bluuebunny/crossref_metadata_2025 huggingface dataset:
166 INFLUENCE OF SEASON ON SEMINAL CHARACTERISTICS OF ALPACAS (VICUGNA PACOS)
Authors: Villanueva J. C. , Huanca W. F. , Turin J. , Huanca W.
Published: December 2016

The study was carried out with the objective to evaluate the influence of season (summer and winter) on seminal characteristics of alpacas. Forty semen samples, 20 for each season, were collected from 4 adult alpaca by artificial vagina technique and evaluated immediately after collection to determine the following semen characteristics: (a) volume, (b) pH, (c) viscosity, (d) color, (e) concentration, (f) motility, (g) viability, (h) abnormalities, and (i) HOST test. Statistical analyses of data were made with least square ANOVA. The males were fed the same diet during both seasons and collection time was standardized in 10 min. The results obtained present differences in viscosity (5.9 ± 0.4 v. 3.7 ± 0.7 mm), motility (58.8 ± 22.1 v. 77.3 ± 11.6%), and concentration (135.9 ± 88.7 × 106 v. 242.4 ± 140.9 × 106; P < 0,05) but not in volume (1.9 ± 0.8 v. 2.1 ± 0.8 mL), pH (7.01 ± 0.3 v. 7.1 ± 0.3), viability (52.9 ± 18.4 v. 64.5 ± 15.9%), HOST test (56.1 ± 15.8 v. 49.3 ± 13.2%), and abnormalities (82.5 ± 7.2 v. 88.1 ± 7.0%) for the summer and winter seasons, respectively. The results suggest differences for some characteristics and possible differences in the thermoregulation scrotal of the alpacas with respect to others species.
200 THE USE OF LONG-ACTING FSH-MAP5 IN SHEEP SUPEROVULATION PROGRAMS
Authors: Fry R.
Published: December 2016

The administration of sustained-release FSH-MAP5 in a 2-injection protocol has been shown to be as effective as a multiple-FSH injection protocol in inducing superovulation in cattle (Bo and Mapletoft 2014 Theriogenology 81, 38) and in sheep (Fry et al. 2016 Reprod. Fertil. Dev. 28, 250); however, the effect on embryo quality in the latter experiment was unclear. The following experiment further investigated the effect of FSH-MAP5 on ovulation rate and embryo quality in a sheep multiple-ovulation embryo transfer (MOET) program conducted in the breeding season. Two hundred sixteen Dohne merino ewes received a 12-day CIDR-S device containing 0.33 g of progesterone (Zoetis, Florham Park, NJ, USA) plus 200 mg of FSH IM (Folltropin-V; Vetoquinol, Belleville, Canada) and 400 IU of eCG IM (Pregnecol: Vetoquinol) in 4 treatment groups. Group 1 (n = 51) received 7 injections (a.m., p.m.) of FSH in saline (2.5, 2.0, 1.5, 1.5, 1.0, 1.0, and 0.5 mL) starting 2.5 days before CIDR withdrawal and 400 IU of eCG in saline at the time of the first FSH injection. Group 2 (n = 53) received 6 injections (a.m., p.m.) of FSH in saline (3.0, 2.0, 1.5, 1.5, 1.0, and 1.0 mL) starting 2.5 days before CIDR withdrawal and 400 IU of eCG in saline at CIDR withdrawal. Group 3 (n = 56) received 3.3 mL of FSH in hyaluronan (50 mg of MAP5, Vetoquinol) and 400 IU of eCG in saline 2.5 days before CIDR withdrawal and 1.7 mL of FSH-MAP5 at 0.5 days before CIDR withdrawal. Group 4 (n = 56) received 3.3 mL of FSH-MAP5 at 2.5 days before CIDR withdrawal, 1.7 mL of FSH-MAP5 at 0.5 days before CIDR withdrawal, and 400 IU of eCG in saline at CIDR withdrawal. Ewes were inseminated with semen collected and pooled from 5 rams at 36 to 40 h after CIDR withdrawal. Donor ewes were slaughtered 6 days after AI, corpora lutea were counted on the ovary, and ova/embryos were collected. Data for corpora lutea, total ova/embryo, and transferable embryo was analysed by the Kruskal–Wallis test and differences between groups were determined by the Dunn’s test. Results are shown in Table 1. There were no differences in the mean number of ovulations or total ova/embryo collected between any group (P > 0.05); however, those receiving eCG at the time of CIDR withdrawal (Groups 2 and 4) had significantly more transferable embryos than those receiving eCG at the time of the first FSH injection (Groups 1 and 3). In conclusion, the administration of the sustained-release FSH-MAP5 in a 2-injection protocol in sheep was as effective as a multiple-FSH injection protocol in inducing multiple ovulations and yielded similar number of transferable embryos, whereas the administration of eCG at the time of CIDR withdrawal resulted in more transferable embryos, possibly by improving fertilisation rates.

