bluuebunny/crossref_metadata_2025 huggingface dataset:
Axiomatic Cloud Computing Architectural Design
Authors: Thomas John , Mantri Pam
Published: December 2019

Modern cloud computing makes available a plethora of scalable cloud computing offerings. The cloud is increasingly becoming the backbone of the highly complex modern knowledge-economy that includes Social, Mobile, IoT, Big-Data and AI. Knowledge-based products and services follow fat-tail distributions such as the power-law that poses major opportunities and challenges for the designer. The Axiomatic Designer is uniquely positioned in designing for the de-novo situations that the fat-tailed distributions expose. Also, the cloud frees-up the architectural decision-making away from the legacy compatibility-burden, and towards various cloud-native (i.e., de-novo/solution-neutral) as well as hybrid (on-prem/cloud & cloud/cloud) architectures. Further more, the competitive landscape around the cloud is not static; it is adaptive and evolving rapidly. Here again, Axiomatic Design (AD) is uniquely positioned in rising upto the various de-novo challenges This, however, requires contributions from frameworks such as Knowledge-as-Heterarchically-Hierarchical (KA|h|H), Stigmergy, Complex Adaptive Systems (CAS), Cynefin, Boyd’s OODA-Loop Theory of asymmetric fast-transients, Axiomatic-Maturity-Diagram (AMD), as well as Weick’s Loose-Coupling approach to help unify and strengthen the Axiomatic approach. This paper unifies the above approaches in order to tackle the architectural challenges of cloud computing.
Das Mikrobiom bei Adipositas und kardiometabolische Erkrankungen: Die Mischung macht es!
Authors: Chakaroun Rima , Blüher Matthias
Published: December 2019

Das Darmmikrobiom ist ein komplexes Ökosystem, das aus einer Vielzahl von hauptsächlich Bakterien, aber auch Archaeen und Viren besteht. In den letzten Jahren konnte ein starker Zusammenhang zwischen Mikrobiom-Veränderungen und der Entstehung von Erkrankungen, zu denen auch Adipositas und verschiedene Stoffwechselkrankheiten gehören, gefunden werden. Darmbakterien sind an der Synthese essentieller Vitaminen, Extraktionen von Nährstoffen sowie den Abbau von Ballaststoffen beteiligt. Zusätzlich spielt das Mikrobiom eine wichtige Rolle in der Reifung des Immunsystems des Wirts, seiner Darmbarriere, der neuronalen intestinalen Aktivität, und der Produktion und Freisetzung von gastrointestinalen Hormonen. Veränderungen der Zusammensetzung des mikrobiellen Ökosystems im Darm sind mit dem Auftreten von Adipositas, Typ 2 Diabetes, arterieller Hypertonie, nicht alkoholischer Lebererkankung (NAFLD) sowie kardiovaskulären Krankheiten assoziiert.

Vor allem tierexperimentelle Untersuchungen weisen auf einen starken Kausalzusammenhang zwischen dem Mikrobiom und diesen Erkrankungen hin. Hierfür können zusätzlich zu wirtspezifischen oder Umweltfaktoren sowohl Zusammensetzung wie auch die Funktion der Darmbakterien eine chronische systemische Inflammation mitverursachen und unterhalten.

Ein möglichst diverses Mikrobiom kann die metabolische Gesundheit des Wirts positiv beeinflussen und stellt somit einen potentiellen therapeutischen Ansatz bei kardiometabolischen Erkrankungen dar. Beispielsweise werden Stuhltransplantationen zur Therapie unterschiedlicher Erkrankungen wie Clostridium difficile-Infektionen eingesetzt und verbessern auch bei Personen mit Metabolischem Syndrom die Insulinsensitivität. Bevor eine Modulation des Mikrobioms in der Zukunft zur Prävention und Therapie von Adipositas und begleitenden Erkrankungen eingesetzt werden kann, müssen noch weitere klinische Studien die Effektivität und Sicherheit einer gezielten Manipulation der Darmbakterien belegen.
Smoldering Myelom
Authors: Arnheim Katharina
Published: December 2019

Patienten mit Smoldering Myelom (SM) profitieren vom präventiven Einsatz von Lenalidomid: Das immunmodulatorisch wirksame Arzneimittel (IMiD®) reduziert das Risiko für eine Progression zum symptomatischen multiplen Myelom (MM) um 72 %.
Gastroösophageale Tumoren
Authors: Arnheim Katharina
Published: December 2019

Wie die Phase-III-Studie GO2 zeigt, kann die Intensität der Chemotherapie bei nicht fitten Patienten mit gastroösophagealen (GO) Karzinomen ohne Wirksamkeitseinbuße deutlich reduziert werden.
Checkpoint-Inhibitoren – Immuneffekte beim NSCLC und Melanom
Authors: Krome Susanne
Published: December 2019

Die Tumorimmuntherapie mit Checkpoint-Inhibitoren verlängert das Überleben von Patienten mit malignen Melanomen und nicht-kleinzelligen Bronchialkarzinomen (NSCLC). Mehrere Studien berichten beim malignen Melanom über eine Assoziation zwischen unerwünschten Immuneffekten und der Therapieresponse. Eine neue Studie belegt dies nun auch für eine anti-PD1-Therapie des NSCLC. Darüber hinaus fanden die Forscher eine pathophysiologische Begründung: Das Tumorgewebe und die Haut wiesen überlappende T-Zellrezeptor-Clonotypen auf.
Steuerung der TKI-Therapie beim EGFR-NSCLC
Authors: Krome Susanne
Published: December 2019

