bluuebunny/crossref_metadata_2025 huggingface dataset:
Metastasiertes Pankreaskarzinom
Authors: Schmale Ine
Published: December 2019

Etwa 4–7 % der Patienten mit metastasiertem Pankreaskarzinom sind Träger einer BRCA1/2-Keimbahnmutation. Diese Patienten profitieren von einer Platin-basierten Chemotherapie. In der 3:2-randomisierten, Placebo-kontrollierten Phase-III-Studie POLO wurde untersucht, ob mit einer PARP-Inhibitor-Erhaltungstherapie der Tumorprogress nach Chemotherapie hinausgezögert werden kann. Primärer Studienendpunkt war das progressionsfreie Überleben (PFS) nach RECIST v1.1-Kriterien. Die Ergebnisse der Studie wurden in der Plenarsitzung beim Jahreskongress der ASCO präsentiert.
Compression Garment-induced Leg Changes Increase Hemodynamic Responses in Healthy Individuals
Authors: Lee Daniel C. W. , Law Helen Ka Wai , Ali Ajmol , Sheridan Sinead E. , Wong Stephen H. S. , Lee Shara Wee Yee
Published: December 2019

This study evaluated the morphological changes of the lower limb and associated hemodynamic responses to different lower-body compression pressures (COMPs) in physically active, healthy individuals at rest. Each of the 32 participants underwent three trials with three different degrees of lower-body compression applied: “Low” (2.2±1.4 mmHg), “Medium” (12.9±3.9 mmHg), and “High” (28.8±8.3 mmHg). In each COMP, a cross-sectional area of leg muscles (CSAmuscle), subcutaneous fat (CSAfat), superficial vessels (SupV), deep arteries (DA), and deep veins (DV) at the calf, knee, and thigh levels were measured using magnetic resonance imaging (MRI). Additionally, blood pressure (BP), heart rate (HR), cardiac output (CO), stroke volume (SV), and systemic vascular resistance (SVR) were measured using Doppler ultrasound (USCOM®). With High COMP, calf CSAmuscle and SupV were smaller (p<0.01), whereas DA and DV were larger (p<0.05). Calf CSAfat, however, was similar among all COMPs. There were no major changes in CSAmuscle and CSAfat at knee and thigh levels. CO (3.2±0.9 L/min) and SV (51.9±16.4 mL) were higher (p<0.05) only with High COMP, but other hemodynamic variables showed no significant changes across different COMPs. The High COMP at the lower limb induces leg morphological changes and increases associated hemodynamic responses of physically active healthy individuals at rest.
Safety and efficacy of endoscopic submucosal dissection for rectal neoplasia: a multicenter North American experience
Authors: Yang Dennis , Aihara Hiroyuki , Perbtani Yaseen B. , Wang Andrew Y. , Aadam Abdul Aziz , Tomizawa Yutaka , Hwang Joo Ha , Zou Baiming , Natov Nikola S. , Siegel Amanda , Khoshknab Milad Pourmousavi , Khashab Mouen A. , Ngamruengphong Saowanee , Khara Harshit S. , Diehl David L. , Maniere Thibaut , Andrawes Sherif , Benias Petros , Kumta Nikhil A. , Ramay Fariha , Kim Raymond E. , Samarasena Jason , Chang Kenneth , Hashimoto Rintaro , Tharian Benjamin , Inamdar Sumant , Lan Gloria , Sethi Amrita , Nosler Michael J. , Tabash Abdalaziz , Othman Mohamed O. , Draganov Peter V.
Published: December 2019

Background and aims Rectal lesions traditionally represent the first lesions approached during endoscopic submucosal dissection (ESD) training in the West. We evaluated the safety and efficacy of rectal ESD in North America.

Methods This is a multicenter retrospective analysis of rectal ESD between January 2010 and September 2018 in 15 centers. End points included: rates of en bloc resection, R0 resection, adverse events, comparison of pre- and post-ESD histology, and factors associated with failed resection.

Results In total, 171 patients (median age 63 years; 56 % men) underwent rectal ESD (median size 43 mm). En bloc resection was achieved in 141 cases (82.5 %; 95 %CI 76.8–88.2), including 24 of 27 (88.9 %) with prior failed endoscopic mucosal resection (EMR). R0 resection rate was 74.9 % (95 %CI 68.4–81.4). Post-ESD bleeding and perforation occurred in 4 (2.3 %) and 7 (4.1 %), respectively. Covert submucosal invasive cancer (SMIC) was identified in 8.6 % of post-ESD specimens. There was one case (1/120; 0.8 %) of recurrence at a median follow-up of 31 weeks; IQR: 19–76 weeks). Older age and higher body mass index (BMI) were predictors of failed R0 resection, whereas submucosal fibrosis was associated with a higher likelihood of both failed en bloc and R0 resection.

