Abstract
Study question
Are the neonatal outcomes normal of babies derived from the transfer of blastocysts derived from mono-pronuclear(1PN) zygotes?
Summary answer
There was no effect on growth or ...increase in congenital anomalies up to 18-months in babies of 1PN-derived births.
What is known already
1PN zygotes are observed in ART, albeit at a low rate. We have previously reported that 80.7% of 1PN zygotes derived from IVF or ICSI had a biparental chromosome using Live Cell imaging techniques, 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. ASRM2020). However, there is some uncertainty about the developmental mechanism of 1PN zygotes, and there is no clear consensus on their clinical utility.
Study design, size, duration
This was a retrospective study which included 55 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 was recorded as well as a physical development survey of 25 children who responded to the 18-months follow-up survey. The time period was 72 months (January 2013 to December 2018).
Participants/materials, setting, methods
Patients seeking fertility treatment at an established private IVF clinic. We compared the birth weight, birth after18-months height and weight of children born to 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 1.8% (1/55). The average birth weight of boys in the 1PN group was 3105.6+/–360.3g, which was not significantly different from 3041.0+/–443.3 g in the 2PN group. In girls, the average birth weight was 3085.7+/–454.9 g in the 1PN group, which was not significantly different from the 2PN group (2938.9+/–311.5 g). The average height at 18-months, was 81.6+/–2.5 cm vs 80.5+/–3.4 cm for boys; 79.0+/–1.8 cm vs 79.0+/–3.4 cm for girls in the 1PN and 2PN groups, respectively. The average body weights of the 1PN and 2PN groups were 11.1+/–1.1 kg vs 10.7+/–1.1 kg for boys; 9.7+/–0.9 kg vs 10.1+/–1.0 kg for girls, respectively. There was no significant difference in average height and weight up-to the 18-months follow-up survey.
Limitations, reasons for caution
The incidence of 1PN derived 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 was around 1.7 to 2%, and the incidence was similar in the 1PN. There was no difference in the birth weight and 18-months follow-up survey of the 1PN compared with the 2PN. We have demonstrated that there is clinical utility of 1PN embryo.
Trial registration number
Not applicable
Abstract
Study question
What is the clinical outcome of oocytes recovered after 39 hours from ovulation inducing drug administration?
Summary answer
Oocytes obtained after 39 hours from follicular ...maturation triggering are equally viable to those obtained at the standard time of 36 hrs.
What is known already
In the clinical setting of ART, ovum pick-up (OPU) is generally performed around 36 hours after the administration of ovulation inducing drugs (OID). However, there are cases where OPU cannot be performed at this time often due to long operating lists. As the time elapsed between the administration of ovulation inducing drugs and OPU becomes longer, there is a concern about time-related oocyte aging. Nevertheless, there are few reports of clinical results of OPU after 36 hours from OID.
Study design, size, duration
We conducted a review of 1187 cycles and 1951 patients in which OPU and embryo transfer was performed in 2017–2018. All cycles underwent a ‘freeze-all’ of embryos and the transfer cycle was in the thawed embryo transfer cycle for all cases.
Participants/materials, setting, methods
The time from the administration of OID to the end of OPU was divided into 36h group and over 39h group and the MII and normal fertilization rate of oocytes obtained from OPU after ovarian stimulation were compared. After confirmation of fertilization, the D3 good-quality embryo and the D5 and 6 good-quality blastocyst rates of embryos that continued to be cultured and the pregnancy and miscarriage rates of cleavage-stage embryos and blastocyst transfers were compared.
Main results and the role of chance
The MII rate in the 36h and >39h groups was 78.1% vs. 80.0%, and the normal fertilization rate was 77.9% vs. 78.1% (ICSI) and 65.4% vs. 67.6% (Conventional-IVF). The D3 good-quality embryo rate (good-quality embryos are embryos with less than 5% fragmentation in 7–9 cells and compaction with more than 50% adhesion between split spheres) was 21.8% vs. 25.3%, the D5 good-quality blastocyst rate (at least 3BB according to Gardner classification) was 33.6% vs. 40.1%, and the D6 good-quality blastocyst rate was 31.1% vs. 37.5%, all of which were not significantly different. The pregnancy rate for cleavage-stage embryo transfer was 26.6% vs. 6.7%, and the miscarriage rate was 25.3% vs. 42.9%, both of which were not significantly different. The pregnancy rate for blastocyst transfer was 45.4% vs. 50.0%, and the miscarriage rate was 22.2% vs. 20.0%, both of which were not significantly different. (The significance difference test was a χ-square test)
Limitations, reasons for caution
The study was a retrospective study.
