bluuebunny/crossref_metadata_2025 huggingface dataset:
Ethanol gas-phase ammoxidation to acetonitrile: the reactivity of supported vanadium oxide catalysts
Authors: Folco F. , Velasquez Ochoa J. , Cavani F. , Ott L. , Janssen M.
Published: December 2016

The gas-phase ammoxidation of ethanol, a bio-based platform molecule, has been investigated as a possible more sustainable route for the production of acetonitrile, using supported vanadium oxide catalysts. The nature of the interaction between the support and the active species greatly affected the catalytic performance.
H3PW12O40/mpg-C3N4 as an efficient and reusable bifunctional catalyst in one-pot oxidation–Knoevenagel condensation tandem reaction
Authors: Wang Hefang , Wang Cunyue , Yang Yongfang , Zhao Meng , Wang Yanji
Published: December 2016

A single-site bifunctional catalyst for oxidation–Knoevenagel condensation tandem reaction was prepared by immobilizing H3PW12O40 on mpg-C3N4via electrostatic interaction.
Electrodeposition of zinc nanoplates from an ionic liquid composed of 1-butylpyrrolidine and ZnCl2: electrochemical, in situ AFM and spectroscopic studies
Authors: Pulletikurthi Giridhar , Shapouri Ghazvini Maryam , Cui Tong , Borisenko Natalia , Carstens Timo , Borodin Andriy , Endres Frank
Published: December 2016

Electrodeposition of Zn nanoplates from an ionic liquid composed of cationic and anionic zinc-chloro complexes as constituent ions.
QSARs for phenols and phenolates: oxidation potential as a predictor of reaction rate constants with photochemically produced oxidants
Authors: Arnold William A. , Oueis Yan , O'Connor Meghan , Rinaman Johanna E. , Taggart Miranda G. , McCarthy Rachel E. , Foster Kimberley A. , Latch Douglas E.
Published: December 2016

One electron oxidation potential predicts the reactivity of phenol and phenolate compounds in a single correlation.
Hydricity, electrochemistry, and excited-state chemistry of Ir complexes for CO2 reduction
Authors: Manbeck Gerald F. , Garg Komal , Shimoda Tomoe , Szalda David J. , Ertem Mehmed Z. , Muckerman James T. , Fujita Etsuko
Published: December 2016

We prepared electron-rich derivatives of [Ir(tpy)(ppy)Cl]+ with modification of the bidentate (ppy) or tridentate (tpy) ligands in an attempt to increase the reactivity for CO2 reduction and the ability to transfer hydrides (hydricity). Density functional theory (DFT) calculations reveal that complexes with dimethyl-substituted ppy have similar hydricities to the non-substituted parent complex, and photocatalytic CO2 reduction studies show selective CO formation. Substitution of tpy by bis(benzimidazole)-phenyl or -pyridine (L3 and L4, respectively) induces changes in the physical properties that are much more pronounced than from the addition of methyl groups to ppy. Theoretical data predict [Ir(L3)(ppy)(H)] as the strongest hydride donor among complexes studied in this work, but [Ir(L3)(ppy)(NCCH3)]+ cannot be reduced photochemically because the excited state reduction potential is only 0.52 V due to the negative ground state potential of −1.91 V. The excited state of [Ir(L4)(ppy)(NCCH3)]2+ is the strongest oxidant among complexes studied in this work and the singly-reduced species is formed readily upon photolysis in the presence of tertiary amines. Both [Ir(L3)(ppy)(NCCH3)]+ and [Ir(L4)(ppy)(NCCH3)]2+ exhibit electrocatalytic current for CO2 reduction. While a significantly greater overpotential is needed for the L3 complex, a small amount of formate (5–10%) generation in addition to CO was observed as predicted by the DFT calculations.
Enrichment of nitrifying bacteria and microbial diversity analysis by high-throughput sequencing
Authors: Yao Renda , Yang Hong , Yu Mengyu , Liu Yi , Shi Huan
Published: December 2016

The nitrifying bacteria community can be enriched by an alternating operation mode.
Approach to high proton/vanadium selectivity in an ionic exchange membrane via a hyperbranching design
Authors: Ma Liying , Li Jing , Xu Guoxiao , Xiong Jie , Cai Weiwei
Published: December 2016

High H+/V selectivity was achieved for the ion exchange membrane due to specially designed hyperbranching structure and intermolecular force.
Electrospun ZIF-based hierarchical carbon fiber as an efficient electrocatalyst for the oxygen reduction reaction
Authors: Liu Chao , Wang Jing , Li Jiansheng , Liu Jizi , Wang Chaohai , Sun Xiuyun , Shen Jinyou , Han Weiqing , Wang Lianjun
Published: December 2016

MOF-based hierarchical carbon fiber is developed as an efficient electrocatalyst for the oxygen reduction reaction.
Surgery for Lung Cancer and the Consequences for the Swallow
Authors: Hales Pippa , Mossey-Gaston Corinne
Published: December 2016

Lung cancer is one of the most commonly diagnosed cancers across Northern America and Europe. Treatment options offered are dependent on the type of cancer, the location of the tumor, the staging, and the overall health of the person. When surgery for lung cancer is offered, difficulty swallowing is a potential complication that can have several influencing factors.

Surgical interaction with the recurrent laryngeal nerve (RLN) can lead to unilateral vocal cord palsy, altering swallow function and safety. Understanding whether the RLN has been preserved, damaged, or sacrificed is integral to understanding the effect on the swallow and the subsequent treatment options available. There is also the risk of post-surgical reduction of physiological reserve, which can reduce the strength and function of the swallow in addition to any surgery specific complications. As lung cancer has a limited prognosis, the clinician must also factor in the palliative phase, as this can further increase the burden of an already compromised swallow.

