scholarly journals Modern Methods for Assessment of the Implantation Potential of Embryos in Assisted Reproductive Programs

Doctor Ru ◽  
2021 ◽  
Vol 20 (8) ◽  
pp. 12-18
Author(s):  
G.V. Savostina ◽  
◽  
S.G. Perminova ◽  
A.V. Timofeeva ◽  
M.A. Veyukova ◽  
...  
Keyword(s):  
Embryo Culture ◽  
Culture Media ◽  
High Sensitivity ◽  
Visual Assessment ◽  
Non Invasive ◽  
Non Coding Rna ◽  
Study Results ◽  
Modern Methods ◽  
Invasive Method ◽  
Aneuploidy Testing

Objective of the Review: To analyse the modern methods for assessment of the implantation potential of embryos in assisted reproductive programs. Key Points. We present the study results for selection of a most optimal embryo for transfer, using visual assessment of embryo quality, preimplantation genetic aneuploidy testing, analysis of metabolomic, proteomic, transcriptomic profiles of culture media and embryo blastocele. We have paid special attention to assessment of small non-coding RNA (sncRNA) in embryo culture medium. Conclusion. Due to the high sensitivity, objectivity and biomarker resistance to degradation, the most promising non-invasive method to assess the implantation potential of an embryo is analysis of the sncRNA profile in embryo culture media. Keywords: aneuploidy, pre-implantation genetic testing, small non-coding RNAs, proteomic analysis, metabolomic analysis.

P-203 Applying artificial intelligence for ploidy prediction: The concentration of IL-6 in spent culture medium, blastocyst morphological grade and embryo morphokinetics as variables under consideration

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
B Aparicio Ruiz ◽  
L Bori ◽  
E Paya ◽  
M A Valera ◽  
A Quiñonero ◽  
...  
Keyword(s):  
Embryo Culture ◽  
Culture Medium ◽  
Culture Media ◽  
General Description ◽  
Ann Model ◽  
Non Invasive ◽  
Morphokinetic Parameters ◽  
Testing Dataset ◽  
Standard Deviations ◽  
The Impact

Abstract Study question Would it be possible to predict embryo ploidy by taking into account conventional morphological and morphokinetic parameters together with IL-6 concentration in spent culture medium? Summary answer Our artificial neural network (ANN) trained with blastocyst morphology, embryo morphokinetics and IL-6 concentration distinguished between euploid/aneuploid embryos in 65% of the testing dataset. What is known already The analysis of spent embryo culture media represents the protein and metabolic state of the embryo and could be a non-invasive method of obtaining information about embryo quality. The impact of the presence/absence of several proteins in embryo culture samples over clinical results has been widely studied. The IL-6 is one of the most mentioned protein for its effect on embryo development, implantation and likelihood of achieving a live birth. In this initial attempt, we examined the predictive value for euploidy of a model that took into account the concentration of IL-6 in the spent culture medium. Study design, size, duration This prospective study included 319 embryos with PGT-A results. Out of the total, 127 were euploid and 192 aneuploid embryos. Concentration of IL-6 in spent embryo culture media (collected on the day of trophectoderm biopsy-fifth/sixth day of development), morphokinetic parameters (division time to 2 cells-t2; to 3 cells-t3, to 4 cells-t4; to 5 cells-t5 and time of blastocyst formation-tB) and blastocyst morphological grade (according to ASEBIR criteria) were considered to predict the embryo ploidy. Participants/materials, setting, methods Embryos were cultured in EmbryoScope. The chromosome analysis was performed using next-generation sequence technology. The concentration of IL-6 was measured in 20µL of spent embryo culture media with ELISA kits. Morphokinetic parameters were automatically annotated and the blastocyst morphology was evaluated by senior embryologists based on blastocele expansion, inner cell mass and trophectoderm quality. All the embryos were divided into 70% for training, 15% for validating and 15% for testing our ANN model with MatLab®. Main results and the role of chance The general description for the euploid embryo population was the following: 2% of the embryos were graded as A, 71% were graded as B and 28% were graded as C; the means and standard deviations were 25.32±2.97 hours (h) for t2, 35.33±5.15h for t3, 37.30±5.43h for t4, 48.24±6.62h for t5 and 103.93±12.8h for tB; and the average of IL-6 concentration was 1.51±0.70 pg/ml. The general description for the aneuploid embryo population was the following: 1% of the embryos were graded as A, 48% were graded as B and 51% were graded as C; the means and standard deviations were 26.13±3.51h for t2, 36.70±4.29h for t3, 38.20±4.24h for t4, 49.86±6.89h for t5 and 107.10±8.29h for tB; and the average of IL-6 concentration was 1.47±0.71 pg/ml. Our ANN model showed a higher general success rate as we increased the variables considered in the final prediction of euploid embryos. The accuracy, sensitivity and specificity for the testing dataset were: 0.60, 0.12 and 0.87 with morphokinetic parameters; 0.63, 0.24 and 0.93 with morphokinetics and IL-6 concentration; and 0.65, 0.16 and 0.96 with morphokinetics, IL-6 concentration and blastocyst morphological grade. Limitations, reasons for caution The low sensitivity and high specificity achieved in our models indicated that they were more capable of detecting aneuploid than euploid embryos. As this was a preliminary study, the small number of embryos included in the test (n = 48) was also a limitation. Wider implications of the findings The results showed that our model tended to classify the embryos as aneuploid. More euploid embryos would be necessary to train our model and achieve better results in the prediction of chromosomally normal embryos. Further studies with large number of embryos and additional variables could improve the non-invasive ploidy prediction. Trial registration number not applicable

