluid levels of vascular endothelial growth factor and its receptors and pregnancy outcome of women participating in intracytoplasmic sperm injection cycles.

PURPOSE: The intracytoplasmic sperm injection (ICSI) outcome is depended mainly on oocyte quality. Cytokines and their receptors play a critical role in oocyte maturation, fertilization, and embryo implantation. The purpose of the study was to study the levels of vascular endothelial growth factors (VEGFA, VEGFR1, VEGFA) in follicular fluids (FF) women participating in ICSI-in vitro fertilization (IVF) cycles in relation to cycle's outcome. MATERIAL AND METHODS: One hundred and fifty three samples of 70 women participating in ICSI cycles were classified in three infertility groups: male factor, female factor, and low responders. For controlled ovarian stimulation in male and female factor group, the long agonist protocol with leuprolide and recombinant follicle stimulating hormone (FSH) was employed, while the antagonist cetrorelix was used in low responders. Cytokines levels were evaluated with enzyme-linked immunosorbent assay (ELISA). RESULTS: In a total of 153 samples, the overall pregnancy rate was 51.6%, the higher one observed in female factor group (59% vs. 37.5% and 28.6% in male a factor and low responders group, p = 0.013. VEGFR2 differed statistically significantly between the two groups, being higher in the pregnancy group [median (IQR): 5,630 (4,870 - 6,651) vs. 4938 (4,068 - 6,020) in the non-pregnancy group, p = 0.003]. There were significant correlations between VEGF receptors, differentiated depending on infertility groups. CONCLUSIONS: The VEGFA/VEGFR2 system is important in human reproduction and the association pattern between VEGFA receptors may serve as a marker for ICSI outcome. Examination for spermatozoa functional defects may increase pregnancy rate in male factor group.

Binovular complexes after ovarian stimulation. A report of four cases.

BACKGROUND: Human ovary is known to contain polyovular follicles at birth but it is considered to be rare in adult ovaries. CASES: Four cases are presented where binovular complexes were found after ovarian stimulation. The age of women was 22 to 33 years and all of them had a good response to ovarian stimulation. Two different types of binovular complexes were found. The mature oocytes of three binovular complexes were fertilized by ICSI. The generated embryos did not develop beyond the eight-cell stage. CONCLUSION: The mature oocytes of binovular complexes can be fertilized but their developmental potential is limited.

Open versus closed vitrification of blastocysts from an oocyte-donation programme: a prospective randomized study.

The use of open carriers for embryo vitrification has raised safety concerns and therefore vitrification in closed systems has been proposed. However, the drop in the cooling rate emerges as a major drawback. The objective of the present study was to compare the efficiency of vitrification in open versus closed conditions. Blastocysts were randomly allocated either to open ultra-rapid vitrification (group I) or closed aseptic vitrification (group II). In group I, blastocysts were exposed to two solutions of ethylene glycol/dimethylsulphoxide (10%/10% and 20%/20%), while in group II, blastocysts were pretreated with a solution of lower concentration (5%/5%). A total of 208 and 224 vitrification-warming cycles were performed for groups I and II, respectively. Both groups were equal in terms of maternal age, sperm parameters and number and quality of blastocysts vitrified, warmed and transferred per cycle. Importantly, there was no significant difference between the groups in the analysed outcomes; embryo survival rate (84.1% versus 82.1%), clinical pregnancy rate (45.9% versus 42.4%), implantation rate (25.6% versus 24.5%), cycle cancellation rate (6.7% versus 8.5%) and live birth rate (41.2% versus 41.0%). These data suggest that ultra-rapid vitrification may be replaced by aseptic vitrification without affecting clinical efficiency.

The effects of benzoylecgonine, oxytocin, ritodrine and atosiban on the contractility of myometrium. An experimental study.