Conclusion Rectal ESD in North America is safe and is associated with high en bloc and R0 resection rates. The presence of submucosal fibrosis was the main predictor of failed en bloc and R0 resection. ESD can be considered for select rectal lesions, and serves not only to establish a definitive tissue diagnosis but also to provide curative resection for lesions with covert advanced disease.
Echocardiographic Assessment of Left Ventricular Remodeling in American Style Footballers
Authors: Moneghetti Kegan James , Singh Tamanna , Hedman Kristofer , Christle Jeffrey W. , Kooreman Zoe , Kobayashi Yukari , Bouajila Sara , Amsallem Myriam , Wheeler Matthew , Gerche Andre La , Ashley Euan , Haddad Francois
Published: December 2019

Several athletic programs incorporate echocardiography during pre-participation screening of American Style Football (ASF) players with great variability in reported echocardiographic values. Pre-participation screening was performed in National Collegiate Athletic Association Division I ASF players from 2008 to 2016 at the Division of Sports Cardiology. The echocardiographic protocol focused on left ventricular (LV) mass, mass-to-volume ratio, sphericity, ejection fraction, and longitudinal Lagrangian strain. LV mass was calculated using the area-length method in end-diastole and end-systole. A total of two hundred and thirty players were included (18±1 years, 57% were Caucasian, body mass index 29±4 kg/m2) after four players (2%) were excluded for pathological findings. Although there was no difference in indexed LV mass by race (Caucasian 78±11 vs. African American 81±10 g/m2, p=0.089) or sphericity (Caucasian 1.81±0.13 vs. African American 1.78±0.14, p=0.130), the mass-to-volume ratio was higher in African Americans (0.91±0.09 vs. 0.83±0.08, p<0.001). No race-specific differences were noted in LV longitudinal Lagrangian strain. Player position appeared to have a limited role in defining LV remodeling. In conclusion, significant echocardiographic differences were observed in mass-to-volume ratio between African American and Caucasian players. These demographics should be considered as part of pre-participation screening.
Interventional Cryoneurolysis: What Is the Same, What Is Different, What Is New?
Authors: Bittman Ross W. , Behbahani Keywan , Gonzalez Felix , Prologo J. David
Published: December 2019

Cryoneurolysis is the deliberate application of cold temperatures to nerves for therapeutic purposes. The idea of treating pain with this technique is thousands of years old and has evolved over time through the application of surgical techniques, nerve stimulation and/or landmark guidance, and through device development. Recent integration of the interventional radiology skill set to this space has unlocked a myriad of opportunities—primarily through a unique ability to percutaneously access deep structures in the body with accuracy and precision, and the capacity to monitor ablation zones. Understanding of the specific neurohistological process that follows targeted cryoneurolysis leads to new options for treating patients in pain without drugs and opens doors for the potential modification of a wide array of disease states.
Lean and Green?: Wie kann der Einkauf die Nachhaltigkeit im Krankenhaus unterstützen?
Authors: Schlüchtermann Jörg , Bächer Franziska
Published: December 2019

Auch Kliniken müssen sich schrittweise zu nachhaltigen Unternehmen wandeln. Im Klinikeinkauf sind Ökologie und Wirtschaftlichkeit heute oft keine Gegensätze mehr. Mit einer verkleinerten Produktpalette und reduziertem Lieferantenportfolio lassen sich Emissionen und Kosten senken. Gerade in der langfristigen Perspektive ergeben sich interessante Weiterentwicklungsmöglichkeiten, Nachhaltigkeit zu einem Wettbewerbsfaktor zu machen.
Flow control on wind turbine airfoil affected by the surface roughness using leading-edge protuberance
Authors: Zhang Yinan , Zhang Mingming , Cai Chang
Published: December 2019

In recent years, leading-edge protuberances have attracted plenty of attention as a passive method to manipulate flow separation on wind turbine blades. In this paper, an experimental investigation is carried out to explore the effect of leading-edge protuberances on the aerodynamic performance of wind turbine airfoils with and without surface roughness. A direct force measurement technique with the application of three-component balance is applied, and a hot-wire anemometer is used to measure the boundary layer flow to understand the governing mechanism for the flow regulation of the leading-edge protuberances. The current results confirm that the surface roughness leads to a decreased lift-to-drag ratio and an increased aerodynamic fluctuation of the baseline airfoil, which may cause severe losses in wind power capture. For the wavy airfoil, the leading-edge protuberance has facilitated the suppression of flow separation and the improvement of the airfoil aerodynamic performance in the poststall region. The results of the velocity measurement indicate that the leading-edge protuberances weaken the vortex shedding on the airfoil, which has a significant impact on the fluctuation of the aerodynamic force. For airfoils with a rough surface, the vortex shedding energy close to the trailing edge is evidently weakened for the wavy airfoil, which contributes effectively to the suppression of the shedding of the separated vortex and improve the stability of the aerodynamic force, compared to the baseline case. The effectiveness of leading-edge protuberances in the improvement of the performance stability may facilitate the service life of the blade and the ability of wind power capture.
Contrasted Sn substitution effects on Dirac line node semimetals SrIrO3 and CaIrO3
Authors: Negishi Masamichi , Hiraoka Naoka , Nishio-Hamane Daisuke , Takagi Hidenori
Published: December 2019

Perovskite-type iridates SrIrO3 and CaIrO3 are a Dirac line node semimetal protected by crystalline symmetry, providing an interesting playground to investigate electron correlation effects on topological semimetals. The effect of Sn doping was examined by growing SrIr1−xSnxO3 and CaIr1−xSnxO3 thin films epitaxially on the SrTiO3(001) substrate using pulsed laser deposition. Upon Sn doping, the semimetallic ground state switches into an insulator. As temperature is lowered, the resistivity, ρ(T), of SrIr1−xSnxO3 above a critical doping level (xc ∼ 0.1) shows a well-defined transition from the semimetal to a weakly ferromagnetic insulator at T = Tc. In contrast, ρ(T) of CaIr1−xSnxO3 with increasing x shows a rapid increase in magnitude but does not show a clear signature of metal-insulator transition in the temperature dependence. We argue that the contrasted behavior of the two closely related iridates reflects the interplay between the effects of electron correlation and disorder enhanced by Sn doping.
Tuning structural, electronic, and magnetic properties of black-AsP monolayer by adatom adsorptions: A first principles study
Authors: Liu Xin , Ni Yu-xiang , Wang Hong-yan , Wang Hui
Published: December 2019