Table 1.
Results1
211 EFFECT OF AMNIOTIC FLUID DERIVED STEM CELL-CONDITIONED MEDIUM ON DEVELOPMENT AND POST-THAW SURVIVAL OF IN VITRO-PRODUCED HOLSTEIN EMBRYOS
Authors: Krautkramer K. , Owen C. M. , Barceló-Fimbres M. , Campos-Chillon L. F.
Published: December 2016

In vitro-produced (IVP) embryos have altered metabolism with lower blastocyst rates and higher lipid accumulation compared with IVP embryos. Culturing embryos with amniotic fluid derived stem cell-conditioned media (AFS) may help mimic in vivo conditions and improve embryo development. We hypothesised that culturing embryos with AFS will improve in vitro culture conditions and developmental competence. The IVP embryos were produced in 5 replicates by aspirating oocytes (n = 1469) of 2 to 8 mm from abattoir ovaries. Oocytes were matured for 23 h, fertilized with semen from 1 of 2 bulls, and cultured in a novel SOF medium (SOF for conventional freezing media) in 38.5°C in 5% O2, 5% CO2, and 90% N2. Media was supplemented with either 0 (control), 5, or 10% AFS. Stage 7 blastocysts were stained with 1 µg mL−1 of Nile Red and 300 nM Mitotracker Red CMX-Rosamine (Molecular Probes Inc., Eugene, OR, USA) to measure lipid content and mitochondrial polarity, respectively. Ten images per stained embryo were acquired by confocal microscopy using a 5 µM step size at 40× magnification. Remaining blastocysts were slow frozen using 2 min of dehydration in 0.6 M sucrose before equilibration for 10 min in a conventional freezing medium (1.5 M ethylene glycol and 0.5 M sucrose). Embryos were thawed and re-expansion was assessed at 24 and 48 h. Data were analysed by general linear model using a one-way ANOVA with means separated by Tukey’s HSD. Results (Table 1) indicated no difference in development, lipid content, mitochondrial polarity, or post-thaw survival (P > 0.01), suggesting that growth factors present in AFS did not improve culture conditions.

Table 1.
Main effect of AFS on embryo developmental competence and post-thaw survival rates
22 DYNAMIC OF SYNCHRONIZED FOLLICULAR WAVE IN EWES SUBJECTED TO DIFFERENT DOSES OF 17β-OESTRADIOL GIVEN AT THE BEGINNING OF THE PROGESTERONE PROTOCOL
Authors: Oliveira M. E. F. , Nociti R. P. , Camela E. S. C. , Padilha-Nakaghi L. C. , Maciel G. S. , Rodriguez M. G. K. , Santos V. J. C. , Fonseca J. F. , Vicente W. R. R.
Published: December 2016

This study was designed to evaluate the effect of different doses of 17β-oestradiol injection at the beginning of the progesterone protocol on follicular wave dynamic in ewes. In a random day of the oestrous cycle (D0), twenty-four Santa Ines ewes received an intravaginal progesterone device (CIDR®) and a injection of 17β-oestradiol (E2, Sincrodiol®, Ourofino, Brazil) in different doses (350, 500, and 1000 µg) for G-350E2, G-500E2, and G-1000E2, respectively (n = 8 per group). Ultrasound examinations were performed daily during the CIDR permanence (10 days) using MyLab 30Vet equipment (Esaote, Italy) connected to transrectal linear transducer (frequency of 7.5 MHz). Follicular wave was defined as a follicle or a group of follicles 2 to 3 mm in diameter that grew to ≥4.5 mm in size before regression or ovulation. The day of wave emergence was based on the day on which the largest follicle of a wave was first detected at 2 or 3 mm (retrospective analysis). Data were analysed by ANOVA followed by a Tukey test (mean ± SEM; P < 0.05) using SAS software. The emergence of a new follicular wave after the beginning of protocols was not detected in 8 out of 24 ewes (33.3%), being 3 from G-350E2, 2 from G-500E2, and 3 from G-1000E2. There was difference (abP = 0.04) for follicular wave emergence day (3.00 ± 0.32b, 4.00 ± 0.45ab and 5.20 ± 0.73a) and averaged day of maximal follicle diameter (8.20 ± 0.58b, 9.50 ± 0.34ab and 9.80 ± 0.20a) for G-350E2, G-500E2 , and G-1000E2, respectively. However, the growing period and the maximum diameter of the largest follicle of this wave did not differ between groups (124.80 ± 8.98 h and 5.42 ± 0.25 mm for G-350E2; 132.00 ± 5.37 h and 5.75 ± 0.23 mm for G-500E2; and 110.40 ± 16.28 h and 5.20 ± 0.73 mm for G-1000E2, respectively). In conclusion, the 17β-oestradiol injection at the beginning of the progesterone protocol was not able to induce the emergence of a new follicular wave in all of females, regardless of dose. However, the synchronized follicular wave occurred earlier in females treated with the lowest dose of the 17β-oestradiol than those that received the highest dose.
The study received financial support from CNPq and FAPESP.
25 PRODUCTION OF TRANSGENIC PIGS WITH CreER-MEDIATED ASTROCYTIC-SPECIFIC RECOMBINATION SYSTEM FOR NEUROLOGICAL DISEASE MODELS
Authors: Hwang S.-U. , Yoon J. D. , Eun K. , Kim H. , Hyun S.-H.
Published: December 2016

