Fertilization in Vitro

The average age of women bearing their first child has increased strongly. This is an important reproductive health problem as fertility declines with increasing female age. Unfortunately, IVF using fresh oocytes cannot compensate for this age-related fertility decline. Oocyte freezing could be a solution. We used the Markov model to estimate the cost-effectiveness of three strategies for 35-year-old women who want to postpone pregnancy till the age of 40: Strategy 1: women undergo three cycles of ovarian hyperstimulation at age 35 for oocyte freezing, then at age 40, use these frozen oocytes for IVF; Strategy 2: women at age 40 attempt to conceive without treatment; and the reference strategy: women at age 40 attempt to conceive and, if not pregnant after 1 year, undergo IVF. Sensitivity analyses were carried out to investigate assumptions of the model and to identify which model inputs had most impact on the results. results: Oocyte freezing (Strategy 1) resulted in a live birth rate of 84.5% at an average cost of E10 419. Natural conception (Strategy 2) resulted in a live birth rate of 52.3% at an average cost of E310 per birth. IVF (the reference strategy) resulted in a cumulative live birth rate of 64.6% at an average cost of E7798. The cost per additional live birth for the oocyte freezing strategy was E13 156 compared to the IVF strategy. If at least 61% of the women return to collect their oocytes, and if there is a willingness to pay E19 560 extra per additional live birth, the oocyte freezing strategy is the most cost-effective strategy. conclusion: Oocyte freezing is more cost effective compared to IVF, if at least 61% of the women return to collect their oocytes and if one is willing to pay E19 560 extra per additional live birth. Our Markov model shows that, considering all the used assumptions, oocyte freezing provides more value for money than IVF.

s of presentations at the Twenty – Ninth Annual American Cytogenet Genome Res Conference held on February 20 ■ 24, 1991 at the Granlibakken Conference Center TahoeCity, CA Organizing Committee: LaDonna Immken (Chairperson) Arthur Brothman Susanne Gollin James Mascarello Stuart Schwartz Urvashi Surti Doris Wurster-Hill

We report on dizygotic (DZ) twins, conceived by IVF and ICSI with assisted hatching, who each had a mixture of 46,XX and 46,XY cells in blood lymphocytes. The female twin had mild genitalia abnormalities but further study revealed anatomically normal reproductive anatomy. Chromosome and fluorescence in situ hybridization studies of buccal, skin and ovarian tissue were normal, as were buccal tissue DNA studies. Fetal ultrasound and fetal membrane pathology were consistent with a monochorionic, diamniotic placenta (MCDAP). These twins thus have blood chimerism but are not chimeric in the other tissues studied. The mechanism for the chimerism could be due to either placental vascular anastamoses (after the development of the haematoblast stem cells) or due to an admixture of trophoblast cells during early blastocyst development. Such trophoblast cell admixtures would be restricted to the extraembryonic tissues so that general physical development in the fetus is normal and without somatic cell chimerism. This case in combination with others previously reported suggests that in IVF conceptions, the prevalence of blood chimerism associated with twinning, and the occurrence of DZ twinning associated with MCDAP, may be higher than previously thought.

Purpose Development of a molecular PGD protocol for a male with an X-linked deletion in the SHOX gene region, located in the pseudoautosomal region of the X/Y chromosomes. Due to excessive recombination in this region, the deletion can be found in male offspring. Methods We developed a 13 marker multiplex fluorescent PCR protocol: 3 markers within the deleted SHOX region, 5 flanking markers, 3 informative markers on chromosome 21 (advanced maternal age) and 2 markers for sex determination. Results Of four embryos, two wild type males, diploid for chromosome 21 were transferred resulting in twin boys. One embryo was an affected female and another embryo was Turner. Amniocentesis confirmed the implanted embryos were males (46XY), with no recombinations. Conclusions While many X-linked disorders can be analyzed by sexing, genes located in the pseudoautosomal regions have high XY recombination rates, requiring multiple markers to enable an accurate diagnosis.

The aim of this study was to analyse the effects of an estradiol (E 2 )-progesterone substitution protocol on the endometrial expression of estrogen-sensitive genes during the peri-implantation period. METHODS: Peripheral blood and endometrial biopsies were obtained from 13 infertile women both in a natural cycle (NC), on days 5 and 7 after ovulation (NC5, NC7), and in an artificial (substituted) cycle (AC), on days 5 and 7 of progesterone addition (AC5, AC7). Estrogen receptor-a (ERa) and progesterone receptor (PR) were assayed by immunohistochemistry. Matrix metalloproteinase-26 (MMP-26) mRNA and tissue inhibitor of metalloproteinase-4 (TIMP-4) mRNA were semiquantitatively assessed in tissue sections using in situ hybridization (ISH) and quantified in tissue extracts using real-time PCR. RESULTS: Levels of both E 2 and progesterone were higher in the peripheral blood in AC than in NC. Also on day AC5, expressions of ERa, PR and MMP-26 mRNA (focally) were increased in the epithelium and TIMP-4 mRNA in the stroma. Expression levels of these genes dropped significantly between AC5 and AC7, but not between NC5 and NC7. Abnormally high levels in AC5 samples suggest overstimulation with E 2 , and the rapid decrease between AC5 and AC7 suggests overstimulation with progesterone. CONCLUSIONS: In ACs, increased levels of E 2 in the blood exaggerate the endometrial expression of estrogen-sensitive genes, whereas higher levels of progesterone in the blood in the secretory phase exaggerate the drop in expression of these genes. Dramatic variations in the gene expression may not be optimal for the implantation process.

