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MTHFR 3′-untranslated region polymorphisms contribute to recurrent pregnancy loss risk and alterations in peripheral natural killer cell proportions.

Eun Sun KIM ; Jung Oh KIM ; Hui Jeong AN ; Jung Hyun SAKONG ; Hyun Ah LEE ; Ji Hyang KIM ; Eun Hee AHN ; Young Ran KIM ; Woo Sik LEE ; Nam Keun KIM

Clinical and Experimental Reproductive Medicine.2017;44(3):152-158. doi:10.5653/cerm.2017.44.3.152

OBJECTIVE: To identify the associations between polymorphisms of the 3′-untranslated region (UTR) of methylenetetrahydrofolate reductase (MTHFR) gene, which codes for an important regulatory enzyme primarily involved in folate metabolism, and idiopathic recurrent pregnancy loss (RPL) in Korean women. METHODS: The study population comprised 369 RPL patients and 228 controls. MTHFR 2572C>A, 4869C>G, 5488C>T, and 6685T>C 3′-UTR polymorphisms were genotyped by polymerase chain reaction-restriction fragment length polymorphism analysis or by TaqMan allelic discrimination assays. Natural killer cell proportions were determined by flow cytometry. RESULTS: The MTHFR 2572-5488-6685 (A-C-T) haplotype had an adjusted odds ratio of 0.420 (95% confidence interval, 0.178–0.994; p=0.048) for RPL. Analysis of variance revealed that MTHFR 4869C>G was associated with altered CD56⁺ natural killer cell percentages (CC, 17.91%±8.04%; CG, 12.67%±4.64%; p=0.024) and folate levels (CC, 12.01±7.18 mg/mL; CG, 22.15±26.25 mg/mL; p=0.006). CONCLUSION: Variants in the 3′-UTR of MTHFR are potential biomarkers for RPL. However, these results should be validated in additional studies of ethnically diverse groups of patients.
Biomarkers ; Discrimination (Psychology) ; Female ; Flow Cytometry ; Folic Acid ; Haplotypes ; Humans ; Killer Cells, Natural* ; Metabolism ; Methylenetetrahydrofolate Reductase (NADPH2) ; Odds Ratio ; Pregnancy*

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The association of follicular fluid volume with human oolemma stretchability during intracytoplasmic sperm injection.

Taketo INOUE ; Yoshiki YAMASHITA ; Yoshiko TSUJIMOTO ; Shuji YAMAMOTO ; Sayumi TAGUCHI ; Kayoko HIRAO ; Mikiko UEMURA ; Kayoe IKAWA ; Kazunori MIYAZAKI

Clinical and Experimental Reproductive Medicine.2017;44(3):126-131. doi:10.5653/cerm.2017.44.3.126

OBJECTIVE: Oocyte degeneration often occurs after intracytoplasmic sperm injection (ICSI), and the risk factor is low-quality oocytes. The follicular fluid (FF) provides a crucial microenvironment for oocyte development. We investigated the relationships between the FF volume aspirated from individual follicles and oocyte retrieval, oocyte maturity, oolemma stretchability, fertilization, and development. METHODS: This retrospective study included data obtained from 229 ICSI cycles. Ovarian stimulation was performed according to a gonadotropin-releasing hormone antagonist protocol. Each follicle was individually aspirated and divided into six groups according to FF volume (<1.0, 1.0 to <2.0, 2.0 to <3.0, 3.0 to <4.0, 4.0 to <5.0, and ≥5.0 mL). Oolemma stretchability during ICSI was evaluated using a mechanical stimulus for oolemma penetration, that is, the stretchability was assessed by oolemma penetration with aspiration (high stretchability) or without aspiration (low stretchability). RESULTS: Oocyte retrieval rates were significantly lower in the <1.0 mL group than in the ≥1.0 mL groups (46.0% [86/187] vs. 67.5%–74.3% [172/255 to 124/167], respectively; p<0.01). Low oolemma stretchability was significantly more common in the <1.0 mL group than in the ≥1.0 mL groups during ICSI (22.0% [13/59] vs. 5.8%–9.4% [6/104 to 13/139], respectively; p=0.018). There was a relationship between FF volume and oolemma stretchability. However, there were no significant differences in the rates of fertilization, cleavage, ≥7 cells at day 3, and blastocyst development among all groups. CONCLUSION: FF volume is potentially associated with the stretchability of metaphase II oolemma during ICSI. Regarding oolemma stretchability, ensuring a uniform follicular size during ovarian stimulation is crucial to obtain good-quality oocytes.
Blastocyst ; Clothing ; Female ; Fertilization ; Follicular Fluid* ; Gonadotropin-Releasing Hormone ; Humans* ; Infertility ; Membranes ; Metaphase ; Oocyte Retrieval ; Oocytes ; Ovarian Follicle ; Ovulation Induction ; Retrospective Studies ; Risk Factors ; Sperm Injections, Intracytoplasmic*

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Addition of interleukin-6 to mouse embryo culture increases blastocyst cell number and influences the inner cell mass to trophectoderm ratio.