Wider implications of the findings: Even if OPU is conducted after 36h of the administration of OID, to the extreme range of 39h–41h, oocyte aging does not seem apparent and pregnancy outcomes are similar to the standard time interval of 36 hours.
Trial registration number
‘not applicable’
Abstract Study question Do embryos that fully developed to blastocyst at day 4 have high aneuploidy rates as assessed by preimplantation genetic analysis? Summary answer Fully developed blastocysts ...at day 4 had a high euploidy rate. What is known already In ART, it is considered important to use good quality blastocysts to increase the pregnancy rate after embryo transfer. Some reports have demonstrated a high pregnancy rate for embryos that fully developed to blastocysts on day 4 after IVF (Ozgur et al., 2021). Furthermore, it has been reported that the speed of embryo development is related to the rate of chromosome aneuploidy (Taylor et al., 2014). There are though, limited reports on the euploidy rate of embryos developed to blastocysts at day 4. Study design, size, duration This retrospective study analyzed 763 embryos (from 184 PGT-A cases) that underwent TE biopsy on day 5 between January to December 2022 to determine their ploidy status. Participants/materials, setting, methods Embryos cultured in a time-lapse incubator were divided into two groups: those that developed to blastocyst (Gardner’s classification: ≥ expansion grade 3) at day 4 (96-100 h) (D4(+) group) and those that did not develop to full blastocyst at day 4 (D4(-) group) and the euploidy rates were compared. Statistical analysis was performed using Fisher’s exact test to determine significant differences (P < 0.05). Main results and the role of chance The euploidy rate of the D4(+) group was 36.5% (80/219), which was significantly higher than the D4(-) group, 23.3% (127/544) (p < 0.01). Moreover, when considering the age of the female, for age ≤39 years, the rates were significantly higher in the D4(+) group (52.1% (61/117) and 35.4% (97/274) p < 0.01 respectively ). For age 40≥ years, it tended to be higher but was not significantly different in the D4(+) group vs D4(-) group (18.6% 19/102 and 11.1% 30/270 respectively). Even when limited to embryos that were graded as good quality blastocysts at day 5, the D4(+) group was significantly higher (p < 0.01) than the D4(-) group (55.0% (60/109) vs 36.1% (90/249)). Limitations, reasons for caution This was a retrospective study limited to day 5 embryos with biopsies taken after vitrification-warming. Wider implications of the findings These results demonstrate that embryos that had already developed to blastocyst at day 4 had a higher rate of euploidy compared to day 5 blastocysts. Thus, when performing PGT-A, biopsy of embryos that have already developed to blastocyst at day 4 are expected to be prioritised for transfer. Trial registration number not applicable
Oxyntomodulin (OXM), glucagon, glucagon-like peptide-1 (GLP-1), and exendin-4 (Ex-4) are peptide hormones that regulate glucose homeostasis in monogastric and ruminant animals. Recently, we reported ...that the insulin-releasing effects of OXM and glucagon in cattle are mediated through both GLP-1 and glucagon receptors. The purpose of this study was to examine the mechanisms of the glucoregulatory actions induced by Ex-4, GLP-1, OXM, and glucagon and the interrelationships among these hormones in cattle. Two experiments were performed in Holstein cattle. In Experiment 1, we initially assessed the effects of intravenous (iv) bolus injection of 0, 0.25, 1, and 2 μg/kg body weight (BW) of Ex-4, GLP-1, and OXM on insulin and glucose concentrations in 3-mo-old intact male Holstein calves. In Experiment 2, we studied insulin and glucose responses to iv coinjection of 0.25 μg of Ex-4 or GLP-1/kg BW with 2 μg of OXM or glucagon/kg BW in 4-mo-old Holstein steers. Administration of peptides and blood sampling were done via a jugular catheter. Plasma was separated and the concentrations of peptides and glucose in plasma were analyzed using radioimmunoassay and enzymatic methods, respectively. Results showed that the potent glucoregulatory action of Ex-4 in 4-mo-old steers was delayed and attenuated when Ex-4 was coinjected with OXM. The decline in plasma glucose concentrations began at 5 min in the Ex-4-injected group (P < 0.05) vs 15 min in the Ex-4 + OXM–injected group (P < 0.05). Plasma concentrations of glucose at 30 min were reduced 26% from basal concentrations in the Ex-4-injected group and 13% in the Ex-4 + OXM–injected group (P < 0.001). Results also showed that the glucose concentrations initially increased in the Ex-4 + glucagon–treated group, but declined to a relatively hypoglycemic condition by 90 to 120 min. In contrast, the glucose concentrations at specific time points between the GLP-1 + OXM–injected group and the OXM-injected group did not differ. Similarly, the glucose concentrations in the GLP-1 + glucagon–injected group did not differ from those in the glucagon-injected group. Because OXM and glucagon mediate glucose concentrations via the glucagon receptor, it is suggested that the potent glucose-lowering action of Ex-4 might include the glucagon receptor antagonistic action of Ex-4.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UL, UM, UPCLJ, UPUK
Abstract
Study question
Can noninvasive preimplantation genetic testing (PGT) using cell free DNA from embryo culture medium discriminate embryonic aneuploidy status?