Tyrosinkinase-Inhibitoren (TKI) haben die Prognose des nicht-kleinzelligen, EGFR-mutierten Bronchialkarzinoms (EGFRmut-NSCLC) substanziell verbessert. Dauerhafte Remissionen sind dabei allerdings selten. Eine retrospektive Studie identifizierte prädiktive genetische Biomarker, welche den Erfolg einer Therapie mit TKI der 1. Generation vorhersagten.
Klinikeinkauf in der Schweiz: Besser werden unter veränderten Rahmenbedingungen
Authors: Rößing Sabine
Published: December 2019

In der Schweiz steigen die Kosten für den medizinischen Sachmittelbedarf, während sich Erlöse und Rahmenbedingungen ändern – etwa mit den Swiss Diagnosis Related Groups (SwissDRG). Um Wirtschaftlichkeit bei höchster medizinischer Qualität zu erzielen, ist in den Spitälern ein Paradigmenwechsel nötig.
Umfrage zum Klinikeinkauf: Die Diskrepanz zwischen Anforderung und Know-how
Authors: Buchhart Susann
Published: December 2019

Was muss ein Klinikeinkäufer können? Welche Kompetenzen fehlen? Und wie bewerten Klinikgeschäftsführer die Qualifikationen in den Einkaufsabteilungen ihrer Klinik? Die Online-Umfrage von consus clinicmanagement zeigt, welche Diskrepanzen es zwischen Anforderungen, Ausbildung und Fortbildungen aktuell gibt.
Deutschland in der Demografischen Krise: Neue Schwerpunkte für das Supply-Chain-Management im Krankenhaus
Authors: Richtzenhain Stephan
Published: December 2019

Krisen kann man nur mit Innovationen begegnen. Einkauf bzw. Supply-Chain-Management bedeutet in der Krise eben auch Innovationsmanagement. Hier müssen zukünftig andere Prioritäten gesetzt werden. Es zählt mehr die Investition in Zukunftsfähigkeit, als der letzte Cent an Einsparung im Einkauf. Und letztendlich hat auch der Einkauf seinen Beitrag zur Entwicklung einer wertschätzenden Unternehmenskultur beizutragen.
Existence and asymptotic stability for the semilinear wave equation with variable-exponent nonlinearities
Authors: Rahmoune Abita
Published: December 2019

In this paper, we review a class of semilinear wave equation with variable-exponent nonlinearities and boundary conditions. This work is devoted to prove the existence of solutions and uniform decay rates of the wave equation with boundary damping and source terms.
Hot carrier dynamics in a dispersionless plasmonic system
Authors: Nayak Banoj Kumar , Prabhu S. S. , Achanta Venu Gopal
Published: December 2019

Hot carrier dynamics in a broadband, dispersionless, plasmonic structure are studied using pump-probe measurements with 45 fs time resolution. The results obtained by simultaneous measurement of differential transmittance and reflectance show the influence of pump generated carriers on the probe generated plasmons. The dependence of decay time on the pump fluence is linear for the interband and intraband carrier excitation near the L-symmetry point in gold. However, 1.908 eV pump excitation, which allows both interband transition at the X-symmetry point and intraband excitation at the L-symmetry point, shows quadratic dependence of decay time on the pump fluence. Degenerate pump-probe measurements at 1.55 eV, which allow L-valley intraband excitations, highlight the difference in surface (reflection) and the bulk (transmission) mechanisms. Hot carrier relaxation time is in the 1–3 ps range for different excitation energies. Rise time, governed by the plasmon to hot carrier conversion and electron-electron scattering processes, is about 200 ± 35 fs for the hot carrier and hot plasmon excitation cases that increased to about 485 ± 35 fs when the pump is resonant with the interband transition at the X-symmetry point. Results show that the hot carrier dynamics are governed by the bulk metal band structure. The dipole matrix element for each of the transitions is estimated by density matrix calculations.
Laser-ion acceleration using mixed compositions: Tailoring the target for each species
Authors: Stark D. J. , Yin L. , Albright B. J.
Published: December 2019

Particle-in-cell simulations of laser- ion acceleration demonstrate marked discrepancies in the acceleration experienced by the different ion species in complex target compositions, especially when the target becomes relativistically transparent to the pulse during irradiation. Beginning with proton contaminants in a carbon target, we show how the higher charge-to-mass ratio of the protons results in species stratification and late-time suppression of the carbon acceleration. The target normal sheath acceleration (TNSA) primarily experienced by the protons can be exploited to mitigate this tamping by using a shaped rear surface of the target, leaving the break-out afterburner-driven carbons to accelerate close to the laser axis and then experience less tamping during a late-time TNSA phase. We then explore preferentially accelerating the lighter species in a mixed composition target, particularly focusing on deuteron beam applications. We examine three different target compositions with the same areal electron density, CD2, CH2, and 7LiD, and propose an alternative shaping of the rear surface of 7LiD to increase the number of high-energy deuterons in the beam.
Demonstration of genuine surface inversion for the p/n-In0.3Ga0.7Sb-Al2O3 MOS system with in situ H2 plasma cleaning
Authors: Millar David A. J. , Peralagu Uthayasankaran , Li Xu , Steer Matthew J. , Fu Yen-Chun , Hurley Paul K. , Thayne Iain G.
Published: December 2019

The results of an investigation into the impact of in situ H2 plasma exposure on the electrical properties of the p/n-In0.3 Ga0.7 Sb-Al2O3 interface are presented. Samples were processed using a clustered inductively coupled plasma reactive ion etching and atomic layer deposition tool. Metal oxide semiconductor capacitors were fabricated subsequent to H2 plasma processing and Al2O3 deposition, and the corresponding capacitance-voltage and conductance-voltage measurements were analyzed quantitatively via the simulation of an equivalent circuit model. Interface state (Dit) and border trap (Nbt) densities were extracted for samples subjected to the optimal process, with a minimum Dit of 1.73×1012 eV−1 cm−2 located at ∼110 meV below the conduction band edge and peak Nbt approximately aligned with the valence and conduction band edges of 3×1019 cm−3 and 6.5×1019 cm−3, respectively. Analysis of the inversion response in terms of the extraction of the activation energy of minority carriers in inversion (p-type) and the observation of characteristics that pertain to minority carriers being supplied from an external inversion region (n-type) unequivocally demonstrate that the Fermi level is unpinned and that genuine surface inversion is observed for both doping polarities.
Control of electrochemical reduction behavior in nonequilibrium Al-doped TiO2 thin films
Authors: Ishii Akihiro , Oikawa Itaru , Takamura Hitoshi
Published: December 2019

