Abstract Study question Is it possible to determine diploid mono-pronuclear (1PN) human embryos by the speed of development to blastocyst formation? Summary answer The developmental time to reach the ...blastocyst stage is a discrimination indicator for diploid 1PN embryos. What is known already We have previously reported that 80.7% of 1PN zygotes had a biparental chromosome using a Live-Cell imaging technique, and some of these developed to the blastocyst stage (Tokoro et al. ASRM 2013). Furthermore, we have reported that these blastocysts can result in a healthy live birth (Tsuji et al. ASRM 2020), and there was no effect on growth up to 5 years of age (Tsuji et al. ASRM 2023). On the other hand, it was also shown that a few of the 1PN embryos that developed into blastocysts were haploid embryos. Study design, size, duration This was a retrospective study that included 28 blastocysts derived from a 1PN zygote. The time period was 72 months (January 2017 to December 2022). These embryos are used for research following obtaining informed consent. Participants/materials, setting, methods The frequency rate of diploid embryos was compared from time of PN fading to blastocyst formation (Gardner’s classification: ≥ expansion grade 3). Two development time range groups were examined and designated: fast-developing embryos (65.7-78.7h) and slow-developing embryos (116.8-139.7h). Ploidy status was determined using whole blastocyst cells and NGS-based SNP analysis. Statistical significance was determined using Fisher’s exact test (level of P<0.05). Main results and the role of chance There was no significant difference in the average age (years) of the two development time groups (fast-developing embryos group; 35.5 +/- 5.5, slow-developing embryos group; 34.1 +/- 2.8). The frequency of diploid status in 1PN embryos with fast developing embryos was 100% (14/14), and in 1PN embryos designated as having slow development, the frequency of diploid embryos was only 14.3% (2/14), haploid embryos 71.4% (10/14), and other was 14.3% (2/14). As a result, the diploid embryo rate was significantly higher in fast-developing embryos than in slow-developing embryos (P<0.001). Limitations, reasons for caution This study was limited to extremely fast or slow-developing embryos to see trends. It is important to study the threshold time between diploidy and haploidy in 1PN embryos. Wider implications of the findings These results confirmed that fast developing 1PN embryos have a high diploid rate and can be used as transferable embryos in addition to 2PN embryos. Conversely, slow developing 1PN embryos should be considered to have a lower priority for transfer because of the high rate of haploid embryos. Trial registration number not applicable
DFN3, an X chromosome-linked nonsyndromic mixed deafness, is caused by mutations in the BRN-4 gene, which encodes a POU transcription factor. Brn-4-deficient mice were created and found to exhibit ...profound deafness. No gross morphological changes were observed in the conductive ossicles or cochlea, although there was a dramatic reduction in endocochlear potential. Electron microscopy revealed severe ultrastructural alterations in cochlear spiral ligament fibrocytes. The findings suggest that these fibrocytes, which are mesenchymal in origin and for which a role in potassium ion homeostasis has been postulated, may play a critical role in auditory function.
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BFBNIB, NMLJ, NUK, PNG, SAZU, UL, UM, UPUK
Abstract
Study question
Do the anti-centromere antibodies (ACA) affect the multi pronuclear formation (MPF) rate?
Summary answer
MPF rate was higher in patients with ACA. The MPF rate of ACA patients ...with antibody titers ≥160-fold was significantly higher than in titers 40-fold.
What is known already
ACA is an anti-nuclear antibody (ANA), and specifically recognizes the centromere. Recently, several studies have reported that the rate of oocyte maturation is lower in patients with ACA. Moreover, in MI oocytes collected from ACA patients, female chromosomes were frequently dispersed in the cytoplasm.
Study design, size, duration
A total of 4756 patients from our clinic were tested for ANA before oocyte retrieval from January 2014 to December 2021. After retrospective ANA testing, patients were classified according to 3 groups; 62 ACA patients (with only ACA), 1134 non-ACA patients (with ANA but not with included ACA) and 3560 non-ANA patients (without ACA and any ANA).We considered ACA-positive levels at titers ≥40. ACA patients were further classified into 6 groups by antibody titer.
Participants/materials, setting, methods
The MPF rate after ICSI was compared in the 3 groups (ACA patients, non-ACA patients, non-ANA patients) and at 6 ACA titers in each group (titer of ACA; 40-fold, 80-fold, 160-fold, 320-fold, 640-fold, ≥1280-fold).MPF rate was calculated by dividing the number of embryos that formed three or more pronuclei by the number of embryos inseminated by ICSI.Ryan's method was used for multiple comparisons of ratios.
