bluuebunny/crossref_metadata_2025 huggingface dataset:
Cicero and the Rise of Deification at Rome
Authors: Cole Spencer
Published: December 2013

This book tells a part of the back-story to major religious transformations emerging from the tumult of the late Republic. It considers the dynamic interplay of Cicero's approximations of mortals and immortals with a range of artifacts and activities that were collectively closing the divide between humans and gods. A guiding principle is that a major cultural player like Cicero had a normative function in religious dialogues that could legitimize incipient ideas like deification. Applying contemporary metaphor theory, it analyzes the strategies and priorities configuring Cicero's divinizing encomia of Roman dynasts like Pompey, Caesar and Octavian. It also examines Cicero's explorations of apotheosis and immortality in the De re publica and Tusculan Disputations as well as his attempts to deify his daughter Tullia. In this book, Professor Cole transforms our understanding not only of the backgrounds to ruler worship but also of changing conceptions of death and the afterlife.
Indian English and the Fiction of National Literature
Authors: George Rosemary Marangoly
Published: December 2013

During the twentieth century, at the height of the independence movement and after, Indian literary writing in English was entrusted with the task of consolidating the image of a unified, seemingly caste-free, modernising India for consumption both at home and abroad. This led to a critical insistence on the proximity of the national and the literary, which in turn, led to the canonisation of certain writers and themes and the dismissal of others. Examining English anthologies of 'Indian literature', as well as the establishment of the Sahitya Akademi (the national academy of letters) and the work of R. K. Narayan and Mulk Raj Anand among others, Rosemary George exposes the painstaking efforts that went into the elaboration of a 'national literature' in English for independent India even while deliberating the fundamental limitations of using a nation-centric critical framework for reading literary works.
Surface-motility induction, attraction and hitchhiking between bacterial species promote dispersal on solid surfaces
Authors: Hagai Efrat , Dvora Reut , Havkin-Blank Tal , Zelinger Einat , Porat Ziv , Schulz Stefan , Helman Yael
Published: December 2013

The ability to move on solid surfaces provides ecological advantages for bacteria, yet many bacterial species lack this trait. We found that Xanthomonas spp. overcome this limitation by making use of proficient motile bacteria in their vicinity. Using X. perforans and Paenibacillus vortex as models, we show that X. perforans induces surface motility, attracts proficient motile bacteria and ‘rides’ them for dispersal. In addition, X. perforans was able to restore surface motility of strains that lost this mode of motility under multiple growth cycles in the lab. The described interaction occurred both on agar plates and tomato leaves and was observed between several xanthomonads and motile bacterial species. Thus, suggesting that this motility induction and hitchhiking strategy might be widespread and ecologically important. This study provides an example as to how bacteria can rely on the abilities of their neighboring species for their own benefit, signifying the importance of a communal organization for fitness.
Biopharmaceutical Evaluation of Various Dosage Forms Intended for Caffeine Topical Delivery
Authors: Briançon S. , Chevalier Y. , Bolzinger M.-A.
Published: December 2013

The efficacy of actives applied by the topical route is limited not just by factors relating to the active ingredient(s) but also formulation design and biological factors such as skin permeability. Topical formulations have to be designed to ensure that a sufficient dose of the active molecule reaches its target site, be it the skin surface, the skin itself or the underlying cutaneous vasculature. In order to achieve this, the active substance has to overcome the natural barrier function of the skin. Several strategies have been studied to enhance skin penetration and so improve cutaneous bioavailability.
Synergistic Effects of Transcutol and Terpenes as Penetration Enhancers: In Vitro and In Vivo ATR-FTIR Spectroscopic Imaging Studies
Authors: Erdal M. S. , Özdin D. , Güngör S.
Published: December 2013

The skin offers an ideal application site to deliver therapeutic agents for both topical and systemic effect. However, the integument is also a well-designed membrane to protect the organism from environmental factors, thus it is an effective obstacle for drug permeation. The main barrier for skin delivery is the stratum corneum (SC), the outermost layer of the epidermis. The SC is formed by dead and keratinized cells which present a unique barrier to passage of drugs across the skin [1, 2]. Several approaches are used to overcome the barrier property of SC in topical and transdermal administration of drugs, the most frequent of which is the inclusion of chemical penetration enhancers in formulations.
Topical Microneedle Drug Delivery Enhanced with Magnetophoresis
Authors: Prow T. W. , Mohammed Y. H. , Ansaldo A. B. , Benson H. A. E.
Published: December 2013

Proteins and peptides are coming of age as targeted biological therapies. A broad range of peptides have now been approved by the FDA and other regulatory agencies for use as therapeutics. Likewise, large proteins such as infliximab and botox are commonly used to treat skin conditions. The most widely used delivery approach for peptides and proteins is parenteral injection, largely due to oral instability and the lack of a suitable alternative. Intense research is being done to improve the short half-lives of these biological therapies and find suitable alternative delivery routes. Transdermal delivery holds promise as an improved delivery route because the skin has low levels of enzymatic activity and can help drugs avoid first-pass metabolism. Further, topical delivery has the potential for direct treatment of skin disease for both therapeutic and cosmetic applications.
Iontophoretic Delivery of Glycomimetics-A New Approach for the Treatment of Inflammatory Skin Diseases
Authors: Gratieri T. , Wagner B. , Kalaria D. , Ernst B. , Kalia Y. N.
Published: December 2013

