Brief Summary of Minutes of Annual Meeting

Brief Summary of Minutes of 2023 Annual Meeting

            University of Florida, Gainesville, FL

March 3^th, 2023

09:03: Dr. Daigneault started the meeting and welcomed the participants.

09:04: Dr. Daigneault presented the agenda.

• There was a discussion about PIs that have left W4171 multistate group, and whether we could remove those that left from the current report.
• Checura pointed out that we were not supposed to remove people from the report without USDA approval.
• Daigneault offered to reach out to Kenneth White, Agriculture Experiment Station coordinator, to verify possible PIs to be excluded from the current group.
• A new participant from the University of Cornell, Dr. Jingyue (Ellie) Duan, introduced herself as a new member of the group.

09:18: Dr. Checura proposed a motion to approve the minutes from last meeting. Dr. Daigneault seconded.

09:18: Dr. Davies reminded everyone that this multistate project approval extends through September, 2024. Therefore, there is a need to restructure the current project and resubmit. 

• Davies also pointed out that the length of the meeting could be increased so participants would have more time to discuss their projects and make collaborations.
• Winger reminded the group that the next IETS meeting will be in the Denver, CO. Dr. Winger offered to organize with the IETS committee so that participants of the W4171 multistate group would have a room for 1 or 2 extra days to be able to participate in our 2024 annual meeting.
• Tian proposed that instead of having a longer group meeting (2 days) we could do a mid-year Zoom meeting to discuss collaborations. Therefore, we would keep a traditional 1-day annual meeting (in person) generally during the IETS annual meeting.
• Daigneault suggested that we make the decision regarding the duration of our annual meeting at the end of today’s meeting.

09:20: Everyone introduced themselves including the participants attending the meeting in person and the ones that joined via Zoom.

09:36: Dr. Davies explained that the summary report should submitted through the NIMSS system. Dr. Checura mentioned that, in the previous year, she submitted the final report directly to Kenneth White.

09:37: Discussion of individual station reports started. It was decided that, in the current year, station reports will be discussed in alphabetical order. For the 2024 annual meeting, it was decided that station reports will be discussed in reverse alphabetical order.

09:40: Connecticut - Dr. Tian started discussing her work on improving maturation conditions. Then she moved to her work generating naïve bovine embryonic stem cells.

• Data mining project: showed widespread intergenic transcription in forms of read-throughs and read-ins in bovine pre-implantation embryos.

10:05: California - Dr. Denicol: Characterization of the bovine ovary by single cell sequencing. Compared ovary from pre-ovulatory and mid-luteal state. Compared fetal and adult bovine ovary using single cell RNA-seq. Provided an update on her work in pre-antral folliculogenesis.

10:25: Colorado - Dr. Winger: Informed everyone that Dr. Jennifer Barfield (Colorado State University) plans on being part of the group. Dr. Barfield works on embryology and bison.

Dr. Winger: presented his model for gene knock down in the trophectoderm of blastocyst using lentivirus. Dr. Winger gave updates on his work with T4 transfer from the mother to the fetus.

11:08: Florida - Dr. Daigneault briefly reported on his work for the characterization of reproductive seasonality in Florida Native sheep, and sperm work on stallions. One of the focuses of Dr. Daigneault lab is to explore how the environment affects post ejaculatory sperm function.

11:30 Dr. Jiang: Discussed his work studying the molecular mechanism of early embryonic development, particularly during pre-implantation development. Highlighted his work aiming to profile the bovine pre-implantation embryo using single cell sequencing. He also presented data on his work on bovine blastoids derived from trophoblast stem cells.

12:00: Illinois – Dr. Wheeler: Dominant follicle removal in Bos indicus cattle and half-blood Gir x Holstein crosses. Dr. Wheeler reported results of a trial feeding rumen-protected methionine on the endometrium and embryos in Holsteins.

• Regarding the biomedical model, Dr. Wheeler reported on the development of a 3D printer cage to be used for fusing damaged spines. This work will move on to clinical trials.

13:35: Iowa - Dr. Youngs: presented some of his work overseas including word done with alpacas in Peru and dairy cattle in Ethiopia.

14:00: Louisiana – Dr. Bondioli: Localization of key proteins involved in meiotic spindle formation. Occyte cryopreservation. Improved methods developed in his lab for ICSI.

14:35: Maryland – Dr. Keefer: Reported on her work with energy metabolism in embryos.

14:45: Mississippi – Dr. Feugang: Reported on studies exploring seasonal variations in extracellular vesicles miRNA in follicular fluid of horses. Characterization of seminal extracellular vesicles of divergent sperm quality.

15:00: Missouri – Dr. Lee: Reported on his work generating GDF9 knockout pigs and work on the identification of off-targeted events introduced by the CRISPR-Cas9 system in gene-edited pigs.

15:21: Nebraska – Dr. White: Transgenic work on GnRH2 using a porcine model.

15:30: Oklahoma – Dr. Moraes: Provided updates on his work investigating the genetic regulation of conceptus elongation in cattle. Collaborative work with Matt. Lucy (University of Missouri) investigating disease programing of dairy cattle infertility. Provided an update on future projects.

15:50: South Carolina – Dr. Checura: Provided updates on her work on photobiomodulation on oocyte competence.

16:00: Utah – Dr. Polejaeva: Laparoscopic OPU derived oocytes associated with improved blastocyst development following SCNT. Development of sheep model for sickle cell disease.

16:20: Dr. Davies reported novel data on the establishment of pregnancy in IFNAR2 knockout sheep.

16:43: New York - Dr. Duan:  Provided an update on her transition from graduate student to assistant professor and setting up her new lab at Cornell. Dr. Duan also reviewed some of the work she performed as a PhD student and during her postdoc training.

