bluuebunny/crossref_metadata_2025 huggingface dataset:
Genome-wide Annotation and Comparative Analysis of Long Terminal Repeat Retrotransposons between Pear Species of P. bretschneideri and P. Communis
Authors: Yin Hao , Du Jianchang , Wu Jun , Wei Shuwei , Xu Yingxiu , Tao Shutian , Wu Juyou , Zhang Shaoling
Published: December 2015

Recent sequencing of the Oriental pear (P. bretschneideriRehd.) genome and the availability of the draft genome sequence of Occidental pear (P. communisL.), has provided a good opportunity to characterize the abundance, distribution, timing and evolution of long terminal repeat retrotransposons (LTR-RTs) in these two important fruit plants. Here, a total of 7247 LTR-RTs, which can be classified into 148 families, have been identified in the assembled Oriental pear genome. Unlike in other plant genomes, approximately 90% of these elements were found to be randomly distributed along the pear chromosomes. Further analysis revealed that the amplification timeframe of elements varies dramatically in different families, super-families and lineages and theCopia-like elements have highest activity in the recent 0.5 million years (Mys). The data also showed that two genomes evolved with similar evolutionary rates after their split from the common ancestor ~0.77–1.66 million years ago (Mya). Overall, the data provided here will be a valuable resource for further investigating the impact of transposable elements on gene structure, expression and epigenetic modification in the pear genomes.
A network medicine approach to quantify distance between hereditary disease modules on the interactome
Authors: Caniza Horacio , Romero Alfonso E. , Paccanaro Alberto
Published: December 2015

We introduce a MeSH-based method that accurately quantifies similarity between heritable diseases at molecular level. This method effectively brings together the existing information about diseases that is scattered across the vast corpus of biomedical literature. We prove that sets of MeSH terms provide a highly descriptive representation of heritable disease and that the structure of MeSH provides a natural way of combining individual MeSH vocabularies. We show that our measure can be used effectively in the prediction of candidate disease genes. We developed a web application to query more than 28.5 million relationships between 7,574 hereditary diseases (96% of OMIM) based on our similarity measure.
Transportation in the Interstitial Space of the Brain Can Be Regulated by Neuronal Excitation
Authors: Shi Chunyan , Lei Yiming , Han Hongbin , Zuo Long , Yan Junhao , He Qingyuan , Yuan Lan , Liu Huipo , Xu Ge , Xu Weiguo
Published: December 2015

The transportation of substances in the interstitial space (ISS) is crucial for the maintenance of brain homeostasis, however its link to neuronal activity remains unclear. Here, we report a marked reduction in substance transportation in the ISS after neuronal excitation. Using a tracer-based method, water molecules in the interstitial fluid (ISF) could be specifically visualized in magnetic resonance (MR) imaging. We first observed the flow of ISF in the thalamus and caudate nucleus of a rat. The ISF flow was then modulated using a painful stimulation model. We demonstrated that the flow of ISF slowed significantly following neuronal activity in the thalamus. This reduction in ISF flow continued for hours and was not accompanied by slow diffusion into the ISS. This observation suggests that the transportation of substances into the ISS can be regulated with a selective external stimulation.
Mating dynamics in a nematode with three sexes and its evolutionary implications
Authors: Chaudhuri Jyotiska , Bose Neelanjan , Tandonnet Sophie , Adams Sally , Zuco Giusy , Kache Vikas , Parihar Manish , von Reuss Stephan H. , Schroeder Frank C. , Pires-daSilva Andre
Published: December 2015

Nematodes have diverse reproductive strategies, which make them ideal subjects for comparative studies to address how mating systems evolve. Here we present the sex ratios and mating dynamics of the free-living nematodeRhabditissp. SB347, in which males, females and hermaphrodites co-exist. The three sexes are produced by both selfing and outcrossing and females tend to appear early in a mother’s progeny. Males prefer mating with females over hermaphrodites, which our results suggest is related to the female-specific production of the sex pheromones ascr#1 and ascr#9. We discuss the parallels between this system and that of parasitic nematodes that exhibit alternation between uniparental and biparental reproduction.
Daily update of motor predictions by physical activity
Authors: Gueugneau Nicolas , Schweighofer Nicolas , Papaxanthis Charalambos
Published: December 2015

Motor prediction, i.e., the ability to predict the sensory consequences of motor commands, is critical for adapted motor behavior. Like speed or force, the accuracy of motor prediction varies in a 24-hour basis. Although the prevailing view is that basic biological markers regulate this circadian modulation, behavioral factors such as physical activity, itself modulated by the alternation of night and day, can also regulate motor prediction. Here, we propose that physical activity updates motor prediction on a daily basis. We tested our hypothesis by up- and down-regulating physical activity via arm-immobilization and high-intensity training, respectively. Motor prediction was assessed by measuring the timing differences between actual and mental arm movements. Results show that although mental movement time was modulated during the day when the arm was unconstrained, it remained constant when the arm was immobilized. Additionally, increase of physical activity, via release from immobilization or intense bout of training, significantly reduced mental movement time. Finally, mental and actual times were similar in the afternoon in the unconstrained condition, indicating that predicted and actual movements match after sufficient amount of physical activity. Our study supports the view that physical activity calibrates motor predictions on a daily basis.
Coaxial carbon fiber/ZnO nanorods as electrodes for the electrochemical determination of dopamine
Authors: Yang Chi , Gu Baoxiang , Zhang Dan , Ge Cunwang , Tao Huimin
Published: December 2015

The adsorption and oxidization mechanism of dopamine on the surface of carbon fiber/ZnO nanorods.
Rh(ii)-Catalyzed formation of pyrrolo[2,3-b]quinolines from azide-methylenecyclopropanes and isonitriles
Authors: Chen Kai , Tang Xiang-Ying , Shi Min
Published: December 2015