Black Arsenic-phosphorus (AsP) monolayer is a novel two-dimensional nanomaterial with the characteristics of modest direct bandgap and superhigh carrier mobility. However, little is known about how the surface adsorption affects the property of AsP monolayer. Motivated by this, we researched systematically the geometry, adsorption energy, magnetic moment and electronic structure of 11 different adatoms adsorbed on AsP monolayer using first-principles calculations. The adatoms used in this study include light nonmetallic (C, N, O) adatoms, period-3 metal (Na, Mg, Al) adatoms, and transition-metal (Ti, V, Cr, Mn, and Fe) adatoms. The adatoms cause an abundant variety of structural, magnetic and electronic properties. This study shows that AsP binds strongly with all adatoms under study and the adsorption energies in all systems are much stronger than that on graphene, SiC, BN, or MoS2. The semiconductor property of AsP is affected by the introduction of adsorbed atoms, which can induce mid-gap states or cause n-type doping. Moreover, the adatom adsorptions cause various spintronic characteristics: N-, Ti-, and Fe-adsorbed AsP become bipolar semiconductors, while the Mn-decorated AsP becomes a bipolar spin-gapless semiconductor. Our results suggest that atomic adsorption on AsP monolayers has potential application in the field of nanoelectronics and spintronics.
Infrared spectroscopy of neutral and cationic pyrrolidine monomer in supersonic jet
Authors: Xie Min , Zhang Zhao-li , Zhang Yu , Sun Xiao-nan , Sun Fu-fei , Hu Yong-jun
Published: December 2019

Pyrrolidine, a five membered heterocyclic molecule, is widely existing in organism. Herein, infrared spectra of pyrrolidine monomer in neutral and cationic states were obtained by vacuum ultraviolet ionization, infrared photodissociation and time of flight mass spectrometry. Both in neutral and cationic states, it is found that their CH stretching vibration bands are red shifted. In the IR spectrum of neutral pyrrolidine, because the electric dipole moment of NH is small, we have not observed the NH stretching vibration bands. However, the NH stretching vibration band of pyrrolidine is greatly enhanced after ionization, and this band red-shifts compared with the previous experiment. The red shifts of CH stretching vibrations in neutral and cationic states are caused by the negative and positive hypercon-jugation, respectively. The enhancement and red shift of the NH stretching band are owing to the ejection of the electrons on the N atom after ionization. Through the calculations, it is found that the acidity of the CH bond is a little stronger than that of NH bond. These kinds of studies would be helpful to understand the intrinsic properties of biomolecules in neutral and cationic states, and to provide reference for the further study of living organic macromolecules.
Self-driving cars face a cloudy future
Authors: Lopatka Alex
Published: December 2019

Better weather predictions and more data on driving could accelerate the widespread deployment of automated and connected vehicles.
Strengthening teamwork capability in allied health: implementation of a team development program in a metropolitan health service
Authors: Wood Angela J. , Grudzinskas Kathy , Ross Julie-Anne , Bailey Sarah , Gordon Gail E. , Burton Christopher , Wishart Laurelie R.
Published: December 2019

ObjectiveCollaborative practice is critical to optimising patient outcomes in contemporary healthcare settings. Evidence suggests interprofessional learning is an effective way to develop teamwork capabilities, yet these skills are traditionally developed in professional silos, or not at all. This study evaluated the implementation of a team development program, the Team Management Systems (TMS) program, for allied health staff within a large metropolitan health service. MethodsA mixed-methods audit–quality improvement study was conducted, using Kirkpatrick’s four-level evaluation model to structure evaluation of the program. Semistructured questionnaire and workforce survey data were retrieved immediately, 6 months and 1–2 years after training and applied to each level of the model (Reaction, Learning, Behaviour, Results). ResultsIn all, 886 staff participated in the TMS program from 2014 to 2018. High satisfaction with the program was observed. Knowledge of what constitutes effective teamwork improved significantly (P=0.008) in TMS participants compared with a matched untrained cohort. Participants reported positive behaviour change and continued engagement with TMS principles 6 months after training. Perceived impact of the program on patient and/or organisational outcomes was evident, although less compelling than the changes to knowledge and behaviour. ConclusionsThe TMS program yielded positive effects on staff satisfaction, knowledge, team dynamics and team behaviours. These findings demonstrate the significant value of such initiatives to enhance the capability and effectiveness of interdisciplinary healthcare teams. What is known about the topic?Complex conditions, increasing comorbidities, specialisation and scarcity of resources mean healthcare workers need to work effectively in teams to achieve quality, safe, person centred patient care. There is some evidence of the effect of teamwork initiatives on knowledge or behaviour in specific clinical specialities, single services or single professions, but limited research is available regarding the effects of teamwork programs across multiple professions, including allied health professions, and on patient and organisational outcomes. What does this paper add?This paper describes the effect of a large-scale teamwork program implemented across multiple professions, including enablers and barriers. It presents outcomes at all four levels of Kirkpatrick’s evaluation model, including the less studied behaviour and results levels. What are the implications for practitioners?This paper supports health service leaders to consider developing and implementing interprofessional teamwork programs to foster essential teamwork capabilities. Learning together about teamwork, across professional silos, will lead to collaborative, patient-centred care, which leads to safe, quality patient outcomes.
In vitro growth of immature bovine follicles and oocytes
Authors: Telfer Evelyn E. , Sakaguchi Kenichiro , Clarkson Yvonne L. , McLaughlin Marie
Published: December 2019