16:50: Dr. Daigneault nominated Dr. Moraes for Chair of the 2024 meeting. Dr. Youngs seconded.

16:53: Dr. Daigneault nominated Dr. Duan for secretary of the 2024 meeting. Dr. Polejaeva seconded.

16:54: Possible dates for the 2024 annual meeting are January 8^th, 9^th or 13^th, depending on the schedule of activities of IETS.

• Once the schedule of activities for IETS has been released, there will be a Doodle Pool to decide the time for the 2024 annual meeting.

17:00: Discussions regarding resubmitting a new project to give continuation to the efforts of the current group. The current project expires in September 2024.

• Daigneault proposed to create a group in Teams to work on the new project.
• White agreed to take the lead coordinating the draft for the new project.
• It was decided that members of this group will work as a team to write the new project.
• For citation, we decided to use DOI to facilitate retrieval of references.
• Daigneault will send out an e-mail to start organizing preparation for next report.

17:25: Dr. Youngs - motion to end the meeting. Dr. Polejaeva seconded. End minutes.

Accomplishments

Objective 1

Understand the biology of gamete development, fertilization and embryogenesis including the underlying cellular and molecular mechanisms.

1. Research on Pre-Synchronization, extended progesterone treatments with 14-day intra-vaginal CIDRs (7&&) and utilization of Gender-Selected semen has proven competitive commercially feasible results.
2. Recent results with the 14-sday CIDR with GnRh at 7 and 6 days had the greatest estrus expression rates and conception rates ever achieved at our experiment stations.
3. Developed an efficient method to isolate preantral follicles from the adult bovine ovary without the need of tissue digestion with enzymes.
4. Demonstrated that the reprogramming of aromatase required for maintenance of an expanded Sertoli cell population is not due to DNA methylation although changes in DNA methylation are related to age-associated changes in aromatase activity.
5. Alteration in expression of CSF1, AR, or SRD5A2 do not appear to be involved in signaling the increase in population size of Sertoli cells. However, macrophages may be involved in the tissue machinery that leads to the enlarged population size following reduced endogenous estrogen signaling.
6. To investigate the impact of CRISPR-cas9-mediated knockout of Keap1 gene in modulating NRF2 activity, three guide RNAs targeting the exon 2 of the Keap1 gene was tested for their efficiency in inducing targeted knockout of the gene in zygotes electroporated with RNP at 10 h post IVF.
7. Supplementation of EVs derived from thermal stress granulosa cells can induce tolerance to oocytes subjected to heat stress by reducing the accumulation of ROS, improving mitochondrial activity, reducing the expression of stress associated genes thereby reducing the severity of stress in oocytes and developing blastocysts.
8. Conducted the first analysis of intergenic transcription activities in natural samples and also the first study on embryos.
9. Established the detailed profiles of hormonal events in follicular fluid in pre-ovulatory follicles, correlated their patterns to other ovarian structures.
10. Characterized the bioenergetic status of bovine sperm when maintained for 24 hr.
11. Optimized bovine sperm conditions for compatibility with SeaHorse assay.
12. Developed a modified semen extender compatible for prolonged storage of fresh and frozen bovine sperm with industry application.
13. Prolonged the maintenance of cooled stallion sperm for up to 6 days
14. Characterized the reproductive parameters of the Florida Native Sheep
15. High-resolution ribosome fractionation and low-input ribosome profiling of bovine oocytes and preimplantation embryos has enabled us to define the translational landscapes of bovine early embryo development.
16. The use of transfer learning together with Kullback-Leibler (KL) divergence to train predictive models and provides completing methylome profiles of bovine early embryos datasets with very low coverage.
17. The H3.3 chaperone Hira complex has a maternal effect function in oocyte developmental competence and embryogenesis, through modulating chromatin condensation and transcriptional quiescence.
18. Understanding the roles of SGK1 in mouse oocytes and establishment of new insights into the regulation of MPF via SGK1 in mammalian oocyte.
19. Oocyte maturation: Results show that performing DFR in half-blood Bos taurus X Bos indicus cattle is beneficial for the development of higher quality COCs and a shorter interval of OPU following DFR in Bos taurus X Bos indicus cattle increases the number of higher quality COCs. Further, new findings suggest that DFR promotes growth of SF into MF, possibly by removing the suppression exerted on SF by the dominant follicle.
20. Embryogenesis: Cows fed supplemental methionine had higher polyunsaturated lipid concentrations in embryos than cows on the control diet. Cows fed supplemental methionine had lower concentrations of un/monounsaturated lipids in uterine tissue at 15 days in milk (DIM) than cows on control diets. Specific lipid classes and lipid unsaturation level of preimplantation embryos and endometrial tissue of cows fed rumen-protected methionine were altered.
21. Using immune staining two key proteins associated with micro tubule organizing center mediated spindle formation were localized in in vitro matured bovine oocytes. Differential localization of these two proteins was observed which is different than observed in other species.
22. Studies looking at the effects of glucose and fructose on lipid droplets and mitochondrial activity in bovine embryos have been completed and the analysis almost finalized. While there appear to be shifts in size and numbers of lipid droplets, these shifts are subtle.
23. Studies involving inhibitors of the mitochondrial pyruvate carrier (UK5099) and the carnitine palmitoyltransferase-1 (CPT-1) (etomoxir) have also been completed and demonstrate that bovine embryos are very metabolically flexible, capable of shifting metabolic pathways depending on substrate availability.
24. MiRNA derived from follicular fluid-extracellular vesicles (FF-EVs) provide greater insights into the stage-specific expression dynamics of the FF EV-miRNAs during equine follicular development, which may play a role during folliculogenesis in mares.
25. Isolated FF-EVs of pre-ovulatory-sized follicles provide greater insight into the seasonal variations of miRNA pools affecting the ovarian activity of mares as seasonal breeders.
26. Near-Infra-Red Spectroscopy (NIRS), seminal plasma-derived EVs, transcriptomics, and proteomics tools were used to define molecular differences between Good and Poor quality semen of commercial boars.
27. Proteome analyses of human sperm and corresponding seminal plasma indicated a rich repertoire of several hundred proteins. Many showed dynamic changes during aging, potentially impacting sperm physiology, structure, motility, metabolism, and fertilization.
28. Production of GDF9 knockout pigs. Detailed analysis indicates GDF9 is essential for folliculogenesis in pigs.
29. Supplementation of IVM medium with E-64 significantly improved the developmental competence of OPU-derived immature oocytes.
30. CTSL supplementation to the IVM medium or IVC medium improved oocyte and embryo developmental competence under normal and heat stress conditions.
31. During the follicular phase of the estrous cycle, concentrations of GnRH-II in follicular fluid did not differ between lines (GnRHR-II knock down (KD) and control).
32. At 24 hours prior to the onset of estrus, GnRH-II levels in the blood were diminished in GnRHR-II KD compared to littermate control females.
33. There was a strong positive correlation between circulating concentrations of 17ß-estradiol and GnRH-II.
34. Protein for GnRHR-II was detected in both porcine granulosa and theca cells.
35. Theca cells from GnRHR-II KD females expressed 40% less GnRHR-II protein than control theca cells or transgenic granulosa cells.
36. Levels of GnRHR-II protein in granulosa cells were similar between lines.
37. Generated spherical, ovoid, tubular, and filamentous bovine conceptuses for sequencing.
38. Generated novel data contributing to the current understanding of how uterine disease postpartum programs long-term infertility in dairy cattle.
39. Generated data on the effects of FGF2, LIF, and IGF1 supplementation on pregnancy success following embryo transfer of bovine IVP embryos.
40. Developed a photobiomodulation system (time, wavelength, and intensity) that affect oocyte competence. The treatment alters ATP concentrations and mitochondria membrane potential by the end of oocyte maturation with lasting impacts into the blastocyst stage.
41. Preliminary data indicate improvement in oocyte maturation and blastocyst development following SCNT using L-OPU in sheep.