To develop resistive switches using TiO2, the control of their switching voltage is a crucial issue. This study shows how acceptor and compensating oxygen vacancies modify the resistive switching behavior from both the experimental and theoretical points of view. Nonequilibrium TiO2 thin films doped with acceptor Al with oxygen vacancies were prepared by pulsed laser deposition. Al doping results in a clear reduction in the switching voltage when the TiO2 thin films are operated as a unipolar-type switch but not when they are operated as bipolar-type switches. Density functional theory calculations revealed that the doped Al and compensating oxygen vacancies ([Al′Ti]=2[VO⋅⋅]) promote the formation of oxygen vacancies compensated by electrons (2[VO⋅⋅]=n) due to the defects association of 2Al′Ti−2VO⋅⋅−2e′. This defect cluster functions like the formation center of conductive nanofilaments for the electrochemical reduction. It was concluded that by promoting the formation of oxygen vacancies compensated by electrons, Al doping results in a decrease in energy consumption of the unipolar-type resistive switching.
Instability of thermocapillary–buoyancy convection in droplet migration
Authors: Hu Kai-Xin , Yan Chen-Yi , Chen Qi-Sheng
Published: December 2019

The instabilities of thermocapillary–buoyancy convection in droplet migration are examined by linear stability analysis. The droplet is flattened by gravity and placed on a unidirectional heated solid surface. The velocity and temperature distributions of basic flow are derived as a function of the migration velocity and the Bond number. The critical Marangoni number is obtained, which depends on the Prandtl number (Pr), the Bond number, and the migration velocity. The preferred modes at small and moderate Pr are oblique waves, which travel either upstream or downstream. For high Pr, the preferred modes include oblique and streamwise waves, while the amplitude of temperature on the surface is much smaller than that of the hot spot in the flow region. The instability mechanism is discussed and comparisons are made with liquid layers.
Enhanced solvation force extrapolation for speeding up molecular dynamics simulations of complex biochemical liquids
Authors: Omelyan Igor , Kovalenko Andriy
Published: December 2019

We propose an enhanced approach to the extrapolation of mean potential forces acting on atoms of solute macromolecules due to their interactions with solvent atoms in complex biochemical liquids. It improves and extends our previous extrapolation schemes by additionally including new techniques such as an exponential scaling transformation of coordinate space with weights complemented by an automatically adjusted balancing between the least square minimization of force deviations and the norm of expansion coefficients in the approximation. The expensive mean potential forces are treated in terms of the three-dimensional reference interaction site model with Kovalenko-Hirata closure molecular theory of solvation. During the dynamics, they are calculated only after every long (outer) time interval, i.e., quite rarely to reduce the computational costs. At much shorter (inner) time steps, these forces are extrapolated on the basis of their outer values. The equations of motion are then solved using a multiple time step integration within an optimized isokinetic Nosé-Hoover chain thermostat. The new approach is applied to molecular dynamics simulations of various systems consisting of solvated organic and biomolecules of different complexity. For example, we consider hydrated alanine dipeptide, asphaltene in toluene solvent, miniprotein 1L2Y, and protein G in aqueous solution. It is shown that in all these cases, the enhanced extrapolation provides much better accuracy of the solvation force approximation than the existing approaches. As a result, it can be used with much larger outer time steps, leading to a significant speedup of the simulations.
Modifications produced on a large magnetized plasma column by a floating end-plate that is partially emissive: Experiment and theory
Authors: Van Compernolle B. , Poulos M. J. , Morales G. J.
Published: December 2019

An experiment is performed on a large plasma device operated by the Basic Plasma Science Facility at the University of California, Los Angeles, in which an electrically floating structure is placed near the end of the 18-m magnetized plasma column. The structure consists of a flat carbon plate that acts as a mask for a smaller, ring-shaped LaB6 emissive surface whose temperature can be externally controlled. This configuration has been previously used to study electron heat transport and pressure-driven avalanches [B. Van Compernolle and G. J. Morales, Phys. Plasmas 24, 112302 (2017)] by biasing the LaB6 ring-cathode with respect to a distant anode in a cold afterglow plasma. In contrast, the present study is performed during the active portion of the steady-state discharge in which the nominal plasma parameters are determined by the injection of an electron beam from a BaO cathode at the opposite end. It is found that, even without an applied bias on the LaB6 cathode, the self-consistent potential and current profiles are modified near the end plate as the LaB6 temperature is increased, resulting in density increases on the field lines in contact with the ring-cathode. In the absence of enhanced ionization, at the largest cathode temperatures, the ambient density can be doubled. A theoretical model is presented that provides a quantitative explanation for the experimental observations.
Spatial distribution of the phononic crystal modes excited by a moving laser source
Authors: Li Junyan , Lomonosov Alexey. M. , Shen Zhonghua , Ni Chenyin , Yuan Ling , Kan Weiwei
Published: December 2019

