Activiteit

Recent years have seen a dramatic rise in the number of frozen-thawed embryo replacement (FER) cycles. Along with the advances in embryo cryopreservation techniques, the optimization of endometrial receptivity has resulted in outcomes for FER that are similar to fresh embryo transfer. However, the question of whether the Freeze all strategy is for all is nowadays a hot topic. This review addresses this issue and describes current evidence based on randomized controlled trials and observational studies. To date, it is reasonable to perform FER in cases with a clear indication for the benefits of such strategy including impending ovarian hyperstimulation syndrome (OHSS) or preimplantation genetic testing for aneuploidy (PGT-A); however, this strategy does not fit for all. This review analyses the pros and cons of the freeze all strategy highlighting the need to follow a personalized plan in embryo transfer, avoiding a freeze all methodology for all patients in an unselected manner.In this paper, the schooling behavior of prey fish population in a predator-prey interaction is investigated. By taking an economical interest which can be elaborated by the presence of nonselective harvesting into consideration, we studied the dynamical behavior. The existence, positivity and boundedness of solution have been established. The analysis of the equilibrium states is presented by studying the local and the global stability. The possible types of local bifurcation that the system can undergoes are discussed. The effect of fishing effort on the evolution of the species is examined. Further, by using Pontryagin’s maximum principle a proper management strategy has been used for avoiding the extinction of the considered species and maximizing the benefits. For the validation of the theoretical result, several of graphical representations have been used.Transcription factors orchestrate complex regulatory networks of gene expression. A better understanding of the common transcription factors, and their shared interactions, among a set of coregulated or differentially expressed genes can provide powerful insights into the key pathways governing such expression patterns. Critically, such information must also be considered in the context of the frequency in which a transcription factor is present in a properly selected background, and in the context of existing evidence of gene and transcription factor interaction. Given the vast amount of publicly available gene expression data that can be further scrutinized by the user-friendly analysis tools described here, many useful insights are assuredly to be revealed. The proceeding methods for application of the analysis tool CiiiDER for transcription factor-binding site identification, enrichment analysis, and coregulatory factor identification should be applicable to any dataset comparing differential gene expression in response to various stimuli and gene coexpression datasets. These methods should assist the researcher in identifying the most relevant regulators within a gene set, and refining the list of targets for future study to those which may share biologically important regulatory networks.Bioinformatics tools and resources are valuable for the analysis of data sets focusing on programmed cell death. This chapter discusses methods for the generation of gene sets as well as enrichment analysis using publicly available databases.CRISPR/Cas9-based gene editing is a recent advance that allows for the knockout or alteration of target genes within mammalian cells. dTRIM24 in vitro Many variations of the technique exist, but here we describe two systems of plasmid-based CRISPR gene knockout which together allow for the selective knockout of virtually any gene target. Compared with other CRISPR-based systems, these plasmids have the advantages of delivering all the necessary components in one plasmid, choice of multiple selectable markers, and choice of route of administration into target cells. In addition, potential off-target effects from one system (dependent upon selection of target gene) can be overcome through use of the second system. Strategies for optimizing the knockout process and selection of finished cell lines are also presented.The rapid, efficient detection of cell death is critical for characterizing the underlying biology of in vitro disease models and, in particular, immunotherapy products used for preclinical therapeutic research. Traditional endpoint assays are laborious to perform for mass screening of therapeutic candidates and may fail to fully capture the kinetics of events surrounding the initiation, duration, and mechanisms of cell death-important events that may affect translational relevance and impact therapeutic decision-making during development. Here, we describe simple, efficient methods to measure apoptosis and immune cell killing in both adherent and nonadherent cell populations using the Incucyte® Live-Cell Analysis system and associated nonperturbing reagents, cells, and protocols. Assays are performed in the user’s own incubator with minimal disturbance and may be readily incorporated into existing workflows. Users may multiplex to maximize data collection from each sample. The integrated, user-friendly software does not require advanced technical training, enabling rapid analysis. Taken together, this method provides essential kinetic insight for greater understanding of cell death and the dynamic interactions between immune cells and their targets.Comprehensive understanding of cellular responses to changes in the cellular environment or by drug treatment requires time-dependent analysis ranging from hours to several days. Here, we describe a sensitive, nonlytic live-cell assay that allows continuous or ‘real-time’ monitoring of cell viability, growth, and cytotoxicity over an extended period of time. We illustrate the use of the assay for small drug molecule and antibody-dependent cytotoxicity studies using cancer cells in 384-well plates. We show that the ability to measure changes in live cells over time provides instantaneous information on the biological status of the cells, information about the mode of action of the drug, and offers an added advantage of preserving the cells for multiplexing with downstream applications.