The limitation in the supply of mature, fertilisable oocytes constitutes a major impediment to increasing the success of assisted reproduction, stem cell derivation and cloning in domestic species. Techniques are being developed to grow immature oocytes invitro that have the potential to increase the supply of oocytes. Mouse oocytes can be cultured from initial stages of development to maturity, and live young have been produced, but for domestic species, such as cows, with long growth periods, invitro systems that allow complete growth of oocytes contained within primordial follicles to maturity is technically challenging and has not yet been achieved. For cows, several culture systems have been developed that support specific developmental stages, but a multistep culture system will be required for complete growth invitro. This review highlights the steps that will be required to achieve the goal of growing oocytes invitro.
Prospects for new oocyte-based assisted reproduction in animals and humans
Authors: Albertini David F.
Published: December 2019

Procuring high-quality oocytes is the rate-limiting step for assisted reproduction technologies intended for embryo production. Although much is known about the intraovarian processes that dictate oocyte growth and maturation, subtleties in the process of oogenesis have yet to be replicated in invitro systems. In contrast with the mouse, in which functional oocytes have been derived from stem cells under ex vivo conditions, the generation of developmentally competent oocytes in other species has yet to be achieved. This paper reviews the principles and practices based on stem cell and organ culture strategies that hold promise for developing a technological base upon which future efforts to recapitulate or augment oogenesis in mammals could be realised.
Livestock pluripotency is finally captured in vitro
Authors: Navarro Micaela , Soto Delia A. , Pinzon Carlos A. , Wu Jun , Ross Pablo J.
Published: December 2019

Pluripotent stem cells (PSCs) have demonstrated great utility in improving our understanding of mammalian development and continue to revolutionise regenerative medicine. Thanks to the improved understanding of pluripotency in mice and humans, it has recently become feasible to generate stable livestock PSCs. Although it is unlikely that livestock PSCs will be used for similar applications as their murine and human counterparts, new exciting applications that could greatly advance animal agriculture are being developed, including the use of PSCs for complex genome editing, cellular agriculture, gamete generation and invitro breeding schemes.
Consequences of assisted reproductive techniques on the embryonic epigenome in cattle
Authors: Rivera Rocío Melissa
Published: December 2019

Procedures used in assisted reproduction have been under constant scrutiny since their inception with the goal of improving the number and quality of embryos produced. However, invitro production of embryos is not without complications because many fertilised oocytes fail to become blastocysts, and even those that do often differ in the genetic output compared with their invivo counterparts. Thus only a portion of those transferred complete normal fetal development. An unwanted consequence of bovine assisted reproductive technology (ART) is the induction of a syndrome characterised by fetal overgrowth and placental abnormalities, namely large offspring syndrome; a condition associated with inappropriate control of the epigenome. Epigenetics is the study of chromatin and its effects on genetic output. Establishment and maintenance of epigenetic marks during gametogenesis and embryogenesis is imperative for the maintenance of cell identity and function. ARTs are implemented during times of vast epigenetic reprogramming; as a result, many studies have identified ART-induced deviations in epigenetic regulation in mammalian gametes and embryos. This review describes the various layers of epigenetic regulation and discusses findings pertaining to the effects of ART on the epigenome of bovine gametes and the preimplantation embryo.
Factors affecting embryo production in superovulated Bos taurus cattle
Authors: Mikkola M. , Hasler J. F. , Taponen J.
Published: December 2019

Despite a long history of bovine superovulation research, significant commercial applications did not start until the early 1970s. For some 20 years thereafter, superovulation represented the primary tool for the production of cattle embryos. In the early 1990s, commercial invitro production (IVP) was initiated in cattle. Although ovum pick-up and IVP are now commercially practiced on a wide scale, superovulation and embryo recovery by flushing remain a widespread and very effective approach to the production of cattle embryos. This review covers both the history and the effects of multiple factors on superovulation in Bos taurus cattle. There are three general protocols for suitable pre-FSH programming of donors so that gonadotrophin-responsive follicles are available. Superovulation protocols vary widely based on the FSH source, the diluent used, the number and timing of FSH injections and the timing and utilisation of various prostaglandins, controlled internal progesterone releasing devices, gonadotrophin-releasing hormone, and other means of controlling follicular development and ovulation. The number of oocytes that can be stimulated to grow and ovulate within any given donor can be estimated by either ultrasound-guided sonography or by measuring concentrations of anti-Müllerian hormone in the blood. Animal-related factors that can influence the efficacy of superovulation include cattle breed, age, parity, genetics, lactational status and reproductive history. In addition, nutrition, stress, season, climate, weather and several semen factors are discussed.
13 Invitro and invivo embryo production during foal heat in a mare: A case report
Authors: Campos-Chillon L. F. , Altermatt J. L.
Published: December 2019