In this paper, we have presented the dispersion curve of Lamb waves in an aluminum plate with a periodic rectangular corrugation experimentally. A mode selection method was applied to generate Lamb waves in the region of the corrugated periodic structure using a moving continuous wave laser, and the signal can be detected at any point in the excitation region. The relationship between the phase velocity and the frequency is observed. It is found that the intensity distribution of the dispersion curves in the experimental results is related to the slope of the dispersion curves as well as the selection of the detection point. Vibrations at a series of locations were detected, reflecting the change in energy that Lamb waves undergo in the process of propagation through the periodic structure, which is different from the existing method of single-point excitation and detection. The spatial distribution of the mode amplitude at the resonances points, in particular induced by the Bragg reflection, has been studied.
Photophysical and electronic properties of bismuth-perovskite shelled lead sulfide quantum dots
Authors: Abdu-Aguye Mustapha , Bederak Dmytro , Kahmann Simon , Killilea Niall , Sytnyk Mykhailo , Heiss Wolfgang , Loi Maria Antonietta
Published: December 2019

Metal halide perovskite shelled quantum dot solids have recently emerged as an interesting class of solution-processable materials that possess the desirable electronic properties of both quantum dots and perovskites. Recent reports have shown that lead sulfide quantum dots (PbS QDs) with perovskite ligand-shells can be successfully utilized in (opto)electronic devices such as solar cells, photoconductors, and field-effect transistors (FETs), a development attributed to the compatibility of lattice parameters between PbS and certain metal halide perovskites that results in the growth of the perovskite shell on the PbS QDs. Of several possible perovskite combinations used with PbS QDs, bismuth-based variants have been shown to have the lowest lattice mismatch and to display excellent performance in photoconductors. However, they also display photoluminescence (PL), which is highly sensitive to surface defects. In this work, we present an investigation of the transport and optical properties of two types of bismuth-based perovskite (MA3BiI6 and MA3Bi2I9) shelled PbS QDs. Our photophysical study using temperature-dependent PL spectroscopy between 5 and 290 K indicates that the PL efficiency of the reference oleic acid (OA) capped samples is much higher than that of the Bi-shelled ones, which suffer from traps, most likely formed at their surfaces during the phase-transfer ligand exchange process. Nevertheless, the results from electrical measurements on FETs show the successful removal of the native-OA ligands, displaying electron dominated transport with modest mobilities of around 10−3 cm2 [V s]−1 – comparable to the reported values for epitaxial Pb-based shelled samples. These findings advance our understanding of perovskite shelled QD-solids and point to the utility of these Bi-based variants as contenders for photovoltaic and other optoelectronic applications.
Influence of a mineral supplement containing calcium, phosphorus and micronutrients on intake, digestibility, performance and mineral status of young Nellore bulls in a feedlot
Authors: Zanetti D. , Godoi L. A. , Estrada M. M. , Engle T. E. , Pacheco M. V. C. , Pereira J. M. V. , Detmann E. , Paulino M. F. , Valadares Filho S. C.
Published: December 2019

We measured nutrient intake and digestibility, performance and mineral status of young Nellore bulls fed three diets with and without supplementation comprising calcium (Ca), phosphorus (P) and micro-minerals during the growing and finishing phases. Five bulls were slaughtered at the beginning of the experiment to measure initial mineral status. Forty-two young Nellore bulls (initial bodyweight (mean ± s.d.) 270.4 ± 36.6 kg, age 8 months) were assigned for 125 days to one of six treatments: sugarcane as a roughage source plus a concentrate based on soybean meal and soybean hull with (SH100) and without (SH0) supplementation; sugarcane as a roughage source plus a concentrate based on soybean meal and ground maize with (SC100) and without (SC0) supplementation; and maize silage as a roughage source plus a concentrate based on soybean meal and maize with (CS100) and without (CS0) supplementation. The experiment was conducted as a completely randomised design with a 3 × 2 factorial arrangement of treatments. Nutrient intake and digestibility, bone and serum parameters related to Ca and P metabolism, and liver mineral concentrations were measured. Nutrient intake, digestibility and performance were not affected (P > 0.10) by the mineral factor. Water intake was higher (P < 0.05) with SH100 than with all other diets. Rib-bone breaking strength and density according to dual-energy X-ray absorptiometry were reduced (P < 0.04) in the absence of supplementation. Metatarsus parameters were not affected (P > 0.10). Liver copper (Cu) content was reduced (P < 0.01) in diets without supplementation. In conclusion, the short-term absence of mineral supplementation did not influence intake and digestibility. However, depending on diet type, absence of Cu may reduce the hepatic Cu content, and a low Ca:P ratio may stimulate bone reabsorption without compromising performance. Therefore, under our conditions, mineral requirements are overestimated by nutritional systems.
125 Transcriptomic changes in bovine ovarian cortex in response to FSH signaling
Authors: Candelaria J. , Rabaglino B. , Denicol A.
Published: December 2019