Context: Increasing evidence suggests that adverse conditions during prenatal life are associated with the development of chronic diseases in adult life. It is still unclear whether in vitro fertilization (IVF) conception could affect the vulnerable developmental processes in humans occurring during early prenatal development with long-term perturbations of developmental pathways. Objective: Our objective was to examine body composition in 8-to 18-yr-old IVF singletons and spontaneously conceived controls born from subfertile parents.

problems are not encountered when IVF is performed in a Netherlands natural cycle. On the other hand, NIVF meets other problems 1 To whom correspondence should be addressed at: Department of such as an increased risk of cancellation of the attempt due to Obstetrics and Gynaecology, IVF Center, PO Box 7057, a luteinizing hormone (LH) surge , poor 1007 MB Amsterdam, The Netherlands follicle development , no oocyte during The results of in-vitro fertilization in natural cycles (NIVF) oocyte retrieval, no fertilization or polyspermic fertilization in women with tubal infertility at our department are with no embryo transfer as a consequence ) presented. The study had a prospective design. We needed and a considerably lower pregnancy rate per started cycle 75 cycles in 50 patients to obtain one oocyte from each versus SIVF . Nevertheless, there is a patient. Successful oocyte recovery rate was 67% per rising interest in NIVF among patients. It feels more comfortstarted cycle and 82% per oocyte retrieval. Thirty-five able to the patient and the physical burden of NIVF is perceived embryos were transferred and resulted in four ongoing to be much lower than that of SIVF. pregnancies (5.3% per cycle, 6.5% per oocyte retrieval, In this report we present our experience with NIVF and the 11.4% per embryo transfer and 11.4% per embryo). Six results from a selected group of 50 patients who underwent, patients who participated in the study made a second in total, 75 monitored natural IVF cycles. attempt at NIVF. Five of them conceived of which four were ongoing. Cumulative ongoing pregnancy rates are 9.8% per cycle, 11.9% per oocyte retrieval, 19.5% per Materials and methods embryo transfer and 19.5% per embryo. We conclude that Fifty patients, aged between 22 and 38 years and with a history of pregnancy for patients with tubal infertility. secondary tubal infertility, underwent in total 75 cycles of NIVF at Key words: HCG/IVF/natural cycle/pregnancy rates our department. The patients were offered a free treatment, as at that time we had no experience with NIVF. All patients had a regular ovulatory cycle between 24 and 35 days. This was confirmed by at least three biphasic basal body temperature charts prior to the treatment cycle and an endometrial biopsy, taken in a previous cycle

Objective: To predict the chance of total fertilization failure (TFF) before the day of ovum pickup with known semen and female variables. Design: A statistical model was constructed to predict TFF by retrospective analysis (2,366 couples) and subsequently tested on a new IVF population (917 couples). Setting: Academic tertiary referral center. Patient(s): Three thousand three hundred eighty-three couples who underwent an IVF-ET treatment. Intervention(s): None. Main Outcome Measure(s): The ability to predict the probability of TFF in IVF. Result(s): Two variables-postwash total progressively motile sperm cell count (postwash TPMC) and number of follicles-were found to be significant. Taking a probability of 25% as an acceptable risk of TFF, we calculated that a postwash TPMC of Ͻ1.1 ϫ 10 6 cells results in a risk of TFF of Ͼ25%. Low responders (Ͻ4 follicles) needed a postwash TPMC of Ͼ2.2 ϫ 10 6 cells to reduce the risk of TFF to Ͻ25%. High responders (Ͼ15 follicles) needed only 0.35 ϫ 10 6 postwash progressively motile spermatozoa. When postwash TPMC and number of follicles are known and an unacceptable TFF outcome is expected, one can propose an ICSI procedure a few days before the day of ovum pickup. (Fertil Steril

Little is known about post-natal growth in IVF offspring and the effects of rates of early post-natal growth on blood pressure and body fat composition during childhood and adolescence. The follow-up study comprised 233 IVF children aged 8 -18 years and 233 spontaneously conceived controls born to subfertile parents. Growth data from birth to 4 years of age, available for 392 children (n ¼ 193 IVF, n ¼ 199 control), were used to study early post-natal growth. Furthermore, early post-natal growth velocity (weight gain) was related to blood pressure and skinfold measurements at follow-up. results: We found significantly lower weight, height and BMI standard deviation scores (SDSs) at 3 months, and weight SDS at 6 months of age in IVF children compared with controls. Likewise, IVF children demonstrated a greater gain in weight SDS (P , 0.001), height SDS (P ¼ 0.013) and BMI SDS (P ¼ 0.029) during late infancy (3 months to 1 year) versus controls. Weight gain during early childhood (1 -3 years) was related to blood pressure in IVF children (P ¼ 0.014 systolic, 0.04 diastolic) but not in controls. Growth during late infancy was not related to skinfold thickness in IVF children, unlike controls (P ¼ 0.002 peripheral sum, 0.003 total sum). Growth during early childhood was related to skinfold thickness in both IVF and controls (P ¼ 0.005 and 0.01 peripheral sum and P ¼ 0.003 and 0.005 total sum, respectively). conclusions: Late infancy growth velocity of IVF children was significantly higher compared with controls. Nevertheless, early childhood growth instead of infancy growth seemed to predict cardiovascular risk factors in IVF children. Further research is needed to confirm these findings and to follow-up growth and development of IVF children into adulthood.

Purpose To compare the expression profiles of Tektin 2 and CatSper 2 motility proteins in the spermatozoa of normozoospermic and oligoasthenozoospermic men and determine its correlation with sperm motility, fertilization rate, embryo quality and pregnancy rate. Methods Tektin 2 and CatSper 2 protein expression was studied using Western Blotting and immunofluorescence. Tektin 2 and CatSper 2 protein levels were quantified by ELISA. Results Oligoasthenozoospermic men were found to have lower fertilization rates, poor embryo quality and lower pregnancy rates as compared to normozoospermic men. The levels of Tektin 2 and CatSper 2 are significantly lower in spermatozoa of oligoasthenozoospermic men as compared to normozoospermic controls; the levels were also lower in immotile fraction as compared to motile fraction of spermatozoa obtained from normozoospermic individuals. The levels of Tektin 2 and CatSper 2 were higher in individuals demonstrating sperm motility >60 % as compared to sperm motility <30 %. Tektin 2 but not CatSper 2 levels were positively associated with fertilization rate, embryo quality and pregnancy rate. Conclusion Levels of Tektin 2 and CatSper 2 proteins are positively associated with sperm motility parameters. Measurements of Tektin 2 levels can be correlated with the clinical outcome of ICSI.

Introduction: Absence of proper access to the sperm during the fertilization is the cause of male infertility problems. Although the freezing of sperm is reduced the critical parameters, sperm cryopreservation technique is effective for this class of problems. Because of the essentiality of the optimality of the environment and freezing procedures, the effect of the using of the L-carnitine in the environment of sperm cryopreservation and its influence on some sperm vital parameters are perused in this study. Methods: 36 Semen samples normally of application of spermogram in the clinical laboratory in Jahrom are collected and after the affiance of being normal, they divided into 3 groups of 12 samples. We added sperm cryopreservation solution, L-carnitine with conectration 3.6 mM to experimental group1 and L-carnitine 100 mM to experimental groupe 2 are added. Sperms are freezed with vitrification technique for 3 months. Then the sperm vital parameters such as morphology, DNA integr...