Summary answer
Aneuploid embryos ...in older patients can be identified by cell free DNA in embryo culture medium.
What is known already
There have been reports of non-invasive PGT by analysing cell free DNA (cfDNA) released from embryos into culture medium, and found to be highly correlated with the analysis results of Trophectoderm (TE)-biopsy (Carmen.Rubio, 2020).We have initiated in 2021, a preliminary research program for the clinical application of niPGT. In aneuploid embryos, we have found a tendency for a high concordance rate between cfDNA analysis and blastocyst analysis results, suggesting the usefulness of cfDNA analysis in terms of identifying aneuploid embryos.Subsequently, we examined the usefulness of cfDNA analysis by comparing the concordance rate with cfDNA analysis and TE biopsy of blastocysts.
Study design, size, duration
After thawing pronuclear-stage frozen embryos for which consent was obtained for research use, they were co-cultured until Day 4, and then individually cultured on in new dishes.40 embryos that reached morula on day 4 or day 5 and blastocyst on day 6 or day 7 were included.
Participants/materials, setting, methods
The biopsied TE, the biopsied blastocyst, and the cfDNA obtained after individual culture were all sent to an external provider (Igenomix: Valencia, Spain) for chromosomal aneuploidy analysis by NGS. The concordance rate of the analysis for blastocysts in cfDNA and TE was calculated by dividing them into euploidy and aneuploidy, and by age. Concordance between the results of the blastocyst analysis and the culture medium were assessed according to the standards of the inspection company.
Main results and the role of chance
For the blastocyst analysis results, the concordance rate with cfDNA was 76.5% (26/34) and the concordance rate with TE was 85.3% (29/34), showing no difference. For the euploid blastocysts analyzed, the matching rate with cfDNA was 61.1% (11/18) and with TE was 77.8% (14/18), showing no difference. Similarly, for aneuploid blastocysts, the concordance rate with cfDNA was 93.8% (15/16) and with TE was 93.8% (15/16), showing no difference. Detailed analysis of aneuploid blastocysts revealed that the concordance rate with cfDNA was 88.9% (8/9) and the concordance rate with TE was 88.9% (8/9) in women under 39 years of age. In addition, the concordance rate with cfDNA was 100.0% (7/7) with TE was 100.0% (7/7) in those aged 40 and over. (Fisher's exact test)
Limitations, reasons for caution
Concordance between cfDNA and blastocyst analyzes in euploid embryos was low, continued follow-up is considered necessary.
Wider implications of the findings
In a detailed analysis of aneuploid blastocysts aged 40 years or older, Similar to TE, cfDNA showed a high concordance rate with the results of blastocyst analysis, demonstrating that cfDNA provides sufficient accuracy for identifying aneuploid embryos in women over 40 years of age.
Trial registration number
not applicable
Predicting the mandible size before the termination of growth of the maxillofacial
bones is essential in pedodontics as well as for the predictions needed for genetic
analysis. Here, Quantitative ...Trait Locus (QTL) analysis was used to detect the
chromosomal regions responsible for the mandible length between the menton and gonion
in an SMXA recombinant inbred strain of mice. Around the region 60 cM from the
centromere in chromosome 10, the logarithm of the odds score showed a higher than
suggestive level. Around the regions 13 cM and 16 cM in chromosome 11, two
significant QTLs were detected. Analysis of genotypes from loci corresponding to
those QTLs revealed a large mandible when the region between the markers
Hba and D11Mit163 and D10Mit70
and D10Mit136 indicated the genotype from the A/J and SM/J alleles,
respectively. These results suggest that the major gene(s) responsible for mandible
length are located in these regions.
