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"Latar Belakang: DNA bebas dalam medium kultur embrio dan kemajuan pemodelan berbasis kecerdasan buatan berpotensi menjadi modalitas uji genetik yang non-invasif. Saat ini, tidak diketahui apakah DNA tersebut dilepaskan oleh sel embrio euploid atau aneuploid, sehingga melemahkan dasar keilmuan penggunaannya. Penelitian ini bertujuan untuk mengetahui sel sumber embrio pelepas DNA bebas dalam medium kultur, validasi potensi klinis penggunaan DNA bebas untuk skrining status ploidi embrio pasien Fertilisasi In-Vitro (FIV), dan konstruksi model pembelajaran mendalam menggunakan gambar embrio tersegmentasi untuk deteksi status ploidi embrio.Metode: Penelitian ini terbagi dalam dua desain penelitian yaitu eksperimental in-vitro menggunakan embrio hewan model dan observasi kohort menggunakan 28 sampel medium kultur embrio dari 21 pasien program FIV di Klinik Morula IVF Jakarta, periode September 2022–Januari 2023. Konstruksi model pembelajaran mendalam menggunakan gambar embrio pasien FIV yang menjalani program bayi tabung periode Januari 2021– Juni 2023. Deteksi sel embrio sumber pelepas DNA bebas dilakukan dengan memapar salah satu embrio (galur DDY atau C57BL) dengan reversin untuk memperoleh blastomer pembawa sel-sel aneuploid. Embrio kontrol dikultur bersamaan tanpa reversin sebagai pembawa sel-sel blastomer euploid. Agregasi membentuk embrio mosaik dilakukan antara embrio pembawa blastomer aneuploid (perlakuan) dan embrio pembawa blastomer euploid (kontrol). Polimorfisme gen GABRA2 antara galur DDY (alel wildtype) dan C57BL (alel delesi) mejadi alel target yang dikuantifikasi dengan metode qPCR. Empat jenis sampel dibuat sebagai berikut: medium kultur tanpa embrio, medium kultur embrio agregasi tanpa pemaparan reversin, rev-DDY, rev-C57BL. Embrio mosaik diwarnai dengan marka apoptosis untuk deteksi mekanisme pelepasan DNA bebas. Analisis status ploidi embrio menggunakan medium kultur embrio pasien FIV dilakukan dengan metode sekuensing. Konstruksi model pembelajaran mendalam menggunakan gambar embrio tersegmentasi yang dipotong urut selama 10 jam sebelum proses biopsi. Variabel yang diamati dalam penelitian adalah konsentrasi alel delesi dan wildtype gen GABRA2, jumlah sel terwarnai marka apoptosis, Pada sampel medium kultur embrio manusia, keberhasilan amplifikasi dan interpretasi hasil sekuensing, serta tingkat kesesuaian uji antara DNA bebas dengan biopsi trofoblas dianalisis. Kemampuan prediksi model berbasis kecerdasan buatan dinilai dengan akurasi dan loss. Analisis data penelitian dan konstruksi model pembelajaran mendalam menggunakan perangkat lunak SPPS versi 21, OpenEpi. pyhton.Hasil: Sebanyak 0,08 ng/reaksi alel wildtype ditemukan pada embrio mosaik dengan pemaparan reversin pada blastomer embrio DDY (rev-DDY, pembawa sel-sel aneuploid) dan 0,01 ng/reaksi alel delesi ditemukan pada embrio mosaik dengan pemaparan reversin pada blastomer embrio C57BL (rev-C57BL, pembawa sel-sel aneuploid). Median jumlah sel embrio terwarnai marka apoptosis antara ketiga group embrio (agregasi kontrol, rev- DDY dan rev-C57BL) tidak berbeda bermakna (nilai p = 0,95 untuk pewarnaan late apoptosis (propidium iodide) dan p = 0,42 untuk early apoptosis (Ann-V) menandakan adanya proses koreksi sel pada kedua group embrio mosaik selama masa perkembangan pra-implantasi. Keberhasilan amplifikasi DNA bebas medium kultur embrio manusia dalah 100%, dengan nilai interpretasi 92,8% (26/28). Nilai kesesuaian DNA bebas dengan biopsi trofoblas adalah rendah sebesar 65,4% (17/26) dengan kesesuaian kromosom seks adalah 61,5% (16/26). Sepuluh dari 11 embrio XY pada biopsi trofoblas terdeteksi XX pada DNA bebas. Seluruh model pembelajaran mendalam mengalami peningkatan akurasi menggunakan gambar embrio tersegmentasi dengan algoritma InceptionV3 mencapai akurasi tertinggi sebesar 0,67 dengan nilai loss sebesar 1,4.Kesimpulan: Sel embrio anueploid adalah sel sember pelepas DNA bebas medium kultur embrio pada embrio mosaik hewan coba mencit yang dilepaskan melalui mekanisme apoptosis. Embrio masik tersebut diperkirakan melakukan self-correction dengan mengeksklusi sel-sel aneploid untuk mempertahankan euploiditasnya. Rendahnya tingkat kesesuaian antara DNA bebas dengan biopsi trofoblas disebabbkan oleh adanya kontaminasi maternal yang ditandai dengan perubahaan koromosm seks yang signifikan. Penggunaan gambar blastosis tersegmentasi meningkatkan akurasi model prediksi pembelajaran mendalam.