Azide-methylenecyclopropanes underwent an intermolecular cyclization with isonitriles catalyzed by a RhIIcomplex to produce a series of pyrrolo[2,3-b]quinolines in good yields. Moreover, synthetic applications to construct useful heterocycles have also been achieved.
From single-point to three-point halogen bonding between zinc(ii) tetrathiocyanate and tetrabromomethane
Authors: Rosokha Sergiy V. , Stern Charlotte L. , Vinakos Michael K.
Published: December 2015

The strengths of three- and two-point halogen bonding in CBr4·[Zn(NCS)4]2−dyads are close to that of single-point CBr4·NCS−interaction.
Monoorganoantimony(v) phosphonates and phosphoselininates
Authors: Ugandhar Uppara , Baskar Viswanathan
Published: December 2015

Synthesis and structural characterization of novel tetra- and dinuclear organoantimony oxo-hydroxo clusters ligated by phosphonates and phosphonates/selininates are reported. Mass spectral studies reveal that the solid state molecular structures of these clusters were retained in solution as well.
Crystal phase competition by addition of a second metal cation in solid solution metal–organic frameworks
Authors: Castillo-Blas C. , Snejko N. , de la Peña-O'Shea V. A. , Gallardo J. , Gutiérrez-Puebla E. , Monge M. A. , Gándara F.
Published: December 2015

Addition of small amounts of a second metal cation results in crystal phase competition during the synthesis of solid-solution MOFs.
Copper(i) complexes with phosphine derived from sparfloxacin. Part II: a first insight into the cytotoxic action mode
Authors: Komarnicka U. K. , Starosta R. , Płotek M. , de Almeida R. F. M. , Jeżowska-Bojczuk M. , Kyzioł A.
Published: December 2015

A first insight into the cytotoxic action mode of copper(i) iodide or copper(i) thiocyanate complexes with a phosphine derivative of sparfloxacin (a 3rdgeneration fluoroquinolone antibiotic agent) and 2,9-dimethyl-1,10-phenanthroline or 2,2′-biquinoline as auxiliary ligands.
A spirobifluorene and diketopyrrolopyrrole moieties based non-fullerene acceptor for efficient and thermally stable polymer solar cells with high open-circuit voltage
Authors: Li Shuixing , Liu Wenqing , Shi Minmin , Mai Jiangquan , Lau Tsz-Ki , Wan Junhua , Lu Xinhui , Li Chang-Zhi , Chen Hongzheng
Published: December 2015

A PCE of 5.16% with a Voc of 1.14 V is achieved with thermally stable P3HT : SF(DPPB)4 solar cells.
Environmentally-safe and transparent superhydrophobic coatings
Authors: Mates J. E. , Ibrahim R. , Vera A. , Guggenheim S. , Qin J. , Calewarts D. , Waldroup D. E. , Megaridis C. M.
Published: December 2015

An environmentally-safe, water-borne coating consisting of titania nanoparticles and fluorine-free polyethylene copolymers, forming a transparent, substrate-independent, superhydrophobic water barrier.
Design, synthesis and antiproliferative activity of the new conjugates of E7010 and resveratrol as tubulin polymerization inhibitors
Authors: Kamal Ahmed , Ashraf Md. , Basha Shaik Thokhir , Ali Hussaini S. M. , Singh Shamshair , Vishnuvardhan M. V. P. S. , Kiran Boppana , Sridhar Balasubramanian
Published: December 2015

A new class of (E)-N-phenyl-3-styrylpyridin-2-amine conjugates were designed and synthesized on the basis of E7010 and resveratrol scaffolds.
Highly efficient synthesis of flavonol 5-O-glycosides with glycosyl ortho-alkynylbenzoates as donors
Authors: Liao Jin-Xi , Fan Nai-Li , Liu Hui , Tu Yuan-Hong , Sun Jian-Song
Published: December 2015

A highly efficient approach to construct the challenging flavonol 5-O-glycosidic linkages is developed, providing a general synthetic route to flavonol 5-O-glycosides.
Effect of heat treatment of magnesium alloy substrates on corrosion resistance of a hybrid organic–inorganic sol–gel film
Authors: Diaz L. , García-Galván F. R. , Llorente I. , Jiménez-Morales A. , Galván J. C. , Feliu Jr S.
Published: December 2015

The influence of heat treatment of magnesium alloy substrates on corrosion resistance of a sol–gel coating has been assessed during immersion tests in 0.6 M NaCl aqueous solution.
The classification and prediction of green teas by electrochemical response data extraction and fusion approaches based on the combination of e-nose and e-tongue
Authors: Wang Jun , Wei ZhenBo
Published: December 2015

Aroma and taste are the most important attributes that influence the pleasantness of tea infusion.
ESIPT blocked CHEF based differential dual sensor for Zn2+ and Al3+in a pseudo-aqueous medium with intracellular bio-imaging applications and computational studies
Authors: Alam Rabiul , Mistri Tarun , Bhowmick Rahul , Katarkar Atul , Chaudhuri Keya , Ali Mahammad
Published: December 2015

A novel H3SAL-NH probe exhibits ESIPT from the phenolic OH group to the azomethine N atom in the excited state. This is blocked in the presence of Zn2+ and Al3+ giving turn on fluorescence which is useful for imaging in live HepG2 cells.
Characterization of fouling layers for in-line coagulation membrane fouling by apparent zeta potential
Authors: Wang Jie , Yang Sasa , Guo Wenshan , Ngo Huu-Hao , Jia Hui , Yang Jun , Zhang Hongwei , Zhang Xinbo
Published: December 2015

This study investigated the apparent zeta potential of fouled membranes for in-line coagulation membrane fouling monitoring in micro-polluted water treatment.
A protocol of self-assembled monolayers of fluorescent block molecules for trace Zn(ii) sensing: structures and mechanisms
Authors: Xu Hongbo , Wang Huanhuan , Zhou Shenghai , Xiao Lili , Yan Yun , Yuan Qunhui
Published: December 2015