Pigs are one of the most suitable alternative laboratory models than other animals, because they have similar cardiovascular, renal and gastrointestinal organs with those of human. However, in the case of genetically engineered animals, early development of embryos is inhibited by expression of foreign genes, there are many cases of miscarriage or birth early mortality. To overcome these problems, we constructed pig glial fibrillary acidic protein (GFAP) promoter-Cre recombinase fused to a mutated ligand-binding domain of the human oestrogen receptor (CreERT2) and enhanced green fluorescent protein (EGFP)-LoxP transgenes for tamoxifen(TM)-inducible CreERT2-mediated recombination. We then established donor transgenic pig fibroblasts with pGFAP-CreERT2; LCMV-EGFPLoxP transgenes for somatic cell nuclear transfer (SCNT). We produced the SCNT embryos using a Cloud male #5 pGFAP-CreERT2+LCMV-EGFPLoxP donor cell line that was verified in vitro. It was transferred into a surrogate mother and then 5 pGFAP-CreERT2; EGFPLoxP TG piglets were born. By immunofluorescence staining and semi-nested PCR analysis, it was proved that CreER-mediated astrocytic-specific recombination system was operated in some cerebral astrocytic cells after TM-administration to TG pig #4. Additionally, we obtained brain magnetic resonance imaging (MRI) images using 3T-tesla MRI. Brain compartment volume (total brain, grey matter, white matter, cerebellum, brainstem, lateral ventricle, thalamus, midbrain, pons, medulla oblongata, hypophysis) was no significant differences between normal pig and pGFAP-CreERT2; EGFPLoxP transgenic (TG) pig. In summary, we verified the pGFAP promoter-driven CreERT2-LoxP recombination system in TG pig generated by SCNT depending on the TM administration. We suggest that this technology will be a useful tool for studying physiology of astrocytes and generating TG pig model of neurological disease such as Huntington’s disease, Alzheimer’s disease and brain tumour.
36 SERIAL SOMATIC CELL NUCLEAR TRANSFER INCREASES PREGNANCY LOSSES IN GOATS
Authors: Yang M. , Hall J. , Meng Q. , Fan Z. , Polejaeva I.
Published: December 2016

Serial cloning by somatic cell nuclear transfer (SCNT) has been successful in several mammalian species. This method can be beneficial for transgenic line expansion or resetting the lifespan of transgenic cells. Previous studies in bovine and porcine have shown a decrease in efficiency over multiple iterations of serial cloning. However, the contradictory data has been reported in mice where no decrease in cloning efficiency was observed after 25 generations of recloning. To our knowledge, no data have been reported investigating the efficiency of serial cloning in goats. The aim of this study was to evaluate whether there is an effect of recloning on goat SCNT efficiency. αMHC-TGF-β1 fetal fibroblast cells (containing transforming growth factor-β under control of a cardiac-specific promoter) were produced by electroporation and used for the first round of SCNT. For serial cloning, we used neonatal fibroblast cells obtained from skin biopsies used as nuclear donors. These cells were collected from the transgenic cloned goats generated by the first round of SCNT. Cumulus-oocyte complexes recovered from abattoir-derived ovaries using slicing technique were matured in vitro for 20 to 24 h. The first polar body and metaphase plate were removed from a cumulus cell-free oocyte, and a donor fibroblast cell was subsequently transferred into the enucleated oocyte. Fused embryos were then activated for 5 min in 5 mM ionomycin followed by 4 h in 2 mM DMAP with 5 mg mL−1 cycloheximide. Activated embryos were cultured in G1 medium with 5 mg mL−1 BSA for 12 h, followed by surgical transfer into the oviducts of recipients synchronized to show oestrus within 12 h of SCNT. In total, 592 and 395 embryos were transferred to 37 and 25 recipient goats, respectively, for the first and second round of SCNT. Pregnancy rate, rate of pregnancy loss, and term rate were analysed by Chi-squared with a 2-tailed P-value. No significant difference was observed in Day 40 pregnancy rates (32.4 v. 36%) and term rates (32.4 v. 20%) between the first round of cloning and the successive recloning. However, the rate of pregnancy losses was significantly greater in recloning group (P < 0.05), with 4 out of 9 pregnancies lost between Day 40 of gestation and term, whereas no pregnancy losses were observed after Day 40 of gestation in the first-round cloning group. The greater pregnancy loss in the recloning procedure might be caused by accumulation of epigenetic errors resulting from incomplete reprogramming. We are assessing the DNA methylation pattern of differentially methylated regions (DMR) of 2 paternally imprinted genes (H19 and IGF2R) in the cloned and recloned goats and expect to see a difference in their imprinted gene DNA methylation pattern, which could explain the greater rate of pregnancy loss in recloned goats.
37 NORMALITY OF NEONATAL REFLEX IN CLONED DOGS
Authors: Setyawan E. M. N. , Kim G. A. , Oh H. J. , Kim M. J. , Taweechaipaisankul A. , Lee S. H. , Choi Y. B. , Lee B. C.
Published: December 2016