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Background: Cell-free DNA and advanced artificial intelligence-based modeling uphold the potential of a non-invasive approach to determining embryo ploidy status. The specific embryonic cells (whether euploid or aneuploid) that release cell-free DNA are largely unknown, causing a weak scientific basis for its use. This study aimed to identify the source of embryonic cells releasing cell-free DNA in culture media, validate the clinical potential of using cell-free DNA to screen embryo ploidy status in an in-vitro fertilization (IVF) program and develop a deep learning model using segmented embryo images to detect embryo ploidy status.

Materials and Methods: This study employed two research designs including an in-vitro experimental study using animal model embryos and an observational cohort study using 28 samples of spent embryo culture media from 21 patients undergoing IVF program at Morula IVF Clinic Jakarta (September 2022 to January 2023). A deep learning model was constructed using images of embryos from IVF patients who participated in IVF program from January 2021 to June 2023. Detection of the source embryonic cells releasing cell-free DNA was achieved by exposing embryos (DDY or C57BL strains) to reversine to induce the formation of blastomeres carrying aneuploid cells. Control embryos were cultured simultaneously without reversine to serve as the source of euploid blastomeres. Mosaic embryo aggregation was performed by combining embryos carrying aneuploid blastomeres (treatment) with those carrying euploid blastomeres (control). The GABRA2 gene polymorphism between the DDY strain (wildtype allele) and the C57BL strain (deletion allele) was the target allele quantified using qPCR. Four types of samples were prepared: culture medium without embryos, culture medium of aggregated embryos without reversine exposure, rev-DDY, and rev-C57BL. Mosaic embryos were stained with an apoptosis marker to detect the mechanism of cell-free DNA release. The ploidy status of embryos using spent embryo culture media from IVF patients was determined using sequencing methods. The deep learning model was constructed using segmented images of embryos captured over 10 hours before the biopsy process. The variables observed in the study included the concentration of deletion and wildtype alleles of the GABRA2 gene, the number of cells stained with apoptosis markers, the success rate of amplification and interpretation of sequencing results from human spent embryo culture medium samples, and the concordance rate between cell-free DNA and trophectoderm biopsy analysis. The predictive ability of the artificial intelligence-based model was evaluated using accuracy and loss metrics. Data analysis and deep learning model construction were performed using SPSS version 21, OpenEpi, and Python.

Results: A total of 0.08 ng/reaction of the wildtype allele was detected in the culture media sample of mosaic embryos exposed to reversine in DDY embryo blastomeres (rev- DDY, carrying aneuploid cells), and 0.01 ng/reaction of the deletion allele was found in the sample exposed to reversine in C57BL embryo blastomeres (rev-C57BL, carrying aneuploid cells). The median number of embryonic cells stained with apoptosis markers among the three groups of embryos (control aggregation, rev-DDY, and rev-C57BL) did not differ significantly (p = 0.95 for late apoptosis staining with propidium iodide and p = 0.42 for early apoptosis with Annexin V), indicating the presence of cell correction processes in both groups of mosaic embryos during pre-implantation development. The success rate of cell-free DNA amplification in human spent embryo culture media was 100%, with an interpretability of 92.8% (26/28). The concordance between cell-free DNA and trophectoderm biopsy was low at 65.4% (17/26), with sex chromosome concordance at 61.5% (16/26). Ten out of eleven XY embryos from the trophectoderm biopsy were detected as XX in cell-free DNA analysis. All deep learning models showed improved accuracy using segmented embryo images with the InceptionV3 algorithm, achieving the highest accuracy of 0.67 with a loss of 1.4.