Objective 2
Refine methods to produce animals by genetic engineering or genome editing for the improvement of livestock production efficiency and development of human biomedical models.

1. Added to data evaluating the likelihood of off-target edits following CRISPR/Cas9 gene editing in porcine embryos.
2. Created a new area of research – investigation of thyroid hormone physiology in growth restricted fetuses induced by knockdown of chorionic somatomammotropin (CSH) using placenta specific RNAi.
3. Established putative naïve bovine embryonic stem cells that have shown in vitro and in vivo differentiation capabilities. Cells are amenable to gene-editing through single cell colony expansion. In vivo same species chimer work in underway.
4. Achieved 100% editing efficiency for zygotic delivery of CRISPR/Cas9 in bovine embryos
5. Establishment of bovine trophoblast stem cells (TSCs). Bovine TSCs exhibit transcriptomic and epigenetic features characteristic of trophectoderm cells from bovine embryos and retain developmental potency to differentiate into mature trophoblast cells.
6. Development of an efficient method to generate bovine blastocyst-like structures (termed blastoids) via the assembly of trophoblast stem cells and expanded potential stem cells. Bovine blastoids resemble blastocysts in morphology, cell composition, single-cell transcriptomes, and represent an accessible in vitro model for studying bovine embryogenesis.
7. Development of Human Biomedical Models: A topology-optimized polycaprolactone cage with BMP-2 is capable of resulting in an intervertebral fusion, similar to a conventional ring-based design of the same bioresorbable material.
8. It was demonstrated that submitting vitrified-warmed, partially denuded bovine oocytes to a post-warming recovery period supplemented with 1µM resveratrol improves vitrification outcomes.
9. A modification in the method used for bovine intra cytoplasmic sperm injection, referred to as vigorous injection, improved subsequent embryo injection.
10. Successfully identified off-targeting events potentially introduced by the CRISPR/Cas9 system in the genome of gene-edited pigs.
11. Generated novel data and insights into how liver responds to and survives oxidative and toxic stresses, using transgenic mice models.
12. Acquisition of a NEPA21 electroporator to knock out genes of interest in bovine embryos using the CRISPR-Cas9 system.
13. Developed an efficient protocol for the introduction of human specific mutations into the sheep fetal fibroblast cells using CRISPR/Cas9 RNP + ssODN
14. Developed a novel sheep model of sickle cell disease, which reproduces human clinical and laboratory parameters.
15. Created a line of type I interferon receptor (IFNAR) KO sheep and are using them to study pregnancy recognition in ruminants.
16. Cas-CLOVER gene editing tool could introduce KO in livestock fetal fibroblasts.

Publications

1. Refereed articles/book chapters

McDonnell SP, Candelaria JI, Morton AJ, Denicol AC. Isolation of Small Preantral Follicles from the Bovine Ovary Using a Combination of Fragmentation, Homogenization, and Serial Filtration. J Vis Exp. 2022 Sep 27;(187). doi: 10.3791/64423.

Pedroza GH, Lanzon LF, Rabaglino MB, Walker WL, Vahmani P, Denicol AC. Exposure to non-esterified fatty acids in vitro results in changes in the ovarian and follicular environment in cattle. Anim Reprod Sci. 2022 Mar;238:106937. doi: 10.1016/j.anireprosci.2022.106937. 

