OBJECTIVE: Stress is known to be an inhibitor of the reproductive hypothalamic-pituitary-gonadal (HPG) axis. However, the neural and molecular connections between stress and reproduction are not yet understood. It is well established that in both humans and rodents, kisspeptin (encoded by the kiss1 gene) is a strong stimulator of the HPG axis. In the present study we hypothesized that endocannabinoids, an important neuromodulatory system in the brain, can act on the HPG axis at the level of kiss1 expression to inhibit reproductive function under stress. METHODS: Adult male Wistar rats were unilaterally implanted with an intracerebroventricular cannula. Afterwards, the animals were exposed to immobilization stress, with or without the presence of the cannabinoid CB1 receptor antagonist AM251 (1 microg/rat). Blood samples were collected through a retro-orbital plexus puncture before and after stress. Five hours after the stress, brain tissue was collected for reverse transcriptase-quantitative polymerase chain reaction measurements of kiss1 mRNA. RESULTS: Immobilization stress (1 hour) resulted in a decrease in the serum luteinizing hormone concentration. Additionally, kiss1 gene expression was decreased in key hypothalamic nuclei that regulate gonadotrophin secretion, the medial preoptic area (mPOA), and to some extent the arcuate nucleus (ARC). A single central administration of AM251 was effective in blocking these inhibitory responses. CONCLUSION: These findings suggest that endocannabinoids mediate, at least in part, immobilization stress-induced inhibition of the reproductive system. Our data suggest that the connection between immobilization stress and the HPG axis is kiss1 expression in the mPOA rather than the ARC.
Adult ; Animals ; Arcuate Nucleus ; Axis, Cervical Vertebra* ; Brain ; Cannabinoids ; Catheters ; Endocannabinoids* ; Gene Expression ; Humans ; Immobilization ; Kisspeptins ; Luteinizing Hormone ; Male* ; Polymerase Chain Reaction ; Preoptic Area ; Punctures ; Rats* ; Rats, Wistar ; Receptor, Cannabinoid, CB1 ; Reproduction ; RNA, Messenger ; Rodentia

OBJECTIVE: To compare the mouse oocyte vitrification outcomes of the CryoLogic vitrification method (CVM) and the conventional open method using a Cryotop. Two CVM methods (original CVM and modified CVM) were tested. METHODS: Mature oocytes obtained from female BDF-1 mice were vitrified by two-step exposure to equilibrium and vitrification solutions. Three vitrification protocols were tested on three groups: the CVM-kit, modified CVM, and Cryotop groups. After exposure to the two solutions, the oocytes were vitrified. After warming, the oocytes were fertilized in vitro, and the embryo development was assessed. Blastomeres positive for caspase were counted using an in situ assay kit. The spindle morphology and chromosome configurations of warmed vitrified oocytes were also assessed. RESULTS: The modified CVM and Cryotop groups showed similar developmental capacities, and similar proportions of cells with intact spindles and chromosome configurations. The modified CVM protocol was superior to the original CVM protocol for developmental competence and intact spindle preservation. However, the CVM group showed a relatively higher number of apoptotic cells in blastocysts. CONCLUSION: Closed vitrification using the modified CVM protocol may be used as an alternative to the conventional open method, but strategies to decrease apoptosis in the blastomere need to be investigated.
Animals ; Apoptosis ; Blastocyst ; Blastomeres ; Embryonic Development ; Female ; Humans ; Mental Competency* ; Methods ; Mice* ; Oocytes* ; Pregnancy ; Vitrification*

Oocyte in vitro maturation (IVM) is an assisted reproductive technology in which oocytes are retrieved from the antral follicles of unstimulated or minimally stimulated ovaries. IVM of human oocytes has emerged as a promising procedure. This new technology has advantages over controlled ovarian stimulation such as reduction of costs, simplicity, and elimination of ovarian hyperstimulation syndrome. By elimination or reduction of gonadotropin stimulation, IVM offers eligible infertile couples a safe and convenient form of treatment, and IVM outcomes are currently comparable in safety and efficacy to those of conventional in vitro fertilization. IVM has been applied mainly in patients with polycystic ovary syndrome or ultrasound-only polycystic ovaries, but with time, the indications for IVM have expanded to other uncommon situations such as fertility preservation, as well as to normal responders. In this review, the current clinical experiences with IVM will be described.
Family Characteristics ; Female ; Fertility Preservation ; Fertilization in Vitro ; Gonadotropins ; Humans ; In Vitro Oocyte Maturation Techniques ; Infertility ; Oocytes ; Ovarian Hyperstimulation Syndrome ; Ovary ; Ovulation Induction ; Polycystic Ovary Syndrome ; Reproductive Techniques, Assisted