Conclusion: Aneuploid embryonic cells were identified as the source releasing cell-free DNA in culture media during embryo animal model experiments, releasing DNA through an apoptotic mechanism. These mosaic embryos were expected to activate embryonic cell correction mechanisms by excluding aneuploid cells to maintain their euploidy. The low concordance rate between cell-free DNA and trophectoderm biopsy was attributed to maternal contamination, as indicated by significant changes in sex chromosomes. The use of segmented blastocyst images improved the model's accuracy.

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"Tujuan: Faktor embrio sangat mempengaruhi hasil dari fertilization in vitro (FIV). Salah satu metode untuk memastikan embrio tidak memiliki kromosom aneuploid adalah Prosedur Pengujian Genetik Praimplantasi untuk Aneuploidi atau Skrining Genetik Praimplantasi, yang melibatkan biopsi blastomer pada fase 8 sel atau trofektoderm pada fase blastokista. Prosedur ini merupakan prosedur yang invasif dan berpotensi membahayakan embrio.Metode: Penelitian adalah penelitian cross-sectional pada pasien program FIV yang dilanjutkan dengan pemeriksaan kromosom dengan NGS di Pusat IVF RS Pondok Indah dan Pusat FIV Morula RS Bunda pada bulan Desember 2021 sampai dengan Desember 2022. Embrio yang mencapai stadium blastokista pada hari ke 5 atau 6 dibersihkan dan dimasukkan ke dalam tabung PCR selama seminggu; dilanjutkan dengan anotasi oleh embriologi untuk menentukan penilaian morfologi dan parameter morfokinetik menggunakan Microscopcopy Time-Lapse. Uji chi-square digunakan untuk menganalisis variabel bivariat.Hasil: Seratus dua puluh empat sampel didapatkan pada hari ke 5 pasien yang menjalani prosedur FIV. Sebanyak 50,8% memiliki kromosom aneuploid, dan 49,2% adalah euploid. Median karakteristik morfokinetik yaitu 3,86 kali lipat. Ditemukan bahwa tingkat ekspansi, time to pro-nuclear fading, dan time to the synchrony of the third cell cycle berhubungan secara signifikan dengan status euploid (p = =0,000; 0,041 dan 0,036).Kesimpulan: Tingkat ekspansi terbukti secara bermakna memiliki pengaruh dalam memprediksi status ploidi embrio.Metode: Penelitian adalah penelitian cross-sectional pada pasien program FIV yang dilanjutkan dengan pemeriksaan kromosom dengan NGS di Pusat IVF RS Pondok Indah dan Pusat FIV Morula RS Bunda pada bulan Desember 2021 sampai dengan Desember 2022. Embrio yang mencapai stadium blastokista pada hari ke 5 atau 6 dibersihkan dan dimasukkan ke dalam tabung PCR selama seminggu; dilanjutkan dengan anotasi oleh embriologi untuk menentukan penilaian morfologi dan parameter morfokinetik menggunakan Microscopcopy Time-Lapse. Uji chi-square digunakan untuk menganalisis variabel bivariat.Hasil: Seratus dua puluh empat sampel didapatkan pada hari ke 5 pasien yang menjalani prosedur FIV. Sebanyak 50,8% memiliki kromosom aneuploid, dan 49,2% adalah euploid. Median karakteristik morfokinetik yaitu 3,86 kali lipat. Ditemukan bahwa tingkat ekspansi, time to pro-nuclear fading, dan time to the synchrony of the third cell cycle berhubungan secara signifikan dengan status euploid (p = =0,000; 0,041 dan 0,036).Kesimpulan: Tingkat ekspansi terbukti secara bermakna memiliki pengaruh dalam memprediksi status ploidi embio.

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Objective : Embryonic factors greatly influence IVF outcomes. One method to ensure the embryo does not have aneuploid chromosomes is Preimplantation Genetic Testing for Aneuploidy or Preimplantation Genetic Screening procedure, which involves undergoing a biopsy of the blastomeres in the 8-cell phase or the trophectoderm in the blastocyst phase. The procedure is invasive and can potentially harm the embryo.

Methods: This study is a cross-sectional that requires patients undergoing IVF followed by chromosome examination with NGS that was conducted at the IVF Center at Pondok Indah Hospital and Morula IVF Center at Bunda Hospital from December 2021 to December 2022. Each embryo that reaches the blastocyst stage on day 5 or 6 will be washed and put into a PCR tube for a week; then, embryologists annotate them to determine morphological assessment and morphokinetic parameters using Time-Lapse Microscopy. The chi-square test was used to analyse bivariate variables.