Berger T, Vanselow J, Conley A, Almand TJ, Nitta-Oda, BS. Multifaceted Epigenetic Regulation of Porcine Testicular Aromatase. Molecular and Cellular Endocrinology. 2022 541: 111526.

Katleba K, Legacki E, Berger T. Expression of CSF1, AR, and SRD5A2 during postnatal development of the boar reproductive tract. Animals 2022  12: 2167.

Kang S, Berger T. Macrophages in juvenile pig testis: Link with increase in Sertoli cells induced by estradiol suppression. Reproduction in Domestic Animals 2023 doi: 10.1111/rda.14322.

Lynch CS, Kennedy VC, Tanner AR, Ali A, Winger QA, Rozance PJ, Anthony RV. Impact of Placental SLC2A3 Deficiency during the First-Half of Gestation. Int J Mol Sci. 2022 Oct 19;23(20):12530

Ali A, Iqbal MA, Abbas MW, Bouma GJ, Anthony RV, Spencer TE, Winger QA. Trophectoderm transcriptome analysis in LIN28 knockdown ovine conceptuses suggests diverse roles of LIN28-let-7 axis in placental and fetal development. Cells. 2022 Apr 5;11(7):1234.

Tanner AR, Kennedy VC, Lynch CS, Hord TK, Winger QA, Rozance PJ and Anthony RV. In Vivo Investigation of Ruminant Placenta Function and Physiology – A Review. J Anim Sci. 2022 Jun 1;100(6).

Sohel MM, Hoelker M, Schellander K, Tesfaye D (2022): The extent of the abundance of exosomal and non-exosomal extracellular miRNAs in the bovine follicular fluid.  Reproduction in Domestic Animals, Vol 57: 1208-1217.

Miskel D, Poirier M, Beunink L, Rings F, Held E, Tholen E, Tesfaye D, Schellander K, Salilew-Wondim D, Blaschka C, Große-Brinkhaus C, Bertram B, Hoelker M (2022): The cell cycle stage of bovine zygotes electroporated with CRISPR/Cas9-RNP affects frequency of Loss-of-heterozygosity editing event. Nature Scientific Report 12 (1), 1-3.

Zhang R, Neuhoff C, Yang Q, Cinar M, Uddin MJ, Tholen E, Schellander K, Tesfaye D (2022): Suforaphane Enhanced proliferation of porcine satelite celkls via epigenetic augumentation of SMAD7. Animals 12 (11), 1365

Ranjitkar S, Duan JE, Srirattana K, Alqahtani F, Tulman ER, Mandoiu I, Venkitanarayanan K, and *Tian XC. 2022. Transcriptomic Responses of Mycoplasma bovis upon Treatments of Plant-derived Antimicrobials. Frontiers in Microbiology, published on June 2, 2022. https://doi.org/10.3389/fmicb.2022.888433.

Liu Z, Cui L, Wang W, Li M, Wang Z, Presicce GA, Tian XC, An L, Du F. 2022. Dynamic and aberrant patterns of H3K4me3, H3K9me3, and H3K27me3 during early zygotic genome activation in cloned mouse embryos. Zygote 30(6): 903-9. DOI: https://doi.org/10.1017/S0967199422000454

Johnson EJ, Duan JE, Srirattana K, Venkitanarayanan K, Tulman ER, *Tian XC. 2022. Effects of intramuscularly injected plant-derived antimicrobials in the mouse model. Scientific Reports 12:5937. doi:10.1038/s41598-022-09705-9

Losano JDA, Daigneault BW. Pharmacological perturbation of peroxisome-proliferator-activated receptor gamma alters motility and mitochondrial function of bovine sperm. Andrology. 2022 Oct 5;. doi: 10.1111/andr.13308.

Daigneault BW. Insights to maternal regulation of the paternal genome in mammalian livestock embryos: A mini-review. Front Genet. 2022;13:909804. doi: 10.3389/fgene.2022.909804. eCollection 2022. Review.

McGraw MS and Daigneault BW. Environment to embryo: intersections of contaminant exposure and preimplantation embryo development in agricultural animals. Biol Reprod. 2022 Jun 11;ioac121. doi: 10.1093/biolre/ioac121. 

Daigneault BW and Losano JA. Tributyltin chloride exposure to post-ejaculatory sperm reduces motility, mitochondrial function and subsequent embryo development. Reproduction Fertility and Development, 2022 May . doi: 10.1071/RD21371.

Wang Y*, Yu L, Li J, Zhu L, Ming H, Pinzon-Arteaga C, Sun H, Wu J^#, Jiang Z^#. Establishment of bovine trophoblast stem cells. bioRxiv. 2022.12.20. doi: https://doi.org/10.1101/2022.12.21.521294.

Pinzon-Arteaga C*, Wang Y*, Wei Y, Li L, Orsi A, Scatolin G, Liu L, Sakurai M, Ye J, Yu L, Li B, Jiang Z^#, Wu J^#. Bovine blastocyst like structures derived from stem cell cultures. bioRxiv, 2022.12.20. doi: https://doi.org/10.1101/2022.12.20.521301.

Zhu L, Zhou T, Iyyappan R, Ming H, Wang Y, Dvoran M, Chen Q, Roberts RM, Susor A, Jiang Z^#. High-resolution ribosome profiling reveals translational selectivity for transcripts in bovine preimplantation embryo development. Development, 2022. PMID: 36227586.

Dodlapati S, Jiang Z, Sun J. Completing single-cell DNA methylome profiles via transfer learning together with KL-divergence. Front Genet. 2022. PMID: 35938031.