P–582 High level of concordance between invasive and non-invasive preimplantation genetic testing for aneuploidies (niPGT-A) at day5 and day6–7

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
A Biricik ◽  
V Bianchi ◽  
F Lecciso ◽  
M Surdo ◽  
M Manno ◽  
...  
Keyword(s):  
Data Analysis ◽  
Embryo Culture ◽  
Culture Media ◽  
Embryo Biopsy ◽  
Non Invasive ◽  
Culture Time ◽  
Ngs Data Analysis ◽  
High Level ◽  
Ngs Data

Abstract Study question To explore ploidy concordance between invasive and non-invasive PGTA (niPGT-A) at different embryo culture time. Summary answer High level (>84%) of concordance rate for ploidy and sex, sensitivity (>88%), and specificity (76%) were obtained for both day6/7 samples and day5 samples. What is known already The analysis of embryo cell free DNA (cfDNA) that are released into culture media during in vitro embryo development has the potential to evaluate embryo ploidy status. However, obtaining sufficient quality and quantity of cfDNA is essential to achieve interpretable results for niPGT-A. More culture time is expected to be directly proportional to the release of more cfDNA. But embryo culture time is limited due to in-vitro embryo survival potential. Therefore, it is important to estimate the duration of the culture that will provide the maximum cfDNA that can be obtained without adversely affecting the development of the embryo. Study design, size, duration A total of 105 spent culture media (SCM) from day5-day7 blastocyst stage embryos have been included in this cohort study. The cfDNA of SCM samples were amplified and analyzed for niPGT-A by NGS analysis. The SCM samples were divided into 2 subgroups according the embryo culture hours (Day5 and Day6/7 group). The DNA concentration, informativity and euploidy results have then been compared with their corresponding embryos after trophectoderm biopsy (TE) and PGT-A analysis by NGS Participants/materials, setting, methods Embryos cultured until Day3 washed and cultured again in 20µl fresh culture media until embryo biopsy on Day5, 6, or 7. After biopsy SCM samples were immediately collected in PCR tubes and conserved at –20 °C until whole genome amplification by MALBAC® (Yicon Genomics). The TE and SCM samples were analyzed by next-generation sequencing (NGS) using Illumina MiSeq® System. NGS data analysis has been done by Bluefuse Multi Software 4.5 (Illumina) for SCM and TE samples Main results and the role of chance Only the SCM samples which have an embryo with a conclusive result were included in this cohort (n = 105). Overall 97.1% (102/105) of SCM samples gave a successful DNA amplification with a concentration ranging 32.4–128.5ng/µl. Non-informative (NI) results including a chaotic profile (>5 chromosome aneuploidies) were observed in 17 samples, so 83.3%(85/102) of SCM samples were informative for NGS data analysis. Ploidy concordance rate with the corresponding TE biopsies (euploid vs euploid, aneuploid vs aneuploid) was 84.7% (72/85). Sensitivity and specificity were 92,8% and 76,7%, respectively with no significant difference for all parameters for day 6/7 samples compared with day 5 samples. The false-negative rate was 3.5% (3/85), and false-positive rate was 11.7% (10/85). Limitations, reasons for caution The sample size is relatively small. Larger prospective studies are needed. As this is a single-center study, the impact of the variations in embryo culture conditions can be underestimated. Maternal DNA contamination risk cannot be revealed in SCM, therefore the use of molecular markers would increase the reliability. Wider implications of the findings: Non-invasive analysis of embryo cfDNA analyzed in spent culture media demonstrates high concordance with TE biopsy results in both early and late culture time. A non-invasive approach for aneuploidy screening offers important advantages such as avoiding invasive embryo biopsy and decreased cost, potentially increasing accessibility for a wider patient population. Trial registration number Not applicable