Since the birth of the world’s first cloned dog, Snuppy, somatic cell nuclear transfer (SCNT) has been a useful tool to propagate the dogs with identical genetic information. However, it is known that cloned animals sometimes exhibit phenotypic instability or abnormality. There have been few investigations about the normality of the neonatal reflex in cloned animals. Therefore, the objective of this study was to evaluate the neonatal reflex in 3 breeds of cloned dogs including shepherd, retriever, and beagle from birth to 28 days of age. Through SCNT, 8 cloned dogs were produced. After birth, 3 types of neonatal reflexes were examined and scored. For examining the flexor dominance reflex, neonatal cloned dogs were held upright and the flexor position of the limb was scored. To evaluate the withdrawal and crossed extensor reflexes, neonates were placed in lateral recumbence and their forelimbs were allowed to relax. Then, the distal forelimbs were pinched and responses were scored according to the frequency and intensity (strong = score 2, variable = score 1, and absent = score 0). The standard responses of neonates were referred from Lindsay et al. (2000 Handbook of Applied Dog Behavior and Training 1, 31–47). Descriptive analysis was used, which was based on the scores from 3 referees who evaluated the videos. The flexor dominance reflex could not be observed (score 0.0) in shepherd by Day 8, in beagle by Day 14 and in retriever by Day 16. Withdrawal reflex began to decrease on Day 22 with score 1.8 for beagle and retriever but decreased in shepherd starting on Day 24 with score 1.8. Crossed extensor reflex for shepherd started to disappear on Day 14 with score 1.5 and completely disappeared (score 0.0) on Day 22; for beagle started to disappear on Day 16 with score 1.8 and was still found until Day 28 with score 1.1; for retriever started to disappear on Day 20 and 28 with score 1.7 and 0.7, respectively. Flexor dominance reflex disappeared in cloned shepherd at a similar time to standard but beagle and retriever seem delayed 6 to 8 days compared with the reference. Withdrawal reflex in all breeds showed normal changes that should persist until adulthood. Cross extensor reflex in shepherd was close to reference but in beagle and retriever was delayed beyond Day 28; this reflex should disappear before adulthood. This study demonstrated that normal neonatal reflexes were identified in the cloned dogs, with some variations among breed. To adapt neonatal reflex as a marker to confirm phenotypic normality in cloned dogs, further investigation using various breeds of cloned dogs and greater numbers of subjects is needed.
This study was supported by IPET (#316002-05-1-SB010), RDA (#PJ010928032016), Research Institute for Veterinary Science, Natural Balance Korea and the BK21 plus program.
56 THE VIABILITY AND LONGEVITY OF OGYE CHICKEN FROZEN SEMEN WITH MITOTEMPOL, MITOCHONDRIA-SPECIFIC ANTIOXIDANT
Authors: Kim S. W. , Kim M. S. , Kim C.-L. , Kim D. , Seong H.-H.
Published: December 2016

Semen from Korean Native Black roosters (Ogye) was cryopreserved with 8% N-Methylacetamide (MA) in HS-1 diluent, of the following composition (per 100 mL): glucose 0.2 g, trehalose dihydrate 3.8 g, l-glutamic acid monosodium salt 1.2 g, potassium acetate 0.3 g, magnesium acetate tetrahydrate 0.08 g, potassium citrate monohydrate 0.05 g, BES 0.4 g, Bis-Tris 0.4 g, and gentamicin sulfate 0.001 g. Ogye semen was collected 2 times a week by dorsal-abdominal massage and cooled into ice slurry. During dilution, semen was diluted by 2 steps. First, HS-1 diluent that was supplemented with respective concentration of MitoTEMPOL, mitochondria-specific antioxidant was added into fresh semen with same volume. Second, after waiting for 10 min, 16% MA containing diluent was added into first diluted semen so the final concentrations of MA were adjusted to 8%. The concentration of the MitoTEMPOL of first diluent was adjusted to 0.1, 1, 5, and 10 μM. After freezing for 30 min by exposure to liquid nitrogen vapor 4 cm above the surface of liquid nitrogen, the semen was thawed 5°C for 2 min 1 to 3 weeks later. The viability and longevity of thawed Ogye semen were analysed by sperm movement methods. Briefly, 5 μL of thawed semen was placed onto a Makler chamber and the movement of spermatozoa was recorded for 10 s by digital camera and saved as movie files on the computer. With digital rewinding for 1 s of saved movies, the non-motile and motile sperm was counted using a manual counter. The resulting numbers of respective spermatozoa were analysed by Student t-test. The 0.1 and 1 μM treated semen showed significant increases in viability compared with control, 5 μM and 10 μM MitoTempol (77.3 and 84.2% v. 65.4, 53.2, and 21.5%; P < 0.05). The longevity of frozen/thawed Ogye semen for 3 h was also higher in 0.1 and 1 μM treated groups than control, 5 μM, and 10 μM (67.5 and 54.2% v. 36, 5.2, and 1.2%; P < 0.05). With these results, utilisation of mitochondria-specific antioxidant for freezing of Ogye spermatozoa could increase the viability and longevity of frozen-thawed semen. However, treatment with concentrations >1 μM showed negative effects on freezing of Ogye chicken semen. These findings could be helpful for cryobanking of rooster semen for preservation of selected breeders from malignant viral avian disease.
60 EFFECTS OF CONCANAVALIN A ON THE PROGESTERONE PRODUCTION BY BOVINE STEROIDOGENIC LUTEAL CELLS IN VITRO
Authors: Destro F. C. , Martin I. , Landim-Alvarenga F. D. C. , Sartori Filho R. , Pate J. L. , Ferreira J. C. P.
Published: December 2016