Llano ED, Iyyappan R, Aleshkina D, Dvoran M, Jiang Z, Kubelka M, Susor A. SGK1 is essential for meiotic resumption in mammalian oocytes. European Journal of Cell Biology. 2022 Feb 25. PMID: 35240557.

Smith R, Susor A, Ming Hao, Tait J, Conti M, Jiang Z^#, Lin CJ^#. The H3.3 chaperone Hira complex orchestrates oocyte developmental competence. Development. 2022 Feb 03. PMID: 35112132

Jackson, K.S.  Fukuda^, M.N., Nowak^, R.A., Wheeler, M.B. 2021. Trophinin expression and regulation in the porcine uterus and embryo during the time of embryo implantation. Adv Anim Biotechnol 1(1):1-8.

Bernadette A. D'Alonzo, Abigail C. Bretzin, Douglas J. Wiebe, Ivy League–BigTen Epidemiology of Concussion Study Investigators: Russell Fiore, MEd, ATC, Bryn    VanPatten, PhD, MSEd, ATC (Brown University) William N. Levine, MD, and Natasha Desai, MD (Columbia University) David C. Wentzel, DO, and Amy Sucheski-Drake, MD (Cornell University) Kristine A. Karlson, MD (Dartmouth College) Frank Wang, MD, and Lars Richardson, MD, PhD (Harvard University) Nicholas L. Port, PhD (Indiana University) Mathew Saffarian, DO, (Michigan State University) Brian Vesci, MA, ATC (Northwestern University) Michael Gay, PhD, ATC (Pennsylvania State University) Carly Day, MD (Purdue University) Margot Putukian, MD (Princeton University) Carrie Esopenko, PhD (Rutgers University) Matthew B. Wheeler, PhD, and Randy A. Ballard, ATC (University of Illinois) Andy Peterson, MD, MSPH (University of Iowa) David Klossner, PhD (University of Maryland) Erin M. Moore, MEd, ATC (University of Minnesota) Art Maerlender, PhD, and Cary R Savage, PhD (University of Nebraska-Lincoln) BrianJ. Sennett, MD (University of Pennsylvania) Stephanie Arlis-Mayor, MD (Yale University, Arthur C. Maerlender, Cary R. Savage. 2022.  The Role of Reported Affective Symptoms and Anxiety in Recovery Trajectories Following Sport-related   Concussion. The American Journal of Sports Medicine (2022), 13 pp. DOI:10.1177/03635465221098112 

Douglas J Wiebe   Abigail C Bretzin, Bernadette A D’Alonzo, the Ivy League–Big Ten Epidemiology of Concussion Study Investigators: Russell Fiore, MEd, ATC, and Bryn VanPatten, PhD, MSEd, ATC (Brown University) William N. Levine, MD, and Natasha Desai, MD (Columbia University) David C. Wentzel, DO, and Amy Sucheski-Drake, MD (Cornell University) Kristine A. Karlson, MD (Dartmouth College) Frank Wang, MD, and Lars Richardson, MD, PhD (Harvard University) Nicholas L. Port, PhD (Indiana University) Mathew Saffarian, DO, (Michigan State University) Brian Vesci, MA, ATC (Northwestern University) Michael Gay, PhD, ATC (Pennsylvania State University) Carly Day, MD (Purdue University) Margot Putukian, MD (Princeton University) Carrie Esopenko, PhD (Rutgers University) Matthew B. Wheeler, PhD, and Randy A. Ballard, ATC (University of Illinois) Andy Peterson, MD, MSPH (University of Iowa) David Klossner, PhD (University of Maryland) Erin M. Moore, MEd, ATC (University of Minnesota) Art Maerlender, PhD, and Cary R Savage, PhD (University of Nebraska-Lincoln) Brian J. Sennett, MD (University of Pennsylvania) Stephanie Arlis-Mayor, MD (Yale University). 2022. Progression through return-to-sport and return-to-academics guidelines for concussion management and recovery in collegiate student-athletes: findings from the Ivy League–Big Ten Epidemiology of Concussion Study. Br J Sports Med 2022; 0:1–12. doi:10.1136/bjsports-2021-104451.

Stella, S.,Velasco-Acosta, D., Ferreira, C., Rubessa, M., Wheeler, M.B., Luchini, D., Cardoso, F. (2022). Effects of Rumen-Protected Methionine on Lipid Profiles of the Preimplantation Embryo and Endometrial Tissue of Holstein Cows. Res. Vet. Sci. RVSC-D-22-00543, http://dx.doi.org/10.2139/ssrn.4152859

Ho, N.C., Hollister, S., Agrawal, V., Flanagan, C. Lee, C., Wheeler, M.B., Wang,H., Ebramzadeh, E., Sangiorgio, S. Evaluation of topology optimization using 3D printing for bioresorbable fusion cages: A biomechanical study in a porcine model. Spine: September 20, 2022 - Volume - Issue - 10.1097/BRS.0000000000004491,doi:10.1097/BRS.0000000000004491

Gutierrez-Castillo, E., F. A. Diaz, S. A. Talbot, and K. R. Bondioli. 2022. Recovery of spindle morphology and mitochondrial function through extended culture after vitrification-warming of bovine oocytes. Theriogenology 189: 192-198.

Girka, E., L. Gatenby, E. J. Gutierrez, and K. R. Bondioli. 2022. The effects of microtubule stabilizing and recovery agents on vitrified bovine oocytes. Theriogenology 182: 9-16.