P–203 Applying artificial intelligence for ploidy prediction: The concentration of IL–6 in spent culture medium, blastocyst morphological grade and embryo morphokinetics as variables under consideration

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
B Aparici. Ruiz ◽  
L Bori ◽  
E Paya ◽  
M A Valera ◽  
A Quiñonero ◽  
...  
Keyword(s):  
Embryo Culture ◽  
Culture Medium ◽  
Culture Media ◽  
General Description ◽  
Ann Model ◽  
Non Invasive ◽  
Morphokinetic Parameters ◽  
Testing Dataset ◽  
Standard Deviations ◽  
The Impact

Abstract Study question Would it be possible to predict embryo ploidy by taking into account conventional morphological and morphokinetic parameters together with IL–6 concentration in spent culture medium? Summary answer Our artificial neural network (ANN) trained with blastocyst morphology, embryo morphokinetics and IL–6 concentration distinguished between euploid/aneuploid embryos in 65% of the testing dataset. What is known already The analysis of spent embryo culture media represents the protein and metabolic state of the embryo and could be a non-invasive method of obtaining information about embryo quality. The impact of the presence/absence of several proteins in embryo culture samples over clinical results has been widely studied. The IL–6 is one of the most mentioned protein for its effect on embryo development, implantation and likelihood of achieving a live birth. In this initial attempt, we examined the predictive value for euploidy of a model that took into account the concentration of IL–6 in the spent culture medium. Study design, size, duration This prospective study included 319 embryos with PGT-A results. Out of the total, 127 were euploid and 192 aneuploid embryos. Concentration of IL–6 in spent embryo culture media (collected on the day of trophectoderm biopsy-fifth/sixth day of development), morphokinetic parameters (division time to 2 cells-t2; to 3 cells-t3, to 4 cells-t4; to 5 cells-t5 and time of blastocyst formation-tB) and blastocyst morphological grade (according to ASEBIR criteria) were considered to predict the embryo ploidy. Participants/materials, setting, methods Embryos were cultured in EmbryoScope. The chromosome analysis was performed using next-generation sequence technology. The concentration of IL–6 was measured in 20µL of spent embryo culture media with ELISA kits. Morphokinetic parameters were automatically annotated and the blastocyst morphology was evaluated by senior embryologists based on blastocele expansion, inner cell mass and trophectoderm quality. All the embryos were divided into 70% for training, 15% for validating and 15% for testing our ANN model with MatLab®. Main results and the role of chance The general description for the euploid embryo population was the following: 2% of the embryos were graded as A, 71% were graded as B and 28% were graded as C; the means and standard deviations were 25.32±2.97 hours (h) for t2, 35.33±5.15h for t3, 37.30±5.43h for t4, 48.24±6.62h for t5 and 103.93±12.8h for tB; and the average of IL–6 concentration was 1.51±0.70 pg/ml. The general description for the aneuploid embryo population was the following: 1% of the embryos were graded as A, 48% were graded as B and 51% were graded as C; the means and standard deviations were 26.13±3.51h for t2, 36.70±4.29h for t3, 38.20±4.24h for t4, 49.86±6.89h for t5 and 107.10±8.29h for tB; and the average of IL–6 concentration was 1.47±0.71 pg/ml. Our ANN model showed a higher general success rate as we increased the variables considered in the final prediction of euploid embryos. The accuracy, sensitivity and specificity for the testing dataset were: 0.60, 0.12 and 0.87 with morphokinetic parameters; 0.63, 0.24 and 0.93 with morphokinetics and IL–6 concentration; and 0.65, 0.16 and 0.96 with morphokinetics, IL–6 concentration and blastocyst morphological grade. Limitations, reasons for caution The low sensitivity and high specificity achieved in our models indicated that they were more capable of detecting aneuploid than euploid embryos. As this was a preliminary study, the small number of embryos included in the test (n = 48) was also a limitation. Wider implications of the findings: The results showed that our model tended to classify the embryos as aneuploid. More euploid embryos would be necessary to train our model and achieve better results in the prediction of chromosomally normal embryos. Further studies with large number of embryos and additional variables could improve the non-invasive ploidy prediction. Trial registration number Not applicable