The objective of this case report is to describe invitro and invivo embryo production during foal heat in a mare. A 16-year-old American Quarter Horse mare was presented for breeding and nonsurgical embryo recovery days post-foaling. Upon transrectal ultrasonographic examination, the donor mare was determined to be in oestrous (foal heat) with presence of moderate uterine oedema, a relaxed cervix, absence of uterine fluid, a 35-mm dominant follicle, and multiple (n=7) subordinate follicles (10-20mm). To increase embryo production, oocytes were retrieved by ultrasound-guided ovum pickup of all subordinate follicles. Upon collection, oocytes (n=4) were placed in embryo holding medium for 18h at room temperature (22°C) and then matured in equine maturation medium (EqMat, Campos-Chillon et al. 2019 J. Equine Vet. Sci. 73, 51-55) for 30h at 38.5°C in 6% CO2, 5% O2, and 89% N2. Matured oocytes (n=3), as assessed by extrusion of the first polar body, underwent intracytoplasmic sperm injection with frozen-thawed semen and a Piezo-driven injection system. Injected oocytes were cultured in SCF1 (Owen et al. 2017 Reprod. Fertil. Dev. 29, 129-130) for 5 days and Dulbecco's modified Eagle medium/F12 plus 5% FBS for 2 days at 38.5°C in 6% CO2, 5% O2, and 89% N2. On Day 10 post-foaling, the dominant follicle was 45mm and the donor mare was inseminated with fresh semen (500×106 progressively motile spermatozoa) and ovulation was induced with 1.8mg of deslorelin IM (SucroMate, Thorn BioScience LLC). The donor mare ovulated within normal limits by 48h post-induction of ovulation (12 days post-foaling) and embryo recovery was attempted 8 days post-ovulation (20 days post-foaling). The donor mare was then administered 250μg of Cloprostenol IM (Estrumate, Merck Animal Health) to induce oestrus and was inseminated again with cooled semen (1×109 progressively motile spermatozoa) for two consecutive cycles. Maturation, cleavage, and blastocyst rates were 57% (4/7), 67% (2/3), and 33% (1/3), respectively. The nonsurgical embryo recovery resulted in one expanded blastocyst grade 1. Both invitro- and invivo-produced blastocysts were transferred into synchronized recipients 5 days after oestrus on Day 16 and 20 post-foaling respectively. The donor mare got pregnant after the second cycle post-embryo recovery attempt. Pregnancy status of recipients and donor mare were evaluated at 14, 21, 30, and 50 days after embryo transfers or insemination with normal development for the three pregnancies. In conclusion, to our knowledge, this is the first case report describing invitro and invivo embryo production performed during foal heat increasing number of pregnancies in a post-foaling mare in a single cycle.
133 CRISPR/Cas9 gene editing of invivo-fertilized bovine embryos via endoscopic oviductal flushing and electroporation of zygotes
Authors: Miskel D. , Beunink L. , Poirier M. , Havlicek V. , Rings F. , Schellander K. , Besenfelder U. , Hölker M.
Published: December 2019

In recent years, CRISPR/Cas9 has been used to efficiently edit the genomes of embryos in many animal models. Due to smaller anatomy, lower costs, and multiple ovulations, it is relatively simple to derive large numbers of invivo fertilized zygotes for gene editing experiments in small mammal models. In cattle, however, harvesting invivo fertilized zygotes generally requires a highly invasive surgical procedure. Here, we use the combination of a minimally invasive endoscopic method for harvesting invivo fertilized zygotes by oviductal flushing of superovulated heifers and the subsequent electroporation of zygotes with CRISPR/Cas9 ribonucleoproteins (RNP). After superstimulation of 21 heifers, on average 12 zygotes were flushed per animal with fetal bovine serum, then stored in synthetic oviductal fluid (SOFaa) before electroporation. Targeting exon 1 of the tyrosinase (Tyr) gene, zygotes were electroporated in 1-mm gap cuvettes (Biorad) in groups of ~20 in 20μL of OptiMEM media containing 3μM Cas9 RNP (IDT Cas9 protein pre-incubated with anti-Tyr guide RNA). Electroporation was performed in 3 replicates of 3 electrical potentials, namely 20, 25, and 30V using a Biojet CF 50. The other electroporation parameters were fixed at 5 repetitions of 2-ms square wave pulses at 100-ms intervals. The zygotes were than cultured under standard embryo culture conditions (SOFaa + 0.3% bovine serum albumin, 5% CO2, 5% O2, 39°C, humidified air). Embryo survival, cleavage, and developmental rates to the blastocyst stage were tracked. Statistical significance between groups was determined by pairwise one-way ANOVA using Sidak correction for multiple comparisons. Electroporation of invivo-derived zygotes using 20V yielded significantly higher survival (83.6% vs. 42.8% vs. 20.7% for 20, 25, and 30V, respectively), cleavage (65.6% vs. 37.9% vs. 40.0%), and developmental rates (47.5% vs. 21.4% vs. 16.5%) than 25 or 30V. There was no statistical difference between 25 and 30V. Subsequently, editing rates were determined using the T7 mismatch assay and verified with Sanger sequencing followed by sequence alignment and analysis using Tracking of Indels by Decomposition (TIDE) software (https://tide.nki.nl/). Although there was high variance between electroporation groups, blastocyst editing rates of up to 80.0% were achieved using 30V. To our knowledge, these are the first confirmed gene-edited bovine embryos produced from invivo fertilized zygotes. This method offers the ability to utilise the embryos of high-value cows or cows with known genotypes for genetic engineering experiments. In addition, given that electroporated bovine zygotes can be transferred back to the oviduct endoscopically, our future attempts will focus on genome editing in bovine embryos developed nearly completely within the physiological invivo environment.
140 The relationship between morphometric characteristics and semen parameters in Bapedi rams
Authors: Maqhashu A. , O'Neill H. A. , Sebei P. J. , Ramukhithi F. V. , Mapholi N. O. , Nephawe K. A. , Nedambale T. L.
Published: December 2019