Preantral follicles serve as a reservoir of female gametes that could be used in assisted reproductive technologies in humans, livestock, and endangered animals. Invitro culture of ovarian cortex is a widely used method to grow preantral follicles. Follicle-stimulating hormone (FSH) is often added to the culture medium as a folliculogenesis-promoting factor. The roles of FSH in antral follicles is well known; however, the effects of FSH in preantral follicles and indirectly in the ovarian cortical cells is largely unknown. The aim of this study was to determine the transcriptomic responses of the ovarian cortex containing preantral follicles to FSH signalling over time. In 3 biological replicates, small strips of bovine ovarian cortex (10×5mm) were dissected from the medulla and evaluated under a stereomicroscope for removal of all visible antral follicles. Resulting cortical strips were cultured in defined medium with human-recombinant FSH or vehicle for 2 or 4 days at 38.5°C and 5% CO2. The RNA was isolated and subjected to cDNA library preparation and 3′-Tag RNA sequencing. Sequencing data analysis was performed using the edgeR and maSigPro packages (Bioconductor-R). Using a time-course analysis, genes up- or downregulated 2-fold or more and associated with an FDR<0.05 were considered differentially expressed (DEG) and were further analysed with NetworkAnalyst software. We found 252 DEG over time in response to FSH. In FSH-treated samples, significantly enriched biological functions from upregulated genes were associated with glycolysis, gluconeogenesis, carbon metabolism, and biosynthesis of amino acids. In contrast, significantly enriched biological functions from downregulated genes found in FSH-treated samples included phagosome and necroptosis. The germ cell markers BMP15, DAZL, DDX4, GDF9, and ZP2/ZP3 were expressed but unchanged by FSH, suggesting the presence of similar numbers of oocytes between samples. The gene B4GALT2, previously reported as a granulosa cell marker, was upregulated in FSH-treated samples at Day 4. The follicular marker RAB23 was expressed in all samples and not changed by FSH. One interesting finding was upregulation of MAPK signalling (represented by the genes MAPKAPK3, ELK4, MKNK2, and TGFB3) in response to FSH signalling, with no change in expression of the cAMP-response element responsive genes CYP19A1 and INHA. Together, these data indicate that FSH stimulates energy metabolism in ovarian cortical cells and represses negative cell function activity. We conclude that these responses are mostly mediated by granulosa cells, because the FSH receptor is not appreciably expressed in the ovarian cortex stroma. Moreover, the data suggest that FSH may utilise alternative signalling pathways, such as MAPK, in early follicles. This information enhances our understanding of FSH signalling pathways in the ovarian cortex, mediated by preantral follicles to create a positive environment for folliculogenesis.
158 Optimal time of AI and changes in vaginal mucus characteristics relative to the onset of standing oestrus in Beetal goats
Authors: Murtaza A. , Irfan-ur-Rehman Khan M. , Abbas M. , Ahmad W. , Tahir M. Z. , Mohsin I.
Published: December 2019

The aim of this study was to determine the optimum time of AI and its relationship with vaginal mucus characteristics relative to the onset of standing oestrus in Beetal goats. For this purpose, goats (n=257) were synchronised for oestrus by administering two doses of prostaglandin F2α (0.075mg intramuscularly, d+ cloprostenol, Dalmazin) 11 days apart during the breeding season (September-December, 2016-2018). The onset of standing oestrus was determined using apronised bucks. Intracervical inseminations were performed at 0 (n=28), 12 (n=84), 24 (n=101), and 36h (n=44) after the onset of standing oestrus using chilled semen (diluted in skim milk at 200×106 spermatozoa per 0.5-mL dose). Color (transparent=1, turbid=2, cheesy/whitish=3), consistency (thin=1, thick=2), and volume (excess=1, scant=2) of the vaginal mucus in goats were observed at the time of inseminations and graded to determine the cumulative mucus score. Resistive index of vaginal mucus (R-value) was monitored at the time of AI using a Draminski heat detector. The intervals to the onset of oestrus among different months of synchronisation, cumulative mucus score, and vaginal electrical resistance at different times of AI were compared using one-way analysis of variance. Pregnancy rates at different times of AI were compared using chi-square analysis, and the Pearson coefficient of correlation was used to correlate mucus color, consistency, and volume with the cumulative mucus score. Following the second prostaglandin F2α administration, the interval to the onset of standing oestrus was longer (P<0.05) in goats synchronised in December than in those synchronised from September to November. The pregnancy rate was lower (P<0.05) in goats inseminated at 0h (28.6%) compared with those inseminated at 12 (58.3%), 24 (56.4%), and 36h (54.5%). However, the pregnancy rates were similar in goats inseminated at 12, 24, and 36h (P>0.05). Odds for pregnancy rate at 12h were 3.5, 1.08, and 1.16 times higher than those at 0, 24, and 36h, respectively. Relative to the time of AI, the cumulative mucus scores varied (P<0.05), and changes in mucus colour and consistency were more highly correlated (r=0.832 and 0.793, respectively; P<0.05) with cumulative mucus score than the mucus volume (r=0.588). Relative to the onset of standing oestrus, the R-value of the mucus was lower at 12h than at 0 and 36h (P<0.05); however, there was no change in R-value of the mucus at 12 and 24h. In conclusion, the optimum time to inseminate Beetal does using chilled semen is 12h following the onset of oestrus, whereas cumulative mucus score and R-value can be used as alternatives if the time of the onset of oestrus is not known.
221 Isolation of turkey spermatogonia
Authors: Tomgorova E. K. , Vetokh A. N. , Volkova L. A. , Volkova N. A. , Ashraf H. V. , Bagirov V. A. , Zinovieva N. A.
Published: December 2019