Results: One hundred twenty four samples were collected on day 5 of patients undergoing the IVF procedure. 50.8% of the samples were aneuploid chromosomes, and 49.2% were euploid. The morphokinetic characteristics median was 3.86 fold. It was found that expansion grade, time to pro-nuclear fading, and time to the synchrony of the third cell cycle were significantly associated with euploid status (p = =0.000; 0.041 and 0.036).

Conclusion: The expansion grade has been proven as the most influential component for accurately predicting the ploidy status of embryos."

"This review article gives a brief history of the classical experiments that led to the development of the embryo culture medium and in vitro embryo culture. It proposes that, in view of the outstanding and significant pioneering contributions of Wesley Kingston Whitten to the development of embryo culture medium, he be considered the “Father of Embryo Culture Medium”. Furthermore, it describes the nutritional requirements of early embryos and how these requirements with specific references to carbohydrates, amino acids, phosphates, growth factors, etc, have been utilized to formulate increasingly more complex embryo culture media. This has led to the development of progressively more efficacious embryo culture media including the formulation of completely defined and synthetic protein-free embryo culture medium. The review also describes physical factors, growth factors, insemination methods for the fertilization of oocytes and culture methods affecting embryo growth, development, metabolism, oxygen embryotoxicity and survival. In procedural terms, the review also summarizes the evolution of embryo culture techniques from tube culture to, microdrop culture under oil to co-culture to ultra microdrop culture techniques. It includes techniques of in vitro maturation and for the selection of potentially viable embryos of various developmental stages. "

"Penilaian morfokinetik embrio dipakai untuk seleksi embrio. Penelitian cohort ini bertujuan untuk evaluasi hubungan antara jumlah salinan mtDNA di cumulus granulosa cells (CGCs) dengan parameter morfokinetik embrio dan status kromosom. Perhitungan jumlah salinan mtDNA menggunakan real-time PCR pada 129 sample CGCs dari 30 pasien yang mengikuti program IVF-IMSI di Morula IVF Jakarta antara Juli-Oktober 2020. Hubungan antara jumlah salinan mtDNA di CGCs dengan semua parameter menggunakan analisa bivariate dan multiple. Terdapat hubungan signifikan antara jumlah salinan mtDNA di CGCs dengan pencapaian blastokista setelah dikontrol variabel usia maternal dan morfologi sperma (coefficient 0.832, p-value = 0.032, RR value 2.299). Hubungan signifikan pada jumlah salinan mtDNA di CGCs dengan fase awal perkembangan embrio M1 (t2-t8), dengan persamaan M1 adalah 5.702-0.271 jumlah salinan mtDNA di CGCs + 0.017 usia maternal + 0.013 motilitas sperma – 0.115 morfologi sperma (p-value = 0.032). Ditemukan hubungan tidak signifikan antara jumlah salinan mtDNA di CGCs dengan parameter morfokinetik lainnya (M2: tC-tEB, M3: t2-tEB, DC, RC, MN dengan P> 0.05), serta dengan status kromosom embrio (euploid: 139.44 ± 133.12, aneuploid: 142.40 ± 111.30, p= 0.806). Penelitian ini menunjukkan bahwa jumlah salinan mtDNA di CGCs merupakan biomarker untuk memprediksi pencapaian blastokista dan fase awal perkembangan embrio, tetapi tidak status kromosom.

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This cohort study evaluates the association between the mtDNA copy number in cumulus granulosa cells (CGCs) with embryo morphokinetic parameters and chromosomal status. mtDNA copy number of 129 CGCs from 30 patients undergoing the IVF-IMSI program at Morula IVF Jakarta between July-October 2020 were analyzed using real-time PCR. Bivariate and multiple analyses were conducted to see its relationship with all parameters. There was a significant correlation between the mtDNA copy number and the blastocyst after adjusting the maternal age and sperm morphology (coefficient 0.832, p-value = 0.032, RR value 2.299). A significant link was observed between mtDNA copy number in CGCs and early embryo developmental phase M1 (t2-t8), with the equation of M1 is 5.702 - 0.271 mtDNA copy number of CGCs + 0.017 maternal age + 0.013 sperm motility -0.115 sperm morphology (p-value = 0.032). No correlation was found between the mtDNA copy number in CGCs with the other morphokinetic parameters (M2: tC-tEB, M3: t2-tEB, DC, RC, MN with p> 0.05), and the chromosomal status (euploid: 139.44 ± 133.12, aneuploid: 142.40 ± 111.30, p= 0.806). mtDNA copy number in CGCs can serve as a useful biomarker for blastocyst status and early embryo developmental phase but not for chromosomal status."