Lugar DJ, Sriram G. Isotope-assisted metabolic flux analysis as an equality-constrained nonlinear program for improved scalability and robustness. PLOS Computational Biology. 2022;18: e1009831. doi:10.1371/journal.pcbi.1009831

Samuel Gebremedhn, Ahmed Gad, Ghassan M. Ishak, Nico G. Menjivar, Melba O. Gastal, Jean M. Feugang, Radek Prochazka, Dawit Tesfaye, Eduardo L. Gastal. Dynamics of extracellular vesicle-coupled microRNAs in equine follicular fluid associated with follicle selection and ovulation. Molecular Human Reproduction. Accepted February 2023.

Dlamini, N.H., Nguyen, T., Gad, A., Tesfaye, D., Liao, S.F., Willard, S.T., Ryan, P.L. and Feugang, J.M. Characterization of Extracellular Vesicle-Coupled miRNA Profiles in Seminal Plasma of Boars with Divergent Semen Quality Status. International Journal of Molecular Sciences, 24(4), p.3194. 2023.

Feugang, Jean M., Ghassan M. Ishak, Matthew W. Eggert, Robert D. Arnold, Orion S. Rivers, Scott T. Willard, Peter L. Ryan, and Eduardo L. Gastal. Intrafollicular injection of nanomolecules for advancing knowledge on folliculogenesis in livestock. Theriogenology 192: 132-140. 2022.

Ishak GM, Feugang JM, Pechanova O, Pechan T, Peterson DG, Willard ST, Ryan PL, Gastal EL. Follicular‐fluid proteomics during equine follicle development. Molecular Reproduction and Development 89 (7): 298-311, 2022.

Balboula, A.Z., M. Aboelenain, M. Sakatani, K. Yamanaka, H. Bai, T. Shirozu, M. Kawahara, A.O. Hegab, S.M. Zaabel, M. Takahashi. 2022. Effect of E-64 Supplementation during In Vitro Maturation on the Developmental Competence of Bovine OPU-Derived Oocytes. Genes, 13 (2), 324-333.

Ho, K.T., A.Z. Balboula, K. Homma, J. Takanari, H. Bai, M. Kawahara, K. Nguyen, M. Takahashi. 2022. Synergistic effect of standardized extract of Asparagus officinalis stem and heat shock on progesterone synthesis with lipid droplets and mitochondrial function in bovine granulosa cells, J Steroid Biochem Mol Biol, 20:106181. doi:10.1016/j.jsbmb.2022.106181.

London-Vasquez D., A. Jurkevich, A.Z. Balboula. 2022. Multi-photon laser ablation of cytoplasmic MTOCs in mouse oocytes, Journal of Visualized Experiments, 11 (189). doi: 10.3791/64439.

Feng J., E.J. Soto-Moreno, A. Prakash, A.Z. Balboula, H. Qiao. 2022. Adverse PFAS effects on mouse oocyte in vitro maturation are associated with carbon chain length and sulfonate group, Cell Proliferation, e13353, doi: 10.1111/cpr.13353.

Vasquez, D.L., K. Rodriguez-Lukey, S.K. Behura, A.Z. Balboula. 2022. Microtubule organizing centers regulate meiotic spindle positioning in mouse oocyte. Developmental Cell, 57 (2), pp. 197-211.

Spooner-Harris, M., K. Kerns, M. Zigo, P. Sutovsky, A. Balboula, A.L., Patterson. 2022. A re-appraisal of mesenchymal-epithelial transition (MET) in endometrial epithelial remodeling. Cell and Tissue Research. 2022 Nov 19. doi: 10.1007/s00441-022-03711-z.

Lenis, YY., J.W., George, S. Lind, A. Balboula, J.M. Teixeira, A.L. Patterson. 2022. The Effects of Periostin Expression on Fibroid-Like Transition of Myometrial Cells. Reproductive Sciences. 2022 Nov 23. doi: 10.1007/s43032-022-01128-1. Online ahead of print.

Rakha, S., M. Elmetwally, H. El-Sheikh, A.Z. Balboula, A. Montaser, S.M. Zaabel. 2022. Lycopene Reduces the In Vitro Aging Phenotypes of Mouse Oocytes by Improving Their Oxidative Status. Veterinary Sciences, 9 (7): 336.

Rakha, S.I., M.A. Elmetwally, A.H. El-Sheikh, A.Z. Balboula, A.M. Mahmoud, S.M. Zaabel. 2022. Importance of Antioxidant Supplementation during In Vitro Maturation of Mammalian Oocytes. Veterinary Sciences, 9 (8): 439.

Chen, P.R., K. Uh, K. Monarch, L.D. Spate, E.D. Reese, R.S. Prather, K. Lee K. Inactivation of growth differentiation factor 9 blocks folliculogenesis in pigs. Biol Reprod. 2023 Jan 16;ioad005. doi: 10.1093/biolre/ioad005. Online ahead of print.

Chen, P., K. Uh, B.K. Bedel, E.D. Reese, R.S. Prather, K. Lee. Production of Pigs From Porcine Embryos Generated in vitro. Front. Anim. Sci., 16 March 2022; https://doi.org/10.3389/fanim.2022.826324

Shearn CT, Anderson AL, Devereux MW, Orlicky DJ, Michel C, Petersen DR, Miller CG, Harpavat S, Schmidt EE, Sokol RJ. The autophagic protein p62 is a target of reactive aldehydes in human and murine cholestatic liver disease. PLoS One. 2022;17(11):e0276879. doi: 10.1371/journal.pone.0276879. eCollection 2022. PubMed PMID: 36378690; PubMed Central PMCID: PMC9665405.

Stancill JS, Hansen PA, Mathison AJ, Schmidt EE, Corbett JA. Deletion of Thioredoxin Reductase Disrupts Redox Homeostasis and Impairs β-Cell Function. Function (Oxf). 2022;3(4):zqac034. doi: 10.1093/function/zqac034. eCollection 2022. PubMed PMID: 35873655; PubMed Central PMCID: PMC9301323.

