6 research outputs found
Expression of both ubiquitin and histone proteins analyzed in blastocoel fluid from IVF embryos is correlated with positive implantation
Introduction:Due to an influx of patients seeking infertility treatments like IVF, the need to identify additional molecular markers, particularly proteins, in preimplantation embryos to predict successful implantation is essential. We propose assessing expression of proteins from the ubiquitin-proteasome family as well as histones in blastocoel fluid-conditioned media from IVF-embryos with known implantation outcomes. These data may provide evidence for embryologists to eventually predict implantation potential of an embryo with these assays.
Methods:Blastocoel fluid conditioned media samples were collected from day-5-IVF embryos at the times of PGT-A biopsy. Media associated with embryos of poor morphology versus good morphology were used in this study. Total RNA and protein concentration were assessed with an Agilent Bioanalyzer. Blastocoel fluid-conditioned media samples were pooled and loaded onto a NuPAGE protein gel to detect total proteins with Colloidal Blue staining and then for Western Blots. Antibodies to detect ubiquitin and histones were used for the blots.
Results:Preliminary results suggest that proteins can be detected in pooled blastocoel fluid conditioned media samples based on Colloidal Blue staining. The initial Western Blot assay detected some expression of ubiquitin.
Conclusions:We hypothesize that the euploid grade AA embryos will present with stronger markers of histone and ubiquitin proteins as well as a higher concentration of protein in the blastocoel fluid, providing evidence that the embryos with stronger markers of said proteins will result in a successful implantation
Bilateral and Unilateral Cochlear Implant Users Compared on Speech Perception in Noise
Objective: Compare speech performance in noise with matched bilateral cochlear implant (ClCl) and unilateral cochlear implant (Cl only) users. Design: Thirty CICI and 30 CI-only subjects were tested on a battery of speech perception tests in noise that use an eight-loudspeaker array. Results: On average, ClCl subject's performance with speech in noise was significantly better than the Cl-only subjects. Conclusion: The CICI group showed significantly better performance on speech perception in noise compared with the CI-only subjects, supporting the hypothesis that CICI is more beneficial than CI only
Expression of signaling pathway genes in IVF embryos that result in miscarriage
Introduction: Approximately 10 to 20 percent of pregnancies end in miscarriage. Infertility is one of the leading causes of recurrent miscarriages and affects approximately 14% of reproductive-age couples. Recurrent miscarriages are presumed to be the result of embryonic chromosomal abnormalities such as aneuploidy. In response to infertility, many couples seek IVF, often without reliable guidelines on which embryo has the best potential for leading to a live birth. Currently, this process is very expensive, costing on average $23,474 per cycle, and emotionally taxing, exacerbating their comorbidities of anxiety, depression, and isolation. Additionally, IVF typically leads to only an approximately 25-50% birth rate. As a result, there are many couples who are unable to conceive through IVF the first time. Due to the high cost of the procedure, many couples may not be able to afford to do multiple rounds of IVF.
Purpose Statement: This study seeks to identify gene expression signatures from blastocoel fluid-conditioned media collected from preimplantation euploid IVF-embryos that resulted in miscarriage.
Methods: Candidate genes were previously identified through whole transcriptome (RNASeq) analysis of blastocoel fluid from euploid IVF-embryos that resulted in successful or unsuccessful implantation. Blastocoel fluid from a new set of euploid IVF-embryos resulting in miscarriage or successful implantation were selected for specific gene expression analysis using RT-qPCR. RNA was first purified from the fluid, then cDNA was synthesized for use in RT-qPCR reactions with gene-specific TaqMan primers. Genes assessed using RT-qPCR were BCL2L12, SHARPIN, CUL2, and DRAP1 with GAPDH as a control. These genes represented pathways involved in apoptosis and protein degradation via ubiquitination.
Results: Gene expression analysis of is ongoing and future studies will continue to assess the expression of these genes in additional samples. Initial studies suggest altered expression patterns in some samples associated with miscarriage. Tested samples associated with positive implantation outcomes showed higher relative expression levels of CUL2. Those embryos with negative implantation outcomes showed higher relative expression levels of SHARPIN.
Conclusion: If genes could be identified that are associated with miscarriage, this additional embryo quality metric may improve the success of each IVF-embryo transfer potentially reducing the number of miscarriages. By reducing the number of miscarriages, fertility treatments may be accessible to a larger population by decreasing the associated costs of having to complete multiple rounds of treating and the psychological burden that may accompany recurrent miscarriage