Across different types of spermatogenic epithelium, researchers have focused on the study and production of spermatogonial stem cells and their use as genetic material for creating gene pool cryobanks of valuable breeds and lines of poultry. The aim of our research was to identify optimal conditions for obtaining and culturing turkey spermatogonia. A series of experiments were carried out to determine the optimal age of males for isolating spermatogonia and optimize conditions for their isolation and cultivation. Age-related features of turkey spermatogenesis were assessed by testis histological studies. Statistical analysis was performed using SPSS v15.0 (t-test; SPSS Inc./IBM Corp.). Turkey testis tissue was mechanically crushed and treated with enzymes: 0.1% collagenase and 0.25% trypsin. Purification of spermatogonia from other types of cells was performed by adhesion and separation. The conditions and duration of spermatogenic cell cultures were selected experimentally. Primary Sertoli cells of turkeys, Sertoli cells of roosters, STO cell line, and transplanted Sertoli cells of pigs (PTP) were used as a feeder layer for the cultivation of spermatogonia. Spermatogonia culture medium consisted of Dulbecco's modified Eagle's medium (DMEM)-high glucose supplemented with 5% fetal calf serum, 2mM α-glutamine, 10µLmL−1 MEM, 100x antibiotics, 10µLmL−1 ITS solution, 5×10−5 M mercaptoethanol, 5mgmL−1 albumin, 1µLmL−1 DL-lactic acid, 20ngmL−1 epidermal growth factor, 10ngmL−1 basic fibroblast growth factor, and 2ngmL−1 leukemia inhibitory growth factor (LIF). The SSEA-1 antibody was used for identification of spermatogonia colonies. Histological study of the turkey testes at different ages showed that in animals under 12 weeks, the population of generative cells was represented mainly by spermatogonia (P<0.05). Thus, spermatogonia were isolated from the testes of 4-wk-old males. Isolated cells were cultured for 24h, after which the supernatant containing non-adherent cells (mainly spermatogonia) was transferred to a new dish for continued culture. Maximal homogeneity of the cell population was detected after transferring non-adherent cells 3 times at 24-h intervals. The best cell growth was observed when spermatogonia were cultivated on turkey Sertoli cells. The number of these cells in suspension reached 81% purity. Formation of spermatogonia colonies was observed on Days 7 to 8 of cultivation, the presence of which was confirmed by immunohistochemical staining with SSEA-1.
This study was supported by the Russian Science Foundation within project no.16-16-04104.
226 Effect of the presence or absence of corpus luteum in females with small antral follicles at the beginning of a progesterone-based oestrus synchronization protocol on superovulatory response and embryo yields in sheep
Authors: Lima M. S. D. , Vergani G. B. , Silva K. M. , Monteiro A. W. U. , Pereira L. M. C. , Ramos L. M. C. , Batista R. I. T. P. , Fonseca J. F. , Vicente W. R. R. , Oliveira M. E. F. , Teixeira D. I. A.
Published: December 2019

The high variability of responses to multiple ovulation and embryo transfer (MOET) programs is related to several factors, including the ovarian population present at the beginning of hormonal protocols. Thus, our objective was to evaluate the effect of the presence or absence of corpus luteum (CL) in females presenting small antral follicles (SF) at the beginning of a progesterone (P4)-based oestrus synchronization protocol on superovulatory response and embryo yields. The study was conducted during September and October in northeast Brazil (03°40′26″S and 40°14′20″W) using 15 adult Santa Inês ewes, in which the ovarian population present were SF (≤3.75mm, n=5) or SF plus CL (SF+CL, n=10) at the beginning of P4-based oestrus synchronization protocol. The females received a new or reused intravaginal device of progesterone (CIDR Eazi-breed, Zoetis) on Day 0, which remained for 9 days. On Day 7, the pFSH (133mg, Folltropin V, Vetoquinol) treatment was begun, with 6 decreasing doses (25, 25, 15, 15, 10, and 10%) injected IM at 12-h intervals. On Day 9, two equal doses of D-cloprostenol were injected at a 12-h interval (37.5μg, Prolise, Agener União). Flunixin meglumine (24.9 mg; Banamine, MSD Animal Health) was administered IM on Days 13 to 15. All ewes showed oestrus and were mated with fertile rams. On Day 16 non-surgical embryo recovery (NSER) was performed after cervical dilation using D-cloprostenol and oestradiol benzoate at 16h and ocytocin at 20min before. Transrectal B-mode ultrasound evaluations (Z5 Vet, Mindray) were performed on Day 0 and just before NSER to evaluate the ovarian population present. Data are showed in mean±s.d. and compared by t-test for independent samples (P<0.05). The number of CL present on NSER day (21.5±4.4 and 4.0±8.5) and the number of viable embryos (11.1±14.9 and 4.0±8.5) differed (P<0.05) between SF+CL ewes and SF ewes, respectively. However, the number of luteinizing unovulated follicles present on NSER day (4.5±2.5 and 12.4±7.8) and the number of recovered structures (i.e. embryos plus unfertilized eggs; 14.2±15.6 and 4.2±8.2) did not differ (P>0.05) between SF+CL ewes and SF ewes, respectively. In conclusion, the presence of a CL at the beginning of the oestrous synchronization protocol used for superovulation treatment affects responses to the MOET program.
Financial support for this study was provided by Embrapa (02.13.06.026.00.02 and 02.13.06.026.00.04) and FAPEMIG (PPM 00201-17).
45 Expression patterns of PRDM family genes in porcine pre-implantation embryos
Authors: Farrell K. , Uh K. , Lee K.
Published: December 2019