DIAGNOSING HIRSCHSPRUNG DISEASE IN CHILDREN: A MULTICENTER RESEARCH TRIAL

2021 ◽  
Vol 100 (2) ◽  
pp. 226-235
Author(s):  
D.A. Morozov ◽  
◽  
E.S. Pimenova ◽  
I.V. Poddoubnyi ◽  
M.Y. Kozlov ◽  
...  
Keyword(s):  
Anorectal Manometry ◽  
Diagnostic Errors ◽  
Hirschsprung Disease ◽  
High Sensitivity ◽  
Intestinal Wall ◽  
Non Invasive ◽  
Invasive Method ◽  
Group A ◽  
Medical Teams ◽  
Spearman’S Correlation

Hirschsprung disease (HD) is a congenital absence of nerve cells (ganglions) in a segment of the intestinal wall, leading to its obstruction. Diagnosis of BG in children is a labor-intensive process, on which surgical treatment tactics depend. The concentration of patients in large surgical centers with equipped diagnostic departments makes it possible to develop the most adequate examination plan while minimizing the risk of diagnostic errors. The aim of the study was to analyze methods for diagnosis of HD in children; a retrospective analysis of the records of patients hospitalized in three medical organizations in Moscow (n=201) in 2017–2019 was carried out. The first group – children with confirmed diagnosis (n=152), the second group – patients with suspicion of BG with unsatisfactory results of previous operative treatment, postoperative complications (n=49). Results: in the 1st group, irrigography was performed before hospitalization in 118 patients (77,6%). In the hospitals, irrigography was performed (repeated) for 109 (71,7%) children. 25 (16,4%) patients showed discrepancies in interpretation of the results. Thus, irrigography was applied to all patients. Intestine biopsy before hospitalization was performed in 79 patients (52%), in the hospitals – performed/repeated in 50 patients (32,9%). There were discrepancies in the interpretation of histological findings in 8 patients (18,6 per cent). Thus, in the diagnosis of HD in children, biopsies were used (before hospitalization and/or during hospitalization) in 89 patients (58,6%). Anorectal manometry was performed to 3 (2%) children. In the 2nd group a histological examination was performed (before and during hospitalization) in 41 patients (84%), anorectal manometry – in 15 patients (31%), irrigography – in all children. Correlation analysis did not reveal any relationship between the HD variant and manifestation symptoms (Spearman's correlation coefficient was –0,232 at p<0,05. Conclusion: all medical teams began examining children with suspected HD with irrigography. Various intestinal biopsies were performed in 58,6% of cases. Anorectal manometry is currently practically not used in the diagnosis of HD in children, although it is a promising non-invasive method with high sensitivity and specificity.

scholarly journals Non-invasive preimplantation genetic testing (niPGT): the next revolution in reproductive genetics?

2019 ◽  
Vol 26 (1) ◽  
pp. 16-42 ◽  
Author(s):  
Megan Leaver ◽  
Dagan Wells
Keyword(s):  
Genetic Testing ◽  
Clinical Diagnosis ◽  
Embryo Culture ◽  
Culture Media ◽  
Genetic Material ◽  
Preimplantation Embryos ◽  
Original Research ◽  
Preimplantation Genetic Testing ◽  
Non Invasive ◽  
The Impact