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CMK, NUK, OILJ, SAZU, UKNU, UL, UM, UPUK
Abstract
Study question
Does the presence of the ACA have any effect on pregnancy and birth?
Summary answer
Presence of ACA did not have an impact on pregnancy and birth parameters measured.
What is ...known already
We have previously shown that patients with high levels of anti-centromere antibody (ACA), one of the anti-nuclear antibodies (ANA), frequently have dispersion of chromosomes in the cytoplasm. Additionally, we reported that it was characterized by a low oocyte maturation rate as observed at ovum pick up, high multiple pronuclear formation after fertilization and a low pregnancy rate after embryo transfer. Thus, the effect of ACA on fertility is clear, but the impact following pregnancy establishment has not yet been reported. Therefore, we followed up on birth outcomes in ACA-positive patients treated with ART.
Study design, size, duration
3379 patients who underwent ANA testing followed by embryo transfer between January 2014 and February 2020 and who gave birth including the results of single births with no abnormal or missing values for week and birth weight were analyzed. The subjects were classified into three groups: ANA-negative (without ACA or any other ANA), ACA-positive (with only ACA) and ANA-positive (with ANA but not ACA).
Participants/materials, setting, methods
We compared mean birth weight (full term), low weight birth infant (less than 2,500g) rate, sex ratio, congenital anomaly rate, preterm birth rate, cesarean section rate, and mean apgar score (AP) in the three groups. We treated “don't know” and “blank” for items other than week and birth weight as non-responses and missing values.
Main results and the role of chance
Of the 3379 cases for analysis, the group numbers were ANA-: 2465, ACA+: 21, ANA+: 893. The mean birth weights were 3099.6g, 3055.3g and 3088.5g respectively. In the 3 groups the rate of low birth weight infants were 8.7% (215/2465), 9.5% (2/21) and 9.2% (82/893), the sex ratio (male/female)-1.03 (1246/1212), 0.62 (8/13) and 0.93 (429/ 460), congenital anomaly rate 2.7% (67/2465), 4.8% (1/21), 3.2% (29/893), preterm birth rate 7.4% (86/2465), 4.8% (1/12), 6.7% (60/893), cesarean rate 40.0% (979/2448), 42.9% (9/21), 41.3% (367/888), and mean AP was 8.3, 8.2, and 8.3 respectively. There was no significant difference between the three groups in any of the measured items.
Limitations, reasons for caution
This study was conducted at a single institution, and only the results of those who responded to the birth survey questionnaire were analyzed. Because of the small number of patients in the ACA group, continued follow-up is considered necessary.
Wider implications of the findings
It was shown that ACA strongly affects maternal fertility, but seems to have no effect on birth parameters. However we will continue to follow up with further long-term prognostic studies.
Trial registration number
not applicable
Human papilloma virus detection in oropharyngeal cancer with gargle samples.
human papilloma virus (HPV) is a major risk factor for oropharyngeal squamous cell carcinoma (OPSCC) and knowledge of a ...patient's HPV status is clinically important in terms of treatment and prognosis. The practicality of using oral gargle samples to reliably detect HPV in patients with OPSCC remains unclear. Therefore, we evaluated the feasibility of HPV detection in gargle samples of OPSCC patients using an HPV-dedicated nucleic acid amplification test (cobas 4800 HPV Test; Roche Diagnostics K.K.).
15 patients with histologically proven OPSCC were evaluated from May 2014 to March 2015. Swab sam- ples served as positive controls and were tested using both the Hybrid Capture II HPV Test (HC-II; Digene Corporation) and the cobas 4800 HPV Test. Oral gargle samples were tested using the cobas 4800 HPV Test. Five of the 15 patients were confirmed to be HPV-positive by a combination of p16 immunohistochemistry, HPV-DNA in situ hybridization and nucleic acid amplification.
the sensitivity and specificity of the gargling method were 60% and 100%, respectively. No false-positives were obtained. Detection of HPV in two very small tumours rising from the base of the tongue was difficult and these cases were overlooked as HPV-negative.
use of the gargling method to determine HPV positivity in OPSCC patients appears feasible, except in patients with very small tumours. Real-time polymerase chain reaction using gargle samples may have greater clinical efficacy than the swabbing method.