Le Gal K, Schmidt EE, Sayin VI. Cellular Redox Homeostasis. Antioxidants (Basel). 2021 Aug 28;10(9). doi: 10.3390/antiox10091377. PubMed PMID: 34573009; PubMed Central PMCID: PMC8469889.

White, B.R., R.A. Cederberg, D.H. Elsken, C.E. Ross, C.A. Lents and A.T. Desaulniers. 2022. Role of gonadotropin-releasing hormone-II and its receptor in swine reproduction. Mol. Reprod. Dev. 1–11. https://doi.org/10.1002/mrd.23662. [Online ahead of print].

Desaulniers, A.T., R.A. Cederberg, C.A. Lents and B.R. White. 2022. Gonadotropin-releasing hormone II and its receptor regulate motility, morphology, and kinetics of porcine spermatozoa. Gen. Comp. Endocrinol. (Submitted).

Davies C.J.*, Z. Fan, K.P. Morgado, Y. Liu, M. Regouski, A.J. Thomas, S-I. Yun, B.-H. Song, J.C. Frank, I. V. Perisse, A. Van Wettere, Y-M. Lee, I.A. Polejaeva* (2022). Development and Characterization of Type I Interferon Receptor Knockout Sheep: A Model for Viral Immunology and Reproductive Signaling. Frontiers in Genetics 13: 986316. PMID: 36246651.

Adams L., Y. Liu, B. Durrant, E. Ruggeri, C. Young, R. Krisher, I. Polejaeva* (2022) Use of Bisection to Reduce Mitochondrial DNA in the Bovine Oocyte. JoVE. PMID: 35876541.

Gao, Y., L. Zhao, J.S. Son, X. Liu, Y. Chen, M. J. Deavilla, M. J. Zhu, G. Murdoch, and M. Du. 2022. Maternal exercise before and during pregnancy facilitates embryonic myogenesis by enhancing thyroid hormone signaling. Thyroid. 32: 581-593.

Zhao, L. X. Liu, N. A. Gomez, Y. Gao, J. S. Son, S. A. Chae, M. J. Zhu, and M. Du. 2023. Stage-specific nutritional management and developmental programming to optimize meat production. Journal of Animal Science and Technology. 14: 2.

2. Books, non-refereed book chapters, proceedings, instructional media, theses/dissertations

Thesis Title: "CRISPR Cas9 Targeted Single Nucleotide Change at the (+15) Loci of the Alpha-Lactalbumin Gene. Sierra Long, M.S., July 2022.

Gutierrez-Castillo, E.  Vitrification of bovine oocytes and embryos. Louisiana State University Dissertation. 2022.

Mustapha A Popoola, Suleiman E Bogoro, Jean M Feugang. Biothermoimaging tools for management of climate smart and precision livestock-assisted reproduction. In “Agricultural Biotechnology, Biodiversity and Bioresources conservation and utilization.” (pp. 315-334). 1st Edition, CRC Press, eBook ISBN 9781003178880. https://doi.org/10.1201/9781003178880, 2022.

3. Abstracts

W E Weber, J G Powell, B P Littlejohn, E B Kegley, N N Noga, C R Looney. 2022 Evaluation of Fixed-time Artificial Insemination and Estrus Expression in Angus-cross Heifers with Female Sex-sorted Semen Using 6-day and 7-day CIDR Protocols at Multiple Time Intervals. Journal of Animal Science, Volume 100, Issue Supplement1, April 2022, Page 10, https://doi.org/10.1093/jas/skac028.018

W E Weber, J G Powell, B P Littlejohn, B W Krumpelman, N T Moss, M S Gadberry, C R Looney.  2022. Evaluation of GnRH Administration day in the 7 & 7 Synch Protocol for Fixed-Timed Artificial Insemination in Multiparous Beef Cows. Journal of Animal Science, Volume 100, Issue Supplement3, October 2022, Page 244, https://doi.org/10.1093/jas/skac247.440

2022. E. Weber, J. G. Powell, B. P. Littlejohn, B. W. Krumplman, J. D. Rivera, W. L. Rook, M. S. Gadberry, N. T. Moss, C. R. Looney^. Evaluation of the 7 & 7 Synch protocol on GnRH administration day for FTAI in multiparous beef cows. AETA-CETA/ACTE Joint Annual Convention, Louisville, KY in October 2022. Meeting Proceedings pg. 96
2023. E. Weber, J. G. Powell, B. P. Littlejohn, B.W. Krumplman, N. T. Moss, M. S. Gadberry, C. R. Looney. Evaluation of GnRH administration day in the 7 & 7 Synch protocol for fixed-timed artificial insemination in multiparous beef cows.^.ARSBC Lauderdale Scholar Poster Presentation, San Antonio, TX, July 2022.

Guiltinan C, Weldon BM, Soto DA, Ross PJ, Denicol AC. Single-cell transcriptome analysis of fetal and adult bovine ovaries reveals developmental progression in cell population composition and function. Reprod, Fert Dev 35(1-2), 2023. https://www.publish.csiro.au/RD/pdf/RDv35n2abs.

Snow A, Donovan A, Winger QA. Investigating the role of thyroid hormone in ovine placenta using trophoblast cell lines. National Veterinary Scholars Symposium, Minneapolis, MN, USA.

Donovan A, Anthony RV, Winger QA. Investigating Transthyretin and Thyroid Hormones in CSH Knockdown IUGR Pregnancies. Society for the Study of Reproduction 55th Annual Meeting, Spokane, WA, USA.