The corpus luteum is a temporary organ that is responsible for progesterone (P4) secretion and is essential for the establishment and maintenance of pregnancy in cattle. Concanavalin A (CONA) is a lectin that was originally extracted from the Jack bean (Canavalia ensiformis) and that interacts with several kinds of cells, including immune cells and luteal cells. The aim of the present study was to evaluate the effects of CONA on the P4 production by bovine steroidogenic luteal cells (LC) in vitro. Luteal cells were collected during the mid-luteal stage (at 10–12 days following ovulation) and processed in the laboratory. Luteal cells were grown for 7 days in a humid atmosphere with 5% CO2, with or without 10% fetal bovine serum (FBS), and were subjected to the following treatments: control: no treatment; CONA (10 μg mL−1); LH (100 μg mL−1); CONA+LH; LH (100 μg mL−1) + prostaglandin F2α (PGF2α; 10 ng mL−1); CONA+LH+PGF2α. Samples of the culture media were collected on Day 1 and Day 7 for P4 quantification. The cells were counted on Day 7 of culture. Differences between treatments were considered statistically significant at P < 0.05. The P4 concentration in the culture media was numerically greater on Day 1 (558.0 ng mL−1) than on Day 7 (25.4 ng mL−1). The P4 concentration in the culture media was numerically greater for treatments with 10% FBS than for the FBS-free treatments, and the presence of CONA decreased LC P4-secreting capacity. This effect required more than 24 h of exposure to CONA to be fully manifested. On Day 1 of culture, CONA had no effect on P4 production of LC cultured in serum-free medium (P > 0.05).The suppressive action of CONA was more pronounced for cultures without FBS. By Day 7 of culture, the effects of CONA on P4 production were readily apparent. In the absence of serum, CONA had a highly significant (P < 0.01) inhibitory effect on basal progesterone production, as well as in the presence of LH or LH + PGF. In the presence of FBS, there was a tendency for decreased P4 in response to CONA in the LH- and the LH + PGF-treated cells (P = 0.090 and 0.085, respectively). The number of the cells present on Day 7 was not affected by the treatments tested (P > 0.05). More studies are required to better understand the effect of CONA on the P4 production of bovine LC.
Financial support from FAPESP is acknowledged: grant no. 2013/00992–3, grant no. 2013/07439–8, and grant no. 2015/01940–2.
73 SPATIAL DIFFERENCES IN METABOLITES AND ENERGY SUBSTRATES IN THE BOVINE OVIDUCT
Authors: Maillo V. , Simintiras C. , Sturmey R. , Lonergan P. , Rizos D.
Published: December 2016

Knowledge of the biochemical composition of the bovine oviduct in the presence of an embryo is lacking. We have recently reported the detection of alterations to the bovine oviduct transcriptome when multiple embryos, but not a single embryo, were present (Maillo et al. 2015 Biol. Reprod. 92, 144). Thus, we hypothesised that the presence of an embryo would modify the concentration of metabolites and energy substrates in the oviducal fluid. Thus, the aim of the present study was to examine how the presence of an embryo affects metabolites and energy substrates within the oviduct. Cross-bred beef heifers were synchronized, and those in standing oestrus were randomly allocated to cyclic, not bred (n = 7), or pregnant, artificially inseminated (n = 11), groups. All heifers were slaughtered on Day 3 after oestrus. The oviducts from each animal were isolated, straightened, and cut, separating ampulla and isthmus. Each portion was flushed with 500 μL of PBS, enabling recovery of any oocyte/embryo. Recovered unfertilized oocytes (cyclic group) and embryos (pregnant group) were located in the isthmus of the oviduct ipsilateral to the corpus luteum. Samples of flushing medium from the isthmus and ampulla from oviducts ipsilateral to the corpus luteum from 5 cyclic and 5 pregnant animals (8-cell embryos) were used for metabolites and energy substrates analysis, together with PBS as blank control. Amino acid content in the oviducal fluid was determined by high performance liquid chromatography. Glucose, lactate, and pyruvate were quantified using microfluorometric enzyme-linked assays. Statistical analysis was carried out by 2-way ANOVA with a Bonferroni post hoc for multiple comparisons. No differences were found either in the concentration of metabolites or in the energy substrates between cyclic and pregnant heifers. However, significant differences were detected between ampulla and isthmus in cyclic and pregnant heifers, which is in agreement with our previous findings on the transcriptome between isthmus and ampulla (Maillo et al. 2016 Reproduction 152, 37–46). In pregnant and cyclic heifers, glycine and alanine were more abundant in the ampulla compared with the isthmus (P < 0.01). In addition, in pregnant heifers, arginine was less abundant in the ampulla than in the isthmus (P < 0.01). Energy substrates were also similar between pregnant and cyclic heifers. However, lactate was significantly different between ampulla and isthmus, being present in the ampulla of cyclic and pregnant heifers and being absent from the isthmus (P < 0.05). In conclusion, the content of metabolites and energy substrates in the oviduct varies between the ampulla and isthmus independent of the presence of an embryo.
This work was funded by the Spanish Ministry of Economy and Competitiveness (AGL-2012–37510 and AGL-2015–70140-R) and Science Foundation Ireland (13/IA/1983).
Egr2 and Egr3 in regulatory T cells cooperatively control systemic autoimmunity through Ltbp3-mediated TGF-β3 production
Authors: Morita Kaoru , Okamura Tomohisa , Inoue Mariko , Komai Toshihiko , Teruya Shuzo , Iwasaki Yukiko , Sumitomo Shuji , Shoda Hirofumi , Yamamoto Kazuhiko , Fujio Keishi
Published: December 2016