Background: The beneficial role of cabergoline as a prophylactic agent to prevent ovarian hyper stimulation syndrome (OHSS) among high-risk patients has been demonstrated in previous studies. But data for its role as a treatment for established severe OHSS is still limited. We represent the treatment results of high dose oral cabergoline in management of six patients after the syndrome is established. Case: High-dose oral cabergoline (1 mg daily for eight days) was prescribed as an adjuvant to symptomatic treatment for six hospitalized patients with established severe OHSS following infertility treatment cycles. In two cases OHSS resolved rapidly despite the occurrence of ongoing pregnancy. Conclusion: Considering the treatment outcomes of our patients, high dose cabergoline did not eliminate the need for traditional treatments, but it was a relatively effective and safe therapy in management of established severe OHSS, and prevented the increase in its severity following the occurrence of pregnancy.

BACKGROUND: The success of IVF treatment is dependent upon embryo quality and coordinated growth and differentiation of the endometrium. Aromatase P450 expression in the human endometrium is thought to be restricted to women with proliferative reproductive tract disorders such as endometriosis, leiomyomas and adenomyosis. METHODS: To determine whether endometrial aromatase P450 mRNA expression is prognostic of IVF outcome, we quanti®ed transcript levels in biopsy specimens from a cohort of subfertile patients awaiting IVF treatment using real-time quantitative PCR. RESULTS: Aromatase P450 transcripts were detected in all endometria examined, although the levels varied considerably between samples, ranging from 0.22 to 486.6 arbitrary units (a.u.). The clinical pregnancy rate in women with high endometrial aromatase P450 mRNA levels (b8.3 a.u.; n = 21) was 9.5% compared with 30.1% in those patients with low expression levels (<8.3 a.u.; n = 101) (P < 0.05). The cycle day of the endometrial biopsy, cause of infertility, age, parity, number of oocytes collected and number of embryos transferred did not differ between patients with high versus low endometrial aromatase P450 mRNA levels (P > 0.1). CONCLUSIONS: Our results indicate that endometrial P450 mRNA levels can identify women at increased risk of IVF treatment failure.

BACKGROUND: Most implantation failures after successful in vitro fertilization-embryo transfer (IVF-ET) result from inadequate uterine receptivity. There is currently no way to predict this receptivity. METHODS: We investigated whether the detection of interleukin-(IL)18 by ELISA in uterine luminal secretions might predict implantation failure. Secretions of 133 patients enrolled in our IVF-ET program were sampled by uterine flushing immediately before oocyte retrieval. We assessed the following outcomes: pregnancy rate, multiple pregnancy rate, and implantation rate per embryo transferred. RESULTS: Interleukin-18 was detected in the flushing fluid of 38 patients (28.6%). Although the two groups were comparable for all other characteristics (age, etiology, ovarian reserve, number of embryos transferred, quality of embryos), all outcome variables differed significantly. The pregnancy rate was 37.9% in the IL-18 2ve group and 15% in the IL-18 1 ve group, the multiple pregnancy rate 27.7% and 0%, and the implantation rate per embryo transferred 19.4% and 6.7% (all comparisons, P 5 0.02). Only embryos meeting good quality criteria were transferred to 65 patients: 50 IL-18 2ve and 15 IL-18 1ve. The pregnancy rate was 51% for the IL-18 2ve group and 20% for the IL-18 1ve group, the multiple pregnancy rate 36% and 0.0%, respectively, and the implantation rate 29% and 8.3% (P 5 0.02). CONCLUSION: This noninvasive and simple method predicted inadequate uterine receptivity, independent of embryo quality.

Objective: To assess intrauterine levels of leukemia inhibitory factor (LIF) by uterine flushing at the time of egg retrieval and to confirm that the procedure has no detrimental effect on pregnancy rates. Design: Prospective study. Setting: Assisted reproductive unit of a university hospital. Patient(s): Uterine flushing was performed in 148 IVF patients. The first 100 patients were compared with a matched control group. Intervention(s): Uterine flushing at the time of egg retrieval. Main Outcome Measure(s): IVF-ET results, pregnancy rates, and intrauterine LIF levels. Result(s): Pregnancy rates were not different in the group of patients with (27%) or without uterine flushing (28%). Leukemia inhibitory factor was detected in 60 patients (46%). Pregnancy rates did not differ between patients' detectable LIF and those in whom LIF was undetectable. Mean levels of LIF were 30.1 Ϯ 49.3 pg/mL and 28.6 Ϯ 51.2 pg/mL in pregnant and nonpregnant patients respectively. The flushing procedure at the time of egg retrieval did not adversely affect pregnancy rates. Leukemia inhibitory factor was detected in 46% of patients at the time of egg retrieval, but no correlation were observed with better pregnancy rates in patients with detectable LIF. Mean LIF levels did not differ in pregnant and nonpregnant women. Access to endoluminal secretions of the endometrium during IVF-ET may represent a new research in human implantation. (Fertil Steril 2003;79:900 -4.

Objective: To investigate the endometrial immunohistochemical staining of interleukin (IL)-12 and IL-18 and to quantify the CD56 bright natural killer (NK) cells in relation to Doppler vascular disorders. Design: Controlled clinical study. Setting: Research unit of a university hospital. Patient(s): Thirty-five women with repeated implantation failure after ET in IVF and 12 fertile control patients. Intervention(s): Ultrasound evaluation and endometrial biopsy on day 20. Main Outcome Measure(s): The balance between IL-12 and IL-18, the number of NK cells, and the vascular status among fertile and implantation failure patients. The control patients displayed normal vascular parameters, a weak anti-IL-12 staining, a consistent moderate stromal anti-IL-18 staining, and fewer than 15 NK cells/field. This pattern was observed among only 17% (6/35) of the implantation failure group. The remaining patients fit into one of two patterns: [1] 37% (13/35) had more than 40 NK cells/field with a strong anti-IL-12 and/or anti-IL-18 staining, and [2] the remaining 46% (16/35) had a marked local depletion of IL-18 and IL-12. Respectively, 85% and 31% of two groups displayed abnormal vascular parameters. Distinctions between the different local dysregulations of the cytokine network may provide clues for further exploration and treatment. (Fertil Steril 2004;81:59 -65.

