Viability of Embryo Sacs and Fruit Set in Different Plum (Prunus domestica L.) Cultivars Grown under Norwegian Climatic Conditions

Compatibility and synchrony between specialized tissues of the pistil, female gametophytes and male gametophytes, are necessary for successful pollination, fertilization, and fruit set in angiosperms. The aim of the present work was to study the development and viability of embryo sacs, as well as fertilization success, in relation to the fruit set of the cultivars ‘Mallard’, ‘Edda’, ‘Jubileum’, and ‘Reeves’, under specific Norwegian climatic conditions. Emasculated, unpollinated, and open-pollinated flowers were collected at the beginning of flowering, and on the 3rd, 6th, 9th, and 12th days after flowering, from all four plum cultivars over two years (2018/2019). Ovaries were dehydrated, embedded in paraffin wax, sectioned, stained, and observed under a light microscope. Results showed the existence of synchronization between successive phases in the development of the embryo sac and individual phases of flowering. All plum cultivars had higher percentages of viable embryo sacs, fertilized embryo sacs, and fruit set in 2018 than in 2019. These differences may be related to the very low temperatures during the post-full-flowering period in 2019, and to the low adaptation of some studied cultivars to unfavorable conditions. In our study, the cultivar ‘Jubileum’ showed the highest percentage of viable embryo sacs, fertilized embryo sacs, and fruit set compared to other cultivars, i.e., the best low-temperature adaptation.

Time Series Prediction of Viable Embryo and Automatic Grading in IVF using Deep Learning

Background: The process of In Vitro Fertilization (IVF) involves collecting multiple samples of mature eggs that are fertilized with sperms in the IVF laboratory. They are eventually graded, and the most viable embryo out of all the samples is selected for transfer in the mother’s womb for a healthy pregnancy. Currently, the process of grading and selecting the healthiest embryo is performed by visual morphology, and manual records are maintained by embryologists. Objectives: Maintaining manual records makes the process very tedious, time-consuming, and error-prone. The absence of a universal grading leads to high subjectivity and low success rate of pregnancy. To improve the chances of pregnancy, multiple embryos are transferred in the womb elevating the risk of multiple pregnancies. In this paper, we propose a deep learning-based method to perform the automatic grading of the embryos using time series prediction with Long Short Term Memory (LSTM) and Convolutional Neural Network (CNN). Methods: CNN extracts the features of the images of embryos, and a sequence of such features is fed to LSTM for time series prediction, which gives the final grade. Results: Our model gave an ideal accuracy of 100% on training and validation. A comparison of obtained results is made with those obtained from a GRU model as well as other pre-trained models. Conclusion: The automated process is robust and eliminates subjectivity. The days-long hard work can now be replaced with our model, which gives the grading within 8 seconds with a GPU.

Comparison of Dydrogesterone and Medroxyprogesterone in the Progestin-Primed Ovarian Stimulation Protocol for Patients With Poor Ovarian Response

ObjectiveTo compare the clinical outcomes of dydrogesterone (DYG) and medroxyprogesterone (MPA) in the progestin-primed ovarian stimulation (PPOS) protocol for patients with poor ovarian response (POR).Patients and MethodsThis was a retrospective cohort study. Women with POR who underwent IVF/ICSI at the Reproductive Center of Third Affiliated Hospital of Zhengzhou University between January 2020 and January 2021 were included. The primary outcome measure of our study was the number of oocytes retrieved. The secondary outcome measures in the present study were the number of 2PN, number of available embryos, oocyte retrieval rate, fertilization rate, viable embryo rate per oocyte retrieved, cancellation rate and pregnancy outcomes of the first embryo transfer cycle, including the biochemical pregnancy, clinical pregnancy and miscarriage rates.ResultsIn total, 118 women underwent hMG +DYG protocols, and 692 women who underwent hMG +MPA met the Bologna criteria for POR. After baseline characteristics were balanced using the PSM model, 118 hMG +DYG protocols were matched to 118 hMG +MPA protocols, and the baseline characteristics were comparable between the two groups. The numbers of oocytes retrieved, 2PN, and available embryos and the oocyte retrieval rate, fertilization rate, viable embryo rate per oocyte retrieved and cancellation rate of the hMG+DYG and hMG+MPA protocols were comparable. Altogether, 66 women in the hMG+DYG group and 87 women in the hMG+MPA group underwent first embryo transfers. In the hMG+DYG group, 81.8% (54/66) of the patients underwent cleavage embryo transfers; similarly, 79.3% (69/87) of patients in the hMG+MPA group had cleavage embryo transfers (P=0.70).The biochemical pregnancy rate of the hMG+DYG group was 42.4%, and this was comparable to the rate in the hMG+DYG group, at 34.5% (P=0.32). The clinical pregnancy rates were similar between the two groups (36.4% vs. 31.0%, P=0.49), and there was no significant difference in the rate of miscarriage between the two groups (12.5% vs. 29.6%, P=0.14).ConclusionFor women with POR, the clinical outcome of the hMG + DYG group was similar to that of the hMG + MPA group, indicating that both combinations can be useful options for PPOS protocols.