Main results and the role of chance
MPF rate was 3.8% (1997/53240) in non-ANA patients, 4.3% (733/17003) in non-ACA patients, and 32.1% (351/1092) in ACA patients, being significantly higher in ACA vs other groups (P<.01). In comparisons between ACA titers, MPN rate was 8.7% (4/46) at 40-fold , 13.0% (3/23) at 80-fold , 36.1% (56/155) at 160-fold , 32.4% (48/148) at 320-fold , 36.0% (111/308) at 640-fold , and 31.3% (129/412) at ≥ 1280-fold, respectively. MPN rate of patients with titers ≥160-fold was significantly higher than with 40-fold.
Limitations, reasons for caution
A potential limitation of the present study is the small sample size. This is because ACA patients account for only 1% of patients who underwent ART treatments.
Wider implications of the findings
MPF rate in ACA patients was significantly higher than in non-ANA and non-ACA patients and was also significantly higher in ACA patients with titers of ≥ 160-fold vs 40-fold. The dispersion of the female chromosome in the cytoplasm of MI oocytes may be a cause of MPN formation.
Trial registration number
None
Background: Smooth muscle cell (SMC)‐rich intima is a morphological feature of atherosclerotic lesions that is observed in eroded plaque and spastic arteries. Arteries with SMC‐rich intima are ...susceptible to vasoconstriction or vasospasm against some vasoactive agents. Objective: The present study evaluates the contribution of SMC‐rich intima to thrombogenic vasoconstriction. Methods: We established SMC‐rich neointima by damaging rabbit femoral arteries using balloons and then measured the isometric tension of the femoral strips against 5‐hydroxytryptamine (5‐HT), adenosine diphosphate, adenosine triphosphate and thrombin. Results: Among these agents, only 5‐HT induced a hypercontractile response of the injured arteries with SMC‐rich neointima, compared with non‐injured arteries. Smooth muscle cells of both the neointima and media expressed 5‐HT2A receptor, and sarpogrelate, a selective 5‐HT2A receptor antagonist significantly inhibited the hypercontraction. Furthermore, 5‐HT induced contraction of separated neointima and hypercontraction of separated media compared with non‐injured media. Sarpogrelate and fasudil, a specific Rho‐kinase inhibitor, significantly suppressed such contraction of both the neointima and media of injured arteries. Conclusions: These results suggest that 5‐HT plays a crucial role in thrombogenic vasoconstriction, and that SMC‐rich intima as well as media directly contributes to the hypercontractile response of atherosclerotic vessels through the 5‐HT2A receptor and the Rho‐kinase pathway.
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FZAB, GEOZS, GIS, IJS, IMTLJ, IZUM, KILJ, KISLJ, NLZOH, NUK, OILJ, PILJ, PNG, SAZU, SBCE, SBJE, SBMB, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
The circulating osteoprotegerin (OPG) level reflects a series of cardiovascular diseases; however, the source(s) of circulating OPG remain(s) to be determined. This study explored whether OPG is ...released in the coronary circulation and whether it is associated with cardiac structure and function. Fifty-six patients (67±10 years old, male 57%, hypertension 73%, coronary artery disease 50%) were enrolled, and blood samples were collected simultaneously from the orifice of the left coronary artery (CA) and the coronary sinus (CS) after angiography. The concentration of OPG was higher in the CS than in the CA (7.7±4.1 vs. 6.7±3.6 pmol/l, p<0.001). The trans-cardiac OPG concentration was significantly (p=0.019) decreased in patients who have been prescribed either an angiotensin converting enzyme inhibitor or an angiotensin II type 1 receptor blocker (ACEI/ARB). In patients subgroup who did not take an ACEI/ARB (n=27), the trans-cardiac OPG level was positively correlated with age (r=0.396, p=0.041) and relative wall thickness of left ventricle (r=0.534, p=0.004). In multivariate linear regression analysis, relative wall thickness remained to be the independent variable for the trans-cardiac OPG level (p=0.004). Moreover, trans-cardiac OPG was significantly (p=0.021) increased in patients with relative wall thickness greater than 0.45 but it did not differ if the left ventricular mass index was increased (≥116 for males, or ≥ 104 for females, g/m(2)) or not (p=0.627). This study suggests that OPG is secreted into the coronary circulation and is associated with concentric remodeling/hypertrophy of LV, possibly in interactions with the renin-angiotensin system.