PURPOSE: To investigate the response of pregnant and non pregnant rat myometrium to benzoylecgonine, a cocaine metabolite, and oxytocin and to investigate the efficiency of ritodrine and atosiban to overcome the effects of benzoylecgonine and oxytocin. METHODS: Isolation of rat myometrial tissue and recording of contractile activity with isotonic muscle transducer. RESULTS: Benzoylecgonine and oxytocin increase myometrial contractility, while atosiban and ritodrine induce myometrial relaxation. Atosiban was able to revoke the action of oxytocin but not the action of benzoylecgonine. Ritodrine was able to induce muscle relaxation in both oxytocin and benzoylecgonine administration. CONCLUSION: Cocaine metabolites seem to act on the myometrium through different pathways compared with oxytocin. After comparing two widely used tocolytic agents: atosiban and ritodrine, it is indicated that only ritodrine, a beta2 adrenergic receptor agonist, can inhibit the action of cocaine metabolites. This finding indicates that the actions of cocaine on adrenergic mechanisms are responsible to a large part for its effects on myometrium contractility. The use of beta2 adrenergic receptor agonists seems to be preferable for the treatment of myometrial contractions induced by cocaine consumption.

Serum pattern of circulating soluble receptor of leptin throughout the menstrual cycle.

AIM: The aim of the study was to investigate the pattern of circulating soluble receptor of leptin (sLeptinR) during the menstrual cycle and the association of sLeptinR to leptin, sex hormones and gonadotropins. METHODS: Fasting blood samples were collected on alternate days throughout a full cycle from fifteen healthy volunteers. Immunoenzymatic assays were employed to record the relevant levels. RESULTS: sLeptinR concentrations throughout the cycle were found to vary negligibly. No significant correlations between sLeptinR and leptin, gonadotropins or progesterone, were established. During the follicular phase, subjects presenting with higher estradiol levels tended to have higher sLeptinR concentrations. CONCLUSION: The little variation of sLeptinR concentrations during the menstrual cycle indicates that the rise of leptin during the luteal phase implies an increase of its bioactivity.

Cytokine profile in cases with premature elevation of progesterone serum concentrations during ovarian stimulation.

The aim of this study was to investigate the concentrations of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), leptin, tumor necrosis factor-alpha, interleukin (IL)-1beta and IL-6, in cycles with a premature rise of serum progesterone. 25 intracytoplasmic sperm injection (ICSI) cycles with (Group 1) and 25 ICSI cycles without a premature progesterone elevation (Group 2) were included. The cut-off value of serum progesterone on the day of human chorionic gonadotropin (hCG) administration was 0.9 ng/ml. The indication for ICSI was male factor infertility exclusively. On the day of hCG injection, serum IL-6, VEGF and bFGF were significantly higher in Group 1 (7.7+/-24.5 pg/ml, 290.2+/-161.4 pg/ml and 15.7+/-8.2 ng/ml respectively) than in Group 2 (1.7+/-0.7 pg/ml, 175.2+/-92.1 pg/ml, and 9+/-1.6 ng/ml respectively). On the day of follicular puncture, serum cytokine concentrations were similar in the two groups. IL-6 intrafollicular concentrations were higher in Group 1 (14.7+/-20.7 pg/ml) than in Group 2 (9+/-9.3 pg/ml, p=0.031). There were no differences regarding the ICSI outcome. Patients with serum progesterone above 0.9 ng/ml, have elevated serum concentrations of IL-6, VEGF, and bFGF, as well as elevated intrafollicular concentrations of IL-6. The outcome of ICSI cycles is not associated with premature elevation of progesterone when the cut-off value is set at 0.9 ng/ml.

Modulatory role of adenosine and its receptors in epilepsy: possible therapeutic approaches.

Adenosine is considered to be the brain's endogenous anticonvulsant as many studies have showed and it is responsible for seizure arrest and postictal refractoriness. Alterations in the adenosinergic system (adenosine and its receptors) have been referred by many previous studies indicating that deficiencies or modifications in the function of this purinergic system may contribute to epileptogenesis. Due to this emerging implication of adenosine in the managing of seizures, a new field of adenosine-based therapies has been introduced including adenosine itself, adenosine receptor agonists and antagonists and adenosine kinase inhibitors. The method with the least side effects (heart rate, blood pressure, temperature or even sedation) is being quested including intracerebral implantation of adenosine releasing cells or devices.