By understanding the surgery and the implications this may have for the swallow, there is the potential to reduce the impact of post-surgical complications and so improve quality of life (QOL) for people with lung cancer.
Personality Traits and Adherence in Head and Neck Cancer Patients—Considerations in Treatment
Authors: Croom Andrea R.
Published: December 2016

Head and neck cancer is a complicated medical condition that results in multiple physical and functional changes (e.g., dysphagia, speech alterations) that leave patients with a poorer quality of life. Many of these symptoms may be effectively reduced through interventions offered by speech language pathologists (SLPs), unfortunately adherence to these treatment recommendations remains low. Personality traits are the characteristics that make each individual unique and play a key role in determining how a person thinks about their life experiences (e.g., a diagnosis of cancer), and the coping strategies that a person uses to manage stressful situations. An awareness of personality traits, illness beliefs, and coping styles can assist SLPs in tailoring interventions and supporting adherence to treatment recommendations.
SatLTP: low extra delay transport protocol for satellite networks
Authors: Li Xiaohui , Wang Junfeng
Published: December 2016

As the exponential growth of traffic patterns with very different characteristics is becoming increasingly common, the high demand for a wide range of services has attracted much attention, e.g. low‐priority (LP) service. To satisfy the requirements of LP service for satellite networks, a LP data transfer protocol called satellite networks (SatLTP) with the adaptation to satellite channels and taking advantage of extra bandwidth without interfering high priority flows is proposed. In order to use extra bandwidth adaptable, SatLTP achieves precise adjustment of the congestion window by dynamically updating indicative parameters and selecting optimal window control policies. Simulation results over satellite networks show that SatLTP enhances the throughput of satellite channels in terms of being friendly and non‐intrusive toward other protocols (e.g. TCP) and able to effectively exploit the available resources over satellite networks.
Convergence of WSN and cognitive cellular network using maximum frequency reuse
Authors: Hassan Farooque , Roy Abhishek , Saxena Navrati
Published: December 2016

The exponential increase in wireless sensor network (WSN) applications and devices crave for the communication resources. Regardless of the access mechanism, the unlicensed industrial, scientific and medical spectrum band is insufficient for massive simultaneous connections. On the other hand, the 4G and the emerging 5G cellular networks provide high data rates of 75–300 Mbps and 7.5 Gbps, respectively. Nevertheless, WSN's extempore inclusion in the cellular network can heavily congest its Random Access Channel (RACH). In this study, the authors investigate a unique concept of maximum frequency reuse in small evolved NodeBs, and subsequently, recommend their deployment in WSN congested areas. They also propose a cognitive radio (CR) algorithm to offload SeNB load over Macrocell. Authors formulate proposed SeNB‐based cellular network in an analytical RACH model, where dynamics of SeNB's and Macrocell users are scrutinised. Observations show that the proposed scheme allocates more users with low collision probability. Simulation results show that the proposed scheme imparts a significant increase of 1–3 Mbps in downlink throughput and culminate a lift of 2–9 Mbps in uplink throughput. The average uplink signal‐to‐interference‐plus‐noise‐ratio (SINR) is also proliferated by approximately 10–15 dB. Moreover, CR‐based SeNB's smart offloading outperforms the conventional offloading in terms of downlink and uplink SINR by about 1–3dB.
Design and parametric investigation of directional antenna for microwave imaging application
Authors: Mahmud M.Z. , Islam Mohammad T. , Samsuzzaman M. , Kibria Salehin , Misran Norbahiah
Published: December 2016

This study presents a new side slotted Vivaldi antenna (SSVA) with size reduction and improve gain with directive featured as compared with traditional Vivaldi antenna. A number of parameters are studied and optimised for breast imaging system over ultra‐wideband frequency (3.1–10.6 GHz). The parameter such as substrate type, radiating fins, cavity diameter, stub radius, stub angle is optimised to reduce the size of the antenna with better antenna performance in terms of bandwidth, gain, efficiency, and directivity. The radiating fins are modified by etching a number of side slot to enhance the gain and electrical length. The optimised size of the antenna is 45 (L) × 37 (W) mm2, 8.5 mm cavity diameter, 9.5 mm stub radius with the angle of 80° by using Rogers 5870 substrate. Results show that the antenna has a bandwidth from 3.9 to 9.15 GHz for reflection co‐efficient <−10 dB with directional radiation pattern. The peak gain of proposed prototype is 6.8 dBi and the radiation efficiency is about 88% on an average over the operating bandwidth. The fidelity factor for face to face is 0.92 and for side by side is 0.62, which prove the directionality and lower distortion of the signal. A number of design iteration is done to achieve the optimum result of the antenna. The prototype of proposed SSVA antenna is successfully fabricated, simulated, and measured. Later, the antenna is used in breast phantom measurement system for microwave imaging application.
Generation of optical picosecond pulses with monolithic colliding‐pulse mode‐locked lasers containing a chirped double‐quantum‐well active region
Authors: Prziwarka Thomas , Klehr Andreas , Wenzel Hans , Fricke Jörg , Bugge Frank , Erbert Götz , Weyers Markus , Knigge Andrea , Tränkle Günther
Published: December 2016