Rebecca L KELLEY ; David K GARDNER

Clinical and Experimental Reproductive Medicine.2017;44(3):119-125. doi:10.5653/cerm.2017.44.3.119

OBJECTIVE: In vitro culture of preimplantation embryos is improved by grouping embryos together in a drop of media. Individually cultured embryos are deprived of paracrine factors; with this in mind, we investigated whether the addition of a single embryo-secreted factor, interleukin-6 (IL-6), could improve the development of individually cultured embryos. METHODS: Mouse embryos were cultured individually in 2 µL of G1/G2 media in 5% oxygen and supplemented with a range of doses of recombinant mouse or human IL-6. RESULTS: Mouse IL-6 increased hatching at doses of 0.01 and 10 ng/mL compared to the control (93% and 93% vs. 78%, p<0.05) and increased the total number of cells at a dose of 0.1 ng/mL compared to the control (101.95±3.36 vs. 91.31±3.33, p<0.05). In contrast, the highest dose of 100 ng/mL reduced the total number of cells (79.86±3.29, p<0.05). Supplementation with human IL-6 had a different effect, with no change in hatching or total cell numbers, but an increase in the percentage of inner cell mass per embryo at doses of 0.1, 1, and 100 ng/mL compared to the control (22.9%±1.1%, 23.3%±1.1%, and 23.1%±1.1% vs. 19.5%±1.0%, p<0.05). CONCLUSION: These data show that IL-6 improved mouse embryo development when cultured individually in complex media; however, an excess of IL-6 may be detrimental. Additionally, these data indicate that there is some cross-species benefit of human IL-6 for mouse embryos, but possibly through a different mechanism than for mouse IL-6.
Animals ; Blastocyst* ; Cell Count* ; Culture Media ; Cytokines ; Embryology ; Embryonic Development ; Embryonic Structures* ; Female ; Fertilization in Vitro ; Humans ; In Vitro Techniques ; Interleukin-6* ; Mice* ; Oxygen ; Pregnancy

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Reduction of the cetrorelix dose in a multiple-dose antagonist protocol and its impact on pregnancy rate and affordability: A randomized controlled multicenter study.

Ayman S DAWOOD ; Adel ALGERGAWY ; Ahmed ELHALWAGY

Clinical and Experimental Reproductive Medicine.2017;44(4):232-238. doi:10.5653/cerm.2017.44.4.232

OBJECTIVE: To determine whether reducing the cetrorelix dose in the antagonist protocol to 0.125 mg had any deleterious effects on follicular development, the number and quality of retrieved oocytes, or the number of embryos, and to characterize its effects on the affordability of assisted reproductive technology. METHODS: This randomized controlled study was conducted at the Fertility Unit of Tanta Educational Hospital of Tanta University, the Egyptian Consultants' Fertility Center, and the Qurrat Aien Fertility Center, from January 1 to June 30, 2017. Patients' demographic data, stimulation protocol, costs, pregnancy rate, and complications were recorded. Patients were randomly allocated into two groups: group I (n=61) received 0.125 mg of cetrorelix (the study group), and group II (n=62) received 0.25 mg of cetrorelix (the control group). RESULTS: The demographic data were comparable regarding age, parity, duration of infertility, and body mass index. The dose of recombinant follicle-stimulating hormone units required was 2,350.43±150.76 IU in group I and 2,366.25±140.34 IU in group II, which was not a significant difference (p=0.548). The duration of stimulation, number of retrieved oocytes, and number of developed embryos were not significantly different between the groups. The clinical and ongoing pregnancy rates likewise did not significantly differ. The cost of intracytoplasmic sperm injection per cycle was significantly lower in group I than in group II (US $494.66±4.079 vs. US $649.677±43.637). CONCLUSION: Reduction of the cetrorelix dose in the antagonist protocol was not associated with any significant difference either in the number of oocytes retrieved or in the pregnancy rate. Moreover, it was more economically feasible for patients in a low-resource country.
Body Mass Index ; Embryonic Structures ; Female ; Fertility ; Follicle Stimulating Hormone ; Gonadotropins ; Humans ; Infertility ; Oocytes ; Parity ; Pregnancy Rate* ; Pregnancy* ; Reproductive Techniques, Assisted ; Sperm Injections, Intracytoplasmic

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Impact of sperm DNA fragmentation on clinical in vitro fertilization outcomes.