Abstract Study question How multinucleation and fragmentation occur in the first mitosis in human embryos? Summary answer The first mitotic spindles had heterogeneity, and the shapes and movements of ...the spindles were strongly associated with multinucleation and fragmentation. What is known already Studies have shown that many problems occur during the first mitosis, such as fragmentation and multinucleation. However, the cell biologic mechanisms whereby these errors occur during the first mitosis in human embryos remain unknown. To clarify this aspect, it is necessary to analysis the dynamics of DNA and cytoskeleton using live human embryos. Although microinjections of DNA or mRNA into mammalian embryos were widely used for live imaging during early mitosis, the method into human poses the risk of ethical issues. Therefore, we established a method used for real-time imaging in human embryos using chemical fluorescent labeling. Study design, size, duration Frozen human two-pronuclear stage embryos were donated for research by couples who had completed fertility treatment. We obtained written informed consent from patients who donated human embryos. The mean age of patients at embryo freezing was 32.00 ± 3.03 (SD) years. We imaged thirty-one living human embryos using chemical fluorescent labeling and confocal microscopy to visualize DNA and tubulin. Participants/materials, setting, methods Thirty-one human embryos were cultured in medium containing the live-cell stain probes SPY555-DNA and SPY650-Tubulin. The Embryos were imaged using an LSM980 laser scanning confocal microscope every 10–20 min for a period of eight hours. Three-dimensional (3D) visualizations of human embryos were performed using the Imaris software. The first spindle shapes were quantified by image analysis and calculated the aspect ratio of the spindles using ImageJ. Main results and the role of chance We discovered that over 81% of two-pronuclear zygotes became multinucleated 2-cell stage embryos. We demonstrated that there was variation in the first mitotic spindles. There was a significant difference between the aspect ratio (AR) of the spindles leading to the mononuclear and multinuclear types at the 2-cell stage (p < 0.005). In addition, the low-AR spindles were unstable and often had more defocused poles than the high-AR spindles, which lead to multinucleation at the 2-cell stage (55 % vs. 0%, p < 0.05). More embryo having high-aspect ratio spindles tend to develop blastocysts than those having low aspect-ratio spindles (p < 0.05). Moreover, the movement of the central spindles varied after chromosome segregation. The embryos with more fragments at two cell stage had a significantly longer period for the central spindles to transform into midbodies compared to those with fewer fragments. The central spindles of embryos with more fragments moved a longer distance from the beginning of furrow ingression to the completion of the first division (p < 0.05). These results suggest that the first mitotic spindle instability and heterogeneity cause many errors such as multinucleation, fragmentation, and embryonic development. Limitations, reasons for caution Imaged human embryos were donated from a fertility clinic, and detailed patient information, such as underlying disease and method of ovarian stimulation, was not available. It is possible that the human zygotes used in this study had a bias in patient characteristics. Wider implications of the findings Our study showed that there were instability and heterogeneity in the first mitotic spindle, which was strongly associated with multinucleation, fragmentation, and embryonic development. Revealing the mechanism underlying spindle formation in cleavage-stage embryos will contribute to improving the success rate of assisted reproduction technologies in humans. Trial registration number not applicable
Background: Handicapped persons living in nursing homes have special risks for oral diseases.
Objective: To investigate the specific factors related to the occurrence of dental caries and tooth ...extraction in patients with severe motor and intellectual disabilities (PSMI) residing in an institution.
Methods: One hundred eighty‐nine PSMI residing in a single institution in Japan were followed for 3 years. Oral examinations were conducted at baseline and 3 years later. The following items were investigated: age of subject at admission, period of institutionalization, age at baseline oral examination, status of rumination, drooling, type of ward, dietary mode, and etiology of the impairment. Logistic regression analyses were conducted to examine factors associated with new dental caries and tooth extraction occurring during the study period.
Results: By multivariate analysis, rumination and tube feeding were identified as significant factors associated with new dental caries. On the other hand, infancy or childhood impairment and drooling were identified as significant factors related to tooth extraction.
Conclusion: Some specific factors in this patient population affect the dental caries and tooth extraction and oral programs targeting these factors may reduce dental degeneration in these patients.
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BFBNIB, CMK, FZAB, GIS, IJS, KILJ, NLZOH, NUK, OILJ, SAZU, SBCE, SBMB, UL, UM, UPUK