In this study, the authors present experimental results on 6 mm long monolithic passively colliding‐pulse mode‐locked (CPM) lasers. The AlGaAs‐based devices with Inx Ga1−x Asy P1−y double‐quantum‐well (DQW) active regions emit between 860 and 880 nm. To optimise the laser performance and to shorten the pulses the optical gain is spectrally broadened to increase the number of longitudinal lasing modes by using a chirped DQW structure where the In‐content of the QWs differs by Δx  = 0.08. Lasers having the chirped DQW with lasers having a conventional, unchirped DQW active region are compared. The mode locking operation is investigated in dependence on gain current, absorber voltage and absorber length. According to the experimental results, the lasers with the chirped DQW exhibit an overall better performance and generate shorter pulses with higher average optical output powers over a wider range of parameters. For an optimum set of parameters, the generated pulses have a full width at half maximum of the autocorrelation function of 1.9 ps at a repetition frequency of 12.9 GHz.
Effect of Screw Perpendicularity on Compression in Scaphoid Waist Fractures
Authors: Swanstrom Morgan , Morse Kyle , Lipman Joseph , Hearns Krystle , Carlson Michelle
Published: December 2016

Background Central and perpendicular (PERP) screw orientations have each been described for scaphoid fracture fixation. It is unclear, however, which orientation produces greater compression.

Questions/Purposes This study compares compression in scaphoid waist fractures with screw fixation in both PERP and pole-to-pole (PTP) configurations. PERP orientation was hypothesized to produce greater compression than PTP orientation.

Methods Ten preoperative computed tomography scans of scaphoid waist fractures were classified by fracture type and orientation in the coronal and sagittal planes. Three-dimensional models of each scaphoid and fracture plane were created. Simulated Acutrak 2 (Acumed, Hillsboro, OR) screws were placed into the models in both PERP and PTP orientations. Engagement length and screw angle relative to the fracture were measured. Compression strength was calculated from the shear area, average density, and angle acuity.

Results The PTP angle between screw and fracture ranged from 36 to 84 degrees. By definition, the PERP screw-to-fracture angle was 90 degrees. Perpendicularity of the PTP screw to the fracture was positively correlated to compression strength. PERP screws had greater compression than PTP screws when the PTP screw-to-fracture angle was < 80 degrees (106 vs. 80 N), but there was no difference in compression when the PTP screw-to-fracture angle was > 80 degrees, approximating the PERP screw.

Conclusion Increasing screw perpendicularity resulted in higher compression when the screw-to-fracture angle of the PTP screw was < 80 degrees. Maximum compression was obtained with a screw PERP to the fracture. The increased compression gained from PERP screw placement offsets the decreased engagement length.

Clinical Relevance These results provide guidelines for optimal screw placement in scaphoid waist fractures.
Lactic Acidosis in Children – A Varied Presentation
Authors: Shah Ira
Published: December 2016

Primary lactic acidemias represent a family of disorders of pyruvate metabolism or defects in the respiratory chain. However, lactic acidosis may also be seen in metabolic disorders such as organic acidemias, urea cycle defects, and fatty acid oxidation defects, which can be easily excluded by serum ammonia estimation, urinary organic acid estimation, and quantification of plasma amino acids. The classical presentation of a patient with primary lactic acidemia is growth retardation, ataxia, stroke, and increased lactic acid levels in the blood and cerebrospinal fluid. Patients may also present with cerebral edema, acute rhabdomyolysis, cardiac arrhythmias, cardiomyopathy, coma, and neurodegeneration. We present three cases of lactic acidemia with varied presentation. The first child presented at 5 years with recurrent hematemesis with hypoglycemia. The second child presented at 11 years of age with recurrent episodes of unconsciousness. The third child presented at one and a half months with convulsions.
Linking macro-level goals to micro-level routines: EHR-enabled transformation of primary care services
Authors: Findikoglu Melike , Watson-Manheim Mary Beth
Published: December 2016

Information and communication technologies are known to be instrumental in the enhancement of healthcare management capabilities in developing countries. Turkey - a developing country - has undergone a major healthcare transformation marked by the redesign of primary care delivery and the implementation of a nation-wide Electronic Health Records (EHR) system. In this research, presenting Turkey's case, we investigate the consequences of EHR implementation in developing countries. We argue that to better understand the consequences, we need to link macro-level healthcare goals with micro-level system usage behaviors that actualize the macro-level goals or alternatively result in unintended negative health outcomes. We posit that this linkage is achieved through the meso-level structures, namely the EHR and the organizational context, in which it is embedded. Hence, we examine the EHR's role in this relationship. Our findings indicate that EHR usage both enables and constrains the achievement of clinicians' professional goals in the context of primary care delivery. Moreover, goal alignment between the government agency as the designer of the system and the clinicians influence the outcomes of the EHR-enabled transformation. When the healthcare goals are aligned, the system enables the clinicians to achieve their professional goals and their system usage behaviors converge, contributing to improvements in health outcomes. Contrarily, when the goals are misaligned, the system constrains goal achievement and the clinicians show divergent usage behaviors, including goal abandonment. In turn, goal abandonment may lead to negative consequences and even adversely affect the achievement of population-level healthcare goals in the long run.
Global and local skin friction diagnostics from TSP surface patterns on an underwater cylinder in crossflow
Authors: Miozzi Massimo , Capone Alessandro , Di Felice Fabio , Klein Christian , Liu Tianshu
Published: December 2016