Establishing proper levels of pluripotency is essential for normal development. The genome of gametes is remodelled upon fertilisation and pluripotency-related genes are expressed in blastocysts. Multiple pluripotency-related genes are involved in the well-orchestrated process; however, detailed mechanistic actions remain elusive. The PRDM family genes are reported to be closely related to the pluripotency. A previous report noted that PRDM14 plays an important role in the maintenance of pluripotency in human embryonic stem cells (ESCs) and potentially murine ESCs; loss of PRDM14 was found to cause abnormalities in genome-wide epigenetic status. Similarly, PRDM15 was found to be a key regulator of pluripotency in mouse ESCs. Structural similarities among the PRDM family suggest that other PRDM family genes may help to establish and maintain pluripotency in embryos. Unfortunately, little is known about the expression profile of PRDM family in porcine embryos. To expand our understanding of the role of PRDM family in porcine embryos, expression patterns of PRDM gene family were investigated using reverse transcription quantitative (RTq)-PCR. Candidate PRDM family genes were selected based on previous RNA-Seq data in porcine oocytes/embryos. To conduct this study, germinal vesicle (GV), MII, zygote, 4-cell, and blastocyst samples were collected. Complementary DNA synthesised from the samples was used for RT-qPCR to analyse the expression pattern of selected PRDM family genes: PRDM2, PRDM4, PRDM6, PRDM14, and PRDM15. The expression of target genes was normalized to the YWHAG level, an internal control. Then, GV stage was used as a control for ΔΔCT analysis. Two technical replications and three biological replications were performed. Analysis of variance was used for statistical analysis and P-values<0.05 were considered significant. There was a significant decrease in PRDM2 expression in 4-cell and blastocyst, PRDM4 expression in 4-cell, and PRDM6 in all stages (MII, zygote, 4-cell, and blastocyst), compared with the GV stage. Because zygotic genome activation occurs at the 4-cell stage in the pig, the significant decrease in gene expression (PRDM2, PRDM4, and PRDM6) indicates they may be maternally originated and involved in the reprogramming process following fertilisation. On the other hand, there was a significant increase in PRDM15 expression in blastocysts and the PRDM14 transcript was only detected in blastocysts in all three biological replicates, suggesting that the genes are most likely involved in pluripotency maintenance, as was found in previous human studies. These results indicate that PRDM family genes are differentially expressed during early embryo development in pigs and may play a role in maintenance of pluripotency. For further study, we intend to evaluate the role of PRDM family genes during early embryo development in pigs.
60 CRISPR-on, a new tool for activation of endogenous gene expression in bovine embryos
Authors: Savy V. , Alberio V. , Canel N. , Ratner L. , Gismondi M. , Taboga O. , Ferraris S. , Rulli S. , Fernandez-Martin R. , Bevacqua R. , Salamone D.
Published: December 2019

The CRISPR-Cas9 system enables precise genome editing in mammalian somatic cells and embryos at a very high efficiency. A modified version of Cas9 (dCas9) was engineered, resulting in a DNA binding protein capable of site-specific target recognition but unable to cut the DNA. By means of dCas9 fusion to heterologous domains, including transcriptional activators or repressors, specific modulation of gene expression has successfully been achieved invitro, making possible the modulation of the cell-differentiation state. However, CRISPR-mediated transcriptional activation (CRISPR-on) has been mainly used invitro, and to our knowledge, there are no reports regarding its use for the activation of endogenous gene expression in mammalian embryos. As a proof of principle, we evaluated the CRISPR-on system in bovine embryos for modulation of endogenous expression of SMARCA4 and TFAP2C, transcription factors implicated in trophoblast lineage commitment. We hypothesised that CRISPR-on may induce SMARCA4 or TFAP2C endogenous expression, enabling the design of strategies to induce trophectoderm proliferation of invitro-derived embryos. To this aim, we designed and synthesised 4 non-overlapping single guide RNAs to target the regulatory region of each of these target genes. Presumptive zygotes were cytoplasmically microinjected with a mix containing dCas9-VP160 mRNA and a pool of 4 single guide RNAs targeting SMARCA4 (dCas9_SM group) or TFAP2C (dCas9_TF group). As control, a non-injected group was also included. Analysis was carried out in pools of 10 early embryos or 5 blastocysts and at least 3 biological replicates were included. Gene expression was assessed by RTqPCR at Days 2, 4, and 7 after microinjection and data were normalized to that obtained for the non-injected group. The CRISPR-on system was efficient to induce expression of SMARCA4 two days after microinjection (dCas9_SM group, Mann-Whitney t-test; P<0.05), but failed to significantly increase TFAP2C expression (dCas9_TF group). Surprisingly, CDX2, which is a downstream effector for trophectoderm maintenance, was induced both in dCas9_SM and dCas9_TF groups, supporting the CRISPR-mediated induction of targeted transcription factors. However, no changes were observed in the endogenous level of NANOG. Additional analysis is currently ongoing to determine whether CRISPR-on mediated induction of SMARCA4 and/or TFAP2C expression affects lineage specification and regulation. To our knowledge, this is the first report on the use of CRISPR-on for modulation of endogenous gene expression in mammalian embryos. Our study lays the foundations for CRISPR-on application in embryos as a useful tool to understand key cell fate decisions and will enable unprecedented studies of significance to embryo development, cell differentiation, and segregation.
73 Cytokine addition does not increase developmental competence of invitro-produced bovine embryos
Authors: Helland C. M. , Barcelo-Fimbres M. , Campos-Chillon L. F.
Published: December 2019

Recently in porcine, the addition of 3 cytokines (FGF2, LIF, and IGF1) improved oocyte maturation, quadrupling the number of piglets born per oocyte collected (Yuan et al. 2017 Proc. Natl. Acad. Sci. USA 114, E5796-E5804). We hypothesised that in the bovine, addition of these cytokines to maturation (MCyt) and culture media (CCyt) would lower lipid content and increase mitochondrial activity, representing an improved developmental competence when compared with standard maturation (MCon) and culture (CCon) conditions. The experimental design was a 2 (MCon and MCyt)×2 (CCon and CCyt) factorial in 8 replicates, testing the interactions of each maturation medium with each culture medium. Invitro-produced embryos produced aspirating cumulus-oocyte complexes (COCs) from 2 to 8mm follicles of abattoir ovaries. The COCs (n=2156) were matured for 23h in MCon or MCyt media at 6% CO2 in air, fertilised with semen from one of two bulls, and cultured in CCon or CCyt media at 38.5°C, 6% CO2, 5% O2, and 89% N2. On Day 7.5 post-fertilisation, blastocyst rates were evaluated and embryos (n=4/replicate/group) were stained with 1µgmL−1 Nile Red for lipid quantification or 300 nM MitoTracker Red CMX-Rosamine for mitochondrial polarity. Images were obtained with confocal microscopy and fluorescent intensity (AFU) was measured by Image J software (National Institutes for Health) adjusted per cell number. Data were analysed by GLM using ANOVA and l.s.d. with SAS (SAS Institute Inc.). Results (Table 1) indicated similar cleavage and blastocyst rates between all groups (P>0.05). The combination of MCon×CCon resulted in higher mitochondrial activity than any other combination (P<0.05). The MCon×CCyt showed the highest lipid levels, whereas MCyt×CCyt showed the lowest lipid levels (P<0.05). Results suggest that the combination of MCyt×CCyt media produces the lowest lipid levels, whereas the MCon×CCon media lead to the highest mitochondrial activity. The addition of cytokines to both maturation and culture media maintains competence and lowers lipid content; however, it also seems to lower mitochondrial activity. Cryopreservation studies and evaluation of pregnancy rates are ongoing.