Significance

Transcription factors early growth response gene 2 (Egr2) and Egr3 have long been regarded as negative regulators of T-cell activation.Egr2is also known as a susceptibility gene for systemic lupus erythematosus characterized by dysregulated humoral immune responses to autoantigens. Previously, we reported that Egr2-expressing CD4+CD25-LAG3+regulatory T cells regulate lupus pathogenesis via production of TGF-β3. However, the role of Egr2 and Egr3 in the regulation of humoral immunity is unclear. Here we report that Egr2 and Egr3 regulate germinal center reactions by promoting TGF-β3 production from regulatory T cells. Egr2 and Egr3 induce the expression of latent TGF-β binding protein 3 (Ltbp3), which is required for TGF-β3 secretion. These findings suggest that Egr2 and Egr3 in T cells may be potential novel therapeutic targets for autoantibody-mediated autoimmune diseases.
O-GlcNAc glycosylation stoichiometry of the FET protein family: only EWS is glycosylated with a high stoichiometry
Authors: Kamemura Kazuo
Published: December 2016

Of the FET (fused in sarcoma [FUS]/Ewing sarcoma protein [EWS]/TATA binding protein-associated factor 15 [TAF15]) family of heterogeneous nuclear ribonucleoprotein particle proteins, FUS and TAF15 are consistently and EWS variably found in inclusion bodies in neurodegenerative diseases such as frontotemporal lobar degeneration associated with FUS. It is speculated that dysregulation of FET proteins at the post-translational level is involved in their cytoplasmic deposition. Here, the O-linked β-N-acetylglucosamine (O-GlcNAc) glycosylation stoichiometry of the FET proteins was chemoenzymatically analyzed, and it was found that only EWS is dynamically glycosylated with a high stoichiometry in the neural cell lines tested and in mouse brain. It was also confirmed that EWS, but not FUS and TAF15, is glycosylated with a high stoichiometry not only in the neural cells but also in the non-neural cell lines tested. These results indicate that O-GlcNAc glycosylation imparts a physicochemical property on EWS that is distinct from that of the other FET proteins in most of cell lineages or tissues.
AGN feedback and the origin of the α enhancement in early-type galaxies – insights from the GAEA model
Authors: De Lucia Gabriella , Fontanot Fabio , Hirschmann Michaela
Published: December 2016

We take advantage of our recently published model for GAlaxy Evolution and Assembly (GAEA) to study the origin of the observed correlation between [α/Fe] and galaxy stellar mass. In particular, we analyse the role of radio-mode active galactic nuclei (AGN) feedback, which recent work has identified as a crucial ingredient to reproduce observations. In GAEA, this process introduces the observed trend of star formation histories extending over shorter time-scales for more massive galaxies, but does not provide a sufficient condition to reproduce the observed α enhancements of massive galaxies. In the framework of our model, this is possible only by assuming that any residual star formation is truncated for galaxies more massive than 1010.5 M⊙. This results, however, in even shorter star formation time-scales for the most massive galaxies, which translate in total stellar metallicities significantly lower than observed. Our results demonstrate that (i) trends of [α/Fe] ratios cannot be simply converted into relative time-scale indicators; and (ii) AGN feedback cannot explain alone the positive correlation between [α/Fe] and galaxy mass/velocity dispersion. Reproducing simultaneously the mass–metallicity relation and the α enhancements observed pose a challenge for hierarchical models, unless more exotic solutions are adopted such as metal-rich winds or a variable initial mass function.
Quality of Life and Sexual Distress in Women With Erosive Vulvovaginal Lichen Planus
Authors: Cheng Harriet , Oakley Amanda , Conaglen John V. , Conaglen Helen M.
Published: December 2016

Objectives

Erosive vulvovaginal lichen planus (EVLP) is a chronic and painful genital dermatosis. Little is published about its impact on quality of life. This study aimed to evaluate quality of life and sexual function in women with EVLP.

Materials and Methods

Women with genital dermatoses were surveyed using the Dermatology Life Quality Index (DLQI) and Hospital Depression and Anxiety Scales. A subgroup completed the Female Sexual Distress Scale and Female Sexual Function Index subscales. Patient characteristics including age, diagnosis, and current treatment were recorded. Results from women with EVLP were compared with other diagnoses.