17alpha-hydroxylase and 17,20-lyase are enzymes encoded by the CYP17A1 gene and are required for the synthesis of sex steroids and cortisol. In 17alpha-hydroxylase deficiency, there are low blood levels of estrogens, androgens, and cortisol, and resultant compensatory increases in adrenocorticotrophic hormone that stimulate the production of 11-deoxycorticosterone and corticosterone. In turn, the excessive levels of mineralocorticoids lead to volume expansion and hypertension. Females with 17alpha-hydroxylase deficiency are characterized by primary amenorrhea and delayed puberty, with accompanying hypertension. Affected males usually have female external genitalia, a blind vagina, and intra-abdominal testes. The treatment of this disorder is centered on glucocorticoid and sex steroid replacement. In patients with 17alpha-hydroxylase deficiency who are being raised as females, estrogen should be supplemented, while genetically female patients with a uterus should also receive progesterone supplementation. Here, we report a case of a 21-year-old female with 17alpha-hydroxylase deficiency who had received inadequate treatment for a prolonged period of time. We also include a brief review of the recent literature on this disorder.
Adrenocorticotropic Hormone ; Amenorrhea ; Androgens ; Corticosterone ; Estrogens ; Female ; Genitalia ; Humans ; Hydrocortisone ; Hypertension ; Male ; Mineralocorticoids ; Progesterone ; Puberty, Delayed ; Steroid 17-alpha-Hydroxylase* ; Steroids ; Testis ; Uterus ; Vagina ; Young Adult

OBJECTIVE: The aim of this study was to evaluate pregnancy outcomes and the live birth rate at 1-year age increments in women aged ≥40 years undergoing fresh non-donor in vitro fertilization (IVF) and embryo transfer (ET), and to identify predictors of success in these patients. METHODS: This retrospective study was performed among women ≥40 years of age between 2004 and 2011. Of the 2,362 cycles that were conducted, ET was performed in 1,532 (73.1%). RESULTS: The clinical pregnancy rate and live birth rate in women ≥40 years significantly decreased with each year of increased age (p<0.001). Maternal age (odds ratio [OR], 0.644; 95% confidence interval [CI], 0.540–0.769; p<0.001), basal follicle-stimulating hormone (FSH) levels (OR, 0.950; 95% CI, 0.903–0.999; p=0.047), the number of high-quality embryos (OR, 1.258; 95% CI, 1.005 –1.575; p=0.045), and the number of transferred embryos (OR, 1.291; 95% CI, 1.064 –1.566; p=0.009) were significant predictors of live birth. A statistically significant increase in live birth rates was seen when ≥3 embryos were transferred in patients 40 to 41 years of age, whereas poor pregnancy outcomes were seen in patients ≥43 years of age, regardless of the number of transferred embryos. Moreover, the cumulative live birth rate increased in patients 40 to 42 years of age with repeated IVF cycles, but the follicle-stimulating hormone in those ≥43 years of age rarely showed an increase. CONCLUSION: IVF-ET has acceptable outcomes in those <43 years of age when a patient's own oocytes are used. Maternal age, basal FSH levels, and the number of high-quality embryos and transferred embryos are useful predictors of live birth.
Embryo Transfer ; Embryonic Structures ; Female ; Fertilization in Vitro* ; Follicle Stimulating Hormone ; Humans ; In Vitro Techniques* ; Infertility ; Live Birth* ; Maternal Age ; Oocytes ; Pregnancy Outcome ; Pregnancy Rate ; Pregnancy* ; Retrospective Studies