Abstract
Study question
TESE is widely used for obstructive azoospermia (OA) as a surgical method for sperm retrieval, but is it benefiting the patient?
Summary answer
Since MESA-ICSI has a very good ...fertilization rate, clinical pregnancy rate and delivery rate, MESA should be employed for the OA subjects, not TESE.
What is known already
TESE and micro-TESE are technically simple, and widely used as a sperm retrieval surgery because they do not require microsurgical skills. Some review articles have shown no significant differences in the rates of cleavage, good-quality embryos, implantation, clinical pregnancy between ICSI with epididymal sperm or testicular sperm. Clinical usefulness of MESA is still controversial according to previous reports.
Study design, size, duration
We studied 110 patients diagnosed with OA and treated with MESA at the Asada Ladies Clinic between April 2004 and December 2021.
Participants/materials, setting, methods
The MESA was performed using a micropipette with a micropuncture technique under operative microscope. When no sperm were present or motility was not observed, additional punctures to the epididymal tubule were performed. Aspirated samples were transferred into modified human tubal fluid and sent to the in vitro fertilization (IVF) laboratory for cryopreservation.
Main results and the role of chance
Motile sperm were recovered in all cases (110 cases). Of these, ICSI using frozen thawed sperm was performed in 101 cases. The rate of normal fertilization rate was 76%. Of the 399 embryo transfer (ET) cycles, 168 had a clinical pregnancy (41% per ET). Of the 101 patients who underwent ART, 94 (93% per case) had clinical pregnancies resulting in 90 (89.1%) deliveries. It should be emphasized that since MESA does not involve incision of the testes, there are fewer postoperative peritoneal irritation symptoms and no concerns about postoperative testicular atrophy or low testosterone levels. Some review articles showed no significant differences in the rates of cleavage, good-quality embryo, implantation, clinical pregnancy between ICSI with epididymal sperm or testicular sperm. Although clinical usefulness of MESA is still controversial, if the ART results of MESA-ICSI and TESE-ICSI are the same, MESA should be performed as it is less invasive on the patient and reduces the burden on the embryologist who have to process the TESE tissue.
Limitations, reasons for caution
Some authors have reported that since MESA specimens contain DNA fragmentated sperm when compared with specimen of TESE, subsequent ICSI results in poorer fertilization and pregnancy rates. We did not evaluate sperm DNA fragmentation.
Wider implications of the findings
A large quantity of uncontaminated sperm can be retrieved using MESA which is less invasive for the patient, and there is no need for special processing before cryopreservation. Additionally, it can reduce the laboratory workload.
Trial registration number
not applicable
Abstract
Study question
Are neonatal outcomes normal up to 3 years of ages from the transfer of blastocysts derived from mono-pronuclear(1PN) zygotes?
Summary answer
There was no effect on growth up ...to 3 years of ages in babies born from 1PN-implanted blastocysts.
What is known already
In human ART, 1PN zygotes are observed at low rates. We have previously reported that 80.7% of 1PN zygotes had a biparental chromosome using a Live-Cell imaging technique, and some of these developed to the blastocyst stage (Tokoro et al. ASRM 2013). Furthermore, we have reported that these blastocysts can result in a viable pregnancy and healthy live birth (Tsuji et al. ASRM 2020), and there was no effect on growth up to 18-months (Tsuji et al. ESHRE 2021). However, there is no information on the long-term prognosis of babies derived from1PN-zygotes.
Study design, size, duration
This was a retrospective study which included 86 cases where there was a live birth after single embryo transfer of a blastocyst derived from 1PN zygote. The incidence of birth defects, birth weight were recorded as well as a physical development survey of 19 children who responded to a 3-years follow-up survey. The time period was 108 months (January 2013 to December 2021).
Participants/materials, setting, methods
Patients seeking fertility treatment at an established private IVF clinic. We compared the birth weight, 3-years old height and weight of children born from 1PN zygotes with data from a control, 2PN group. Statistical significance was determined using the t-test (level of P < 0.05).