O-049 How is AI changing the way we make babies?

text ART is one of the fastest-growing fields in medicine. Besides the advancements in the hormonal stimulation and treatment regimes, the laboratory aspect of IVF showed emerged development in science and especially technology. The emphasis of future IVF treatments and laboratory procedures is on the non-invasive methods of embryo evaluation and selection. The application of Artificial Intelligence (AI) in daily life prompts us to evaluate this technology in this context. The AI is one of the main candidates to drive future IVF not only in the lab but also in all aspects of the IVF procedures. The benefit of AI technology involves “big data” analysis, objectivity, standardization, and precision. The most advancement of AI application is in the IVF lab where technology can be used as a tool to enhance our ability to select the “best” viable embryo objectively. As a new technology, it is important to evaluate it on the large scale. The AI technology will also enable us to standardize clinical parameters of patient reproductive potential and objectively predict each patient reproductive outcome per cycle. Ultimately, AI will help us to achieve the personalized and precision medicine needed in our field.

A comparative study on the reproductive success of two rewarding Habenaria species (Orchidaceae) occurring in roadside verge habitats

Background Most orchid species have been shown to be severely pollination limited, and the factors affecting reproductive success have been widely studied. However, the factors determining the reproductive success vary from species to species. Habenaria species typically produce nectar but exhibit variable fruit set and reproductive success among species. Here, we investigated the influence of the flowering plant density, inflorescence size, breeding system, and pollinator behaviour on the reproductive success of two rewarding Habenaria species. Results Our observations indicated that Habenaria limprichtii and H. petelotii co-occur in roadside verge habitats and present overlapping flowering periods. Both species were pollination limited, although H. limprichtii produced more fruits than H. petelotii under natural conditions during the 3-year investigation. H. petelotii individuals formed distinct patches along roadsides, while nearly all H. limprichtii individuals clustered together. The bigger floral display and higher nectar sugar concentration in H. limprichtii resulted in increased attraction and visits from pollinators. Three species of effective moths pollinated for H. limprichtii, while Thinopteryx delectans (Geometridae) was the exclusive pollinator of H. petelotii. The percentage of viable seeds was significantly lower for hand geitonogamy than for hand cross-pollination in both species. However, H. limprichtii may often be geitonogamously pollinated based on the behaviours of the pollinators and viable embryo assessment. Conclusions In anthropogenic interference habitats, the behaviours and abundance of pollinators influence the fruit set of the two studied species. The different pollinator assemblages in H. limprichtii can alleviate pollinator specificity and ensure reproductive success, whereas the more viable embryos of natural fruit seeds in H. petelotii suggested reducing geitonogamy by pollinators in the field. Our results indicate that a quantity-quality trade-off must occur between species with different breeding strategies so that they can fully exploit the existing given resources.

Assessment of the pregnancy rates using sequential day 3 and day 5 embryo transfer in IVF/ ICSI patients

Background: To assess the pregnancy rates using sequential day 3 and day 5 embryo transfer in IVF/ ICSI patients.Methods: This prospective study was conducted in Aarogya Hospital and test tube baby Centre, Raipur from 1st January 2013 to 30th November 2019. Total 100 patients undergoing IVF/ICSI in the study period were offered sequential transfer.Results: Our fertilization rates were 80% with 85.7% grade I embryos on day 3. Blastocyst formation rate was 71.42%. Cycle cancellation rates were nil. Clinical pregnancy rates per retrieval cycle were 50% and implantation rates were 24% with acceptable multiple pregnancy rates of 12%.Conclusions: We advocate that this technique is useful in all patients having good quality embryos in adequate number for double transfer as this optimizes the chance of selection of the most viable embryo for transfer which is probably the key for a successful IVF program.