Abstract BACKGROUND Preimplantation genetic testing (PGT) encompasses methods that allow embryos to be tested for severe inherited conditions or for chromosome abnormalities, relevant to embryo health and viability. In order to obtain embryonic genetic material for analysis, a biopsy is required, involving the removal of one or more cells. This invasive procedure greatly increases the costs of PGT and there have been concerns that embryo viability could be compromised in some cases. The recent discovery of DNA within the blastocoele fluid (BF) of blastocysts and in spent embryo culture media (SCM) has led to interest in the development of non-invasive methods of PGT (niPGT). OBJECTIVE AND RATIONALE This review evaluates the current scientific evidence regarding non-invasive genetic assessment of preimplantation embryos. The success of different PGT methodologies in collecting and analysing extra-embryonic DNA is evaluated, and consideration is given to the potential biological and technical hindrances to obtaining a reliable clinical diagnosis. SEARCH METHODS Original research and review papers concerning niPGT were sourced by searching PubMed and Google Scholar databases until July 2019. Searches comprised the keywords: ‘non-invasive’; ‘cell-free DNA’; ‘blastocentesis’; ‘blastocoel fluid’; ‘spent culture media’; ‘embryo culture medium’; ‘preimplantation genetic testing’; ‘preimplantation genetic diagnosis’; ‘preimplantation genetic screening’; and ‘aneuploidy’. OUTCOMES Embryonic DNA is frequently detectable in BF and SCM of embryos produced during IVF treatment. Initial studies have achieved some success when performing cytogenetic and molecular genetic analysis. However, in many cases, the efficiency has been restricted by technical complications associated with the low quantity and quality of the DNA. Reported levels of ploidy agreement between SCM/BF samples and biopsied embryonic cells vary widely. In some cases, a discrepancy with respect to cytogenetic data obtained after trophectoderm biopsy may be attributable to embryonic mosaicism or DNA contamination (usually of maternal origin). Some research indicates that aneuploid cells are preferentially eliminated from the embryo, suggesting that their DNA might be over-represented in SCM and BF samples; this hypothesis requires further investigation. WIDER IMPLICATIONS Available data suggest that BF and SCM samples frequently provide DNA templates suitable for genetic analyses, offering a potential means of PGT that is less expensive than traditional methods, requires less micromanipulation skill and poses a lower risk to embryos. Critically, DNA isolation and amplification protocols must be optimised to reproducibly obtain an accurate clinical diagnosis, whilst minimising the impact of confounding factors such as contamination. Further investigations are required to understand the mechanisms underlying the release of embryonic DNA and to determine the extent to which this material reflects the true genetic status of the corresponding embryo. Currently, the clinic al potential of niPGT remains unknown.

scholarly journals APPLICATION OF LASER PHOTONICS METHODS IN ORTHOPEDIC DENTISTRY FOR NON-INVASIVE NON-CONTACT EXAMINATION OF BULK MICRO-DEFORMATIONS OF BONE TISSUE

2018 ◽  
Vol 7 (2) ◽  
pp. 13-18
Author(s):  
M. A. Grossmann ◽  
A. I. Larkin
Keyword(s):  
Relaxation Time ◽  
Bone Tissue ◽  
High Sensitivity ◽  
Medical Problem ◽  
Speckle Interferometry ◽  
Non Invasive ◽  
Mini Implants ◽  
Modern Methods ◽  
Volumetric Objects ◽  
Medical Recommendations

Modern methods of laser photonics, which make it possible to measure the dynamics of micro-deformations of volumetric objects without contact and with high sensitivity, find increasing application in a wide variety of fields. Particularly attractive is the use of such a delicate, non-perturbing technique in medicine. The article reports on experiments using laser photonics methods in dental implantology, which allowed to give practical medical recommendations. In the work, a study was made of the possibilities of holographic and speckle interferometry methods for measuring the deformation of the jaw bone tissue with a step-by-step installation of cylindrical and conical mini implants. The possibility of determining the relaxation time of deformations after the installation of mini implants was demonstrated. The purpose of this work is to justify the possibility and feasibility of using laser photonics  to solve this medical problem.

Non-invasive preimplantationsexingusingRT-PCRon the spent embryo culture media

2019 ◽  
Vol 38 ◽  
pp. e35-e36
Author(s):  
Mahnaz Esmaeili ◽  
Masood Bazrgar ◽  
Hamid Gourabi ◽  
Bita Ebrahimi ◽  
Parnaz Borjian-Borujeni
Keyword(s):  
Embryo Culture ◽  
Culture Media ◽  
Non Invasive
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