"Latar belakang. Metode penilaian viabilitas embrio yang masih kurang akurat menjadi tantangan dalam peningkatan keberhasilan fertilisasi in vitro. Kualitas embrio dipengaruhi oleh umur kronologis dan regulator ekspresi gen. Salah satu regulator ekspresi gen tersebut merupakan MicroRNA-135b. MicroRNA-135b sangat potensial untuk dikembangkan lebih lanjut sebagai biomarker kualitas embrio fertilisasi in vitro yang bersifat non-invasif. MicroRNA-135b terekspresi secara stabil di medium kultur embrio. Selain itu, terjadi peningkatan ekspresi MicroRNA-135b di medium kultur embrio aneuploidi dibandingkan medium kultur embrio euploidi pada fertilisasi in vitro. Kondisi aneuploidi pada embrio memiliki korelasi positif dengan umur kronologis dari pasien fertilisasi in vitro. Oleh karena itu, akan diteliti apakah terdapat korelasi antara umur kronologis dan ekspresi MicroRNA-135b pada medium kultur embrio pasien fertilisasi in vitro. Tujuan. 1) Mengetahui sebaran umur kronologis dan ekspresi MicroRNA-135b pada pasien fertilisasi in vitro. 2) Mengetahui korelasi umur kronologis dan ekspresi MicroRNA-135b di medium kultur embrio pasien fertilisasi in vitro. Metode. Studi ini merupakan sebuah studi cross-sectional yang dilakukan pada pasien fertilisasi in vitro Klinik Yasmin RSCM Kencana. Data umur kronologis pasien diperoleh dari data rekam medis pasien. Sampel medium kultur embrio diambil di hari ke-5 prosedur kultur embrio. Selain itu, dilakukan juga pengambilan sampel medium basal sebagai kelompok kontrol. Sampel yang telah diambil akan diperiksa nilai ekspresi MicroRNA-135b dengan analisis kuantitatif real time PCR. Analisis data dilakukan dengan IBM SPSS Statistics 25. Hasil. Total sampel penelitian adalah sebanyak 31 medium kultur embrio dari 11 orang pasien. Umur kronologis dan ekspresi MicroRNA-135b tersebar secara tidak normal. Dari penelitian ini, ditemukan bahwa terdapat korelasi positif bermakna dengan kekuatan statistik sedang antara umur kronologis dengan ekspresi MicroRNA-135b di medium kultur embrio pasien fertilisasi in vitro. Selain itu, ditemukan pula peningkatan relatif ekspresi MicroRNA-135b sebesar 4,9 fold pada medium kultur embrio dibandingkan dengan medium basal fertilisasi in vitro. Kesimpulan. Umur kronologis yang semakin meningkat diikuti dengan peningkatan ekspresi MicroRNA-135b di medium kultur embrio pasien fertilisasi in vitro.

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Background. The lack of accuracy in embryo viability assessment methods still become a challenge to increase the in vitro fertilization (IVF) success rate. The quality of embryo influenced by the chronological age and gene expression regulator. One of the gene expression regulator is MicroRNA-135b. MicroRNA-135b is very potential to become a noninvasive biomarker of IVF embryo quality. MicroRNA-135b express stably in the spent embryo media. There is an increasement of MicroRNA-135b expression in aneuploidy spent embryo media than euploidy spent embryo media of IVF. Aneuploidy embryo has positive correlation with the IVF patient’s chronological age. Therefore, in this study, we will determine whether chronological age has correlation with MicroRNA- 135b expression in spent embryo media of IVF patient.

Objectives. 1) To determine the chronological age and MicroRNA-135b expression distribution at IVF patient. 2) To determine the correlation between chronological age and MicroRNA-135b expression in spent embryo media of IVF patient.

Methods. This study is a cross-sectional study which was done to IVF patient of Yasmin Clinic in RSCM Kencana. The chronological age data were collected from the medical records of the patient. The spent embryo media sample were taken at the 5th day of the spent embryo media procedure. We also collected the basal media sample as the control group. The MicroRNA-135b expression were analysed using quantitative real time PCR analysis. The data analysis was using IBM SPSS Statistics 25.