IGF-I and epidermal growth factor levels in follicular fluid of women undergoing controlled ovarian hyperstimulation using the multidose GnRH-antagonist protocol or the long GnRH-agonist protocol.

The intrafollicular levels of IGF-I and epidermal growth factor (EGF) were studied in women undergoing controlled ovarian hyperstimulation using the multidose GnRH-antagonist protocol or the long agonist protocol, in an attempt to elucidate whether GnRH-antagonists affect the levels of the two growth factors. The follicular fluid concentration of IGF-I, EGF, estradiol and progesterone were detected in 68 women undergoing ovarian hyperstimulation for intracytoplasmic sperm injection (ICSI) cycles. There were no differences in intrafollicular concentrations of EGF and IGF-I in the two studied groups. Additionally, we found no correlation between the intrafollicular levels of IGF-I or EGF and the ICSI outcome. The intrafollicular levels of IGF-I were positively correlated with those of progesterone. In conclusion, the intrafollicular levels of IGF-I and EGF do not seem to be influenced by the stimulation protocol. The intrafollicular levels of both growth factors can not serve as prognostic markers for the ICSI outcome.

The role of basic-fibroblast growth factor (b-FGF) in cyclosporine-induced nephrotoxicity.

BACKGROUND: The effect of the b-fibroblast growth factor (b-FGF) on cyclosporine A (CsA)-induced nephrotoxicity in the rat kidney was investigated. MATERIALS AND METHODS: The rats were divided into six groups: A (control), B (b-FGF-treated), C, D: (CsA-treated and sacrificed on days 14 or 21), E, F (Cs A- and b-FGF- treated and sacrificed on days 14 or 21). The antibody mouse anti-rat CD31 was used to evaluate the kidney vessels present in histological preparations. RESULTS: The kidney vessels in group B were increased in comparison with the control group (p<0.05). Reduction of kidney vessels in groups C and D (p<0.05) in comparison with the controls was observed, while in groups E and F they were increased when compared to group C (p<0.05) and D (p<0.05), respectively. CONCLUSION: The angiogenic role of b-FGF was confirmed in normal rats and a possible "protective" role of b-FGF was shown in rat kidney with CsA-induced nephrotoxicity.

Angiogenic effect of intramuscular administration of basic fibroblast growth factor in atrophied muscles: an experimental study in the rat.

BACKGROUND: Although angiogenic therapy using recombinant growth factors holds much hope for the treatment of ischaemic diseases, there are still many unanswered questions, including its effectiveness on atrophic muscles. OBJECTIVE: To evaluate the angiogenic effects of intramuscularly administered basic fibroblast growth factor (b-FGF) on normal gastrocnemius muscles of rats and atrophic gastrocnemius muscles after tenotomy. METHODS: Forty rats were divided into groups as follows: group A, controls; group B, injected with 1 microg b-FGF; group C, tenotomy performed on the right gastrocnemius muscle; group D, tenotomy and 1 microg b-FGF. Mouse anti-rat CD31 antibody was used to evaluate the number of blood vessels present in histological preparations. RESULTS: There was a significant (p<0.01) decrease in the number of blood vessels compared with the controls in the atrophic muscles of group C. This was similar to the decrease in muscle weight in this group. However, there was a significant (p<0.01) increase in the number of blood vessels compared with the controls in groups B and D. Similarly, there was a significant (p<0.01) increase in the number of blood vessels in group D compared with the atrophic muscles in group C. CONCLUSION: Intramuscular administration of b-FGF increases angiogenesis in both normal and atrophic rat gastrocnemius muscles at the injection area.

Angiogenic effect of intramuscular administration of basic and acidic fibroblast growth factor on skeletal muscles and influence of exercise on muscle angiogenesis.