Bapedi is a fat-tailed, hardy, and disease-tolerant breed of sheep that is indigenous to South Africa. The Bapedi sheep breed is facing genetic degradation due to nonselective cross-breeding and irregular mating; therefore, breeding strategies for urgent conservation of Bapedi sheep are crucial. The aim of the present study was to investigate the relationship between body measurements and semen traits of Bapedi rams conserved in situ (conserved in their natural habitat) and ex situ invivo (live animals conserved outside their natural habitat). Semen was collected twice per week from 33 rams aged 2-4 years (four ejaculates per ram) using an electroejaculator. Body measurements (rump height, body length, scrotum circumference (SC), scrotal width, and scrotum length (cm)), body condition score (BCS; scale of 1-5), bodyweight (BW; kg), rectal temperature (°C), semen volume (mL), spermatozoa concentration (billionsmL−1), and motility parameters were measured using a computer-aided semen analysis system. Data were analysed using analysis of variance and the general linear models of SAS (SAS Institute Inc.). The Pearson correlation coefficient in SAS was used to determine the relationship among variables. There was uniformity in all body measurements of Bapedi sheep regardless of the conservation method (P>0.05). There were no significant differences in body temperature during semen collection, SC, semen volume, semen pH, semen concentration, spermatozoa total motility, and kinematics in Bapedi rams between methods of conservation (P>0.05). Pearson correlation revealed moderate significant relationships between semen volume and BW, BCS, and SC (r=0.315, 0.638, and 0.381, respectively). Rump length had a moderate correlation (0.566) with normal spermatozoa (P<0.05). It was concluded that Bapedi sheep are still a uniform breed regardless of their decreasing numbers. The BW, BCS, and SC must be considered in the selection criteria for improving the reproductive performance of Bapedi breeding rams because they were positively correlated with semen volume. It is recommended that more studies be done to correlate body measurements, testicular morphometry, and semen parameters in this breed.
145 ProAKAP4 concentrations in semen as a predictive tool of bull fertility: A preliminary study
Authors: Ruelle I. , Seregeant N. , Bencharif D. , Charreaux F. , Thorin C. , Michaud S. , Dordas-Perpinyà M. , Jouy N. , Audry S. , Maurage C. , Delehedde M. , Briand-Amirat L.
Published: December 2019

Recently, ProAKAP4 has been described as a pertinent indicator of sperm quality in humans, pigs, and stallions. In knockout mouse models lacking AKAP4 expression, the male mice were infertile. As high proAKAP4 levels were significantly correlated with a lower proportion of abortions in intrauterine insemination settings in human reproduction, proAKAP4 could be considered a pertinent new sperm parameter for assessing embryo quality. Our main goal was to assess the proAKAP4 concentrations in Holstein bull semen for comparison with the motility sperm parameters and fertility outcomes in post-thawed conditions. Straws issued from 52 ejaculates from 13 bulls, retrospectively identified with known nonreturn rates (NRR) as a fertility indicator, were provided by Evolution XY. Expression of ProAKAP4 and AKAP4 was assessed using enzyme-linked immunosorbent assay, western blotting, flow cytometry, and microscopy methods. Using the Bull 4MID kit (4BioDx), striking variations in proAKAP4 concentrations were observed independently of the classic sperm parameters that were measured using computer-assisted semen analysis. A mean proAKAP4 concentration of 44.42ng per 10 million spermatozoa was obtained through all our series. Interestingly, the variations in proAKAP4 concentrations were positively correlated with progressive motility and with the linearity coefficient parameter. Furthermore, the post-thawed concentrations of proAKAP4 were significantly higher in bulls with a higher NRR in a field study of more than 190 000 AI. We then demonstrated for the first time a correlation between the semen concentration of proAKAP4 and NRR (P=0.05) in bulls. Threshold values of proAKAP4 were then determined, with good values being between 25 and 60ngmL−1. Below 25ngmL−1, the sperm were of poor quality. The proportion of functional spermatozoa (i.e. spermatozoa expressing proAKAP4 in ejaculates) was assessed using flow cytometry. We observed that the cell debris and dead spermatozoa were never immunolabeled with proAKAP4 antibodies. On testis tissue sections, proAKAP4 was expressed only from the spermatids stages up to the ejaculated spermatozoa, being influenced by external factors and reflecting good spermatogenesis. Our preliminary study highlighted the pertinence of proAKAP4 in assessing sperm quality in bulls. It could be interesting to further analyse the effect of proAKAP4 level of expression on capacitation and IVF. As high levels of proAKAP4 were significantly correlated with fertility rates and with progressive motility, proAKAP4 could be proposed as a predictive marker of bull fertility and could be further investigated to evaluate the quality of invitro-produced embryos.
152 An effective method of inducing oestrus in superovulation-treated Japanese Black donor cows after egg collection
Authors: Aoyagi Y. , Takeuchi M. , Oono T. , Hayama K. , Urakawa M. , Oono Y. , Koiwa M.
Published: December 2019