Results. There were 31 spent embryo media from 11 patients. The chronological age and MicroRNA-135b expression distribute abnormal. We also found that there was a positive significant correlation with moderate statistical power between chronological age and MicroRNA-135b expression in spent embryo media of IVF patient. We also found that MicroRNA-135b expression increased 4,9 fold in spent embryo media than basal media of IVF.

Conclusion. The increase of chronological age followed by the increase of MicroRNA-135b expression in spent embryo media of IVF patient."

"ABSTRAKLatar Belakang: Mendapatkan satu embrio dengan potensi implantasi yang tertinggi merupakan hal penting dari teknologi fertilisasi invitro. Metode yang banyak digunakan selama ini untuk seleksi embrio adalah melalui penilaian morfologi. Namun morfologi embrio belum dapat menggambarkan potensi embrio dengan baik. Teknologi metabolomik merupakan metode yang memiliki potensi yang cukup baik karena memiliki beberapa kelebihan yaitu pengukuran yang cepat dan tidak bersifat invasif. Viabilitas hasil konsepsi dipengaruhi oleh proses biologis intrasel yang menghasilkan metabolom. Embrio dengan morfologi yang baik ternyata memiliki gambaran metabolomik yangberbeda. Dari penelitian ini diharapkan dapat menghasilkan model prediksi dari pola metabolomik medium kultur embrio untuk memprediksi kemampuan pembentukan blastokista. Jika kita dapat memprediksi potensi keberhasilan pembentukan blastokista dengan cara yang nir invasif dan cepat, diharapkan dapat meningkatkan proses pemilihan embrio dengan potensi implantasi yang tinggi, tanpa memperpanjang masa kultur embrio hingga hari kelima.Tujuan: Mengembangkan metode nir invasive dalam memprediksi kemampuan embrio berkembang menjadi blastokista.Metode: Penelitian kohort terhadap data spektrum FTIR medium kultur embrio hari pertama dan hari ketiga dalam memprediksi keberhasilan pembentukan blastokista, dengan membuat model prediksi dari spektrum tersebut.Hasil: Didapatkan blastokista dengan kualitas baik sebanyak 16 dari 44 embrio yang diteliti. Didapatkan nilai AUC 0,752 pada model prediksi analisis FTIR medium kultur embrio hari pertama. dengan sensitifitas 0,727 dan akurasi 72,7%. Pada analisis spektrum hari ke 3 didapatkan model prediksi dengan AUC 0,674, dengan sensitifitas 0,614 dan akurasi 0,614.Kesimpulan: Pola metabolomik medium kultur embrio dapat membedakan antara embrio yang berhasil menjadi blastokista kualitas baik dan tidak menjadi blastokista kualitas baik. Perbedaan ini dapat dideteksi dengan menggunakan analisis dengan FTIR yang kemudian dibuat model prediksinya.

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ABSTRACTBackground: To get an embryo with the highest implantation potential is an important aspect of in vitro fertilization. The most widely used method for embryo selection is morphological assessment. However, embryo morphology has not been able to describe the potential of an embryo properly. Metabolomic technology is a method that has goodpotential because it has several advantages, namely rapid measurement and not invasive. Viability of the results of conception is influenced by intracellular biological processes that produce the metabolome. Embryos with good morphology have a different metabolomic profilling. From this research it is expected to produce a predictive model of the metabolomic profilling of embryo culture medium to predict the successful of good quality blastocyst formation. If we can predict the potential success of blastocyst formation in a non-invasive and fast way, it is hoped that it can improve the process of selecting embryos with high implantation potential, without extending embryo culture tothe fifth day.Objective: To develop a non-invasive method for predicting the ability of an embryo to develop into a good quality blastocyst.Method: A cohort study of FTIR spectrum data of first and third day embryo culture medium in predicting the success of good quality blastocyst formation, by making a prediction model of the spectrum.Results: There were good quality blastocysts from 16 of the 44 embryos studied. AUC value of 0.752 was obtained from the FTIR analysis model prediction for the first day of embryo culture medium. with a sensitivity of 0.727 and an accuracy of 72.7%. In the spectrum analysis of day 3was obtained predictive models with AUC of 0.674, with a sensitivity of 0.614 and an accuracy of 0.614.Conclusion: The metabolomic profilling of embryo culture medium can distinguish between an embryo that succeeds in becoming a good quality blastocyst and not a good quality blastocyst. This difference can be detected by using analysis with FTIR."

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