A systematical method is formulated for extracting skin-friction fields from Temperature Sensitive Paint (TSP) images in the sense of time-averaging and phase-averaging. The method is applied to an underwater cylinder in crossflow at two subcritical regimes (Re = 72 000 and 144 000). TSP maps are decomposed in a time-averaged, a phase-averaged, and a random component. The asymptotic form of the energy equation at the wall provides an Euler-Lagrange equation set that is solved numerically to gain the relative skin friction time- and phase-averaged fields from the TSP surface temperature maps. The comparison of the time averaged relative skin-friction profiles with the literature data shows an excellent agreement on the whole laminar boundary layer up to the laminar separation line. Downstream of separation, time averaged results identify the secondary reattachment/separation events, which are lost in the available literature data. The periodic behavior of the skin-friction is taken, describing how the laminar separation bubble evolves by providing the time history of the laminar separation line and of the secondary reattachment/separation over the entire vortex shedding period. Instantaneous skin friction maps reveal the existence of coherent structures by capturing their footprint on the cylinder’s surface. An array of Π-shaped traces marks the existence of counter-rotating, streamwise-oriented vortices just before the laminar separation line. Their interaction with the laminar boundary layer and with the separation line is briefly described. An example of the intermittent excerpt of their influence through the laminar separation line is reported.
Method for reconstructing nonlinear modes with adaptive structure from multidimensional data
Authors: Gavrilov Andrey , Mukhin Dmitry , Loskutov Evgeny , Volodin Evgeny , Feigin Alexander , Kurths Juergen
Published: December 2016

We present a detailed description of a new approach for the extraction of principal nonlinear dynamical modes (NDMs) from high-dimensional data. The method of NDMs allows the joint reconstruction of hidden scalar time series underlying the observational variability together with a transformation mapping these time series to the physical space. Special Bayesian prior restrictions on the solution properties provide an efficient recognition of spatial patterns evolving in time and characterized by clearly separated time scales. In particular, we focus on adaptive properties of the NDMs and demonstrate for model examples of different complexities that, depending on the data properties, the obtained NDMs may have either substantially nonlinear or linear structures. It is shown that even linear NDMs give us more information about the internal system dynamics than the traditional empirical orthogonal function decomposition. The performance of the method is demonstrated on two examples. First, this approach is successfully tested on a low-dimensional problem to decode a chaotic signal from nonlinearly entangled time series with noise. Then, it is applied to the analysis of 250-year preindustrial control run of the INMCM4.0 global climate model. There, a set of principal modes of different nonlinearities is found capturing the internal model variability on the time scales from annual to multidecadal.
A post-fabrication selective magnetic annealing technique in standard MEMS processes
Authors: Mohammadi A. , Karmakar N. C. , Yuce M. R.
Published: December 2016

A selective electrothermal magnetic annealing technique is introduced that provides programming capabilities for mechanical micro-resonators. In the proposed approach, the magnetic properties of resonators can be locally tuned in a post-fabrication batch-compatible process step. A prototype is implemented in a standard microfabrication process, where resonating ferromagnetic elements are suspended on top of a polysilicon resistive heater. The ferromagnetic elements consist of electroplated Nickel (Ni) with minor Iron (Fe) impurities. The electro-thermo-mechanical heating phenomenon is simulated for design purposes. The magnetization of micro-resonators with and without magnetic annealing is measured. The resulting magnetic property enhancement is illustrated by hysteresis (M-H) loop variations.
Self-assembled dual-sided hemispherical nano-dimple-structured broadband antireflection coatings
Authors: Lin Cheng-Yen , Lin Kun-Yi , Tsai Hui-Ping , He Yi-Xuan , Yang Hongta
Published: December 2016

A non-lithography-based approach is developed in this study for assembling monolayer close-packed hemispherical nano-dimple arrays on both sides of a PET film by a scalable Langmuir-Blodgett technology. The resulting gratings greatly suppress specular reflection and therefore enhance specular transmission for a broad range of visible wavelengths, resulting from a gradual change in the effective refractive index at air/PET interface. The experimental results reveal that the antireflection properties of the as-fabricated coatings are affected by the size of the nano-dimples. Moreover, both optical performances of single-sided and dual-sided nano-dimple-structured coatings have been investigated in this study.
Imaging photocurrent collection losses in solar cells
Authors: Huhn Vito , Pieters Bart E. , Augarten Yael , Gerber Andreas , Hinken David , Rau Uwe
Published: December 2016

A method is proposed that enables the imaging of the photocurrent collected by a solar cell under arbitrary operating conditions. The method uses a series of luminescence images under varying illumination to derive the total photocurrent collection efficiency at a given voltage bias. The resulting total photocurrent collection image directly relates to the difference between the dark and illuminated current-voltage characteristics of the cell. A crystalline silicon solar cell is used to test the method, and the images of the total photocurrent collection efficiency are used to quantify the influence of a crack on the total collected photocurrent of the solar cell.
Physics in 2116: Megatelescope releases its first image
Authors: Austin Robert
Published: December 2016

With its construction just beginning, the Asteroid Belt Astronomical Telescope has already photographed an exoplanet with unrivaled clarity.
Physics in 2116: Emergent consciousness decoded
Authors: LaSala John
Published: December 2016

SAMI 9000, one of the world’s most sophisticated artificial intelligences, has allowed human colleagues to probe his brain as he drifts in and out of consciousness.
100 FACTORS AFFECTING PREGNANCY RATES IN THE TRANSFER OF IN VITRO-PRODUCED JAPANESE BLACK CATTLE EMBRYOS
Authors: Nishisouzu T. , Abe A. , Matoba S. , Dochi O. , Okamura K.
Published: December 2016