The fluorescent behavior of the SAM of TPE-C4-L2 block molecules on HOPG surface has been revealed using STM.
Micellization and adsorption behaviour of bile salt systems
Authors: Mukherjee Bedachhanda , Dar Aijaz Ahmad , Bhat Parvaiz Ahmad , Moulik Satya Priya , Das Akhil Ranjan
Published: December 2015

Variation of enthalpy change with concentration of binary bile salt mixture.
Selective oxidation of 1,2-propanediol to lactic acid catalyzed by hydroxyapatite-supported Pd and Pd–Ag nanoparticles
Authors: Feng Yonghai , Xue Wuping , Yin Hengbo , Meng Minjia , Wang Aili , Liu Shuxin
Published: December 2015

Selective catalytic oxidation of 1,2-propanediol is an alternative, effective, and environmentally benign method for producing lactic acid.
Percolative silver/alumina composites with radio frequency dielectric resonance-induced negative permittivity
Authors: Shi Zhi-cheng , Mao Fan , Wang Jing , Fan Run-hua , Wang Xin
Published: December 2015

Radio frequency dielectric resonance-induced negative permittivity was observed in silver/alumina composites beyond the percolation threshold.
Titanium aminophosphates: synthesis, characterization and crystal violet dye degradation studies
Authors: Anumula Rajini , Adepu Ajay Kumar , Chirra Suman , Siliveri Suresh , Narayanan Venkatathri
Published: December 2015

Titanium aminophosphates are prepared using titanium tetraisopropoxide, orthophosphoric acid and aliphatic amines. Their detailed characterization and catalytic crystal violet dye degradation were studied.
Dip- and spin-assisted stereocomplexation-driven LbL self-assembly involving homochiral PVA-g-OLLA and PVA-g-ODLA copolymers
Authors: Bahloul Mohamed , Pruvost Sébastien , Fleury Etienne , Portinha Daniel , Charlot Aurélia
Published: December 2015

Layer by Layer (LbL) thin films stemming from the formation of stereocomplex between oligolactate with opposite chirality (OLLA and ODLA) covalently anchored onto poly(vinyl alcohol) (PVA) chains is described herein for the first time.
Ammonolysis of gallium phosphide GaP to the nanocrystalline wide bandgap semiconductor gallium nitride GaN
Authors: Drygas Mariusz , Sitarz Maciej , Janik Jerzy F.
Published: December 2015

Ammonolysis of microcrystalline powders of gallium phosphide GaP afforded nanopowders or nanowires of hexagonal gallium nitride GaN.
FeS2-sensitized ZnO/ZnS nanorod arrays for the photoanodes of quantum-dot-sensitized solar cells
Authors: Wang Mudan , Qin Haiying , Fang Youtong , Liu Jiabin , Meng Liang
Published: December 2015

FeS2-sensitized ZnO/ZnS nanorod arrays were fabricated and used as the photoanodes for quantum-dot-sensitized solar cells (QDSSCs).
CPL emission of chiral BINOL-based polymers via chiral transfer of the conjugated chain backbone structure
Authors: Li Fei , Wang Yuxiang , Sheng Yuan , Wei Guo , Cheng Yixiang , Zhu Chengjian
Published: December 2015

Chiral BINOL-based polymers incorporating phenothiazine moieties can exhibit strong mirror image Cotton effects and emit green color CPL signals via effective chiral transfer of the conjugated polymer chain backbone.
Size and distribution control of surface plasmon enhanced photoluminescence and SERS signal in Ag–GaN hybrid systems
Authors: Thakur Varun , Siddhanta Soumik , Narayana C. , Shivaprasad S. M.
Published: December 2015

In the present experiment, two GaN nanowall network (NWN) samples with different porosity were grown on c-sapphire substrates using plasma assisted molecular beam epitaxy (PA-MBE).
Redox-ligand sustains controlled generation of CF3 radicals by well-defined copper complex
Authors: Jacquet Jérémy , Blanchard Sébastien , Derat Etienne , Desage-El Murr Marine , Fensterbank Louis
Published: December 2015

Cooperative catalysis involving copper(ii) complex bearing redox ligands allows generation of CF˙3 radicals through ligand-based SET while metallic oxidation state is preserved.
A diversity-oriented synthesis of bioactive benzanilides via a regioselective C(sp2)–H hydroxylation strategy
Authors: Sun Yong-Hui , Sun Tian-Yu , Wu Yun-Dong , Zhang Xinhao , Rao Yu
Published: December 2015

A diversity-oriented synthesis of bioactive benzanilidesviaC(sp2)–H hydroxylation has been studied. The reaction demonstrates excellent regioselectivity, good tolerance of functional groups, and high yields.
Concave and duck web-like platinum nanopentagons with enhanced electrocatalytic properties for formic acid oxidation
Authors: Lai Jianping , Niu Wenxin , Li Suping , Wu Fengxia , Luque Rafael , Xu Guobao
Published: December 2015

Pt-branched structures featuring concave and duck web-like nanopentagons with high-energy {110} and {554} facets, multiple twin boundaries, duck web-like edges and inherent anisotropic branches are prepared.
Direct electrochemical formation of nanostructured amorphous Co(OH)2on gold electrodes with enhanced activity for the oxygen evolution reaction
Authors: Sayeed Md Abu , Herd Tenille , O'Mullane Anthony P.
Published: December 2015

The rapid and direct electrochemical formation of amorphous nanostructured Co(OH)2on gold electrodes under room temperature conditions was found to be a highly active, yet stable, precursor for the OER.
Phosphorus-doped carbon–carbon nanotube hierarchical monoliths as true three-dimensional electrodes in supercapacitor cells
Authors: Patiño Julián , López-Salas Nieves , Gutiérrez María C. , Carriazo Daniel , Ferrer M. Luisa , Monte Francisco del
Published: December 2015