Objective: To document in the endometrium the correlation among the interleukin (IL)-12, -15, and -18 mRNA and the correlation between cytokine levels, vascular status, and endometrial natural killer (NK) cell count in the context of recurrent implantation failure. Design: A pilot study. Setting: Department of Reproductive Immunology. Patient(s): Women who failed to become pregnant after repeated IVF-embryo transfer and fertile control subjects. Intervention(s): Ultrasonic evaluation and endometrial biopsy in luteal phase. Main Outcome Measure(s): Uterine artery Doppler, count of uterine CD56 bright cells/field, and quantification by real-time polymerase chain reaction (PCR) to monitor IL-12 family (IL-12p40, IL-12p35, EBI3, IL-23), the IL-18 system (IL-18, IL-18R, IL18BP), and the IL-15 mRNA ratio. The uterine artery Doppler and the CD56 bright cell counts were significantly different in fertile and infertile patients. The mean uterine artery pulsatility index correlated significantly negatively with the IL-18/actin ratio suggesting a defect of the cytokine-dependent vascular remodeling pathway. The number of uterine CD56 bright cells was significantly correlated with the IL-15/actin and IL-18/IL-18BP ratios. Thus, IL-18 and IL-15 seems to be involved in the local recruitment and the activation of uterine natural killer (uNK) cells. IL-18 was itself correlated with IL-15 and IL-12, suggesting a local control of uNK cells activation. The assessment of the tripod IL-12/-15/-18 shows distinct immune-related mechanisms that are involved in the broader context of inadequate uterine receptivity.

In the current review the main fundamental biological differences in reproductive function between Bos taurus and Bos indicus cattle are discussed. Breed differences regarding puberty, estrous cycle patterns, estrous behavior, acquisition of ovulatory capacity, ovarian structures and reproductive hormones are presented. The main physiological differences that Bos indicus cattle present relative to Bos taurus cattle include: delayed age at puberty; higher circulating concentrations of hormones such as estradiol, progesterone, insulin and IGF-I, despite having smaller ovulatory follicle size and corpora lutea; greater population of small follicles and smaller size of the dominant follicle at deviation; and greater sensitivity of follicles to gonadotropins. Knowledge of the differences between Bos indicus and Bos taurus breeds help explain different management procedures and responses to hormonal treatments associated with artificial insemination, ovarian superstimulation, and in vivo and in vitro embryo production.

A 37-year-old woman receiving in vitro fertilisation (IVF) treatment presented with lethargy, fevers and anuria. Her background included complex surgically managed Crohn&#39;s disease and mild right-sided hydronephrosis (with no evidence of obstruction on MAG3 renogram). On examination, she had a distended abdomen with generalised tenderness. Blood tests revealed an acute kidney injury and confirmed sepsis. She was found to have bilateral ureteric obstruction with worsening of the right-sided hydronephrosis and new-onset left-sided hydronephrosis secondary to bilateral ovarian masses, diagnosed as mild ovarian hyperstimulation syndrome (OHSS). She was initially managed with fluid resuscitation, bilateral nephrostomies, antibiotics and supportive management of mild OHSS. She made a good recovery and was discharged after placement of antegrade stents and removal of nephrostomies. On follow-up, following resolution of ovarian hyperstimulation, she has been diagnosed with a right-sided ...

Spermiogenesis is a complex process consisting of three main phases: the round, elongating, and elongated spermatid phases. Although the germ cells acquire a haploid set of paternal chromosomes after meiosis, how functional these male haploid germ cells are as male gametes at various differentiation stages has remained unclear. We selectively injected specific steps of haploid male germ cells into oocytes and assessed the function of the zygotes. Applying the transillumination technique using acrosin-green fluorescent protein transgenic mice, we succeeded in selecting four types of haploid male germ cells for microinsemination: early round spermatids (steps 2-3), late round spermatids (steps 7-8), elongating spermatids (steps 9-10), and elongated spermatids (step 16). The microinsemination technique revealed that the early and late round spermatids had similar developmental abilities in producing progeny, indicating that the nuclear status of newly generated round spermatids was similar to that of late round spermatids. An increased birthrate of progeny was first observed in steps 9-10 of elongating spermatids, but the frequency was slightly lower than that of the elongated spermatids. These results indicated that the transition from steps 7-8 of round spermatids to steps 9-10 of elongating spermatids is a key step in changing the nuclear status of male gametes in producing progeny.

Somatic cell nuclear transfer to an enucleated oocyte is used for reprogramming somatic cells with the aim of achieving totipotency, but most cloned embryos die in the uterus after transfer. While modifying epigenetic states of cloned embryos can improve their development, the production rate of cloned embryos can also be enhanced by changing other factors. It has already been shown that abnormal chromosome segregation (ACS) is a major cause of the developmental failure of cloned embryos and that Latrunculin A (LatA), an actin polymerization inhibitor, improves F-actin formation and birth rate of cloned embryos. Since F-actin is important for chromosome congression in embryos, here we examined the relation between ACS and F-actin in cloned embryos. Using LatA treatment, the occurrence of ACS decreased significantly whereas cloned embryo-specific epigenetic abnormalities such as dimethylation of histone H3 at lysine 9 (H3K9me2) could not be corrected. In contrast, when H3K9me2 was normalized using the G9a histone methyltransferase inhibitor BIX-01294, the Magea2 gene-essential for normal development but never before expressed in cloned embryos-was expressed. However, this did not increase the cloning success rate. Thus, non-epigenetic factors also play an important role in determining the efficiency of mouse cloning.

Several lines of evidence indicate that the formation of a transcriptionally repressive state during the two-cell stage in the preimplantation mouse embryo is superimposed on the activation of the embryonic genome. However, it is difficult to determine the profile of newly synthesized (nascent) RNA during this phase because large amounts of maternal RNA accumulate in maturing oocytes to support early development. Using 5-bromouridine-5′-triphosphate labeling of RNA, we have verified that nascent RNA synthesis was repressed between the two-cell and four-cell transition in normally fertilized but not in parthenogenetic embryos. Moreover, this repression was contributed by sperm (male) chromatin, which we confirmed by studying androgenetic embryos. The source of factors responsible for repressing nascent RNA production was investigated using different stages of sperm development. Fertilization with immature round spermatids resulted in a lower level of transcriptional activity than wit...