Hwa Young CHOI ; Seul Ki KIM ; Seok Hyun KIM ; Young Min CHOI ; Byung Chul JEE

Clinical and Experimental Reproductive Medicine.2017;44(4):224-231. doi:10.5653/cerm.2017.44.4.224

OBJECTIVE: We studied the association between sperm DNA fragmentation (SDF) and several clinical in vitro fertilization outcomes. METHODS: We retrospectively analyzed 169 consecutive fresh IVF cycles. Semen was collected on the day of oocyte retrieval, and we assessed standard semen parameters and the SDF level (by terminal deoxynucleotidyl transferase dUTP nick-end labeling). Poor ovarian response (POR) was defined as the collection of three or fewer mature oocytes. Oocytes were inseminated by the conventional method or intracytoplasmic sperm injection. RESULTS: SDF did not affect the fertilization or pregnancy rate, but did have a significant effect on the miscarriage rate. In the miscarriage group (n=10), the SDF level was significantly higher (23.9% vs. 14.1%) and number of mature oocytes was significantly lower (4.3 vs. 7.6) than in the live birth group (n=45). Multiple regression analysis showed that SDF was an independent predictor of miscarriage (odds ratio, 1.051; 95% confidence interval, 1.001–1.104). The cutoffs for the SDF level and number of mature oocytes that could predict miscarriage were >13% and ≤3, respectively. In the low-SDF group (≤13%), the miscarriage rate was similar in POR patients and those with a normal ovarian response (NOR; 14.2% vs. 4.3%). In the high-SDF group (>13%), the miscarriage rate was significantly higher in the POR group than in the NOR group (60.0% vs. 13.3%, p=0.045). CONCLUSION: Our study demonstrated that a high SDF level (>13%) was associated with a high miscarriage rate, and that it mainly contributed to miscarriage in the POR group. The results suggest that SDF measurements should be considered in couples with POR in order to predict the prognosis of the pregnancy.
Abortion, Spontaneous ; DNA Fragmentation* ; DNA Nucleotidylexotransferase ; DNA* ; Family Characteristics ; Female ; Fertilization ; Fertilization in Vitro* ; Humans ; In Vitro Techniques* ; Live Birth ; Methods ; Oocyte Retrieval ; Oocytes ; Pregnancy ; Pregnancy Rate ; Prognosis ; Retrospective Studies ; Semen ; Sperm Injections, Intracytoplasmic ; Spermatozoa*

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T helper cell subsets and related cytokines in infertile women undergoing in vitro fertilization before and after seminal plasma exposure.

Marziyeh AZAD ; Sara KESHTGAR ; Bahia Namavar JAHROMI ; Zahra KANANNEJAD ; Behrouz GHARESI-FARD

Clinical and Experimental Reproductive Medicine.2017;44(4):214-223. doi:10.5653/cerm.2017.44.4.214

OBJECTIVE: In vitro fertilization (IVF) is a well-known method for the treatment of infertility. The present study aimed to compare the differences between infertile women with successful and unsuccessful IVF outcomes regarding the expression of T helper (Th) cell transcription factors and a group of related cytokines before and after exposure to their husbands' seminal plasma. METHODS: This study was performed on 19 couples with unexplained infertility undergoing IVF treatment. Among the studied group, nine and 10 couples had successful and unsuccessful IVF outcomes, respectively. This study was carried out using real-time polymerase chain reaction. RESULTS: Before seminal plasma exposure, the expression levels of T-bet (p < 0.007), interferon-γ (p=0.013), and tumor necrosis factor (TNF)-α (p=0.017) were higher in the infertile women with IVF failure than in those with successful IVF outcomes, while those of GATA3 (p < 0.001), Foxp3 (p=0.001), and interleukin (IL)-35 (p < 0.003) were lower. After seminal exposure, the expression of T-bet (p=0.02), Rorc (p < 0.001), TNF-α (p=0.001), Foxp3 (p=0.02), and interferon-γ (p=0.001) increased in the unsuccessful IVF group, while the expression of Foxp3 (p=0.02), Rorc (p < 0.001), IL-23 (p=0.04), IL-17 (p=0.02), IL-6 (p < 0.001), transforming growth factor-β (p=0.01), and IL-35 (p < 0.001) increased in the successful IVF group. CONCLUSION: In summary, IVF failure was associated with imbalanced Th1/Th2/Th17/Treg responses. Moreover, our results show that seminal plasma might have a positive effect on IVF outcomes via changes in peripheral blood T cell subsets.
Cytokines* ; Family Characteristics ; Female ; Fertilization in Vitro* ; Humans ; In Vitro Techniques* ; Infertility ; Interleukin-17 ; Interleukin-23 ; Interleukin-6 ; Interleukins ; Methods ; Real-Time Polymerase Chain Reaction ; Semen* ; T-Lymphocyte Subsets ; T-Lymphocytes, Helper-Inducer* ; Transcription Factors ; Tumor Necrosis Factor-alpha