Results

Data from 77 women who participated between March 2013 and March 2014 were analyzed. Of these, 17 had EVLP. Comparator groups included women with vulval lichen sclerosus (n = 48) and vulval dermatitis (n = 12). In women with EVLP, 59% reported at least moderate impact on quality of life; mean DLQI scores: EVLP, 7.18; lichen sclerosus, 3.79; dermatitis, 8.67; p = .008. Overall, scores suggested depression in 14% and anxiety in 16% of participants. Sexual distress scores 11 or higher were recorded by 69% of women with EVLP, 63% of women with lichen sclerosus, and 56% of women with dermatitis. In those completing all sections of the survey (n = 40), DLQI was significantly correlated with depression (p = .004), sexual distress (p = .001), and sexual satisfaction (p = .01).

Conclusions

Sixty-nine percent of women with EVLP reported sexual distress. Women with EVLP reported lesser quality of life than those with lichen sclerosus. Quality of life, anxiety and depression, sexual distress, and sexual function were all related in these participants.
Understanding Interrater Reliability and Validity of Risk Assessment Tools Used to Predict Adverse Clinical Events
Authors: Siedlecki Sandra L. , Albert Nancy M.
Published: December 2016

Purpose:

This article will describe how to assess interrater reliability and validity of risk assessment tools, using easy-to-follow formulas, and to provide calculations that demonstrate principles discussed.

Description:

Clinical nurse specialists should be able to identify risk assessment tools that provide high-quality interrater reliability and the highest validity for predicting true events of importance to clinical settings. Making best practice recommendations for assessment tool use is critical to high-quality patient care and safe practices that impact patient outcomes and nursing resources. Optimal risk assessment tool selection requires knowledge about interrater reliability and tool validity.

Outcome:

The clinical nurse specialist will understand the reliability and validity issues associated with risk assessment tools, and be able to evaluate tools using basic calculations.

Conclusion:

Risk assessment tools are developed to objectively predict quality and safety events and ultimately reduce the risk of event occurrence through preventive interventions. To ensure high-quality tool use, clinical nurse specialists must critically assess tool properties. The better the tool's ability to predict adverse events, the more likely that event risk is mediated. Interrater reliability and validity assessment is relatively an easy skill to master and will result in better decisions when selecting or making recommendations for risk assessment tool use.
Safety Profile of Children in an Enclosure Bed
Authors: Sherburne Eileen , Snethen Julia A. , Kelber Sheryl
Published: December 2016

Purpose:

Pediatric nurses care for many children in the hospital setting whose behavioral challenges can impact safety. One intervention utilized to prevent injury and improve safety when caring for this population of children has been the enclosure bed. Scant information was found in the literature that discusses the use of the enclosure bed in the pediatric setting. The purpose of this article is to examine the use of the enclosure bed in the pediatric setting. A secondary aim is to identify the population of children where an enclosure bed was implemented and the safety factors associated with use of the enclosure bed.

Methods:

An exploratory retrospective chart review was conducted of 208 pediatric enclosure bed encounters in an acute care setting over a 2-year period. Variables included demographics, length of stay and bed use, behavioral medications, restraint and sitter usage, skin breakdown, fall risk, and falls.

Results:

Three categories of children based on cognitive function, no cognitive impairment, new cognitive impairment, and congenital cognitive impairment, were extracted from the chart review. Significant differences were found between groups of children and between safety variables observed with enclosure bed use. Children with new-onset cognitive impairment were more likely to incur falls, skin breakdown, and injury during use of the enclosure bed.

Conclusion:

Use of the enclosure bed is a reasonable intervention in certain children to ensure their safety in the hospital setting. This information will enhance nursing knowledge related to providing safe and optimal care of this challenging population of children.
Reduced Length of Stay and Adverse Events Using Bier Block for Forearm Fracture Reduction in the Pediatric Emergency Department
Authors: Fauteux-Lamarre Emmanuelle , Burstein Brett , Cheng Adam , Bretholz Adam
Published: December 2016

Objectives

Forearm fractures are among the most common pediatric injuries. Procedural sedation is frequently used for analgesia during fracture reduction but requires a prolonged recovery period and can be associated with adverse events. Bier block is a safe alternative for fracture reduction analgesia. This study sought to compare Bier block and procedural sedation for forearm fracture reduction.

Methods

We performed a retrospective study of patients aged 6 to 18 years, presenting with forearm fractures requiring closed reduction from June 2012 to March 2014. Outcomes assessed were emergency department length of stay, reduction success rates, adverse events, and unscheduled return visits.

Results

Overall, 274 patients were included (Bier block, n = 109; procedural sedation, n = 165). Mean length of stay was 82 minutes shorter for Bier block patients (279 vs 361 minutes, P < 0.001). Subanalysis revealed a reduced length of stay among Bier block patients with forearm fractures involving a single bone (286 vs 388 minutes, P < 0.001) and both bones (259 vs 321 minutes, P < 0.05). Reduction success did not differ between Bier block and procedural sedation (98.2% vs 97.6%, P = 0.74). There were no major adverse events in either group, but Bier block patients experienced fewer minor adverse events (2.7% vs 14.5%, P < 0.001). Return visit rates were similar between Bier block and procedural sedation (17.6% vs 16.9%, P = 0.92).