A key event in the inflammatory response is the recruitment of leukocytes from the microcirculation to the inflamed tissue through endothelial-dependent mechanisms that include leukocyte tethering and rolling, activation, firm adhesion and diapedesis to the interstitium. Leukocyte tethering and rolling along the endothelial surface is mediated by a family of carbohydrate-binding proteins (E-, P- and L-selectins). While L-selectin is constitutively expressed on the surface of leukocytes, expression of E- and P-selectin is upregulated on the endothelial surface during inflammation.
Topical Bioequivalence: A Comprehensive Approach Using Multiple Surrogate Methods
Authors: Lehman P. A. , Raney S. G. , Franz T. J.
Published: December 2013

At the present time, establishing the bioequivalence (BE) of topical drug products is not a simple process: in contrast to oral medications, costly and time-consuming clinical trials are required for most topical products. In the United States there are provisions within the regulations that allow other methods (i.e. surrogates for clinical trials) to be used to establish BE. The stated preference of the Food and Drug Administration (FDA) is for pharmacodynamics effect studies, but in vivo animal studies as well as in vitro studies are potentially acceptable. Although there is currently only one approved surrogate test within the topical products area (the “skin blanching” or “vasoconstrictor assay” for glucocorticoids), FDA activity over the past twenty five years through sponsored scientific meetings and research, as well as in-house research and initiatives, has led to the development of several alternative test methods for topical products.
Classification of Irritant/Non-Irritant and Corrosive Chemicals Using Excised Human Skin In Vitro
Authors: Stewart C. A. , Turner R. B. , Brown M. B. , Traynor M. J.
Published: December 2013

Any chemical that is applied to the skin has the potential to induce a skin reaction. These reactions are complicated and are difficult to fully replicate in vitro and may exhibit species variations when reproduced in vivo. Skin irritation is the observed response (erythema, oedema, pain itching and heat) to chemicals that result in inflammation at the site of application. Skin irritation is defined as reversible damage of the skin following application of a chemical for up to 4 h. By contrast, skin corrosion is defined as ‘'irreversible damage to the skin, namely visible necrosis through the epidermis and into the dermis, following the application of a test substance for a period of 3 min up to 4 h”.
Influence of Receptor Media on GD Skin Penetration In Vitro
Authors: Dalton C. H. , Graham S. J. , Payne O. J. , Chipman J. K. , Chilcott R. P. , Jenner J.
Published: December 2013

When designing in vitro static diffusion studies the choice of receptor media is important as it may influence measured penetration rates. OECD guidelines regarding suitable receptor fluids state that “The use of a physiologically conducive receptor fluid is preferred although others may also be used provided that they are justified”. Further to this is the statement, “Adequate solubility of the test chemical in the receptor fluid should be demonstrated so that it does not act as a barrier to absorption.” Guidance for acceptable receptor fluids for non-viable skin preparations is split between the evaluation of water soluble compounds and lipohilic test substances. For the former saline receptor solutions are preferred, whilst for lipophilic test substances it is suggested that “the receptor fluid can contain organic solvents such as 1:1 ethanol: water or 6% polyethylene glycol 20 oleyl ether in water.”
Evaluation of Methylphenidate Permeation from Daytrana™ Patches Across Silicone and Human Epidermal Membranes
Authors: Edwards A. , Liu F. , Brown M. B. , Mcauley W. J.
Published: December 2013

Methylphenidate (MPH) is a mild central nervous system stimulant and has been a common drug used for treating attention deficit hyperactivity disorder (ADHD) for numerous years. MPH inhibits re-uptake of dopamine into the pre-synaptic nerve by blocking the dopamine transporter, leading to an increase in dopamine within the synapse and further stimulation of the dopamine receptors on the post-synaptic nerve. In patients with ADHD this has a beneficial effect on behaviour, cognition, short-term memory, reaction time, vigilance and learning. MPH is currently available in a range of dosage forms from oral controlled or immediate release formulations through to Daytrana™ a controlled release transdermal matrix patch.
A T7 exonuclease-assisted cyclic enzymatic amplification method coupled with rolling circle amplification: a dual-amplification strategy for sensitive and selective microRNA detection
Authors: Cui Liang , Zhu Zhi , Lin Ninghang , Zhang Huimin , Guan Zhichao , Yang Chaoyong James
Published: December 2013

A T7 exonuclease-assisted cyclic enzymatic amplification method (CEAM) was combined with rolling circle amplification (RCA) to develop a RCA–CEAM dual amplification method for ultrasensitive detection of microRNA with excellent selectivity.
Experimental and theoretical analysis of lp⋯π intermolecular interactions in derivatives of 1,2,4-triazoles
Authors: Shukla Rahul , Mohan T. P. , Vishalakshi B. , Chopra Deepak
Published: December 2013

The calculations performed on the evaluation of the electrostatic potential provide deeper insights into the nature of lp⋯π interactions.
Ultralong H2V3O8 nanowire bundles as a promising cathode for lithium batteries
Authors: An Qinyou , Sheng Jinzhi , Xu Xu , Wei Qiulong , Zhu Yaqin , Han Chunhua , Niu Chaojiang , Mai Liqiang
Published: December 2013

In situ transport measurement of a single nanowire demonstrates that the H2V3O8 nanowire possesses good electrical transport properties.
Exploring the interaction of cisplatin with β2-microglobulin: new insights into a chemotherapeutic drug
Authors: Zhang Ningbo , Cui Meng , Du Yonggang , Liu Zhiqiang , Liu Shuying
Published: December 2013