Somatic cell nuclear transfer to an enucleated oocyte is used for reprogramming somatic cells with the aim of achieving totipotency, but most cloned embryos die in the uterus after transfer. While modifying epigenetic states of cloned embryos can improve their development, the production rate of cloned embryos can also be enhanced by changing other factors. It has already been shown that abnormal chromosome segregation (ACS) is a major cause of the developmental failure of cloned embryos and that Latrunculin A (LatA), an actin polymerization inhibitor, improves F-actin formation and birth rate of cloned embryos. Since F-actin is important for chromosome congression in embryos, here we examined the relation between ACS and F-actin in cloned embryos. Using LatA treatment, the occurrence of ACS decreased significantly whereas cloned embryo-specific epigenetic abnormalities such as dimethylation of histone H3 at lysine 9 (H3K9me2) could not be corrected. In contrast, when H3K9me2 was normalized using the G9a histone methyltransferase inhibitor BIX-01294, the Magea2 gene-essential for normal development but never before expressed in cloned embryos-was expressed. However, this did not increase the cloning success rate. Thus, non-epigenetic factors also play an important role in determining the efficiency of mouse cloning.

Spermiogenesis is a complex process consisting of three main phases: the round, elongating, and elongated spermatid phases. Although the germ cells acquire a haploid set of paternal chromosomes after meiosis, how functional these male haploid germ cells are as male gametes at various differentiation stages has remained unclear. We selectively injected specific steps of haploid male germ cells into oocytes and assessed the function of the zygotes. Applying the transillumination technique using acrosin-green fluorescent protein transgenic mice, we succeeded in selecting four types of haploid male germ cells for microinsemination: early round spermatids (steps 2-3), late round spermatids (steps 7-8), elongating spermatids (steps 9-10), and elongated spermatids (step 16). The microinsemination technique revealed that the early and late round spermatids had similar developmental abilities in producing progeny, indicating that the nuclear status of newly generated round spermatids was similar to that of late round spermatids. An increased birthrate of progeny was first observed in steps 9-10 of elongating spermatids, but the frequency was slightly lower than that of the elongated spermatids. These results indicated that the transition from steps 7-8 of round spermatids to steps 9-10 of elongating spermatids is a key step in changing the nuclear status of male gametes in producing progeny.

The utility of cryopreserved mouse gametes for reproduction of transgenic mice depends on development of assisted reproductive technologies, including vitrification of unfertilized mouse oocytes. Due to hardening of the zona pellucida, spermatozoa are often unable to penetrate vitrified-warmed (V-W) oocytes. Laser-assisted in vitro fertilization (LAIVF) facilitates fertilization by allowing easier penetration of spermatozoa through a perforation in the zona. We investigated the efficiency of V-W C57BL/6NTac oocytes drilled by the XYClone laser, compared to fresh oocytes. By using DAP213 for cryoprotection, 83% (1,470/1,762) of vitrified oocytes were recovered after warming and 78% were viable. Four groups were evaluated for two-cell embryo and live offspring efficiency: 1) LAIVF using V-W oocytes, 2) LAIVF using fresh oocytes, 3) conventional IVF using V-W oocytes and 4) conventional IVF using fresh oocytes. First, the groups were tested using fresh C57BL/6NTac spermatozoa (74% motile, 15 million/ml). LAIVF markedly improved the two-cell embryo efficiency using both V-W (76%, 229/298) and fresh oocytes (69%, 135/197), compared to conventional IVF (7%, 12/182; 6%, 14/235, respectively). Then, frozen-thawed C57BL/6NTac spermatozoa (35% motile, 15 million/ml) were used and LAIVF was again found to enhance fertilization efficiency, with two-cell embryo rates of 87% (298/343) using V-W oocytes (P,0.05, compared to fresh spermatozoa), and 73% (195/266) using fresh oocytes. Conventional IVF with frozen-thawed spermatozoa using V-W (6%, 10/168) and fresh (5%, 15/323) oocytes produced few two-cell embryos. Although live offspring efficiency following embryo transfer was greater with conventional IVF (35%, 18/51; LAIVF: 6%, 50/784), advantage was seen with LAIVF in live offspring obtained from total oocytes (5%, 50/1,010; conventional IVF: 2%, 18/908). Our results demonstrated that zona-drilled V-W mouse oocytes can be used for IVF procedures using both fresh and frozen-thawed spermatozoa, producing live pups. The ability to cryopreserve mouse gametes for LAIVF may facilitate management of large-scale transgenic mouse production facilities.

Background. Thyroid autoimmunity (TAI) affects approximately 13–19% of those experiencing infertility and is often associated with unexplained infertility, recurrent miscarriage, polycystic ovary syndrome, lower antral follicular count, reduced ovarian reserve and embryo quality.

Objective of the review: to consolidate and interpret recent studies investigating the role of TAI in ART success.

Materials and methods. Following several key principles of the PRISMA guidelines the following databases were searched for the last 5 years: MEDLINE (PubMed), Embase, Scopus, Lilacs, Web of Science, Clinical Trials, and SciELO.

Analysis of literary data. While some evidence links TAI with impaired reproductive outcomes, such as reduced fertilization and implantation rates and increased miscarriage, other studies propose that intracytoplasmic sperm injection may alleviate these issues by avoiding immune interference during fertilization. TAI-positive women tend to develop hypothyroidism during ovarian stimulation with human chorionic gonadotropin trigger, which may further compromise pregnancy outcomes. Data supporting the same risk under alternative stimulation protocols are currently not well established. Lastly, transitional hyperthyroidism in cases of postpartum thyroiditis, presents an added challenge for women initiating ART soon after miscarriage.

Conclusions. Early identification of TAI in women undergoing ART is crucial. Screening programs and personalized treatment strategies play a key role in improving fertility outcomes.

Existing data highlights potential areas for further study. These involve the influence of postpartum thyroiditis on ART results, as well as the role of alternative ovulation triggers in TAI-positive patients in order to get better obstetric outcomes. In addition, upcoming research should concentrate on optimizing diagnostic models and developing personalized treatment plans.