BACKGROUND: Angiogenic factors which control the angiogenic process represent a promising strategy for restoration of blood flow, but require further evaluation before clinical use. Exercise has also been reported to induce neovascularisation in muscles. OBJECTIVES: To evaluate the angiogenic effects of basic fibroblast growth factor (b-FGF) and acidic fibroblast growth factor (a-FGF) on rat gastrocnemius muscle, when administered intramuscularly, and to compare them with those obtained by daily exercise. METHODS: Forty nine rats were allotted to the following groups: A, controls; B, exercise by swimming; C1 and C2, intramuscular injection of b-FGF and a-FGF respectively; D1 and D2, b-FGF and a-FGF injection in combination with exercise. The antibody mouse anti-rat CD31 was used to evaluate the numbers of blood vessels present in histological preparations of gastrocnemius muscle. RESULTS: Significant increases in the numbers of blood vessels of the right gastrocnemius muscles in groups C1 and D1 were observed compared with controls (p<0.05). There was only a slight increase in angiogenesis in the left gastrocnemius muscle of groups C1 and D1 compared with controls (p>0.05), and there was a decrease in angiogenesis in the gastrocnemius muscle of the swimming group compared with controls. CONCLUSION: The intramuscular administration of b-FGF, but not a-FGF, induced significant local angiogenesis in gastrocnemius muscle at the site of injection.

Clinicopathologic and prognostic significance of cyclooxygenase-2 expression in endometrial carcinoma.

BACKGROUND: Endometrial carcinoma is the most common malignancy of the female genital tract in the Western world. COX-2 is highly expressed in endometrial carcinoma, but there is controversy regarding its clinical role and its possible prognostic role. COX-2 expression was determined by immuno-histochemistry and was correlated to standard clinico-pathologic variables in a series of primary untreated endometrial carcinoma patients. COX-2 as an accurate predictor of the disease was also analyzed. METHODS: One-hundred and ten cases of primary untreated endometrial carcinoma hosts who were admitted to the Department of Obstetrics and Gynecology, University General Hospital of Alexandroupolis, were investigated. Immunohistochemistry was performed using rabbit polyclonal antiserum against human COX-2. RESULTS: Twenty-eight patients (25.5%) were scored as COX-2 positive. A statistically significant association was found between COX-2 overexpression and FIGO stage (p=0.010). A positive correlation was also found with histological grade (p=0.019) and myometrial invasion (p=0.026). No significant association was found with histologic type of the tumor (p=0.164). COX-2 positive patients had a significant association with sort survival (p=0.028). CONCLUSIONS: COX-2 expression is an independent clinicopathologic factor and an independent prognostic factor in endometrial carcinoma. It could be used to plan treatment modalities for hosts.

Follicular fluid levels of vascular endothelial growth factor and leptin are associated with pregnancy outcome of normal women participating in intracytoplasmic sperm injection cycles.

Cytokines play a critical and multifarious role in follicular maturation. Consequently, they may influence the pregnancy outcome in cycles of assisted reproduction. The aim of this study was to measure the levels of tumor necrosis factor-alpha (TNFalpha), vascular endothelial growth factor (VEGF) and leptin in serum and follicular fluids (FFs) from women undergoing controlled ovarian hyperstimulation (COH) for intracytoplasmic sperm injection cycles (ICSI). We tried to investigate their interrelationships and to evaluate them as predictive markers for the cycle's outcome. Seventeen women participated in this study. Male factor infertility was the only indication for ICSI cycles. For COH, the long agonist protocol with triptorelin and recombinant FSH was employed. Cytokines levels were evaluated by ELISA. Serum cytokine levels did not differ between pregnant and non-pregnant women. FF-VEGF levels were significantly elevated in non-pregnant women (722.2+/-1093.2 pg/ml) as compared to pregnant women (290.3+/-259.8 pg/ml). Leptin concentrations were also significantly higher in FFs of non-pregnant women (682.6+/-625.1 ng/ml) than those of pregnant women (231.6+/-286.5 ng/ml). There were significant positive correlations between FF-leptin and age, as well as between FF-leptin and FF-VEGF concentrations. It was concluded that elevated FF-leptin and VEGF levels are associated with failure of conception in IVF cycles and may serve as markers in clinical practice.