Main results and the role of chance
The incidence of birth defects in 1PN embryo-derived infants was 1.2% (1/86). The average birth weight of boys in the 1PN group was 3147.6+/-408.7 g, which was not significantly different from 3070.5+/-479.5 g in the 2PN group. In girls, the average birth weight was 3048.1+/-510.7 g in the 1PN group, which was not significantly different from the 2PN group (2966.2+/-462.7 g). The average height at 18-months, was 81.3+/-2.5 cm vs 80.9+/-3.4 cm for boys; 79.5+/-2.2 cm vs 79.5+/-3.0 cm for girls in the 1PN and 2PN groups, respectively. The average body weights of the 1PN and 2PN groups were 11.0+/-1.2 kg vs 10.8+/-1.1 kg for boys; 9.8+/-1.0 kg vs 10.2+/-1.1 kg for girls, respectively. The average height at 3-years, was 95.2+/-3.1 cm vs 93.8+/-4.5 cm for boys; 91.5+/-2.9 cm vs 92.5+/-4.9 cm for girls in the 1PN and 2PN groups, respectively. The average body weights of the 1PN and 2PN groups were 14.7+/-1.2 kg vs 14.1+/-1.5 kg for boys; 13.2+/-1.9 kg vs 13.6+/-1.5 kg for girls, respectively. There was no significant difference in average height and weight up-to the 3-years follow-up survey.
Limitations, reasons for caution
The incidence of 1PN- zygotes that develop to blastocysts resulting in births is low and the study was limited to cases of single blastocyst embryo transfer.
Wider implications of the findings
The incidence of congenital anomalies in Japan is around 1.7 to 2%, and the incidence was similar in the 1PN group. Also, there was no difference in 3-years follow-up survey of the 1PN compared with the 2PN. These data are reassuring for the clinical utility of 1PN derived embryos.
Trial registration number
not applicable
Abstract
Study question
Does Day 7 blastocyst-transfer affect prognosis of babies born?
Summary answer
There was no effect on growth up to 3 years of age in babies from Day 7 blastocyst-derived ...births.
What is known already
Culture medium and technology have advanced, and the efficiency to culture up to the blastocyst stage in order to obtain viable embryos has increased dramatically. As a result, blastocyst transfer is associated with higher clinical pregnancy outcomes than cleavage transfer. In 1998, it was first reported that some embryos may develop into blastocysts on Day 7. However, there are few reports on neonatal outcomes from Day 7 blastocyst transfer. Therefore, the utility of Day 7 blastocysts is controversial.
Study design, size, duration
This was retrospective study which included 23 cases where there was a live birth after single embryo transfer of a Day 7-derived blastocyst. The incidence of birth defects, birth weight was recorded as well as a physical development survey. The height and weight were recorded in 10 children aged 18-months and 4 children aged 3 who responded to follow-up survey. The time period was from January 2013 to December 2021.
Participants/materials, setting, methods
Patient seeking fertility treatment at an established private IVF clinic. We compared the birth weight, and birth after 18-months and 3 years height and weight of children born to a Day 7 blastocyst with data from babies born from Day 5 and Day 6 derived blastocysts. Statistical significance was determined using Kruskal-Wallis test.
Main results and the role of chance
The incidence of birth defects was 2.4% (115/4828), 1.7% (16/954) and 0% (0/23) in the Day 5, Day 6 and Day 7 groups, respectively. The average birth weight of the Day 7 group was 3057+/-532 g, which was not significantly different from 3024+/-472 g and 3041+/-480 g in the Day 5 and Day 6 groups. The average height at 18-months, was 80.0+/-5.3 cm, 79.8+/-5.1 cm, 77.8+/-6.9 cm in the Day 5, Day 6 and Day 7 groups, respectively. The average weight at 18-months, was 10.5+/-1.2 kg, 10.4+/-1.1 kg, 10.5+/-1.4 kg. The average height at 3 years old, was 93.2+/-4.8 cm, 92.9+/-4.7 cm, 93.4+/-5.9 cm. The average weight at 3 years old, was 13.9+/-1.6 kg, 13.8+/-1.4 kg, 14.1+/-2.6 kg, respectively. There was no significant difference in average height and weight up-to the 3 years old follow-up survey.
Limitations, reasons for caution
The incidence of Day 7 blastocyst-transfer is low and the study was limited to cases of single blastocyst embryo transfer. The study was not separated by sex, because sample size was small.
Wider implications of the findings
There was no difference in growth up to a 3-year-old follow-up survey of the Day 7 blastocyst compared with the Day 5 and Day 6 groups. Transfer of a Day 7-derived blastocyst does not appear to affect infant development and should be considered in cases where blastocyst development is delayed.
Trial registration number
not applicable
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