Despite the rapid expansion of in vitro embryo production (IVP) technology for genetic improvement in the cattle industry in the last decades, pregnancy rates by the transfer of IVP embryos are still lower than those derived from in vivo-produced embryos. The objective of this study was to analyse factors affecting pregnancy rates after the transfer of IVP Japanese Black cattle embryos under farm conditions. Holstein heifers (n = 4,475) and cows (n = 8,541) were selected as recipients. Most cows (80%) were managed in tie-stall barns and most heifers (80%) were managed in pens. Embryo transfers were performed for 9 years, from 2004 to 2012. The embryos were produced from oocytes derived from a local abattoir and semen from 14 proven bulls by the Livestock Improvement Association of Japan (Hamano and Kuwayama 1993 Theriogelogy 39, 703–712). The fresh IVP embryos (quality; IETS code 1) that reached the blastocyst stage after 7 to 8 days (insemination = Day 0) were transported by an airplane (2 h) and subsequently by a car (1.5 h). Embryos were non-surgically transferred to each recipient on Day 7 to 9 of their natural oestrous cycle on farms. Pregnancy was diagnosed on Day 40 to 60 after oestrus. Pregnancy results were statistically analysed using the GLM procedures of SAS. The following variables were included in the model: recipient parity (0, 1, 2, or 3), day (7, 8, or 9) of the oestrous cycle at the time of embryo transfer, oestrus behaviour (increased activity observed by farmers), presence of mucus at oestrus, presence of blood after oestrus, and year (1, 2, 3, 4, 5, 6, 7, 8, or 9) and season (April–June as spring, July–September as summer, October–December as fall, or January–March as winter) of embryo transfer. The Bonferroni correction was used to counteract the problem of multiple comparisons. Heifers had significantly higher pregnancy rates than cows (51.0% v. 37.9%, respectively; P < 0.01), and first parity cows had higher pregnancy rates than third parity cows (42.9% v. 35.7%, respectively; P < 0.01). Pregnancy rates were significantly higher in recipients that received an embryo transfer on Day 8 of their oestrous cycle, than on Day 7 (46.6% v. 42.4%, respectively; P < 0.01). Recipients without oestrus behaviour had higher pregnancy rates than those with oestrus behaviour (46.3% v. 43.4%, respectively; P < 0.01). The presence of mucus and/or blood after oestrus and the season of transfer were not found to significantly affect pregnancy rates. The results of this study indicated that performing IVP embryo transfers on Day 8 of a recipient’s oestrous cycle will improve the pregnancy rate, season does not have an effect on pregnancy rate, and the detection of oestrus by monitoring increased activity is not always reliable and instead should be determined by multiple symptoms on farm conditions.
118 EXOSOME-MEDIATED OXIDATIVE STRESS RESPONSE IN BOVINE GRANULOSA CELLS
Authors: Saeed-Zidane M. , Salilew-Wondim D. , Linden L. , Held E. , Neuhoff C. , Hoelker M. , Schellander K. , Tesfaye D.
Published: December 2016

Exosomes are nano-sized (30–100 nm) extracellular membrane vesicles released through exocytosis process in most cells and biological fluids. They contain a cargo of nucleic acids, proteins, lipids and play a vital role in cell-cell communications. Various cell types have been shown to release exosomes into extracellular space as a response to various environmental stress conditions. However, little is known about the response of granulosa cells to oxidative stress, with regard to release of exosomes that may carry mRNA and protein molecules related to cellular oxidative stress response. Here we aimed to investigate the potential release of stress elements by granulosa cells to culture media through exosomes under oxidative stress conditions. For that, bovine granulosa cells from small follicles were aspirated and cultured in DMEM/F-12 media supplemented with exosome free fetal bovine serum (Exo-FBS) and treated with 5 µM H2O2 for 40 min. Granulosa cells were collected 24 h post-treatment to quantify the expression of antioxidants (Nrf2, Keap1, SOD1, CAT1, PRDX1, HOMOX1, TXN1, and NQO1), cell proliferation (PCNA and CNND2), cell differentiation (CYP11A1 and STAR), apoptosis (Casp3), and antiapoptosis (BCL2L1) genes. Reactive oxygen species accumulation, mitochondrial distribution, cell viability, and cell cycle assays were performed in cultured granulosa cells, and the culture medium was used to isolate exosomes using ultracentrifugation procedure. The identity of exosomes was confirmed by immunoblotting of Alix and CD63 proteins, and the expression level of antioxidant was analysed in mRNA isolated from exosomes. Data from 3 independent biological replicates were statistically analysed using the 2-tailed t-test. Results showed that H2O2 treatment increased mRNA and protein level of antioxidants (Nrf2, PRDX1, and TXN1), as well as cell differentiation and apoptosis-related genes compared to untreated controls. However, granulosa cells treated with H2O2 showed lower expression of cell proliferation marker genes (PCNA and CNND2). Cells treated with H2O2 showed increases in reactive oxygen species level, inadequate mitochondrial distribution, and lower cell viability. Cell cycle assay revealed a reduction in G0/G1 proportion and increase in G2 phase in cells treated with H2O2. Higher levels of antioxidant (Nrf2, CAT1, and TXN1) transcripts were detected in exosomes isolated from media with cells under oxidative stress conditions compared to the controls. Labelling and co-transfection of exosomes from stressed cell culture medium with untreated treated recipient granulosa cells resulted in increased abundance of cellular mRNA and protein of Nrf2 and CAT1 in those cells. In conclusion, granulosa cells exposed to oxidative stress could release exosomes that carry molecules related to oxidative stress response, which can be up taken by recipient cells.
120 ANTRAL FOLLICLE COUNT OF 2-YEAR-OLD MARES IS HIGHLY CORRELATED WITH ANTI-MÜLLERIAN HORMONE CONCENTRATIONS AT 24 to 28 WEEKS OF AGE
Authors: Scarlet D. , Kuhl J. , Wulf M. , Ille N. , Aurich C.
Published: December 2016