Mammalian fertilization is a complex and highly regulated process that relies on multiple interactions among the proteins present on the membranes of sperm and oocyte. This intricate interplay ultimately leads to the fusion of sperm and oocyte; however, the mechanisms underlying gametic membrane recognition and fusion remain largely elusive. Previous studies have identified the ligand-receptor pair IZUMO1 and JUNO as essential for sperm-oocyte fusion. Nevertheless, it has been observed that the interaction between IZUMO1 and JUNO does not facilitate membrane fusion in somatic cells, indicating that additional molecules are required for successful gamete membrane fusion. In this study, we demonstrated significant inhibition of rat sperm penetration into oocytes following treatment with anti-PTCH1 monoclonal antibodies targeting both gamete types. The hedgehog receptor PTCH1 is expressed in rat sperm and oocytes during metaphase of meiosis II (MII) of meiosis, engages in physical interactions with IZUMO1, JUNO, and PDIA3; specifically, the extracellular domain ECD1 of PTCH1 selectively interacts with JUNO, but displays no detectable interaction with either IZUMO1 or PDIA3; conversely ECD2 shows broader binding to IZUMO1, JUNO, and PDIA3. Furthermore, PTCH1 colocalizes with IZUMO1 on the surface of the acrosome-reacted sperm heads and with PDIA3 on the surface of the oocyte. Our research reveals that PTCH1 plays a crucial role in mediating sperm-oocyte fusion through its interaction with IZUMO1 and PDIA3. These findings provide new insights into the molecular mechanisms governing mammalian sperm-oocyte adhesion and fusion.

Follicular fluid (FF) is biological fluid rich in nutrients, growth factors, hormones and may affect the sperm quality. Sperm washing has been done using conventional media in laboratory procedure so far. This study aimed to investigate the effects of FF on survival and maintenance of chromatin integrity post swim up. Each washed semen sample was divided into two parts; the control group was incubated in the media, and the experimental groups incubated in the media containing 10% follicular fluid. Smears were prepared after 20 min, 180 min, 24 hours and no incubation times. Sperm chromatin changes like protamine, histone, DNA denaturation, sperm chromatin stability and motility were evaluated at different times. Incubation of sperm in the follicular fluid increased sperms with normal histone, normal chromatin protamine and sperm with normal head size (p &lt; 0.05). Administration of follicular fluid into the culture media of the sperms that had been separated by swim up method could...

Background Hepatitis B virus (HBV) infection is a major global health issue, affecting millions of individuals worldwide. Chronic HBV infection is associated with various systemic health complications, including liver disease with its impact on reproductive health in male fertility. Methods This study investigated the impact of HBV on sperm quality in a cohort of 1566 male infertile patients undergoing assisted reproductive treatment from 2021 to 2024. Patients aged 23 to 40 years (mean age 34.2 ± 3.62 years) were categorized into HBV-positive (783 cases) and HBV-negative (783 cases) groups based on serological tests for hepatitis B surface antigen (Hbs Ag). Results HBV-positive and HBV-negative groups exhibited significantly lower progressive motility (41.3% vs. 43.0%; P = 0.006), sperm concentration (53.6 vs. 59.8 × 10 6 /mL; P = 0.009), and normal morphology (4.5% vs. 5.0%; P < 0.001), with reduced viability (P = 0.027). No differences were observed in semen volume (P = 0.807) or DNA fragmentation index (P = 0.970). Conclusion HBV infection is associated with impaired sperm motility, concentration, and morphology in infertile men.
Highlights • HBV seropositivity exhibits negative associations with progressive motility, sperm concentration, and normal morphology in infertile men. • The absence of association with DNA fragmentation indicates that HBV may impair spermatogenesis through pathways independent of direct genetic damage. • HBV exhibits associations with poorer sperm quality.

Background Low efficiency of Somatic Cell Nuclear Transfer (NT) has been widely addressed with high incidence of placental abnormalities due to genetic and epigenetic modifications. MiRNAs are shown to be major regulators of such modifications. The present study has been carried out to identify the expression patterns of 377 miRNAs, their functional associations and mechanism of regulation in bovine placentas derived from artificial insemination (AI), in vitro production (IVP) and NT pregnancies. Results This study reveals a massive deregulation of miRNAs as chromosomal cluster or miRNA families without sex-linkage in NT and in-vitro derived IVP placentas. Cell specific localization miRNAs in blastocysts and expression profiling of embryos and placentas at different developmental stages identified that the major deregulation of miRNAs exhibited in placentas at day 50 of pregnancies is found to be less dependent on global DNA methylation, rather than on aberrant miRNA biogenesis mole...

Objective: To review the mechanisms by which endometriosis may affect reproductive function. Design: Review of the English literature from 1986 to 2007 after searching Medline, EMBASE, Cochrane, and BIOSIS, as well as relevant meeting abstracts. Setting: Fertility research center and obstetrics and gynecology department in a tertiary care hospital. There is compelling evidence in the literature that endometriosis has detrimental effects on ovarian and tubal function and uterine receptivity, resulting in female infertility. The mechanisms of infertility associated with endometriosis remain controversial and include abnormal folliculogenesis, elevated oxidative stress, altered immune function, and hormonal milieu in the follicular and peritoneal environments, and reduced endometrial receptivity. These factors lead to poor oocyte quality, impaired fertilization, and implantation. Through unraveling the mechanisms by which endometriosis leads to infertility, researchers are sure to find a nonsurgical means to diagnose endometriosis, most likely through serum and peritoneal markers. Cytokines, interleukins, oxidative stress markers, and soluble cellular adhesion molecules all show potential to be used as a reliable marker for diagnosing endometriosis. After analyzing the pathogenic mechanisms of endometriosis, it seems that the future treatment of this entity may include cyclo-oxygenase-2 inhibitors, immunomodulators, or hormonal suppressive therapy to eliminate the need for surgical treatment of endometriosis.