Phosphorus-doped carbon–carbon nanotube hierarchical monoliths exhibiting energy densities of around 22.6 W h kg−1 at power densities of up to 10 kW kg−1 were capable of functioning as true three-dimensional electrodes in supercapacitor cells.
From Maya blue to biomimetic pigments: durable biomimetic pigments with self-cleaning property
Authors: Zhang Yujie , Zhang Junping , Wang Aiqin
Published: December 2015

Durable and self-cleaning biomimetic superhydrophobic pigments of various colors were fabricated.
Beyond efficiency: scalability of molecular donor materials for organic photovoltaics
Authors: Po Riccardo , Roncali Jean
Published: December 2015

The preparation of various molecules taken as representative examples of some of the main classes of molecular donors for organic solar cells is discussed in order to assess the complexity and possibilities of scaling-up their synthesis.
Graduate Student Experiences Using Easy Onsets
Authors: Hughes Charles , Palasik Scott , Ellis Kellie
Published: December 2015

This qualitative study explored reports from 18 graduate students related to their experiences using easy onsets in three speaking situations for a total of 54 speaking situations recorded across all participants. Thematic analysis revealed three major themes and seven minor themes. Major Theme 1 was related to the influence that the listener and situation had on self-perceptions. The 3 minor themes that contributed to this major theme were: anxiety related to listener reactions, the telephone being an easier speaking situation, and nature of the situation having an influence. Major Theme 2 was related to the uncomfortable nature of using the skills with 2 minor themes contributing and consisting of: mental effort and energy to use the skill, and the feeling of unnatural sounding and difficulty in using the skill. Major Theme 3, the final major theme found, related to previous success and comfort using the skill. Two minor themes contributed to this same major theme and included: previous experience using the skill had an impact on self-perceptions and more comfort using the skill over multiple attempts.
An estimate of above-ground carbon stock in tropical rainforest on Manus Island, Papua New Guinea
Authors: Arihafa Arison , Dalgarno Sebastian , Neale Ezra
Published: December 2015

Forest carbon emission mitigation schemes seek to protect tropical forest, combat effects of climate change, and offer potential cash and development opportunities. Reducing emissions from deforestation and degradation (REDD+) projects based on a foundation of accurate carbon stock assessment provide such an opportunity for Papua New Guinea. The objective of this study was to quantify the carbon stock of the central forests of Manus Island, Papua New Guinea, and identify factors that underpin any observed variation within it. We employed the Winrock Standard Operating Procedures for Terrestrial Carbon Measurement for plots and associated measurements. In 75 variable-radius nested plots (total area = 14.4 ha), we assessed above-ground and total carbon stock of stems ≥5 cm diameter at breast height via general linear models in a model-selection framework. The top models described variation in average carbon stock at 95% lower and upper confidence interval in above-ground biomass solely in terms of forest type: primary hill forest 165.0 Mg C ha–1 (148.3–183.7, n = 48), primary plain forest 100.9 Mg C ha–1 (78.0–130.6, n = 10) and secondary hill forests 99.7 Mg C ha–1 (80.9–122.9, n = 17). To a lesser extent, above-ground carbon stock increased with slope and varied idiosyncratically by the nearest village. Our estimates are comparable with published studies for Papua New Guinea and the wider tropical region. These data should strengthen pre-existing knowledge and inform policies on carbon accounting for REDD+ projects in the region.
Livestock in biomedical research: history, current status and future prospective
Authors: Polejaeva Irina A. , Rutigliano Heloisa M. , Wells Kevin D.
Published: December 2015

Livestock models have contributed significantly to biomedical and surgical advances. Their contribution is particularly prominent in the areas of physiology and assisted reproductive technologies, including understanding developmental processes and disorders, from ancient to modern times. Over the past 25 years, biomedical research that traditionally embraced a diverse species approach shifted to a small number of model species (e.g. mice and rats). The initial reasons for focusing the main efforts on the mouse were the availability of murine embryonic stem cells (ESCs) and genome sequence data. This powerful combination allowed for precise manipulation of the mouse genome (knockouts, knockins, transcriptional switches etc.) leading to ground-breaking discoveries on gene functions and regulation, and their role in health and disease. Despite the enormous contribution to biomedical research, mouse models have some major limitations. Their substantial differences compared with humans in body and organ size, lifespan and inbreeding result in pronounced metabolic, physiological and behavioural differences. Comparative studies of strategically chosen domestic species can complement mouse research and yield more rigorous findings. Because genome sequence and gene manipulation tools are now available for farm animals (cattle, pigs, sheep and goats), a larger number of livestock genetically engineered (GE) models will be accessible for biomedical research. This paper discusses the use of cattle, goats, sheep and pigs in biomedical research, provides an overview of transgenic technology in farm animals and highlights some of the beneficial characteristics of large animal models of human disease compared with the mouse. In addition, status and origin of current regulation of GE biomedical models is also reviewed.
110 TREATMENT WITH GnRH ON DAY 5 REDUCES PREGNANCY LOSS IN HEIFERS RECEIVING IN VITRO-PRODUCED EXPANDED BLASTOCYSTS
Authors: Garcia-Guerra A. , Sala R. V. , Baez G. M. , Fosado M. , Melo L. F. , Motta J. C. L. , Leffers L. , Walleser E. A. , Ochoa J. C. , Moreno J. F. , Wiltbank M. C.
Published: December 2015