Four binding sites for cisplatin to β2M have been identified.
Stable phase domains of the TiO2–Ti3O5–Ti2O3–TiO–Ti(CxOy)–TiC system examined experimentally and via first principles calculations
Authors: Kim Jiwoong , Kang Shinhoo
Published: December 2013

The stable phase domains of the TiO2–Ti3O5–Ti2O3–TiO–Ti(CxOy)–TiC system were examined experimentally and using first principles calculations.
Hierarchical core–shell α-Fe2O3@C nanotubes as a high-rate and long-life anode for advanced lithium ion batteries
Authors: Gu Xin , Chen Liang , Liu Shuo , Xu Huayun , Yang Jian , Qian Yitai
Published: December 2013

Hierarchical core–shell α-Fe2O3@C nanotubes, prepared by a mild hydrothermal reaction, presented excellent cycling stability and rate capability for lithium ion batteries.
High performance hybrid white and multi-colour electroluminescence from a new host material for a heteroleptic naphthyridinolate platinum complex dopant
Authors: Poloek Anurach , Chen Chin-Ti , Chen Chao-Tsen
Published: December 2013

A new heteroleptic platinum complex FPtmND doped in a conventional CBP or new 4P-NPD host has been explored for high efficiency greenish-yellow or orange-red phosphorescent OLEDs, and high colour rendering index hybrid white OLEDs with simple device configuration.
Deep atomic force microscopy
Authors: Barnard H. , Drake B. , Randall C. , Hansma P. K.
Published: December 2013

The Atomic Force Microscope (AFM) possesses several desirable imaging features including the ability to produce height profiles as well as two-dimensional images, in fluid or air, at high resolution. AFM has been used to study a vast selection of samples on the scale of angstroms to micrometers. However, current AFMs cannot access samples with vertical topography of the order of 100 μm or greater. Research efforts have produced AFM scanners capable of vertical motion greater than 100 μm, but commercially available probe tip lengths are still typically less than 10 μm high. Even the longest probe tips are below 100 μm and even at this range are problematic. In this paper, we present a method to hand-fabricate “Deep AFM” probes with tips of the order of 100 μm and longer so that AFM can be used to image samples with large scale vertical topography, such as fractured bone samples.
Self-consistent continuum solvation (SCCS): The case of charged systems
Authors: Dupont C. , Andreussi O. , Marzari N.
Published: December 2013

The recently developed self-consistent continuum solvation model (SCCS) [O. Andreussi, I. Dabo, and N. Marzari, J. Chem. Phys. 136, 064102 (2012)] is applied here to charged species in aqueous solutions. Describing ions in solution represents a great challenge because of the large electrostatic interactions between the solute and the solvent. The SCCS model is tested over 106 monocharged species, both cations and anions, and we demonstrate its flexibility, notwithstanding its much reduced set of parameters, to describe charged species in solution. Remarkably low mean absolute errors are obtained with values of 2.27 and 5.54 kcal/mol for cations and anions, respectively. These results are comparable or better than the state of the art to describe solvation of charged species in water. Finally, differences of behavior between cations and anions are discussed.
Velocity gradient dynamics in compressible turbulence: Characterization of pressure-Hessian tensor
Authors: Suman Sawan , Girimaji Sharath S.
Published: December 2013

Pressure-Hessian tensor produces the most significant difference between incompressible and compressible velocity gradient dynamics in turbulent flows. Characterization of pressure-Hessian tensor as a function of the level of compressibility is therefore of much interest. Using direct numerical simulation results, we demonstrate that the pressure-Hessian tensor behavior can be most exclusively characterized in terms of the compressibility parameter δ which is defined to be the growth-rate of dilatation-rate. A key compressibility effect is the distinct change in the alignment between pressure-Hessian and velocity gradient tensors with increasing δ. In incompressible turbulence, the pressure-Hessian eigenvectors exhibit a mild tendency to align at 45° angle with the principal directions of strain rate. With increasing δ, the pressure-Hessian tensor shows progressively stronger tendency to align along principal directions of the local strain-rate tensor. We show that this change in pressure-Hessian orientation causes the compressible velocity gradient dynamics to significantly differ from its incompressible counterpart. In incompressible turbulence, pressure mildly moderates the inherent gradient-steepening tendencies of the nonlinear inertial term. On the other hand, in highly compressible turbulence (extreme values of δ), pressure effects can lead to intense gradient steepening or smoothing depending upon the growth-rate of dilatation rate, thereby profoundly altering the cascade process.
Barrier height formation in organic blends/metal interfaces: Case of tetrathiafulvalene-tetracyanoquinodimethane/Au(111)
Authors: Martínez José I. , Abad Enrique , Beltrán Juan I. , Flores Fernando , Ortega José
Published: December 2013