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Effects of infections with five sexually transmitted pathogens on sperm quality.

Sung Jae KIM ; Doo Jin PAIK ; Joong Shik LEE ; Hyo Serk LEE ; Ju Tae SEO ; Mi Seon JEONG ; Jae Ho LEE ; Dong Wook PARK ; Sangchul HAN ; Yoo Kyung LEE ; Ki Heon LEE ; In Ho LEE ; Kyeong A SO ; Seon Ah KIM ; Juree KIM ; Tae Jin KIM

Clinical and Experimental Reproductive Medicine.2017;44(4):207-213. doi:10.5653/cerm.2017.44.4.207

OBJECTIVE: This study investigated the prevalence of infections with human papillomavirus, Chlamydia trachomatis, Ureaplasma urealyticum, Mycoplasma hominis, and Mycoplasma genitalium in the semen of Korean infertile couples and their associations with sperm quality. METHODS: Semen specimens were collected from 400 men who underwent a fertility evaluation. Infection with above five pathogens was assessed in each specimen. Sperm quality was compared in the pathogen-infected group and the non-infected group. RESULTS: The infection rates of human papillomavirus, C. trachomatis, U. urealyticum, M. hominis, and M. genitalium in the study subjects were 1.57%, 0.79%, 16.80%, 4.46%, and 1.31%, respectively. The rate of morphological normality in the U. urealyticum-infected group was significantly lower than in those not infected with U. urealyticum. In a subgroup analysis of normozoospermic samples, the semen volume and the total sperm count in the pathogen-infected group were significantly lower than in the non-infected group. CONCLUSION: Our results suggest that infection with U. urealyticum alone and any of the five sexually transmitted infections are likely to affect sperm morphology and semen volume, respectively.
Chlamydia trachomatis ; Family Characteristics ; Fertility ; Humans ; Male ; Mycoplasma genitalium ; Mycoplasma hominis ; Prevalence ; Semen ; Semen Analysis ; Sexually Transmitted Diseases ; Sperm Count ; Spermatozoa* ; Ureaplasma urealyticum

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Sperm DNA fragmentation and sex chromosome aneuploidy after swim-up versus density gradient centrifugation.

Sung Woo KIM ; Byung Chul JEE ; Seul Ki KIM ; Seok Hyun KIM

Clinical and Experimental Reproductive Medicine.2017;44(4):201-206. doi:10.5653/cerm.2017.44.4.201

OBJECTIVE: The aim of this study was to compare the efficacy of swim-up and density gradient centrifugation (DGC) for reducing the amount of sperm with fragmented DNA, sex chromosome aneuploidy, and abnormal chromatin structure. METHODS: Semen samples were obtained from 18 healthy male partners who attended infertility clinics for infertility investigations and were processed with swim-up and DGC. The percentages of sperm cells with fragmented DNA measured by the sperm chromatin dispersion test, normal sex chromosomes assessed by fluorescence in situ hybridization, and abnormal chromatin structure identified by toluidine blue staining were examined. RESULTS: The percentage of sperm cells with fragmented DNA was significantly lower in the swim-up fraction (9.7%, p=0.001) than in the unprocessed fraction (27.0%), but not in the DGC fraction (27.8%, p=0.098). The percentage of sperm cells with normal X or Y chromosomes was comparable in the three fractions. The percentage of sperm cells with abnormal chromatin structure significantly decreased after DGC (from 15.7% to 10.3%, p=0.002). The swim-up method also tended to reduce the percentage of sperm cells with abnormal chromatin structure, but the difference was not significant (from 15.7% to 11.6%, p=0.316). CONCLUSION: The swim-up method is superior for enriching genetically competent sperm.
Aneuploidy* ; Centrifugation, Density Gradient* ; Chromatin ; DNA Fragmentation* ; DNA* ; Fluorescence ; Humans ; In Situ Hybridization ; Infertility ; Male ; Methods ; Semen ; Sex Chromosomes* ; Spermatozoa* ; Tolonium Chloride ; Y Chromosome

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Effects of laser-assisted hatching and exposure time to vitrification solution on mouse embryo development.