Success of assisted reproductive techniques, as determined by the response to hormonal treatments and embryo quality, can successfully be predicted by the concentration of anti-Müllerian hormone (AMH) in plasma of several species. Being able to predict ovarian follicular reserve of prepubertal female horses (fillies) would help to select fertile broodmares and reduce costs associated with animal upkeep. The objectives of this work were to (1) assess AMH dynamics in female horses during the first year of life and (2) determine whether AMH concentrations detected in plasma of prepubertal fillies are correlated with AMH concentrations and antral follicle count (AFC) after puberty. Warmblood fillies (n = 14) born from February to May of 1 year in the same stud were used. Blood samples for AMH determinations were collected from birth onward every 4 weeks up to the age of 1 year. At 2 years, blood samples were collected and AFC was determined by transrectal ultrasonography. The AMH concentrations were determined by ELISA (AL-115, Ansh Laboratories, Webster, TX, USA). Transrectal ultrasonography was used to determine the AFC, which corresponds to the total number of antral follicles detected with ultrasound. Statistical analysis was done with the SPSS Statistics 24 software (SPSS Inc., Chicago, IL, USA). The AMH was detectable in the plasma of all animals from birth onward. At birth, mean AMH concentration was 4.5 ± 1.2 ng mL−1. The AMH concentration increased and peaked between 24 weeks (8.7 ± 4.4 ng mL−1) and 28 weeks (6.7 ± 2.1 ng mL−1) and subsequently decreased again (52 weeks: 3.9 ± 0.9 ng mL−1). Very high variation among individuals during the first year was lost at 2 years of age. The AMH concentration at 2 years was highly correlated with AMH concentration at birth (r = 0.62, P < 0.05) and with AFC (r = 0.78, P < 0.001). Also, AMH concentration (r = 0.73, P < 0.01) and AFC (r = 0.6, P < 0.05) at 2 years were highly correlated with AMH concentrations at 24 and 28 weeks. Gestational length (337 ± 1 days), parity of the dam (4.6 ± 0.8), and placental weight (6983 ± 352 g) did not influence AMH concentrations at any time. Our results demonstrate that AMH is detectable in blood of female horses from birth onward. Despite its high variability between individuals up to 52 weeks, strong correlations were observed during the first 2 years of life. High correlations to AFC at 2 years suggest that determination of AMH in prepubertal female horses helps to predict the ovarian reserve and fertility in postpubertal life.
122 OVARIAN FOLLICULAR DYNAMICS IN CROSS-BRED EWES DURING THE RAINY SEASON UNDER TROPICAL CONDITIONS
Authors: Reyes E. A. , Bergfelt D. R. , Ratto V. F. , Valderrama X. P. , Arcelay E. , Ratto M. H.
Published: December 2016

Small ruminants are an important livestock species on many island nations of the Eastern Caribbean region and serve as a vital source of food and income. This study was designed to characterise follicular development during the inter-ovulatory interval (interval between 2 consecutive ovulations) under tropical conditions. Non-lactating, 2- to 4-year-old crossbred Barbados-Black Belly × White Virgin Island ewes weighing a mean 30.4 kg with mean body condition score 2.9 (score 1–5) were used during September–November for the study. Ewes were maintained in a paddock of 100 m2 at the research farm in St. Kitts and Nevis (17.3°N, 62.7°W). Animals were fed daily with 100 g of concentrate (17% crude protein) and had free access to elephant grass (Pennisetum purpureum), mineral salt, water, and shade. Ovaries of 19 females were examined daily for 46 days by transrectal B-mode ultrasonography using a 7.5-MHz lineal array transducer (Honda HS-2200UV, MA, USA) to detect ovulations and growth and regression of antral follicles. Data were analysed using chi-square, Student t-test, or one-way ANOVA. Proportion of females with 2, 3, and 4 follicular waves was 2/19 (10.5%), 10/19 (52.6%), and 7/19 (36.8%), respectively. Mean (± SEM) length of the inter-ovulatory intervals did not differ (P = 0.9) among females with 2 (16.5 ± 0.5 days), 3 (16.3 ± 0.4 days), and 4 (16.6 ± 0.2 days) follicular waves, respectively. Mean diameter of ovulatory follicles was 4.4 ± 0.1 mm, proportion of multiple ovulations was 11/19 (57.9%), and mean number of follicular waves was 3.3 ± 0.1. Wave emergence was normalized to the first ovulation of the inter-ovulatory interval (ovulation = Day 0). Mean days of wave emergence for 2 follicular waves were Days 1.0 ± 0 and 10.0 ± 0.0; for 3 follicular waves were Days 0.7 ± 0.1, 5.6 ± 0.5, and 10.4 ± 0.3; and 4 follicular waves were Days 0.8 ± 0.3, 4.7 ± 0.6, 8.4 ± 0.5, and 11.7 ± 0.3, respectively. The inter-wave intervals were 9.0 ± 0.0 days for 2 waves; 5.1 ± 0.5 and 4.8 ± 0.3 days for 3 waves; and 3.8 ± 0.5, 3.7 ± 0.3, and 3.3 ± 0.4 days for 4 waves. Growth rate (mm/day) of the largest follicle/wave did not differ (P = 07) among animals with 2 follicular waves (1.2 ± 0.7 v. 1.1 ± 0.0) or 3 follicular waves (0.9 ± 0.1 v. 0.9 ± 0.1 v. 1.2 ± 0.2). However, in animals with 4 follicular waves, the largest follicle grew faster (P < 0.05) for wave 1 (1.3 ± 0.2) than that of waves 2 (0.5 ± 0.2), 3 (0.6 ± 0.1), and 4 (0.6 ± 0.1). In conclusion, length of inter-ovulatory intervals, emergence of antral follicles, growth, and regression follicles in a wave-like pattern, inter-wave intervals, and diameter of ovulatory follicles in cross-bred ewes under tropical conditions are comparable to that described in sheep from temperate climates.
128 NEURONATIN TRANSCRIPT ANALYSES AS MARKER TO CONFIRM SUCCESSFUL FERTILIZATION IN BOVINE BLASTOCYSTS
Authors: Stöhr J. , Maurer L. , Zimmer B. , Wrenzycki C.
Published: December 2016