Background: Endometrium disorders cause significant failure in fertility. Our study aimed to investigate the cellular, histological, and molecular properties of patients with thin endometrium compared to healthy endometrium without pathological conditions (n = 6). Methods: This study was a randomized control (n = 6) in which samples of thin endometrium were obtained from IVF patients enrolled in the IVF clinic in the National Hospital Ciptomangunkusumo, Jakarta, Indonesia, from 2018 to 2019. The typical samples were taken from patients who underwent tubectomy with no pathological findings in the endometrium. Endometrial cells were isolated using an enzymatic method transported in medium, PCR samples were used in an RNA solution, and histology samples were stored in neutral-buffered formalin 10% (v/v). Results: The number of cells, presented as density of cells mg -1 wet weight during endometrial biopsy, was about 10,190 cells mg -1 wet weight in the healthy-endometrium group, while the thin-endometrium group had 3305 cells mg -1 wet weight. A flow cytometry analysis of the endometrial in the healthy group showed all panels were significantly higher compared to the thin group: BCRP1/Breast Cancer-Resistant Protein (74.85 ± 15.63% vs. 15.58 ± 7.19%), ICAM-1/Intercellular Adhesion Molecule or CD54 (35.76 ± 6.23% vs. 7.24 ± 1.65%), PECAM/platelet endothelial cell adhesion molecule-1 or CD31 (29.42 ± 2.16% vs. 4.19 ± 2.03%), and cKit/type III receptor tyrosine kinase or CD117 (27.29 ± 1.93% vs. 5.32 ± 2.38%). The exception was SUSD-2/Sushi Domain Containing Protein-2, although these results were insignificant (1.33 ± 0.39% vs. 0.26 ± 0.39%, respectively) (mean ± 95% C.l). Endometrial receptivity, indicated by HOXA10/Homeobox protein-10, LIF/Leukemia inhibitory factor, SPP1/osteopontin, and VEGF/Vascular Endothelial Growth Factor, was significantly higher in the healthy group compared to the thin group. Immunohistochemical labelling showed that E-cadherin, estrogen/ER-α, and progesterone receptors/PR were found in healthy-endometrium patients while they were less expressed among thin-endometrium patients. Conclusions: Patients with healthy endometrium had significantly higher cell numbers per mg, significantly higher endometrial and endothelial cells, and expressed higher receptivity genes than patients with thin endometrium, although the SUSD-2-positive cells showed no difference.

Background: Endometrium disorders cause significant failure in fertility. Our study aimed to investigate the cellular, histological, and molecular properties of patients with thin endometrium compared to healthy endometrium without pathological conditions (n = 6). Methods: This study was a randomized control (n = 6) in which samples of thin endometrium were obtained from IVF patients enrolled in the IVF clinic in the National Hospital Ciptomangunkusumo, Jakarta, Indonesia, from 2018 to 2019. The typical samples were taken from patients who underwent tubectomy with no pathological findings in the endometrium. Endometrial cells were isolated using an enzymatic method transported in medium, PCR samples were used in an RNA solution, and histology samples were stored in neutral-buffered formalin 10% (v/v). Results: The number of cells, presented as density of cells mg -1 wet weight during endometrial biopsy, was about 10,190 cells mg -1 wet weight in the healthy-endometrium group, while the thin-endometrium group had 3305 cells mg -1 wet weight. A flow cytometry analysis of the endometrial in the healthy group showed all panels were significantly higher compared to the thin group: BCRP1/Breast Cancer-Resistant Protein (74.85 ± 15.63% vs. 15.58 ± 7.19%), ICAM-1/Intercellular Adhesion Molecule or CD54 (35.76 ± 6.23% vs. 7.24 ± 1.65%), PECAM/platelet endothelial cell adhesion molecule-1 or CD31 (29.42 ± 2.16% vs. 4.19 ± 2.03%), and cKit/type III receptor tyrosine kinase or CD117 (27.29 ± 1.93% vs. 5.32 ± 2.38%). The exception was SUSD-2/Sushi Domain Containing Protein-2, although these results were insignificant (1.33 ± 0.39% vs. 0.26 ± 0.39%, respectively) (mean ± 95% C.l). Endometrial receptivity, indicated by HOXA10/Homeobox protein-10, LIF/Leukemia inhibitory factor, SPP1/osteopontin, and VEGF/Vascular Endothelial Growth Factor, was significantly higher in the healthy group compared to the thin group. Immunohistochemical labelling showed that E-cadherin, estrogen/ER-α, and progesterone receptors/PR were found in healthy-endometrium patients while they were less expressed among thin-endometrium patients. Conclusions: Patients with healthy endometrium had significantly higher cell numbers per mg, significantly higher endometrial and endothelial cells, and expressed higher receptivity genes than patients with thin endometrium, although the SUSD-2-positive cells showed no difference.

Background: Endometrium disorders cause significant failure in fertility. Our study aimed to investigate the cellular, histological, and molecular properties of patients with thin endometrium compared to healthy endometrium without pathological conditions (n = 6). Methods: This study was a randomized control (n = 6) in which samples of thin endometrium were obtained from IVF patients enrolled in the IVF clinic in the National Hospital Ciptomangunkusumo, Jakarta, Indonesia, from 2018 to 2019. The typical samples were taken from patients who underwent tubectomy with no pathological findings in the endometrium. Endometrial cells were isolated using an enzymatic method transported in medium, PCR samples were used in an RNA solution, and histology samples were stored in neutral-buffered formalin 10% (v/v). Results: The number of cells, presented as density of cells mg -1 wet weight during endometrial biopsy, was about 10,190 cells mg -1 wet weight in the healthy-endometrium group, while the thin-endometrium group had 3305 cells mg -1 wet weight. A flow cytometry analysis of the endometrial in the healthy group showed all panels were significantly higher compared to the thin group: BCRP1/Breast Cancer-Resistant Protein (74.85 ± 15.63% vs. 15.58 ± 7.19%), ICAM-1/Intercellular Adhesion Molecule or CD54 (35.76 ± 6.23% vs. 7.24 ± 1.65%), PECAM/platelet endothelial cell adhesion molecule-1 or CD31 (29.42 ± 2.16% vs. 4.19 ± 2.03%), and cKit/type III receptor tyrosine kinase or CD117 (27.29 ± 1.93% vs. 5.32 ± 2.38%). The exception was SUSD-2/Sushi Domain Containing Protein-2, although these results were insignificant (1.33 ± 0.39% vs. 0.26 ± 0.39%, respectively) (mean ± 95% C.l). Endometrial receptivity, indicated by HOXA10/Homeobox protein-10, LIF/Leukemia inhibitory factor, SPP1/osteopontin, and VEGF/Vascular Endothelial Growth Factor, was significantly higher in the healthy group compared to the thin group. Immunohistochemical labelling showed that E-cadherin, estrogen/ER-α, and progesterone receptors/PR were found in healthy-endometrium patients while they were less expressed among thin-endometrium patients. Conclusions: Patients with healthy endometrium had significantly higher cell numbers per mg, significantly higher endometrial and endothelial cells, and expressed higher receptivity genes than patients with thin endometrium, although the SUSD-2-positive cells showed no difference.