The interface between the tetrathiafulvalene/tetracyanoquinodimethane (TTF-TCNQ) organic blend and the Au(111) metal surface is analyzed by Density Functional Theory calculations, including the effect of the charging energies on the molecule transport gaps. Given the strong donor and acceptor characters of the TTF and TCNQ molecules, respectively, there is a strong intermolecular interaction, with a relatively high charge transfer between the two organic materials, and between the organic layer and the metal surface. We find that the TCNQ LUMO peak is very close to the Fermi level; due to the interaction with the metal surface, the organic molecular levels are broadened, creating an important induced density of interface states (IDIS). We show that the interface energy level alignment is controlled by the charge transfer between TTF, TCNQ, and Au, and by the molecular dipoles created in the molecules because of their deformations when adsorbed on Au(111). A generalization of the Unified-IDIS model, to explain how the interface energy levels alignment is achieved for the case of this blended donor/acceptor organic layer, is presented by introducing matrix equations associated with the Charge Neutrality Levels of both organic materials and with their intermixed screening properties.
Double shock front formation in cylindrical radiative blast waves produced by laser irradiation of krypton gas
Authors: Kim I. , Quevedo H. J. , Feldman S. , Bang W. , Serratto K. , McCormick M. , Aymond F. , Dyer G. , Bernstein A. C. , Ditmire T.
Published: December 2013

Radiative blast waves were created by irradiating a krypton cluster source from a supersonic jet with a high intensity femtosecond laser pulse. It was found that the radiation from the shock surface is absorbed in the optically thick upstream medium creating a radiative heat wave that travels supersonically ahead of the main shock. As the blast wave propagates into the heated medium, it slows and loses energy, and the radiative heat wave also slows down. When the radiative heat wave slows down to the transonic regime, a secondary shock in the ionization precursor is produced. This paper presents experimental data characterizing both the initial and secondary shocks and numerical simulations to analyze the double-shock dynamics.
High multi-photon visible upconversion emissions of Er3+ singly doped BiOCl microcrystals: A photon avalanche of Er3+ induced by 980 nm excitation
Authors: Li Yongjin , Song Zhiguo , Li Chen , Wan Ronghua , Qiu Jianbei , Yang Zhengwen , Yin Zhaoyi , Yang Yong , Zhou Dacheng , Wang Qi
Published: December 2013

Under 980 nm excitation, high multi-photon upconversion (UC) emission from the 2H11/2/4S3/2 (green) and 4F9/2 (red) levels of Er3+ ions were observed from Er3+ singly doped BiOCl microcrystals. These high-energy excited states were populated by a three to ten photon UC process conditionally, which depended on the pump power density and the Er3+ ion doping concentration, characterizing as a hetero-looping enhanced energy transfer avalanche UC process. UC emission lifetime and Raman analysis suggest that the unusual UC phenomena are initiated by the new and intense phonon vibration modes of BiOCl lattices due to Er3+ ions doping.
Note: Hollow cathode lamp with integral, high optical efficiency isolation valve: A modular vacuum ultraviolet source
Authors: Sloan Roberts F. , Anderson Scott L.
Published: December 2013

The design and operating conditions of a hollow cathode discharge lamp for the generation of vacuum ultraviolet radiation, suitable for ultrahigh vacuum (UHV) application, are described in detail. The design is easily constructed, and modular, allowing it to be adapted to different experimental requirements. A thin isolation valve is built into one of the differential pumping stages, isolating the discharge section from the UHV section, both for vacuum safety and to allow lamp maintenance without venting the UHV chamber. The lamp has been used both for ultraviolet photoelectron spectroscopy of surfaces and as a “soft” photoionization source for gas-phase mass spectrometry.
Near-field heat transfer between gold nanoparticle arrays
Authors: Phan Anh D. , Phan The-Long , Woods Lilia M.
Published: December 2013

The radiative heat transfer between gold nanoparticle layers is presented using the coupled dipole method. Gold nanoparticles are modelled as effective electric and magnetic dipoles interacting via electromagnetic fluctuations. The effect of higher-order multipoles is implemented in the expression of electric polarizability to calculate the interactions at short distances. Our findings show that the near-field radiation reduces as the radius of the nanoparticles is increased. Also, the magnetic dipole contribution to the heat exchange becomes more important for larger particles. When one layer is displayed in parallel with respect to the other layer, the near-field heat transfer exhibits oscillatory-like features due to the influence of the individual nanostructures. Further details about the effect of the nanoparticles size are also discussed.
Size-dependent shifts of the Néel temperature and optical band-gap in NiO nanoparticles
Authors: Thota Subhash , Shim J. H. , Seehra M. S.
Published: December 2013

Bulk NiO is a well-known antiferromagnet with Neel temperature TN(∞) = 524 K and an optical band-gap Eg = 4.3 eV. With decrease in particle size D from 40 nm to 4 nm of NiO, systematic changes of TN and Eg are observed and discussed here. From magnetic measurements, the changes in TN with D are found to fit finite-size scaling equation TN(D) = TN(∞) [1 − (ξo/D)λ] with λ = 3.2 ± 0.5 and ξo = 3.2 ± 0.2 nm, in good agreement with the predictions for a Heisenberg system. The observed blue shifts of Eg with decrease in D reaching Eg = 5.12 eV for D ∼ 4 nm are likely due to quantum confinement and non-stoichiometry.
Robust adaptive variable speed control of wind power systems without wind speed measurement
Authors: Song YongDuan , Zhang ZhenYu , Li Peng , Wang WenLiang , Qin Ming
Published: December 2013