In humans, intracytoplasmic sperm injection (ICSI) has become widely applied to overcome infertility. In addition to its clinical usefulness, ICSI is also a valuable research tool for studying fundamental aspects of gamete interaction during fertilization. Oocyte activation is a prerequisite for decondensation of the sperm head and formation of the male pronucleus regardless of whether the spermatozoon enters the oocyte by injection or by fertilization. In several species, the injection procedure itself is sufficient to activate the oocyte. However, the bovine oocyte is rarely activated by the mechanical stimulation of injection alone. Therefore, additional stimuli are required during ICSI, which may also initiate parthenogenetic development. Because blastocyst origin, either via IVF or after parthenogenetic activation, could not be distinguished at the morphological level via microscopic evaluation, the paternal genome needs to be determined, employing a molecular approach. Therefore, the aim of this study was to validate the expression of neuronatin (NNAT) mRNA, a paternally expressed gene, via RT-PCR, to verify whether bovine blastocysts were generated by fertilization or parthenogenesis. Bovine cumulus-oocyte-complexes were recovered from abattoir-derived ovaries by the slicing method. After 24 h of maturation, fertilization was achieved using a standard protocol. Presumptive zygotes were cultured in SOFaa for a period of 8 days. To generate parthenogenetic blastocysts, oocytes were activated, employing the following protocol. After 24 h of maturation, the oocytes were exposed to 5 µM ionomycin for 5 min and were then incubated in 1.9 mM 6-dimethylaminopurine for 4 h. Afterwards, treated oocytes were cultured in SOFaa for 8 days. Blastocysts from both experimental groups (IVF-derived and parthenogenetic) were stored at −80° until analysis. RT-PCR was performed as described previously (Stinshoff et al. 2014 Reprod. Fertil. Dev. 26, 502–510) for 6 embryos stemming from the IVF-group and 3 embryos out of the chemical-activated group. The NNAT mRNA was expressed in 5 blastocysts stemming from the IVF group; one blastocyst did not show NNAT mRNA expression, suggesting that it was derived from parthenogenesis. Conversely, NNAT mRNA could not be detected in all blastocysts derived from chemical activation, indicating that only the maternal genome was present. These results clearly demonstrate that NNAT mRNA expression is a reliable method to prove the paternal participation in early bovine embryo development.
148 EFFECTS OF CAFFEINE SUPPLEMENTATION ON BOVINE OOCYTE DEVELOPMENTAL CAPACITY
Authors: Bernal-Ulloa S. M. , Lucas-Hahn A. , Aldag P. , Herrmann D. , Baulain U. , Hadeler K.-G. , Niemann H.
Published: December 2016

Oocyte culture in the presence of the nonspecific competitive phosphodiesterase inhibitor caffeine has been reported to increase developmental capacity of oocytes in different mammalian species. Here, we evaluated the effects of caffeine supplementation during the final phase of in vitro maturation (IVM) on developmental rates and blastocyst cell numbers. Bovine ovaries were collected from a local abattoir. A total of 1142 cumulus-oocyte-complexes were obtained by slicing. Cumulus-oocyte complexes were either in vitro matured for 24 h (Standard) or matured for 20 h followed by additional culture for 6 h in fresh IVM medium supplemented with 10 mM caffeine (Caffeine 6 h). In vitro fertilization was performed for 19 h using frozen-thawed sperm from 2 different bulls. After IVF, presumptive zygotes were cultured in vitro for 8 days until the blastocyst stage. Cleavage and blastocyst rates were evaluated 3 and 8 days after IVF, respectively. Expanded blastocysts from the different treatments were submitted to differential staining. SAS/STAT software (SAS Institute Inc., Cary, NC, USA) was used to evaluate cleavage and blastocyst rates using the Glimmix procedure and blastocyst cell numbers were compared using the linear model procedure. Cleavage rates were lower using caffeine for bull B and blastocyst production decreased for bull A. Caffeine treatment increased inner cell mass (ICM) number for bull B and decreased trophectoderm (TE) and total cell numbers for bull A. However, similar TE and total cells were obtained for bull B (Table 1; P < 0.05). Results show that developmental competence can be affected by caffeine supplementation at the final phase of IVM probably due to oocyte-sperm interaction changes.

Table 1.