A borderline form of empty follicle syndrome. Case report.

BACKGROUND: Empty follicle syndrome is known as the failure of oocyte retrieval despite the adequate response to ovarian stimulation. It is a rare phenomenon in in-vitro fertilization and borderline forms of this syndrome have also been described. MATERIALS AND METHODS: Two cycles in the same patient were stimulated with GnRH agonist/hMG and recFSH; the first followed the long and the second followed the short protocol. RESULTS: There was a sudden drop in estradiol levels while the ovaries contained a large number of small and medium sized follicles. hCG was administered and oocyte retrieval was performed 36 hours later. There was no indication of low hCG levels. For the first cycle two oocytes were collected: one degenerated and one of poor quality. The second cycle resulted in total failure of oocyte retrieval. CONCLUSION: The two cycles were classified as borderline forms of empty follicle syndrome. The possible aetiology is discussed.

Triptorelin versus cetrorelix in intracytoplasmic sperm injection cycles in women with a single ovary.

OBJECTIVE: To compare the response to ovarian stimulation with either the long protocol of a GnRH-agonist or the multiple protocol of a GnRH-antagonist, in women with a single ovary who underwent intracytoplasmic sperm injection (ICSI) cycles. STUDY DESIGN: Retrospective study including 75 ICSI cycles from 26 women. Sixty-three cycles were stimulated with triptorelin/hMG or rFSH, whereas 12 cycles were stimulated with cetrorelix/hMG or rFSH. RESULTS: There was not found any statistical significant difference between the two groups regarding the days of stimulation, the number of gonadotropins' ampoules, the peak estradiol levels, the number of aspirated follicles and the number of retrieved oocytes. The fertilization rate, the number of transferred embryos as well as the cumulative embryo score were also similar in both groups. CONCLUSION: The multiple stimulation protocol of cetrorelix is equally effective with the long protocol of triptorelin in the ovarian stimulation of women with a single ovary.

Gonadotropin releasing hormone antagonists and cryopreservation outcome: a review.

REVIEW: The outcome of freeze-thaw cycles with pronuclear stage oocytes or embryos, derived from collecting cycles stimulated with gonadotropin-releasing hormone (GnRH)-antagonists' protocols, was reviewed. CONCLUSION: The viability of cryopreserved pronuclear stage oocytes and embryos, the quality of transferred embryos and the pregnancy rates of the freeze-thaw cycles seem to be satisfactory regardless of the type and dose of GnRH-antagonist.

Father-to-son sperm donation. A report of three cases.

Sperm donation is a common practice in assisted reproduction. In cases of azoospermia and negative results of testicular sperm extraction, it appears as the only solution. Sperm donation entails a complete genetic dissociation between husband and offspring, which brings psychological stress for the couple arising from ethical and existential dilemmas. Faced with such dilemmas, some couples prefer father-to-son donation as an alternative solution. Here, three cases of non-obstructive azoospermia are presented where intracytoplasmic sperm injection was performed with father-to-son sperm donation.

A comparison of the ovarian response to external stimulation between women with right or left ovary participating in programs of ICSI/ET.

INTRODUCTION: Single ovary women compose a group of special interest in assisted reproduction technologies. The aim of the present study was to investigate whether the response to external stimulation and the outcome in women with a single ovary was affected by the location of the ovary. METHODS: Sixty cycles from 24 women, which stimulated with either hMG or rFSH and a GnRH agonist, were retrospectively studied. Twelve women had right and 12 had left ovary. In all cases, the fertilization was performed by intracytoplasmic sperm injection. RESULTS: There was not found any statistical significant difference between the two groups regarding the days of stimulation, the peak estradiol levels, the number of aspirated follicles, the number of retrieved oocytes, the fertilization rate and the number of transferred embryos. The cumulative embryo score (CES) was also similar in both groups. Six pregnancies in the group of the right ovary and seven in the group of left ovary were achieved. CONCLUSION: According to these results, right and left ovary, in single ovary women under external stimulation, are equally active and potent for reproduction.