The objective of this study was to evaluate the effect of inducing oestrus in superovulation-treated Japanese Black donor cows after egg collection using a controlled internal drug release (CIDR) device and prostaglandin F2α treatments. The test animals were 50 biparous or triparous Japanese Black cows that had been treated for superovulation during the period of 75-110 days post-calving and from each of which 10 or more ova or embryos had been recovered using a uterine reflux method. In the 25 cows of the control group that received the conventional treatment, a CIDR was not introduced into the vagina, the number of days from egg harvesting to standing oestrus was recorded, and AI was performed using frozen semen. At the time of AI, 50µg of gonadotrophin-releasing hormone (GnRH; Consultan, Aska Animal Health) was administered intramuscularly. The cows that did not conceive in the first AI attempt were again given the same AI and GnRH treatments in the next oestrus cycle. In the 25 cows of the treatment group, a CIDR (CIDR 1900, Zoetis) device was inserted in the vagina after egg harvesting and was kept there for 12 days. At the time of insertion and removal of the CIDR, 150µg of d-cloprostenol formulation (Dalmazin, Kyoritsu Seiyaku) was administered intramuscularly. The number of days from egg harvest to standing oestrus was recorded, and GnRH was administered at the time of AI. The cows that did not conceive in the first AI attempt were given the same AI and GnRH treatments in the next oestrus cycle. The control and treatment groups were compared for the number of days from egg harvesting to oestrus (t-test) and the conception rate after AI (chi-square test). The mean number of days from egg collection to standing oestrus in the control group was 38.6±11.9, and the conception rate was 60% (15/25). Among the animals that failed to conceive in the first AI attempt, the second standing oestrus occurred 60.7±13.1 days after egg collection, and 70% (7/10) of the cows conceived after the second AI. In the treatment group, on the other hand, the mean number of days from egg collection to standing oestrus was 15.8±3.9, and 72% (18/25) of the cows conceived after the first AI. Among the cows that failed to conceive, the second standing oestrus occurred 37.4±4.7 days after egg harvest, and 71% (5/7) of the cow conceived after the second AI. There was a significant difference between the two groups in the number of days from egg collection to the first standing oestrus (P<0.05). The results described above suggest that keeping a CIDR in the vagina for 12 days and intramuscularly administering prostaglandin F2α at the time of insertion and removal of the CIDR in superovulated cows are effective for early onset of standing oestrus after egg collection and thus can reduce the number of days to conception.
204 Discriminating intrauterine growth-restricted piglets through near-infrared spectroscopy of amniotic fluids
Authors: Feugang J. M. , Santos-Rivera M. , Grant A. E. , Park S. B. , Devos-Burnett D. , Lemley C. O. , Vance C. K.
Published: December 2019

The occurrence of intrauterine growth restriction (IUGR) is a great concern in animal production systems, leading to substantial economic losses. The multifactorial causes of IUGR make their prediction before or during pregnancy a challenging task. The amniotic fluid, containing chemical secretions of both the developing fetus and the amnion-allantois membrane, may offer a unique opportunity to investigate the causes of IUGR in piglets. Here we applied near-infrared spectroscopy (NIRS) as a rapid tool to discriminate the chemical composition of the amniotic fluid from IUGR fetuses at different gestational ages. The uteri of gravid sows were dissected at 60% (n=6 sows) and 90% (n=6 sows) of gestational age. Amniotic fluids were collected with a sterile 10-mL syringe connected to an 18-G needle under visual guidance, and samples without blood contamination were kept at 4°C while the fetus was measured. Samples were selected according to fetal weight and were classified as small (mean − 2 SD), medium (mean), and large (mean + 2 SD) sizes in each litter. Selected samples were kept at −80°C until NIRS transmittance spectra collection (350-2500nm). At 60% of gestational age, male fetuses weighed 96±1.2, 133±1.7, and 159±1.9g and female fetuses weighed 117±1.1, 137±1.8, and 152±1.8g for small, medium, and large groups, respectively. The fetal weights at 90% of gestational age averaged 517±0.6, 757±0.6, and 912±0.7g for males and 532±0.6, 718±0.9, and 911±0.6g for females for small, medium, and large sizes, respectively. Chemometric analysis of the amniotic fluid spectra was applied to the wavelength range 1300-1600nm using a balanced data set (n=425) with an 80/20 calibration/validation distribution for linear discriminant analysis (LDA-PCA). Overall, we were able to discriminate samples according to fetal size and gestational age; LDA-PCA showed the highest size classification occurring with spectra derived from the small fetuses with an accuracy of 94.5-100% and 100% and 70.6% for the calibration and validation processes from 60% and 90% of gestational age, respectively. By contrast, when the analysis considered each sex at 60% of gestational age, the medium and small fetuses presented the highest size classification for females and males correspondingly. At 90% of gestational age, the large fetuses presented the best discrimination for both sexes. In conclusion, we were able to detect and discriminate chemical changes of the amniotic fluid from different fetuses using NIRS, indicating that this fluid contains molecules that may characterise IUGR in piglets. These observations will permit future investigations to identify specific molecules associated with IUGR that would lead to possible real-time corrective actions before birth.
This work was supported by USDA-ARS grant no. 58-6402-3-018.
228 Effect of seminal plasma on the interval to application of equine chorionic gonadotrophin for the recovery of cumulus-oocyte complexes in alpacas (Vicugna pacos)
Authors: Huanca W. F. , Palomino J. M. , Villanueva J. C. , Malaga J. , Huanca W.
Published: December 2019