OBJECTIVE: The purpose of this study was to evaluate the efficacy of frozen mixed double-embryo transfer (MDET; the simultaneous transfer of day 3 and day 5 embryos) in comparison with frozen blastocyst double-embryo transfer (BDET; transfer of two day 5 blastocysts) in patients with repeated implantation failure (RIF). METHODS: A total of 104 women with RIF who underwent frozen MDET (n=48) or BDET (n=56) with excellent-quality embryos were included in this retrospective analysis. All frozen embryo transfers were performed in natural cycles. The main outcome measures were the implantation rate, clinical pregnancy rate, multiple pregnancy rate, and miscarriage rate. These measures were compared between the patients who underwent MDET or BDET using the chi-square test or the Fisher exact test, as appropriate. RESULTS: The implantation and clinical pregnancy rates were significantly higher in patients who underwent MDET than in those who underwent BDET (60.4% vs. 39.3%, p=0.03 and 52.1% vs. 30.4%, p=0.05, respectively). A significantly lower miscarriage rate was observed in the MDET group (6.9% vs. 10.7%, p=0.05). In addition, the multiple pregnancy rate was slightly, but not significantly, higher in the MDET group (27.1% vs. 25.0%). CONCLUSION: MDET was found to be significantly superior to double blastocyst transfer. It could be regarded as an appropriate approach to improve in vitro fertilization success rates in RIF patients.
Abortion, Spontaneous ; Blastocyst ; Embryo Transfer ; Embryonic Structures ; Female ; Fertilization in Vitro ; Humans ; Outcome Assessment (Health Care) ; Pregnancy ; Pregnancy Rate ; Pregnancy, Multiple ; Retrospective Studies

OBJECTIVE: This study aimed to compare the clinical outcomes between in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) in sibling oocytes. Additionally, we evaluated whether the implementation of split insemination contributed to an increase in the number of ICSI procedures. METHODS: A total of 571 cycles in 555 couples undergoing split insemination cycles were included in this study. Among them, 512 cycles (89.7%) were a couple's first IVF cycle. The patients were under 40 years of age and at least 10 oocytes were retrieved in all cycles. Sibling oocytes were randomly allocated to IVF or ICSI. RESULTS: Total fertilization failure was significantly more common in IVF cycles than in ICSI cycles (4.0% vs. 1.4%, p<0.05), but the low fertilization rate among retrieved oocytes (as defined by fertilization rates greater than 0% but <30%) was significantly higher in ICSI cycles than in IVF cycles (17.2% vs. 11.4%, p<0.05). The fertilization rate of ICSI among injected oocytes was significantly higher than for IVF (72.3%±24.3% vs. 59.2%±25.9%, p<0.001), but the fertilization rate among retrieved oocytes was significantly higher in IVF than in ICSI (59.2%±25.9% vs. 52.1%±22.5%, p<0.001). Embryo quality before embryo transfer was not different between IVF and ICSI. Although the sperm parameters were not different between the first cycle and the second cycle, split insemination or ICSI was performed in 18 of the 95 cycles in which a second IVF cycle was performed. CONCLUSION: The clinical outcomes did not differ between IVF and ICSI in split insemination cycles. Split insemination can decrease the risk of total fertilization failure. However, unnecessary ICSI is carried out in most split insemination cycles and the use of split insemination might make ICSI more common.
Embryo Transfer ; Embryonic Structures ; Family Characteristics ; Fertilization ; Fertilization in Vitro* ; Humans ; In Vitro Techniques* ; Insemination* ; Oocytes ; Siblings ; Sperm Injections, Intracytoplasmic* ; Spermatozoa

OBJECTIVE: We investigated whether the insemination method (in vitro fertilization [IVF] or intracytoplasmic sperm injection [ICSI]) affected morphokinetic events and abnormal cleavage events in embryonic development. METHODS: A total of 1,830 normal fertilized embryos were obtained from 272 IVF and ICSI cycles that underwent ovum retrieval culture using a time-lapse system (Embryoscope) from June 2013 to March 2015. All embryos were investigated by a detailed time-lapse analysis that measured the developmental events in the hours after IVF or ICSI insemination. RESULTS: No significant differences were observed between the two groups regarding clinical outcomes (p>0.05). ICSI-derived embryos showed significantly faster morphokinetics than those derived from conventional IVF, from the time to pronuclear fading to the time to 6 cells (p<0.05). However, no significant differences were found from the time to 7 cells to the time to expanded blastocyst (p>0.05). There were no differences in abnormal cleavage events between the two groups (p>0.05); they showed the same rates of direct cleavage from 1 to 3 cells, 2 multinucleated cells, 2 uneven cells, and reverse cleavage. CONCLUSION: The morphokinetics of embryo development was found to vary between IVF- and ICSI-fertilized oocytes, at least until the 6-cell stage. However, these differences did not affect the clinical outcomes of the embryo. Additionally, no significant differences in abnormal cleavage events were found according to the fertilization method.
Blastocyst ; Embryonic Development* ; Embryonic Structures* ; Female ; Fertilization ; Fertilization in Vitro ; Humans* ; In Vitro Techniques* ; Insemination ; Methods ; Oocytes* ; Ovum ; Pregnancy ; Sperm Injections, Intracytoplasmic ; Spermatozoa* ; Time-Lapse Imaging