The hypothesis was that GnRH on Day 5 of a synchronized cycle in embryo transfer recipients would increase progesterone (P4) concentrations, embryo size, and fertility. Holstein and cross-bred Holstein heifers (n = 1562) were synchronized using a modified 5-day CIDR Co-Synch as follows: Day –8 CIDR inserted; Day –3 CIDR removed; prostaglandin F2α treatment; Day –2 second prostaglandin F2α; Day 0 gonadotropin-releasing hormone (G1, 100 μg of gonadorelin acetate) to induce ovulation. On Day 5.5, heifers were assigned in a completely randomised design to 1 of 2 treatments: Control (untreated) or GnRH (200 μg of gonadorelin acetate). Transfer of fresh in vitro-produced embryos was performed between d 6 and 8 after G1. Data collected from each heifer included embryo stage and quality, body condition score, technician, interval from G1 to transfer, and number of previous transfers. All heifers were evaluated by transrectal ultrasonography on Day 5, 33, and 62 and a subset of heifers was scanned on Day 12 (n = 718; to determine ovulation to treatment) and another subset on Day 33 (n = 296; 16-s video to determine embryo and amniotic vesicle size). Serum P4 was determined from a subset of heifers on Day 12 (n = 467). Fertility data were analysed by logistic regression (LOGISTIC procedure, SAS 9.4), whereas continuous outcomes were analysed by ANOVA (MIXED procedure). Ovulation to Day 5.5 gonadotropin-releasing hormone was 83.9% (302/360) in GnRH-treated heifers v. 3.3% (12/358) in Control (P < 0.001). Progesterone on Day 12 was greater in GnRH-treated heifers 7.2 ± 0.1 ng mL–1 v. Controls 6.0 ± 0.1 ng mL–1 (P < 0.001). There was an effect of embryo stage at Day 33 and 60 of pregnancy, with Stage 7 having greater P/ET than Stage 6 embryos. Treatment with GnRH did not alter pregnancy per embryo transfer with either embryo stage but decreased pregnancy loss in Stage 7 embryos, as shown in Table 1. Embryo size measured as crown-rump length (CRL) did not differ, as shown in Table 1. Similarly, amniotic vesicle volume (AVV) was not different between GnRH (549.1 ± 16 mm3) and Control (543.5 ± 14 mm3; P = 0.86), nor was there an interaction between treatment and embryo stage (P = 0.71). In addition, neither AVV (P = 0.22) nor CRL (P = 0.41) were associated with pregnancy loss between Day 33 and 60. In conclusion, treatment with GnRH on Day 5 resulted in increased P4 and a reduction in pregnancy loss in heifers receiving a Stage 7 embryo without changing conceptus size.

Table 1.Pregnancies per embryo transfer (P/ET), crown-rump length (CRL), and pregnancy loss in embryo recipients receiving gonadotropin-releasing hormone (GnRH) on Day 5.5 v. control
112 PRESENCE OF BACTERIA IN THE ENDOMETRIUM AND OVIDUCT OF COWS WITH PYOMETRA AS DETECTED BY FLUORESCENCE IN SITU HYBRIDIZATION
Authors: Karstrup C. C. , Karstrup C. C. , Knudsen L. , Knudsen L. , Jensen T. K. , Jensen T. K. , Schou K. , Schou K. , Agerholm J. S. , Agerholm J. S. , Pedersen H. G. , Pedersen H. G.
Published: December 2015

The objective of the study was to identify the location of the present bacteria in the uterus and oviducts of cows with pyometra. Pyometra is one of the postpartum infectious diseases in cattle that can result in infertility and thereby affect reproduction performance. Reproductive tracts (n = 21) were collected at a slaughterhouse in Denmark and send to The University of Copenhagen for examination and sampling. The uteri were included in the study when the following criteria were met: the cow was more than 21 days postpartum, the uterus was distended with pus, the cervix was closed, and a corpus luteum was present in one or both ovaries. A full thickness uterine tissue sample from the previous pregnant horn and both oviducts were sampled and then fixed in formalin. The tissues were trimmed, processed by routine methods, embedded in paraffin, sectioned at 3 microns, and prepared for fluorescence in situ hybridization using a probe targeting the 16S ribosomal RNA of the domain bacteria (i.e. targeting all bacteria regardless of species). Using fluorescence microscopy, the presence of bacteria within or on the surface of the endometrium and in the oviducts were noted. The endometrial biopsies from all cows (n = 21) contained bacteria, while 75% (16/21) of the cows had bacteria in one or both oviducts. The bacteria were located on the luminal surface and in the lamina propria in 38.1% (8/21) of the uterine biopsies. In the remaining 62% of the uterine biopsies, the bacteria were only located above the basal membrane. Regarding the oviduct biopsies, the bacteria were located on the luminal surface and in lamina propria in 9.5% (2/21) of the biopsies, whereas the bacteria were located only above the basal membrane in 90.5% of the biopsies. In conclusion, 1) bacteria are present in the uteri and oviducts of cows with pyometra and 2) the bacteria are primarily located on the luminal epithelia surface above the basal membrane. Further analyses will investigate which specific species of bacteria that are located in the lamina propria of the uterine and oviduct biopsies.
114 ENDOPLASMIC RETICULUM (ER) STRESS IN HYPOXIA-INDUCED DIABETES MELLITUS MODEL
Authors: Ahn C. , Lee D. , Kim K. P. , Lee M. H. , Jeung E.-B.
Published: December 2015