Alpacas as other camelids are induced ovulators and require an external stimulus-mounting by the male-for ovulation. NGF-β, protein present in the seminal plasma (SP) is reported as being responsible for stimulating ovulation. However, limited information exists on the effect of ovulation inducers in the new follicular wave emergence with application on protocols of superstimulation. The aim of the study was to determine the effect of two ovulation induced techniques and two different times on the interval to application of equine chorionic gonadotrophin (eCG) for the ovarian superstimulation for the recovery of COCs by ovum pick- up (OPU). Alpacas were evaluated by transrectal ultrasonography with an Aloka SSD 500 ultrasound and transducer (7.5MHz) to determine the presence of a dominant follicle=7mm. A 2×2 experimental design was used with adult female alpacas, 6 to 8 years old, which were assigned to one of the treatments when a dominant follicle=7mm was present. Alpacas were assigned randomly 1 of 2 groups (Day 0) in which follicular ovulation was induced by seminal plasma 1mL IM (SP, n=13; group 1) with application of 650IU of eCG at 36h (n=6) or 48h (n=7) post-application of SP, or by GnRH with 0.008mg of Buserelin IM (GnRH, n=10; group 2) with application of 650IU eCG at 36h (n=5) or 48h (n=5) post-application of GnRH. On Day 7, COCs were counted and the OPU procedure was performed in every group. Data were analysed using ANOVA. Results are presented in Table 1. In conclusion, both inducers of ovulation and both times of application of eCG were effective for producing follicles of 7mm and COCs acceptable for recovery in Alpacas.

Table 1.Number of follicles=7mm (top) and number of COCs recovered (mean±s.e.m.)

Outcome and group
36 h
48 h
Total

Follicles

GnRH
6.8±1.93
13.4±4.34
10.10±2.50a

SP
5.5±2.23
5.29±1.15
5.38±1.25b

Total
6.09±1.44
8.67±2.18
7.43±1.33

COCs

GnRH
5±0.97
3±1.26
4.33±0.74a

SP
1±0.41
0.33±0.22
0.67±0.25b

Total
3.29±0.67x
1.4±0.6y
2.5±0.53

a,bValues within columns with different letters differ significantly (P=0.05).
x,yValues within columns with different letters differ significantly (P=0.05).

This research funded by CIENCIACTIVA-CONCYTEC as part of the project title “Role of seminal plasma in reproductive physiology and application of biotechnologies in camelids” (149-2017).
71 Comparison of sexed semen ULTRA-4M with conventional semen for the invitro production of bovine embryos
Authors: Alvarez H. , Kjelland M. , Villaseñor F. , Pérez M. , Romo S.
Published: December 2019

The first commercial production of sexed semen was at the Cogent company in the United Kingdom. Since then millions of offspring have been born using sexed semen produced by flow cytometry. Sexed semen technology has recently been modernized to what is now known as ULTRA (ST Genetics), completely modifying the technique, medium, and sperm concentration. In field trials using AI, there was no difference between conventional semen (CONV) and ultra-sexed semen at a concentration of 4 million sperm per straw (ULTRA-4M). The combination of invitro embryo production (IVP) and sexed semen technologies has been successful for intensity selection in the cattle industry. The objective of this work was to compare the CONV and ULTRA-4M semen for bovine IVP. The research was carried out in the reproduction laboratory at the Facultad de Estudios Superiores Cuautitlán-Universidad Nacional Autónoma de México (FESC-UNAM). The IVP was performed with a continuous invitro culture (IVC) system. Ovaries (n=213) were collected from a slaughterhouse (Querétaro, México) and transported to the laboratory within 2h (FESC-UNAM) in physiological saline solution (0.9% NaCl) supplemented with penicillin G (100IUmL−1) and streptomycin sulfate (100µgmL−1). For IVP, VITROGEN media were used for IVM, IVF, and IVC. For the IVM, the cumulus-oocyte complexes were selected (only grades 1 and 2) and matured for 24h at 38.5°C in 5% CO2, 95% air, and 100% humidity. Matured oocytes (n=1000) were divided into two groups, the CONV group and the ULTRA-4M group. The IVF process was developed with CONV and ULTRA-4M semen from the same bull (Holstein) at a concentration of 2×106 and 0.5×106 spermmL−1, respectively, for 18h in 38.5°C, 5% CO2, 95% air, and 100% humidity. The presumptive zygotes were denuded by pipetting and left in IVC until Day 7 at 38.5°C, 5% CO2, 5% O2, and 90% N2 at 100% humidity. The cleavage rate, embryos of more than 6 cells, and blastocysts on Day 7 of culture were evaluated. The statistical analysis was carried out with the GLM procedure of the SAS software (version 9.3; SAS Institute Inc.) to evaluate the results of CONV vs. ULTRA-4M (α level=0.05). Percent cleavage for CONV was 72.2%±2.53 and 75.6%±2.53 for ULTRA-4M. For embryos with more than 6 cells, the results for CONV and ULTRA-4M were 59.8%±5.61 and 62.8%±5.61, respectively. The percentage of blastocysts on Day 7 was 37.8%±5.39 for CONV and 43.6%±5.39 for ULTRA-4M. There were no significant differences between the groups (P>0.05) for all variables analysed. Although the number of blastocysts on Day 7 were numerically higher in the ULTRA-4M, differences were not significant. In conclusion, under the conditions of this research the ULTRA-4M had similar results as CONV for bovine IVP.