OBJECTIVE: The aim of this study was to explore the effects of the insemination method on the outcomes of elective blastocyst culture. METHODS: We retrospectively analyzed the outcomes of elective blastocyst culture performed between January 2011 and December 2014. RESULTS: There were 2,003 cycles of conventional in vitro fertilization (IVF) and 336 cycles of intracytoplasmic sperm injection (ICSI), including 25,652 and 4,164 embryos that underwent sequential blastocyst culture, respectively. No significant differences were found in the female patients' age, basal follicle-stimulating hormone level, basal luteinizing hormone level, body mass index, number of oocytes, maturity rate, fertilization rate, or good-quality embryo rate. However, the blastocyst formation rate and embryo utilization rate were significantly higher in the conventional IVF group than in the ICSI group (54.70% vs. 50.94% and 51.09% vs. 47.65%, respectively, p<0.05). The implantation/pregnancy rate (IVF, 50.93%; ICSI, 55.10%), miscarriage rate (IVF, 12.57%; ICSI, 16.29%), and live birth rate (IVF, 42.12%; ICSI, 44.08%) were similar (p>0.05). No cycles were canceled due to the formation of no usable blastocysts. CONCLUSION: Although the fertilization method had no effect on clinical outcomes, the blastocyst formation rate and embryo utilization rate in the ICSI group were significantly lower than those observed in the conventional IVF group. Therefore, more care should be taken when choosing to perform blastocyst culture in ICSI patients.
Abortion, Spontaneous ; Blastocyst* ; Body Mass Index ; Embryonic Structures ; Female ; Fertilization ; Fertilization in Vitro ; Follicle Stimulating Hormone ; Humans ; Insemination* ; Live Birth ; Luteinizing Hormone ; Methods* ; Oocytes ; Pregnancy ; Pregnancy Rate ; Retrospective Studies ; Sperm Injections, Intracytoplasmic

OBJECTIVE: Optimizing in vitro maturation (IVM) media to achieve better outcomes has been a matter of interest in recent years. The aim of this prospective clinical trial was to investigate the effects of different media on the IVM outcomes of immature oocytes at the germinal vesicle (GV) stage. METHODS: A total of 400 immature oocytes at the GV stage with normal morphology were retrieved from 320 infertile women aged 31±4.63 years during stimulated intracytoplasmic sperm injection (ICSI) cycles. They were divided into groups of homemade IVM medium (I, n=100), cleavage medium (II, n=100), blastocyst medium (III, n=100), and Sage IVM medium (IV, n=100) and cultured for 24 to 48 hours at 37℃. ICSI was performed, and the rates of fertilization and embryo formation were compared across the four groups. RESULTS: In the 400 retrieved GV oocytes, the total maturation rates showed significant differences in groups I to IV (55%, 53%, 78%, and 68%, respectively, p<0.001). However, there were no significant differences in the fertilization, embryo formation, or arrest rates of metaphase II oocytes across these groups. In all groups, GV maturation was mostly completed after 24 hours, with fewer oocytes requiring 48 hours to mature (p<0.01). Moreover, the rate of high-quality embryos was higher in group IV than in the other groups (p=0.01). CONCLUSION: The quality of the IVM medium was found to affect clinical IVM outcomes. Additionally, blastocyst medium may be a good choice in IVM/ICSI cycles as an alternative IVM medium.
Blastocyst ; Embryonic Structures ; Female ; Fertilization ; Humans* ; In Vitro Techniques* ; Metaphase ; Oocytes* ; Prospective Studies ; Sperm Injections, Intracytoplasmic*