Endoplasmic reticulum (ER) regulates calcium ion concentration as a reservoir in the cell. ER stress is a cellular stress response related to the endoplasmic reticulum. At the initial stage of ER stress, ER tries to restore normal function by halting protein translation, degrading misfolded proteins, and increasing production of chaperones involved in protein folding. If ER fails to restore ER stress, ER stress can lead cells to apoptosis. To study the signaling between ER stress and calcium channels under ER-stressed circumstances, we designed a hypoxia-induced diabetic model. Nine-week-old male mice were chosen, maintained under hypoxic condition under 10% O2, 5% CO2 for 10 days, and the expression of ER stress markers and calcium channel gene expression were examined by real-time PCR. By maintaining hypoxic condition, the mice showed high glucose levels. Under this diabetic condition, in pancreatic beta cells, ER stress markers were elevated. This tendency showed an increase in calbindin-D9k KO mice. Chaperones such as calreticulin and calnexin were decreased, but in calbindin-D9k KO mice chaperone calnexin was not decreased. Interestingly, the calbindin-D9k KO normoxia mice showed increased glucose level compared with wild-type normoxia mice. Also, calnexin expression of pancreas was decreased in calbindin-D9k KO normoxia mice. This result indicates that pancreas cells were under endoplasmic reticulum stress. Taken together, calbindin may play an important role in endoplasmic reticulum stress in pancreas.
This work was supported by the National Research Foundation of Korea (NRF) grant of Korean government (MEST) (No. 2013-010514).
13 EFFECT OF 6 TIMES REUSING OF CONTROLLED INTERNAL DRUG RELEASE (CIDR) FOR SHORT TERM (6 DAYS) ON PROGESTERONE LEVEL AND REPRODUCTIVE PERFORMANCE OF AWASSI EWES
Authors: Swelum A. , Mouamen A. , Alowaimer A.
Published: December 2015

This study was carried out using 120 multiparous Awassi ewes during breeding season to compare the effect of 6 times reusing of CIDR short term (6 days) on hormonal and reproductive performance. Ewes were equally and randomly allotted into 6 consecutive oestrus synchronization treatments. For group CIDR6×1, the ewes (n = 20) received new CIDR for 6 days with 300 IU of eCG at withdrawal time. For the other 5 groups, the same protocol was used, differing only by the use of the same CIDR for a second time in CIDR6×2, for a third time in CIDR6×3, for a fourth time in CIDR6×4, for a fifth time in CIDR6×5, and for a sixth time in CIDR6×6. Oestrus was detected using a vasectomized ram starting 12 h after progestagen withdrawal and repeated every 12 h up to 84 h. Blood samples were collected at withdrawal time (Day 6). Progesterone serum concentrations were measured using commercial ELISA kits and micro-titrimetric plates. Timed insemination was performed 48 h after CIDR withdrawal. Pregnancy and number of fetuses were diagnosed by ultrasonography at Day 23 after insemination and confirmed at Day 35 and 60. Comparisons among groups was evaluated using chi-square (χ2) test in all parameters except hormones levels, which was analysed by repeated-measures ANOVA, using SAS (SAS Institute Inc., Cary, NC, USA, 2000). The results revealed that there is no significant difference between the percentages of ewes detected in heat in groups CIDR6×1, CIDR6×2, and CIDR6×3 (55, 70, and 55%, respectively). While, the percentages of ewes detected in heat in group CIDR6×2 was significantly (P ≤ 0.05) higher than in groups CIDR6×4, CIDR6×5, and CIDR6×6 (35, 35, 31.58%, respectively). On the other hand, pregnancy rate was significantly (P ≤ 0.05) higher in CIDR6×2 (65%; 13/20) than CIDR6×3, CIDR6×4, CIDR6×5, and CIDR6×6 (30, 30, 30, and 15.79, respectively). No significant difference was detected in pregnancy rate between CIDR6×1 and CIDR6×2. No significant differences were detected in progesterone between CIDR6×1 and CIDR6×2, CIDR6×3, CIDR6×4, CIDR6×5, and CIDR6×6 (1.73, 1.67, 1.87, 1.57, 1.60, and 1.36 ng mL–1, respectively). These results indicated that reusing of CIDR devices for short term are efficient in synchronizing oestrus in ewes.
132 BLASTOCYST PRODUCTION FROM BOVINE OVARIAN CORTEX FRAGMENTS XENOGRAFTED UNDER THE BACK SKIN OF MICE
Authors: Alves T. J. M. , Dias M. T. , Assuncao C. M. , Camargo L. S. A. , Viana J. H. M. , Campos Junior P. H. A.
Published: December 2015

Chemo- or radiotherapy negatively affects the fertility of female patients undergoing oncological treatments. Ovaries are sensitive to such treatments, resulting in an increasing number of premature ovarian failures. Graft techniques are a promising alternative to preserve the fertility of such patients. So far, 35 birthed from human ovarian cortex autografts were reported in the literature; however, in this approach there is a risk of neoplastic reincidence. The aim of the present study was to evaluate the feasibility of ectopic ovarian cortex xenograft (using the bovine model) under the back skin of immunodeficient mice. Female SCID mice (~60 days, n = 38) were anesthetized with ketamine/xilazine and were placed on ventral decubitus. Ovarian cortex fragments from 8 cows (1.5 mm3; n = 152) were grafted through incisions made in the dorsal region (4 grafts per mouse). Ten days after ovarian fragments xenograft, the recipients were killed and the xenografts were harvested. The mice and grafts were weighed before and after the transplant. From the xenografts recovered, 88 were either routinely processed for histology (n = 26), to evaluate the progression of folliculogenesis, or sliced (n = 62) to recover the cumulus‐oocyte complexes, which were morphologically classified and used for in vitro embryo production, using standard procedures (in vitro maturation, fertilization, and embryo culture). The remaining grafts recovered (64) were stored in liquid nitrogen for future studies. Differences between means were compared using Student’s t-test. There was no difference between the body weight of recipient mice before and after xenograft (20.5 ± 0.4 v. 21 ± 0.8 g, respectively; P > 0.05). On the other hand, the grafts increased weight (11.6 ± 3.4 v. 14.8 ± 5.2 mg before and after transplant; P < 0.01). Histological analysis of the slices showed primordial, primary, multilaminar, antral, and atretic follicles, indicating the progression of folliculogenesis and neo-angiogenesis in the grafts. Twenty-four viable cumulus‐oocyte complexes were recovered from ovarian xenografts, from which 2 blastocysts were produced in vitro 8 days later (8.3% blastocyst rate). In summary, this study showed that ovarian xenografts were (i) healthily maintained under the back skin of immunodeficient mice, (ii) responsive to murine gonadotrophins, and (iii) able to produce viable cumulus-oocyte complexes that, (iv) by in vitro fertilization, can originate blastocysts. In general, our findings show the feasibility of the ovary xenograft as an alternative technique to fertility preservation in oncogenic patients, avoiding the risk of neoplastic re-incidence.
Study approved by Animal Experimentation Ethics Committee/FUSJ-009/15. Financial support was received from Fapemig and CNPq.
138 LIVER RECEPTOR HOMOLOG 1 INFLUENCES BLASTOCYST HATCHING IN PORCINE
Authors: Guo J. , Kim T. , Kim N.-H. , Cui X.-S.
Published: December 2015