Background: Endometrium disorders cause significant failure in fertility. Our study aimed to investigate the cellular, histological, and molecular properties of patients with thin endometrium compared to healthy endometrium without pathological conditions (n = 6). Methods: This study was a randomized control (n = 6) in which samples of thin endometrium were obtained from IVF patients enrolled in the IVF clinic in the National Hospital Ciptomangunkusumo, Jakarta, Indonesia, from 2018 to 2019. The typical samples were taken from patients who underwent tubectomy with no pathological findings in the endometrium. Endometrial cells were isolated using an enzymatic method transported in medium, PCR samples were used in an RNA solution, and histology samples were stored in neutral-buffered formalin 10% (v/v). Results: The number of cells, presented as density of cells mg-1 wet weight during endometrial biopsy, was about 10,190 cells mg-1 wet weight in the healthy-endometrium group, while the thin-endometrium group had 3305 cells mg-1 wet weight. A flow cytometry analysis of the endometrial in the healthy group showed all panels were significantly higher compared to the thin group: BCRP1/Breast Cancer-Resistant Protein (74.85 ± 15.63% vs. 15.58 ± 7.19%), ICAM-1/Intercellular Adhesion Molecule or CD54 (35.76 ± 6.23% vs. 7.24 ± 1.65%), PECAM/platelet endothelial cell adhesion molecule-1 or CD31 (29.42 ± 2.16% vs. 4.19 ± 2.03%), and cKit/type III receptor tyrosine kinase or CD117 (27.29 ± 1.93% vs. 5.32 ± 2.38%). The exception was SUSD-2/Sushi Domain Containing Protein-2, although these results were insignificant (1.33 ± 0.39% vs. 0.26 ± 0.39%, respectively) (mean ± 95% C.l). Endometrial receptivity, indicated by HOXA10/Homeobox protein-10, LIF/Leukemia inhibitory factor, SPP1/osteopontin, and VEGF/Vascular Endothelial Growth Factor, was significantly higher in the healthy group compared to the thin group. Immunohistochemical labelling showed that E-cadherin, estrogen/ER-α, and progesterone receptors/PR were found in healthy-endometrium patients while they were less expressed among thin-endometrium patients. Conclusions: Patients with healthy endometrium had significantly higher cell numbers per mg, significantly higher endometrial and endothelial cells, and expressed higher receptivity genes than patients with thin endometrium, although the SUSD-2-positive cells showed no difference.

In fish with external fertilization, sperm must reach the oocyte through the micropyle to enter the cytoplasm. Fertilization success is then influenced by characteristics of oocytes or sperm. In this study, we evaluated oocyte morphology and sperm motility parameters and their effects on the inseminating dose in a teleost fish Astyanax altiparanae. Interestingly, we found one of the lowest yet described inseminating doses in teleosts (2390 spermatozoa oocyte-1 ml-1). Such a fertilization efficacy may be explained by the long duration of sperm motility (&gt;75 s), the small oocyte diameter (695.119 µm), large micropyle diameter (7.57 µm), and the presence of grooves on the oocyte surface that guides spermatozoon to the fertilization area. Additionally, we have described for the first time a structure that combines grooves on the chorion surface and a ridge in the micropylar area.

Purpose: The presence of a clear half-moon-like zone of cytoplasm in oocytes is called “halo effect.” The prognostic value of this effect is not yet determined. Aligned nucleoli in pronuclei (PN) represent a further polarization phenomenon and a marker for implantation potential. Aim of the prospective study was to evaluate the influence of the halo effect on IVF outcome and

background: Debate exists regarding the effect of raised BMI on the outcome of pregnancies after assisted reproduction technology. We assessed the effect of BMI on the risk of miscarriage in women conceiving following single blastocyst transfer (SBT) after controlling for confounding factors. methods: Fresh and cryo-thawed cycles of SBT that resulted in a pregnancy between January 2006 and March 2010 were included. Patients with BMI , 18.5 kg/m 2 or older than 40 years were excluded. Patients were grouped according to their BMI at the start of treatment cycle. The main outcome measure was the miscarriage rate before 23 weeks gestation. Confounding variables examined included female age, duration and cause of infertility, previous miscarriage, smoking status and quality of blastocyst replaced. results: A total of 413 women conceived following SBT in fresh (n ¼ 325) or cryo-thawed (n ¼ 88) IVF cycles, of whom 244 had a normal BMI (18.5-24.9) and 169 had a raised BMI of ≥25. Overall, 27% (113/413) of women miscarried before 23 weeks gestation. Women with a BMI of ≥25 had more than double the risk of miscarriage compared with women who had normal BMI [38 versus 20%, odds ratio (OR): 2.4, 95% confidence interval (CI) 1.6 -3.8, P , 0.001, respectively]. After adjusting for confounding variables, having a BMI of ≥25 significantly increased the risk of clinical miscarriage before 23 weeks gestation in both fresh (adjusted OR ¼ 2.7, 95% CI 1.5 -4.9, P ¼ 0.001) and cryo-thawed IVF cycles (OR ¼ 6.8, 95% CI 1.5-31.1, P ¼ 0.012). conclusions: Raised BMI is independently associated with higher miscarriage rate after IVF treatment.

drug.'3 A recent review of experience with azathioprine in 27 patients over a median duration of seven years suggested that side effects were few, and treatment had had to be discontinued in only four patients'4; all of the others improved and were able to take lower dosages of anticholinesterase drugs and corticosteroids. It has been argued that corticosteroids and immunosuppressants do not diminish the already low mortality from myasthenia gravis,j but there is little doubt that they improve the quality of life. Despite the lack of evidence from rigorous controlled trials the conclusion must be that the outlook for patients with myasthenia gravis has improved considerably over the past