Hye Jin KIM ; Sung Baek PARK ; Jung Bo YANG ; Young Bae CHOI ; Ki Hwan LEE

Clinical and Experimental Reproductive Medicine.2017;44(4):193-200. doi:10.5653/cerm.2017.44.4.193

OBJECTIVE: This study was conducted to investigate the efficacy of laser-assisted hatching (LAH) and various vitrification times for embryonic development and blastocyst cell numbers. METHODS: First, 2-cell and 8-cell embryos were collected by flushing out the oviducts. In the control groups, they were vitrified for 8 or 10 minutes without LAH. The LAH groups underwent quarter laser zona thinning-assisted hatching before vitrification (4, 6, and 8 minutes or 4, 7, and 10 minutes, respectively). After incubation, double-immunofluorescence staining was performed. RESULTS: The hatched blastocyst rate 72 hours after the 2-cell embryos were thawed was significantly higher in the 2LAH-ES8 group (33.3%) than in the other groups (p < 0.05). In the control-8 group (22.1±4.6), the cell number of the inner cell mass was higher than in the LAH groups (p < 0.05). The number of trophectoderm cells was higher in the 2LAH-ES6 group (92.8±8.9) than in the others (p < 0.05). The hatched blastocyst rate 48 hours after the 8-cell embryos were thawed was higher in the 8LAH-ES4 group (45.5%) than in the other groups, but not significantly. The inner cell mass cell number was highest in the 8LAH-ES7 group (19.5±5.1, p < 0.05). The number of trophectoderm cells was higher in the 8LAH-ES10 group (73.2±12.1) than in the other groups, but without statistical significance. CONCLUSION: When LAH was performed, 2-cell embryos with large blastomeres had a lower hatched blastocyst rate when the exposure to vitrification solution was shorter. Conversely, 8-cell embryos with small blastomere had a higher hatched blastocyst rate when the exposure to vitrification solution was shorter.
Animals ; Blastocyst ; Blastomeres ; Cell Count ; Embryonic Development* ; Embryonic Structures* ; Female ; Flushing ; Herpes Zoster ; Mice* ; Oviducts ; Pregnancy ; Vitrification*

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Fertility preservation for patients with hematologic malignancies: The Korean Society for Fertility Preservation clinical guidelines.

Dong Yun LEE ; Seul Ki KIM ; Miran KIM ; Kyung Joo HWANG ; Seok Hyun KIM

Clinical and Experimental Reproductive Medicine.2017;44(4):187-192. doi:10.5653/cerm.2017.44.4.187

Although the survival rate of hematologic malignancies in young patients is very high, cytotoxic therapies such as chemotherapy and total body irradiation therapy can significantly reduce a patient's reproductive capacity and cause irreversible infertility. Early ovarian failure also commonly occurs following additional cancer treatment, bone marrow transplantation, or autologous transplantation. Because the risk of early ovarian failure depends on the patient's circumstances, patients with a hematologic malignancy must consult health professionals regarding fertility preservation before undergoing treatments that can potentially damage their ovaries. While it is widely known that early menopause commonly occurs following breast cancer treatment, there is a lack of reliable study results regarding fertility preservation during hematologic malignancy treatment. Therefore, an in-depth discussion between patients and health professionals about the pros and cons of the various options for fertility preservation is necessary. In this study, we review germ cell toxicity, which occurs during the treatment of hematologic malignancies, and propose guidelines for fertility preservation in younger patients with hematologic malignancies.
Autografts ; Bone Marrow Transplantation ; Breast Neoplasms ; Drug Therapy ; Female ; Fertility Preservation* ; Fertility* ; Germ Cells ; Health Occupations ; Hematologic Neoplasms* ; Humans ; Infertility ; Menopause ; Ovary ; Survival Rate ; Transplantation, Autologous ; Whole-Body Irradiation

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