The stochastic and immeasurable nature of wind imposes significant challenge to control design for wind turbine (WT) systems. This work presents a robust adaptive approach to achieve variable speed tracking control of WT wind turbine without the need for wind speed measurement. The adverse effect arisen from wind is accommodated by the robust and adaptive control algorithms based on virtual parameter concept. A hill-climb searching algorithm is exploited to provide desired rotor speed signal, avoiding wind speed measurement. The stability of the proposed control scheme is authenticated by Lyapunov stability theory. The effectiveness and the benefits of the proposed approach are validated and demonstrated by numerical simulation.
Partial cationic inversion-induced magnetic hardening of densely packed 23-nm-sized nanocrystallite-interacting nickel ferrite electrospun nanowires
Authors: Li Jian-Min , Zeng Xian-Lin , Xu Zhu-An
Published: December 2013

Although the average crystallite size (23 nm) is very close to a critical radius of superparamagnetism, a magnetic hardening (coercive field and saturation magnetization) was observed in interacting single-domain nanogranular nickel ferrite electrospun nanowires as compared with the bulk. The phenomena can be attributed to a small change in the cation occupancy on the spinel sites from (Fe3+)A(Ni2+Fe3+)BO4 to [F1−δ3+Niδ2+] A[Ni1−δ2+Fe1+δ3+] BO4 (δ = 3.67%), and an additive interaction of the “chain of spheres” type. The larger shape anisotropy contribution (Keff = 8.24 × 104 erg/cm3) is also as a factor leading to enhanced coercive field.
Permeation of low-Z atoms through carbon sheets: Density functional theory study on energy barriers and deformation effects
Authors: Huber Stefan E. , Mauracher Andreas , Probst Michael
Published: December 2013

Energetic and geometric aspects of the permeation of the atoms hydrogen to neon neutral atoms through graphene sheets are investigated by investigating the associated energy barriers and sheet deformations. Density functional theory calculations on cluster models, where graphene is modeled by planar polycyclic aromatic hydrocarbons (PAHs), provide the energies and geometries. Particularities of our systems, such as convergence of both energy barriers and deformation curves with increasing size of the PAHs, are discussed. Three different interaction regimes, adiabatic, planar and vertical, are investigated by enforcing different geometrical constraints. The adiabatic energy barriers range from 5 eV for hydrogen to 20 eV for neon. We find that the permeation of oxygen and carbon into graphene is facilitated by temporary chemical bonding while for other, in principle reactive atoms, it is not. We discuss implications of our results for modeling chemical sputtering of graphite.
Mobilising the Child Victim: The Localisation of Human Trafficking in Singapore through Global Activism
Authors: Yea Sallie
Published: December 2013

In this paper I examine the local mobilisation of the dominant global framing of the problem of human trafficking through ‘the female child victim’ in child sex trafficking advocacy campaigns. The child victim is a symbolic and emotive frame embodied in the Third World female child and enacted through her helplessness and experiences of extreme violence and (sexual) abuse in trafficking situations across diverse contexts globally. I use The Body Shop's (TBS's) 2009–12 global campaign against child sex trafficking as my site for discussion of the way frames in global human rights activism move into local contexts, often coming to define the ways contemporary human rights problems are understood and reproduced locally. I draw on ethnographic research on human trafficking in Singapore to explore the ways in which the child victim frame is mobilised in a specific locale through the involvement of a local nongovernmental organisation and university student actors as part of TBS's campaign strategy. Although recent geographical scholarship on social movements has embraced a networked approach, I argue for heightened attention to the geographies of scaled (re)iteration, or local mobilisation that occurs as transnational activism connects with particular places. The role of framing in embedding global human rights issues locally in transnational activism is central to this process.
Genetic control of reproduction in dairy cows
Authors: Butler Stephen T.
Published: December 2013

The advent of AI has markedly improved the production potential of dairy cows in all systems of production and transformed the dairy industry in many countries. Unfortunately, for many years breeding objectives focused solely on milk production. This resulted in a major decline in genetic merit for fertility traits. In recent years, the underlying physiological mechanisms responsible for this decline have started to be unravelled. It is apparent that poor genetic merit for fertility traits is associated with multiple defects across a range of organs and tissues that are antagonistic to achieving satisfactory fertility performance. The principal defects include excessive mobilisation of body condition score, unfavourable metabolic status, delayed resumption of cyclicity, increased incidence of endometritis, dysfunctional oestrus expression and inadequate luteal phase progesterone concentrations. On a positive note, it is possible to identify sires that combine good milk production traits with good fertility traits. Sire genetic merit for daughter fertility traits is improving rapidly in the dairy breeds, including the Holstein. With advances in animal breeding, especially genomic technologies, to identify superior sires, genetic merit for fertility traits can be improved much more quickly than they initially declined.
104 FAILURE TO REMOVE BLUETONGUE SEROTYPE 8 VIRUS (BTV-8) FROM IN VITRO-PRODUCED BOVINE EMBRYOS
Authors: Penido A. O. , De Clerq K. , Haegeman A. , Vandaele L. , Nauwynck H. , Van Soom A.
Published: December 2013