Tanner A.R., V. C. Kennedy, C. S. Lynch, Q. A. Winger, P. J. Rozance, R. V.  Anthony. Maternal Hyperglycemia Increases Uteroplacental Oxygen Utilization but Fails to Restore Fetal Glucose Concentrations in Chorionic Somatomammotropin RNA Interference Pregnancies. Reprod Sci 2022; Society for Reproductive Investigation 69th Annual Scientific Meeting, Denver, CO.

Tanner A.R., V. C. Kennedy, C. S. Lynch, Q. A. Winger, P. J. Rozance, R. V.       Anthony. Varying Degrees of Fetal Growth Restriction in Chorionic Somatomammotropin RNA Interference Pregnancies. Reprod Sci 2022 Society for Reproductive Investigation 69th Annual Scientific Meeting, Denver, CO.

Lynch C.S., A. Ali, V. C. Kennedy, A. R. Tanner, Q. A. Winger, P. J. Rozance,  R. V.   Anthony. Assessing the Impact of Placental SCL2A3 RNA Interference at Mid-Gestation. Reprod Sci 2022; Society for Reproductive Investigation 69th Annual Scientific Meeting, Denver, CO.

Hord T.K., A. R. Tanner, C.S. Lynch, V. C. Kennedy, Q. A. Winger, P. J.    Rozance, R. V. Anthony. Placental Expression of the IGF Axis Following Chorionic Somatomammotropin RNA Interference. Reprod Sci 2022; Society for Reproductive Investigation 69th Annual Scientific Meeting, Denver, CO.

Menjivar N.G., Gad A., Thompson R.E., Meyers M.A., Hollinshead F.K., Tesfaye D. (2022) Bovine Oviductal Organoids: Cellular and Extracellular Response to Heat Stress. Graduate Student Showcase 2022; Page XX. Fort Collins, CO, USA.

Tesfaye D., Menjivar N.G., Gad A. Next generation miRNA loading into extracellular vesicles: Potential application for EV-coupled miRNA functional studies. BMS Retreat 2022; Page 29. Estes Park, CO, USA.

Menjivar N. G., Gebremedhn S., Gad A., Tesfaye D. (2022) Emerging role of extracellular vesicles as messengers for protection against heat stress in bovine oocytes. Society for the Study of Reproduction 2022 (SSR); 55^th Annual Meeting. Spokane, WA, USA.

Gad A., Gebremedhn S., Menjivar N., Feugang J., Ishak G., Prochazka R., Tesfaye D., Gastal E. (2022) Dynamics of Extracellular vesicle-coupled microRNAs in equine follicular fluid associated with follicular selection. Society for the Study of Reproduction 2022 (SSR); 55^th Annual Meeting. Spokane, WA, USA.

Kamryn Joyce, Ahmed Gad, Nico  G Menjivar, Daniella C Heredia, Camila Santos Rojas, Dawit Tesfaye, Angela M Gonella, PSIII-17 Transcriptome Response of Granulosa Cells to Seasonal Heat Stress in Beef Cows, Journal of Animal Science. American Society of Animal Sciences 2022 (ASAS-CSAS-SSASAS) Annual Meeting & Trade Show; Volume 100,Issue Supplement3, October 2022, Pages 242-243, Oklahoma City, OK, USA. https://doi.org/10.1093/jas/skac247.438

Menjivar N. G., Gebremedhn S., Tesfaye D. (2022) Impact of supplementation of granulosa cell-derived extracellular vesicles during bovine oocyte maturation under thermal stress conditions. International Congress on Animal Reproduction 2022 (ICAR); 19^th International Conference. Bologna, Italy.

Menjivar N. G., Gebremedhn S., Gad A., Tesfaye D. Next generation miRNA loading system into extracellular vesicles: Potential application for EV-coupled miRNA functional studies. Rocky Mountain Reproductive Sciences Symposium 2022 (RMRSS); 15^th Annual Conference: Page 25. Fort Collins, CO, USA.

Gad A., Joyce K., Menjivar N., Heredia D., Rojas C., Gonella-Diaza A., Tesfaye D. Extracellular vesicle-coupled microRNAs in the follicular fluids of heat-stressed beef cows. Rocky Mountain Reproductive Sciences Symposium 2022 (RMRSS); 15^th Annual Conference: Page 14. Fort Collins, CO, USA.

Menjivar N. G., Gebremedhn S., Gad A., Krisher R., Yuan Y., Tesfaye D. (2022) Interspecies Translation: Bovine extracellular vesicles impact on human cumulus cells. CSU Demo Day 2022; Annual Showcase. Fort Collins, CO, USA.

Ranjitkar S, Shiri M, Sun J, Liu G, Tian XC. 2023 Transcription read-through in in vivo developed bovine oocytes and pre-implantation embryos. Poster at the 49th annual meeting of the International Embryo Technology Society, January 16-19, 2023, Lima, Peru.

Salman S, Gungor O, Ranjitkar, S, Zhang D, Oberhaus E, Tian XC. 2023. Hormonal profiles of bovine follicular fluid during the window of in vivo oocyte maturation. Poster at the 49th annual meeting of the International Embryo Technology Society, January 16-19, 2023, Lima, Peru.

Losano JDA, Daigneault BW. Environmental modulation of sperm energetics by non-canonical PPARʏ signaling. Society for the Study of Reproduction 55th Annual Meeting, Spokane, WA 2022.

P- Losano JDA, Daigneault BW. Detection of novel bioenergetic traits in bovine sperm for improved semen quality analyses. Proceedings of the Eighth UF/IFAS Animal Sciences Symposium, St. Augustine, FL 2022.

G-McGraw M, Daigneault BW. Functional characterization of PPAR