In vitro developmental competence of oocytes cultured with caffeine at the end of IVM
156 OVIDUCTAL FLUID SUPPLEMENTATION DURING MATURATION AND FERTILIZATION IMPROVES IN VITRO EMBRYONIC DEVELOPMENT IN PIGS
Authors: Goldacker A. , Winn E. , Current J. Z. , Whitaker B. D.
Published: December 2016

Oviducal fluid has a major role in the maturation of gametes and the process of fertilization. The objective of this study was to determine the effects of oviducal fluid supplementation in vitro, during oocyte maturation and IVF on fertilization characteristics and early embryonic development rates. Oocytes from aspired aspirated mature follicles (3–6 mm diameter) were obtained from a local abattoir. During the last 24 h of maturation, oocytes (n = 1303) were placed into maturation media supplemented either 1% (vol/vol) or 5% (vol/vol) thawed snap-frozen oviducal fluid. Fertilization was performed using pooled frozen-thawed semen from 3 different boars. During IVF, the fertilization medium was supplemented with 1% (vol/vol) or 5% (vol/vol) oviducal fluid. Fertilization characteristics were evaluated 12 h after IVF and rates of embryonic cleavage and blastocyst development were observed at 48 and 144 h after IVF, respectively. Data were analysed using ANOVA with the main effects including treatment, well, and replicate. Chi-square analysis was used to determine percentages of embryos reaching the different developmental stages for each treatment. There were no significant differences in the percentages of oocytes that reached metaphase II by the end of maturation or in sperm penetration rates after IVF. However, oocytes treated with 1% (vol/vol) oviducal fluid during the end of maturation and IVF (33.33 ± 2.61) and 5% (vol/vol) oviducal fluid during maturation (33.33 ± 2.66) or IVF (39.53 ± 3.78) had significantly less (P < 0.05) incidence of polyspermic penetrations and a significantly higher (P < 0.05) incidence of male pronuclear formation (87.50 ± 4.01; 86.67 ± 4.83; 86.05 ± 3.19, respectively) compared with no oviducal fluid supplementation. Oocytes supplemented with 5% (vol/vol) oviducal fluid during maturation and IVF had significantly lower (P < 0.05) incidences of polyspermic penetration (27.91 ± 2.50) and significantly higher (P < 0.05) percentages of embryos reaching the 2-cell stage (81.76 ± 3.72) and blastocyst stage of development (37.74 ± 1.09) by 48 and 144 h, respectively, compared with all other groups. The results of this study suggest that supplementing 5% (vol/vol) oviducal fluid during maturation and IVF improves the success rates of in vitro embryo development in pigs.
194 EFFECT OF SMALL MOLECULE INHIBITORS ON Oct-4, Nanog, AND Sox-2 EXPRESSION IN BOVINE FETAL FIBROBLAST CELLS
Authors: Gaspard L. W. C. , Bondioli K. R.
Published: December 2016

Successful reprogramming of somatic cells towards pluripotency requires the epigenetic marks characteristic of the differentiated cell type first be erased in order to inactivate the somatic cell program and activate the embryonic program. However, the majority of cells that undergo reprogramming become trapped in a partially reprogrammed state that is characterised by the down-regulation of somatic cell marker genes, incomplete reactivation of pluripotency genes, maintenance of viral expression, and the inability to form chimeras. Several small molecules, which act on specific signaling pathways or chromatin modifications, have been shown to improve both the kinetics and efficiency of reprogramming. These chemical modifiers aid in overcoming the roadblocks encountered during the reprogramming process by inducing the necessary epigenetic modifications needed to silence the somatic cell genome and reactivate the embryonic stem cell genome. Chemical treatment of cells before reprogramming can remodel the epigenetic landscape to be more like that of embryonic stem cell by removing the repressive epigenetic marks and relaxing chromatin structure to allow the reprogramming factors easier access to target genes. In the present study, we assessed the effect of pretreatment with small molecules on the expression of Oct-4, Nanog, and Sox-2 in bovine fetal fibroblast cells. Chemical treatment consisted of 3 small molecules: PD0325901, a mitogen-activated protein kinase inhibitor; CHIR99021, a glycogen synthase kinase-3 inhibitor; and NuP0178 (NuPotential, Baton Rouge, LA, USA), a G9a histone methyltransferase inhibitor. Cells were seeded at a density of 0.7 × 106 and expanded before being divided evenly into 2 groups. Control cells were cultured in complete culture medium (DMEM with 10% fetal bovine serum), whereas treatment cells were cultured in complete culture medium containing 0.5 mM PD0325901, 3 mM CHIR99021, and 1.8 mM NuP0178. Messenger RNA was isolated from cell cultures using Dynabeads® mRNA DIRECT™ Kit (Dynal Inc., Lake Success, NY, USA) on Day 7 and 14, and the resulting RNA products were transcribed into cDNA using Bio-Rad iScript™ cDNA Synthesis Kit (Bio-Rad, Hercules, CA, USA). Quantitative RT–PCR was performed to measure transcript levels of Oct-4, Nanog, Sox-2, and poly adenylate polymerase in treated and untreated cells at Days 7 and 14. Transcript levels were quantified by relative quantification using the ΔΔCt method and expressed as ratios of target genes (Oct-4, Nanog, Sox-2) to the reference gene (PAP) and normalized against a calibrator consisting of untreated bovine fetal fibroblast cells. No difference in expression levels between untreated and treated cells was detected at either Day 7 or 14. Currently, we are utilising chromatin immunoprecipitation to examine chromatin and DNA methylation patterns around the promoters of Oct-4, Nanog, and Sox-2 to further elucidate the effects of treatment with this combination of small molecule inhibitors.