Liver receptor homolog 1 (LRH1, NR5A2) belongs to the orphan nuclear receptor superfamily and has diverse functions in development, differentiation, metabolism, and cell death. The LRH1 gene is expressed in embryonic stem cells and regulates the expression of OCT4, which is the key factor required to maintain pluripotency. However, the function of LRH1 in early porcine embryo development is still unknown. In the present study, we examined the localization and mRNA level of LRH1 in porcine parthenotes by immunofluorescence and real-time RT-qPCR. To explore the function of LRH1, the embryos were treated with the specific antagonist of LRH1, 505601, which is a cell-permeable pyrazolylbiphenylethanone compound that targets LRH-1/NR5A2 ligand binding domain (LBD) via direct affinity interaction, preventing LRH-1 from assuming an active conformation. Every group contained 20 embryos. The results showed that the immunofluorescence signal of LRH1 was only observed in nuclei of blastocyst cells. Inhibition of LRH1 by antagonist significantly (P < 0.05) reduced the blastocyst rate in the 50 µM (37.30 ± 3.67%) and 100 µM (38.02 ± 5.12%) treatment groups compared with the control group (53.43 ± 3.67%). The hatching rate of blastocysts was also reduced (P < 0.01) by 50 µM (12.66 ± 3.13%) and 100 µM (20.10 ± 2.81%) antagonist compared with control (40.59 ± 0.59%), which resulted from the decreased (P < 0.05) expression of hatching-related genes, such as FN1, ITGA5, and COX2. Inhibition of LRH1 also increased (P < 0.05) the number of apoptotic cells in blastocysts. Moreover, inhibition enhanced expression of apoptotic genes, BAX and CASP3, and reduced the anti-apoptosis gene BCL2 (P < 0.05). Messenger RNA and protein level of OCT4 were sharply decreased in blastocysts after LRH1 inhibition. All data were analysed with a one-way ANOVA, and differences between treatment groups were assessed by the least significant difference (LSD) test using SPSS software (SPSS Inc., Chicago, IL, USA). In conclusion, LRH1 affects blastocyst formation and hatching by regulating OCT4 expression and cell apoptosis.
144 DOSAGE COMPENSATION AND X-LINKED GENE EXPRESSION IN BOVINE IN VIVO AND IN VITRO EMBRYOS
Authors: Duan J. E. , Jue N. K. , Jiang Z. , O'Neill R. , Wolf E. , Blomberg L. A. , Dong H. , Zheng X. , Chen J. , Tian X. C.
Published: December 2015

In human and mouse diploid cells and gametes, expression levels of X-linked genes are hypothesised to balance with those of autosomal genes (Ohno’s “dosage compensation”). Such a phenomenon, however, has not been systematically studied in cattle or compared between in vivo and in vitro embryos. Using RNA-seq data, we compared dosage compensation and expression differences of X-linked genes in bovine in vitro and in vivo oocytes and embryos. RNA-seq datasets GSE59186 and GSE52415 were non-uniquely (paralogs included) mapped to the bovine reference genome assembly UMD3.1 using tophat2. Cufflinks v1.0.3 was used to estimate fragments per kb of exon per million fragments (FPKM), which were then log2-transformed. In order to assess overall patterns of chromosomal gene expression without bias, statistical outliers were removed. A total of 12 928 X-linked transcripts were used to calculate the relative X to autosomal gene (A) expression (RXE): log2 (X expression) – log2 (A expression) for dosage compensation. Values ≥0 indicate dosage compensation (or X : A ratio ≥ 1); values <0 indicate incomplete dosage compensation; value = –1 indicates no dosage compensation (or X : A ratio = 0.5). Cuffdiff was used to identify differentially expressed genes between stages and the 2 datasets. Cuffnorm normalized FPKM for expression patterns, which were further clustered and graphed in R. Expression pattern distributions across regions on X were calculated by merging Cuffnorm output genes.attr_table to the expression pattern lists. The RXE values were higher than –1 in all embryonic stages studied, with in vitro embryos having higher RXE, suggesting some but incomplete dosage compensation and in vitro embryos exhibiting higher levels of X-gene expression. In vitro-produced immature oocytes and 4-cell embryos had RXE of 0, suggesting that both may be completely compensated. Additionally, embryos of the 2 sources exhibited significant differences in levels of X-gene expression at 4-cell to 8-cell and 16-cell to blastocyst stages. All expressed X-linked genes fell in 5 groups (patterns 1–5): increased, decreased, increased and then decreased, constant, decreased and then increased. Patterns 1 to 4 were seen in both datasets, whereas pattern 5 was only present in vitro, which may be why in vitro embryos had higher dosage of X. We further found that these expression patterns correlated with the genes’ proximity to the X inactivation centre (Xic: ~82 Mb): the highly dynamic patterns 3 and 5 were associated with proximity to Xic, supporting the notion that the Xist RNA spreads along the X through the Xic. In vivo and in vitro bovine oocytes and embryos have undergone some degree but incomplete dosage compensation. Expression of X-linked genes is correlated with their proximity to Xic. Globally, in vivo and in vitro embryos exhibit major differences in levels of X-gene expression.
154 MOLECULAR CHARACTERIZATION OF CAT PRIMORDIAL GERM CELLS
Authors: Galiguis J. , Pope C. E. , Dumas C. , Wang G. , MacLean R. A. , Gómez M. C.
Published: December 2015