Bluetongue virus serotype 8 (BTV-8) causes some unique characteristics compared with other BTV strains, such as transplacentary transmission, infertility, and diminished health of the offspring (De Clercq et al. 2008 Transboundary and Emerging Diseases 55, 352–359), and concerns exist about the risk of the transmission of the disease via embryo transfer (Vandaele et al. 2012). It is known that most pathogenic agents can be eliminated by washing and trypsin treatment of intact embryos according to the IETS guidelines, but some viruses adhere strongly to the zona pellucida and are not removed by this process (Ali al Ahmad et al. 2011 Theriogenology 76, 126–132). The aim of this study was to investigate decontaminating methods for bovine in vitro embryos that had been infected in vitro with BTV-8, which were earlier shown to be effective in goat embryos (REF). In vitro bovine blastocysts (n = 105) were placed in 800 μL of minimal essential medium (MEM), containing 104.9 50% tissue culture infectious doses (TCID50) of BTV-8 (Bel 2006/2 P5, VAR, Brussels, Belgium) and incubated for 1 h at 39°C in 5% CO2 in air (Vandaele et al. 2011 Vet. Res. 42, 14–21). The embryos were exposed to trypsin either at 37°C [Group 1 (G1)] or at room temperature [Group 2 (G2)], with 3 treatments per group (5 embryos/treatment), consisting of 5 washes in PBS without BSA; 2 washes in 0.25% trypsin for 45 s each [treatment 1 (T1)], 2 washes in 0.25% trypsin-EDTA for 60 s each [treatment 2 (T2)], or 2 washes in 0.25% trypsin for 90 s each [treatment 3 (T3)]; and 10 washes in PBS + 0.4% BSA. All the treatments were done in triplicate. The efficiency of the different washing techniques and trypsin temperature for virus removal was evaluated by RT-quantitative PCR (qPCR) on embryos and washes. Virus isolation was performed on embryonated chicken eggs as described by Vandaele et al. (2011 Vet. Res. 42, 14–21) for the first and last washing fluids and for the embryos. Room temperature was 24.9°C. Viral BTV RNA was detected by RT-PCR in the first 5 washes in all groups and treatments. After the trypsin wash, all samples remained negative until the last wash procedure. Viral isolation was positive in the first 3 washes and negative in the 10th wash. The embryos were positive on RT-PCR in at least 2 replicates of each treatment, but all samples remained negative on virus isolation. The results show that the wash procedure is efficient to remove the virus from the wash media, but it failed to remove the virus from bovine embryos produced in vitro. The temperature (37°C or room temperature) did not influence the efficiency of the trypsin treatment.
111 REGULATORY microRNA ENRICHMENT AND DEGRADATION IN GRANULOSA CELLS DURING BOVINE FOLLICULAR RECRUITMENT AND DOMINANCE
Authors: Salilew-Wondim D. , Ahmad I. , Gebremedhn S. , Sahadevan S. , Hoelker M. , Rings F. , Udin J. , Tholen E. , Looft C. , Schellander K. , Tesfaye D.
Published: December 2013

Follicular development is a result of complex hormonal and biochemical synergies that could be activated or deactivated in a spatiotemporal manner in oocytes and surrounding cells including theca, granulosa, and cumulus cells. The microRNA (miRNA), 19 to 22 nucleotides noncoding RNA, are one of the molecular cues that could play a role in posttranscriptional regulation of genes involved in follicular development. Here we aimed to understand the availability and abundance of miRNA in bovine granulosa cells (GC) derived from subordinate (SF) and leading or dominant (DF) follicles during bovine follicular recruitment and dominance at Day 3 and 7 of the oestrus cycle, respectively. For this, Simmental heifers (n = 15) were oestrus synchronized and slaughtered at 3 (n = 6) and 7 (n = 7) days after the onset of the oestrous. The SF and DF were retrieved from each animal to obtain the corresponding GC, which were subjected to miRNA-enriched total RNA isolation using the miRNeasy mini kit (Qiagen GmbH, Hilden, Germany). The integrity and quality of RNA was determined using Agilent 2100 Bioanalyzer (Agilent Technologies Inc., Santa Clara, CA, USA) and Nanodrop 8000 Spectrophotometer (Thermo Fisher Scientific Inc., DE, USA), respectively. The RNA was then subjected to miRNA deep sequencing using the Illumina HISEqn 2000. Raw sequence data were further processed and analysed using miRDeep2 software package. Quantification of differentially expressed (DE) miRNA was done using R software and DESEqn 2 package. MiRNA with log2 fold change difference ≥1, P-value ≤0.05, and false discovery rate ≤1 were considered to be significant. Data analysis revealed that 291 and 311 miRNA were detected in GC of SF and 312 and 314 were detected in GC of DF at Days 3 and 7, respectively. A total of 17 miRNA were DE in GC from SF compared with the DF at Day 3, of which 15 miRNA were enriched and the remaining 2 were down-regulated in SF. Similarly, at Day 7 a total of 136 miRNA was altered with 51 miRNA showed to be enriched, whereas 85 others remained low in SF compared with the DF. Nine miRNA (bta-miR-21–3p, bta-miR-221, bta-miR-708, bta-miR-214, bta-miR-335, bta-miR-155, bta-miR-199a-5p, bta-miR-21–5p and bta-miR-222) were commonly differentially expressed both at Day 3 and 7 between SF and DF. Interestingly, all of the commonly DE miRNA, except bta-miR-335, were enriched in GC of SF at both days. Gene ontological analysis indicated that majority of the DE miRNA were found to be involved in regulation of programmed cell death, cell projection morphogenesis, regulation of cell proliferation, and macromolecule biosynthesis. Therefore, the temporal abundance of mature miRNAs in GC during bovine follicular development may suggest their potential role in regulation of follicular development in general and follicular recruitment and dominance in particular.
114 PROGESTERONE CONCENTRATIONS DURING IVM AFFECT BOVINE OOCYTE QUALITY AT THE MOLECULAR LEVEL
Authors: Schlüter N. , Kassens A. , Stinshoff H. , Knauer K. , Wilkening S. , Wrenzycki C.
Published: December 2013