Systematic comparison of developmental GRNs explains how novelty is incorporated in early development

SUMMARYThe impressive array of morphological diversity among animal taxa represents the product of millions of years of evolution. Morphology is the output of development, therefore phenotypic evolution arises from changes to the topology of the gene regulatory networks (GRNs) that control the highly coordinated process of embryogenesis1. While genetic variation can arise anywhere in the genome and affect any part of an individual GRN, the need to form a viable embryo provides a constraint on the types of variation that pass the filter of selection. A particular challenge in understanding the origins of animal diversity lies in determining how GRNs incorporate novelty while preserving the overall stability of the network, and hence, embryonic viability. Here we assemble a comprehensive GRN, consisting of 42 genes (nodes) and 84 interactions (edges), for the model of endomesoderm specification in the sea star from zygote through gastrulation that corresponds to the GRN for sea urchin development of equivalent territories and stages2. Using these detailed models we make the first systems-level comparison of early development and examine how novelty is incorporated into GRNs. We show how the GRN is resilient to the introduction of a transcription factor, pmar1, the inclusion of which leads to a switch between two stable modes of Delta-Notch signaling. Signaling pathways can function in multiple modes and we propose that GRN changes that lead to switches between modes may be a common evolutionary mechanism for changes in embryogenesis. Our data additionally proposes a model in which evolutionarily conserved network motifs, or kernels, may function throughout development to stabilize these signaling transitions.

Embryoscope - An Advanced Embryo Monitoring Biomedical Instrument

Embryoscope is a machine which is used to monitor embryo from the time of conception. It allows fertility specialist to select the most viable embryo during in-vitro fertilization. This instrument reduces the risk of taking embryo from the incubator for observation. It can monitor 12 embryos at once and take photos of each embryo every 5 to 10 minutes during the entire incubation period. With the help of this instrument, fertility expert can also monitor the abnormality in embryo. The embryoscope is an incubator with an integrated camera. The detection of abnormality can be done without harming the embryo.

Reducing time to pregnancy and facilitating the birth of healthy children through functional analysis of embryo physiology†

An ever-increasing number of couples rely on assisted reproductive technologies (ART) in order to conceive a child. Although advances in embryo culture have led to increases in the success rates of clinical ART, it often takes more than one treatment cycle to conceive a child. Ensuring patients conceive as soon as possible with a healthy embryo is a priority for reproductive medicine. Currently, selection of embryos for transfer relies predominantly on the morphological assessment of the preimplantation embryo; however, morphology is not an absolute link to embryo physiology, nor the health of the resulting child. Non-invasive quantitation of individual embryo physiology, a key regulator of both embryo viability and health, could provide valuable information to assist in the selection of the most viable embryo for transfer, hence reducing the time to pregnancy. Further, according to the Barker Hypothesis, the environment to which a fetus is exposed to during gestation affects subsequent offspring health. If the environment of the preimplantation period is capable of affecting metabolism, which in turn will affect gene expression through the metaboloepigenetic link, then assessment of embryo metabolism should represent an indirect measure of future offspring health. Previously, the term viable embryo has been used in association with the potential of an embryo to establish a pregnancy. Here, we propose the term healthy embryo to reflect the capacity of that embryo to lead to a healthy child and adult.

Isolation of Sperm and Egg Cells from Pepper

Pepper (Capsicum annuum) pollen is bicellular and contains a vegetative cell and a generative cell, which divides in pollen tubes to form two sperm cells. Sperm cells of pepper were isolated using an in vivo–in vitro method. Hand-pollinated styles were first grown in vivo for several hours, then cut from their base and cultured in vitro until pollen tubes grew from the cut end. When the pollen tubes were transferred to a breaking solution, sperm cells were released from broken tubes. Viable embryo sac cells of pepper were isolated using enzymatic digestion and mechanical dissection. Isolated ovules were digested using cellulase and pectinase for 40 minutes and then transferred to an enzyme-free solution for mechanical dissection. Three cells of the egg apparatus and a central cell were released from a cut at the chalazal end of each ovule by pressing on the micropylar area of the ovule with a microneedle. Optimal isolation conditions included 11% mannitol, 0.04% CaCl2, 1% bovine serum albumin (BSA), 1% cellulase, 1% pectinase, and 0.3% pectolyase. Using this protocol, populations of pepper egg cells, synergids, and central cells were isolated.