As precursors to germline stem cells and gametes, there are many potential applications for primordial germ cells (PGC). Primordial germ cell-like cells have been generated from mouse embryonic stem cells and induced pluripotent stem cells, which subsequently were used to produce functional spermatozoa, oocytes, and healthy offspring (Hayashi et al. 2012 Science 338(6109), 971–975). Applying this approach to generate sperm and oocytes of endangered species is an appealing prospect. Detection of molecular markers associated with PGC is essential to optimizing the process of PGC induction. In the current study, in vitro-derived domestic cat embryos were assessed at various developmental stages to characterise the expression of markers related to the specification process of cat PGC. In vivo-matured, IVF oocytes were cultured until Days 7, 9, and 12 post-insemination. Then, embryos were assessed by RT-qPCR to determine relative transcript abundance of the pluripotency markers NANOG, POU5F1, and SOX2; the epiblast marker DNMT3B; the primitive endoderm marker GATA4; the PGC marker PRDM14; and the germ cell marker VASA; RPS19 was used as the internal reference gene. To validate the qPCR results, fibroblasts served as the negative control cells, whereas spermatogonial stem cells (SSC) served as the positive control cells for GATA4, PRDM14, and VASA. Total mRNA were isolated using the Cells-to-cDNA™ II Kit (Ambion/Thermo Fisher Scientific, Waltham, MA, USA) from either pools of 2 to 6 embryos or ~25 000 fibroblasts/SSC. A minimum of 2 biological replicates for each sample type was analysed, with transcript abundance detected in 2 technical replicates by SYBR Green chemistry. Student’s t-tests were performed on the ΔCts for statistical analysis. PRDM14, specific to the germ cell lineage, was detected as early as Day 7, suggesting the presence of PGC precursor cells. Compared with their levels at Day 7, PRDM14 expression was 0.34-fold lower in SSC (P < 0.05), whereas expression of VASA and GATA4 were 1964-fold and 144-fold higher, respectively (P < 0.05). This seems to emphasise the relative importance of PRDM14 in pre-germ cell stages. In general, all genes analysed were up-regulated from Day 7 to Day 9. This up-regulation was statistically significant for SOX2 and GATA4 (P < 0.05). Relative to that at Day 9, all transcripts were relatively less abundant at Day 12 (P < 0.05 for NANOG, POU5F1, SOX2, DNMT3B, and PRDM14). The data suggest that PGC specification takes place near Day 9, with peak specification activity concluding by Day 12. Although much needs be explored about PGC specification in the cat before applying induction and in vitro germ cell production techniques, these findings represent the first step towards a new potential strategy for preserving endangered and threatened felids.
156 EFFECT OF 2-METHOXYESTRADIOL (2-ME) ON HUMAN UTERINE LEIOMYOSARCOMA
Authors: Kim K. P. , Lee J.-S. , Ahn C. , Choi Y.-K. , Jeung E.-B.
Published: December 2015

2-Methoxyestradiol (2-ME) is an endogenous metabolite of 17β-oestradiol (E2) and has affinity for oestrogen receptors. It was reported as a promising antitumour drug due to antiproliferative activity in many tumour cell types. It has also been used in several preclinical and clinical studies for treatment of solid tumours. Thus, several studies have been conducted to investigate the cytotoxic effect of 2-ME on tumour cell lines in which it induced G2/M cell cycle arrest and subsequent apoptosis. Uterine leiomyosarcoma is a rare form of malignant smooth muscle cell tumour in the uterus muscle layer. This tumour is also influenced by oestrogen, which acts as a promoter. The purpose of this study was to examine the antiproliferative effect of 2-ME in vitro and in vivo. To perform in vitro experiments, we evaluated the antiproliferative effect of 2-ME on SK-LMS-1 cells using MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay and Western blot. In vivo experiment also investigated the effect of 2-ME on uterine leiomyoma by measuring tumour size and Western blotting. Employing the mouse xenograft model using human leiomyosarcoma SK-LMS-1 cell line compared with the positive control flavopiridol for 2 weeks, mice were divided into 4 groups (6 mice per each group); VE, 2-ME (75 or 150 mg kg–1), or flavopiridol (7.5 mg kg–1). All experimental procedures were performed in triplicate. P-values were calculated using one-way ANOVA, followed by Tukey’s test for multiple comparisons of columns. Data were considered statistically significant at P < 0.05. In vitro, we confirmed that 2-ME at 10–5 M and flavopiridol have an antiproliferative influence on SK-LMS-1 cells, but the same effect at 10–7 M or 10–6 M dose of 2-ME were not detected. These 2 doses showed little proliferative response in the MTT assay. Also, BAX/BCL-2 and LC3 expression were increased by 2-ME (10–5 M) treatment in Western blot analysis. In the in vivo xenograft study, application of 2-ME (75 mg kg–1) resulted in an increased tumour size but 150 mg kg–1 of 2-ME and flavopiridol decreased the size. In a previous study, we found that 2-ME has 2 types of action in an in vitro and an in vivo model. The 2-ME, which is used as a therapeutic agent for solid cancers, has not only apoptotic but also a proliferative effect, depending on the dose. We expect that 2-ME may be a potential therapeutic reagent for human uterine leiomyoma, but the appropriate dose of 2-ME should be used for oestrogen-response tumour treatment.