Progesterone (P4) is important for the developmental competence of cumulus–oocyte complexes (COC) used for in vitro maturation (IVM). In a recent study, we were able to show that circulating P4 concentrations had an effect on the molecular quality of COC recovered during repeated ovum pickup sessions, which might affect further development. (Schlüter et al. 2013 Reprod. Fertil. Dev. 25, 250). The aim of the present study was to determine the influence of different P4 concentrations during IVM on the molecular quality of bovine COC. The COC were collected from slaughterhouse ovaries and cultured as described recently (Stinshoff et al. 2011 Theriogenology 76, 1433–1441). The IVM medium was supplemented with 0, 50, 150, 300, and 450 ng mL–1 P4. Ethanol served as vehicle control. After IVF with a bull of proven fertility, the presumptive zygotes were cultured in SOF under 5% oxygen (Stinshoff et al. 2011). Cleavage and developmental rates were determined at Day 3 and Day 7/8 (Day 0: IVF). Additionally, maturation rates were assessed. For mRNA analysis, immature and matured denuded COC (n = 5) were individually frozen at –80°C to analyse the relative transcript abundance using RT-qPCR. The transcripts studied play important roles during oocyte development [growth differentiation factor 9 (GDF9), bone morphogenetic protein 15 (BMP15), glucose transporter 1 (SCL2A1), hypoxia inducible factor 2α (HIF2α), nuclear progesterone receptor (PGR), progestin and adipoQ receptor 5 (PAQR5), progesterone receptor membrane component 1 and 2 (PGRMC1, PGRMC2)]. Data were tested using ANOVA followed by multiple pairwise comparisons using Tukey's test. A P-value of <0.05 was considered significant. The percentage of oocytes that reached the MII stage was similar in oocytes from all treatment groups (82.2–88.1%). Despite similar cleavage rates across all groups, the developmental rates did show significant differences. More embryos developed to the blastocyst stage stemming from oocytes cultured without any supplement or cultured only with alcohol compared to oocytes stemming from the group cultured with less than 50 ng mL–1 P4 (25.4 ± 5.7 and 27.9 ± 7.2 v. 15.8 ± 2.6). The relative abundance of SCL2A1, BMP15, and PGRMC1transcripts in single oocytes did not show differences related to the supplementation of the IVM medium, whereas GDF9, HIF2α, and PAQR5 mRNA was reduced in oocytes of all groups compared with immature ones. The PGR and PGRMC2 transcripts were increased in matured oocytes of the control group and the vehicle control group (PGRMC2). In summary, supplementation of the IVM medium with different P4 concentrations had an effect on the molecular quality of oocytes after IVM, which might affect further development.
The financial support of the FBF (Förderverein Biotechnologieforschung) e.V. is gratefully acknowledged.
121 EXPRESSION OF STEROID RECEPTORS IN THE CUMULUS - OOCYTE COMPLEX AROUND OVULATION IN THE BITCH
Authors: Tahir M. Z. , Reynaud K. , Thoumire S. , Chastant-Maillard S. , Saint-Dizier M.
Published: December 2013

In the bitch, oocytes are ovulated at an immature stage (prophase I) and resume meiosis in the oviduct, 3 to 4 days after ovulation while they are still surrounded by 2 to 3 layers of cumulus cells. Canine cumulus-oocyte complexes (COC) are exposed to high and changing plasma concentrations of 17β-oestradiol (E2) and progesterone (P4) during the periovulatory period. In order to explore whether oocyte maturation may be regulated by steroids in this species, the expression of E2 (ERα, ERβ) and P4 (nuclear: PR; membrane: PGRMC1, PGRMC2, mPRα, mPRβ, mPRγ) receptors was studied in COC at precise times around ovulation. Ovaries were collected from Beagle bitches during anestrus (n = 4), after the beginning of proestrus, and before the LH peak (Pre-LH, n = 7), after the LH peak and before ovulation (Pre-ov, n = 8), and at Day 1 (n = 11) and Day 4 (n = 8) post-ovulation. Anoestrus COC were recovered from follicles smaller than 1 mm in diameter. The COC at the Pre-LH and Pre-ov stages were aspirated from preovulatory follicles (4.5–6 mm in diameter). Such follicular COC were partially denuded to leave the 2 to 3 innermost cumulus layers firmly attached to the zona pellucida. Post-ovulatory COC, naturally surrounded by 2 to 3 cumulus layers, were recovered by oviductal flushing. Total RNA was extracted from 3 batches of 10 COC per stage, then reverse transcribed. The expression of steroid receptors was assessed in duplicate by qPCR (LightCycler® 480, Roche Diagnostics) using the relative standard curve method and normalized by the geometric mean value of the two most stable reference genes (BGLR and RPS5; NormFinder software) chosen among four genes previously tested. Relative amounts of mRNA levels were compared between stages by ANOVA followed, when necessary, by a Tukey test. The ERα and ERβ expression did not vary significantly with the stage. In contrast, a significant variation between stages in nuclear and 4 membrane P4 receptor expression was observed (P < 0.0001 for PR; P < 0.001 for PGRMC1 and mPRβ; P < 0.05 for PGRMC2 and mPRγ). The PR mRNA levels were significantly higher at Pre-ov than at any other stage. PGRMC1 expression was significantly higher at Pre-ov and Day 4 compared with anestrus and Pre-LH, and was at an intermediate level at Day 1. The expression of PGRMC2, mPRβ, and mPRγ remained low from anestrus to Day 1 and increased significantly at Day 4. Lastly, mRNA levels of mPRα were below the detection limit at all stages. This is the first report of steroid receptor expression in canine COC at precise times around ovulation. The stage-specific variation in expression of nuclear and of several membrane P4 receptors around ovulation suggests a role for P4 in canine oocyte maturation. The exact localisation of these receptors in cumulus cells, oocytes, or both remains to be determined.
123 CHANGES IN EXPRESSION OF GENES ASSOCIATED WITH GENETIC VARIATION IN PRE-IMPLANTATION DEVELOPMENT OF THE BOVINE EMBRYO
Authors: Ortega M. S. , Cole J. B. , Sonstegard T. S. , Hansen P. J.
Published: December 2013