Table 1.Oocyte and embryo developmental competence matured and cultured in control and cytokine-added media

Treatment

Oocytes (n)
Cleavage (%)
Blastocysts per oocyte (%)
Nile Red per cell (AFU)
MitoTracker per cell (AFU)

Maturation main effects
MCon
1000
96.8±0.4
29.9±2.7
36.1±2.1a
385.1±65.8a

MCyt
1156
96.0±0.4
26.2±2.7
30.0±2.1b
209.1±65.8b

Culture main effects
CCon
1036
96.2±0.4
28.0±2.7
33.1±2.1
392.0±65.8a

CCyt
1120
97.7±0.4
28.1±2.7
33.0±2.1
202.2±65.8b

Interactions
MCon×CCon
461
96.6±0.8
27.1±3.7
33.6±3.0ab
559.0±91.1a

MCon×CCyt
539
95.3±0.9
29.5±2.7
38.6±3.0a
211.2±91.1b

MCyt×CCon
575
95.4±0.6
27.4±2.1
32.7±3.0bc
224.9±91.1b

MCyt×CCyt
581
95.6±0.9
23.2±2.9
27.4±3.0c
193.1±91.1b

a-cValues with different superscript in the same column differ (P<0.05).
99 Short-term storage of equine embryos at 5 or 20°C does not cause lipid peroxidation
Authors: Gastal G. D. , Scarlet D. , Aurich C.
Published: December 2019

Maintaining the integrity of equine embryos during storage for transportation is essential for successful conception after transfer. During storage, reactive oxygen species may originate from embryo metabolism, causing lipid peroxidation and increasing its end product malondialdehyde (MDA); MDA is one of the important biomarkers for oxidative stress. This study aimed to evaluate the temperature curve, pH and lipid peroxidation of equine embryos stored in holding medium after 24h at 5 or 20°C. Embryos (n=33) were collected on Day 7 (n=21, 7 embryos per group) or Day 8 (n=12) after ovulation and assigned to four groups: Day 7 control (D7, fresh); Day 7, 24h at 5°C (E5C); Day 7, 24h at 20°C (E20C); and Day 8 control (D8, fresh 24h time control). After collection, embryos were washed and kept in holding medium (Minitube) for morphological classification and measurements. For pH and lipid peroxidation measurement, embryos were kept in a fixed volume of holding medium (150µL) within a microtube (200µL); the microtube was kept within a falcon tube (50mL) inside of an Equitainer (Hamilton Biovet). The temperature was recorded by a data logger (Testo 175, Testo) every 10min for 24h. The pH was assessed by a pH meter using a microelectrode (InLab Ultra-Micro-ISM, Mettler Toledo) for small sample volumes. Lipid peroxidation was assessed using the MDA assay kit (catalog number MAK085, Sigma-Aldrich) according to the manufacturer's instructions. Statistical analyses were performed using the Kruskal-Wallis nonparametric test and Mann-Whitney U to compare differences among groups. Embryo size differed (P<0.05) between D7 (383±41 µm) and D8 (1044±131 µm). Storage temperature (E5C or E20C) did not affect embryo size (382±47 and 553±99µm, respectively; P>0.05). The temperature curve was similar (P>0.05) among embryos within the treatment groups during the storage period. The pH (7.22±0.07 and 7.22±0.09) did not differ (P>0.05) between E5C and E20C. Lipid peroxidation levels were below the limit of quantification (0.04 nmol) in all groups. The present findings suggest that holding temperature does not affect the size, pH, or lipid peroxidation of equine embryos stored for 24h in holding medium. However, our previous studies (Gastal et al. 2018 J. Equine Vet. Sci. 66, 185) have shown that holding temperature influences the expression of genes involved in equine embryo development. In conclusion, variations regarding embryo development and conception rate after transfer of embryos stored at different temperatures might be related to factors other than lipid peroxidation.
Influence of right-wing authoritarianism and the values proposed by Schwartz on the perception of psychological abuse as gender violence: an exploratory study / Influencia del autoritarismo de derechas y de los valores propuestos por Schwartz en la percepción del abuso psicológico como violencia de género: un estudio exploratorio
Authors: Canto Jesús M. , Álvaro Jose-Luis , Da Costa-Silva Khalil , Maribondo Iara , Torres Ana-Raquel
Published: December 2019

Psychological abuse in relationships is an extremely important social problem and has serious consequences for the victims. The vast majority of women who have suffered from physical violence have also been victims of psychological violence. There is a line of research that seeks to analyse the role played by certain ideologies in the way the different forms of violence against women are perceived. This study analyses the relationship between right-wing authoritarianism and the values proposed by the Schwartz model in the perception of psychological abuse suffered by women in relationships as gender violence. A total of 308 university students participated in the research (206 women and 102 men). The results indicate that right-wing authoritarianism acts as a risk factor and the value of self-transcendence as a protective factor in the perception of psychological abuse as gender violence. In addition, a moderating effect of the value of self-transcendence was found in the relationship between right-wing authoritarianism and the perception of psychological abuse. The data confirm the importance of conservative ideology and values in the perception of gender violence.