The objective was to identify patterns of expression during the pre-implantation period of several genes associated with genetic variation in fertility (CWC15) or development to the blastocyst stage (C1QB, MON1B, PARM1, PCCB, PMM2, TBC1D24, and WBP1). These genes are involved in cellular processes such as mRNA splicing, immune protection, fatty acid oxidation, resistance to apoptosis, glycoprotein synthesis, and intracellular transport. Embryos were produced in vitro from slaughterhouse oocytes and semen using a mix of Bos taurus and Bos indicus cows and bulls. Pools of 40 matured oocytes or embryos at the 2-cell [27–31 h post-insemination (hpi)], 3- to 4-cell (46–52 hpi), 5- to 8-cell (49–59 hpi), 9- to 16-cell (72–75 hpi), morula (120–123 hpi), and blastocyst (168–171 hpi) stages were collected. The RNA was purified and synthesised into cDNA for real-time qPCR analysis. The YWHAZ, GAPDH, and SDHA were used as steady-state controls of expression. A total of 5 pools were analysed for each of the 6 stages. The C1QB was not detected at any stage; however, transcript amounts for the other genes were affected by stage of development (P < 0.05). The WBP1 remained low from the oocyte to the 5- to 8-cell stage (fold-change relative to matured oocytes: 1.0 ± 0.2 v. 1.4 ± 0.2), increased at the 9- to 16-cell stage (14.8 ± 0.2), and decreased to the blastocyst stage (7.1 ± 0.2). The expression pattern of PARM1 was similar, with greatest expression at the 9- to 16-cell stage. In contrast, expression of PMM2 and TBC1D24 was highest at the 2-cell stage and decreased at the morula and blastocyst stages. Expression of CWC15, MON1B, and PCCB decreased steadily from the oocyte to the blastocyst stage. Given that the major round of embryonic genome activation occurs at the 8- to 16-cell stage, it is possible that PARM1 and WBP1 play important roles around this time. The PMM2 and TBC1D24 may represent genes activated before the 8- to 16-cell stage. The CWC15 has been identified as a lethal gene; results suggest lethality occurs after the blastocyst stage. Further research will clarify the role and importance of these genes in the early development of the bovine embryo.
The authors acknowledge support from AFRI Grant No. 2013–68004–20365 from USDA NIFA.
135 RECOMBINANT BOVINE SOMATOTROPIN ON THE IN VITRO PRODUCTION OF BOVINE EMBRYOS
Authors: Berté L. , Vasconcelos L. , Hatamoto-Zervoudakis L. , Yamazaki W. , Yamazaki L. , Santos G.
Published: December 2013

Bovine growth hormone (bGH) has been used to improve the results for in vitro production of bovine embryos. Inclusion of bGH in the maturation medium increases both rate of cleavage and frequency of blastocyst development. Thus, the purpose of the present study was to evaluate the effect of recombinant bovine somatotropin (rBST) on cleavage and blastocyst development of bovine embryos when included during in vitro maturation (IVM) only (Group 1), during both IVM and in vitro culture (IVC; Group 2), during IVC only (Group 3), or not included during either IVM or IVC (Group 4). Specifically, in Group 1, oocytes were matured in TCM 199 (Earle's salts) supplemented with 10% FCS, LH, FSH, oestradiol, and amikacin (IVM medium), plus 100 ng mL–1 of rBST and cultured in SOFaaci supplemented with essential amino acids, tri-sodium citrate, myo-inositol, and 5% FBS. In group 2, oocytes were matured in IVM medium containing 100 ng mL–1 of rBST and cultured in SOFaaci supplemented with essential amino acid, tri-sodium citrate, myo-inositol, 5% FBS; on Day 5, rBST (50 ng mL–1) was added. In Group 3, oocytes were matured in IVM medium without rBST; on Day 5, rBST (50 ng mL–1) was added. Group 4 was the control, without rBST supplementation. The treatment groups were analysed using the SAS® (SAS Institute Inc., Cary, NC, USA) in a completely randomised design (P < 0.05). Somatotropin has receptors in cumulus cells and in the zona pellucida acting directly in the oocyte; however, the increase in cleavage rate seen in previous studies after rBST treatment was not observed in the present study. Supplementation of culture medium with rBST during Day 2 to 6 of IVC has been shown to increase the number of trophoblast and subsequent pregnancy rate following transfer. However, in the present study, addition of 50 and 100 ng mL–1 of rBST to the maturation or culture medium did not affect the cleavage rate of embryos and blastocyst production.

Table 1.Analysis of the meaning and percentages related to cleavage and production of embryos