Conclusions

Compared with procedural sedation, forearm fracture reduction performed with Bier block was associated with a reduced emergency department length of stay and fewer adverse events, with no differences in reduction success or return visits.
Optic neuritis: the eye as a window to the brain
Authors: Jenkins Thomas M. , Toosy Ahmed T.
Published: December 2016

Purpose of review

Acute optic neuritis is a common clinical problem, requiring a structured assessment to guide management and prevent visual loss. The optic nerve is the most accessible part of the central nervous system, so optic neuritis also represents an important paradigm to help decipher mechanisms of damage and recovery in the central nervous system. Important developments include the advent of optical coherence tomography as a biomarker of central nervous system axonal loss, the discovery of new pathological antibodies, notably against aquaporin-4 and, more recently, myelin oligodendrocyte protein, and emerging evidence for sodium channel blockade as a novel therapeutic approach to address energy failure in neuroinflammatory disease.

Recent findings

We will present a practical approach to assessment of optic neuritis, highlighting the role of optical coherence tomography, when to test for new antibodies and the results of recent trials of sodium channel blockers.

Optic neuritis remains a clinical diagnosis; increasingly optical coherence tomography is a key ancillary investigation. Patients with ‘typical’ optic neuritis, commonly a first presentation of multiple sclerosis, must be distinguished from ‘atypical’ optic neuritis, who require testing for new pathological antibodies and require more aggressive-targeted treatment. Sodium channel blockade is an emerging and novel potential therapeutic pathway in neuroinflammatory disease.
I3S Classic and Insect Species Identification of Diptera and Hymenoptera (Mosquitoes and Bees)
Authors: Vyas-Patel Nayna , Mumford John D
Published: December 2016

A number of image recognition systems have been specifically formulated for the individual recognition of large animals. These programs are versatile and can easily be adapted for the identification of smaller individuals such as insects. The Interactive Individual Identification System, I3S Classic, initially produced for the identification of individual whale sharks was employed to distinguish between different species of mosquitoes and bees, utilising the distinctive vein pattern present on insect wings. I3S Classic proved to be highly effective and accurate in identifying different species and sexes of mosquitoes and bees, with 80% to100% accuracy for the majority of the species tested. The sibling speciesApis melliferaandApis mellifera carnicawere both identified with100% accuracy.Bombus terrestris terrestrisandBombus terrestris audax; were also identified and separated with high degrees of accuracy (90% to 100% respectively for the fore wings and 100% for the hind wings). When bothAnopheles gambiaesensu stricto andAnopheles arabiensiswere present in the database, they were identified with 94% and 100% accuracy respectively, allowing for a morphological and non-molecular method of sorting between these members of the sibling complex. Flat, not folded and entire, rather than broken, wing specimens were required for accurate identification. Only one wing image of each sex was required in the database to retrieve high levels of accurate results in the majority of species tested. The study describes how I3S was used to identify different insect species and draws comparisons with the use of the CO1 algorithm. As with CO1, I3S Classic proved to be suitable software which could reliably be used to aid the accurate identification of insect species. It is emphasised that image recognition for insect species should always be used in conjunction with other identifying characters in addition to the wings, as is the norm when identifying species using traditional taxonomic keys.
Valence biases factual and counterfactual learning in opposite directions
Authors: Palminteri Stefano , Lefebvre Germain , Kilford Emma J. , Blakemore Sarah-Jayne
Published: December 2016

Previous studies suggest thatfactuallearning, that is, learning from obtained outcomes, is biased, such that participants preferentially take into account positive, as compared to negative, prediction errors. However, whether or not the prediction error valence also affectscounterfactuallearning, that is, learning from forgone outcomes, is unknown. To address this question, we analysed the performance of two cohorts of participants on reinforcement learning tasks using a computational model that was adapted to test if prediction error valance influences learning. Concerning factual learning, we replicated previous findings of a valence-induced bias, whereby participants learned preferentially from positive, relative to negative, prediction errors. In contrast, for counterfactual learning, we found the opposite valence-induced bias: negative prediction errors were preferentially taken into account relative to positive ones. When considering valence-induced bias in the context of both factual and counterfactual learning, it appears that people tend to preferentially take into account information that confirms their current choice
Recurring functional interactions predict network architecture of interictal and ictal states in neocortical epilepsy
Authors: Khambhati Ankit N. , Bassett Danielle S. , Oommen Brian S. , Chen Stephanie H. , Lucas Timothy H. , Davis Kathryn A. , Litt Brian
Published: December 2016

Human epilepsy patients suffer from spontaneous seizures, which originate in brain regions that also subserve normal function. Prior studies demonstrate focal, neocortical epilepsy is associated with dysfunction, several hours before seizures. How does the epileptic network perpetuate dysfunction during baseline periods? To address this question, we developed an unsupervised machine learning technique to disentangle patterns of functional interactions between brain regions, or subgraphs, from dynamic functional networks constructed from approximately 100 hours of intracranial recordings in each of 22 neocortical epilepsy patients. Using this approach, we found: (i) subgraphs from ictal (seizure) and interictal (baseline) epochs are topologically similar, (ii) interictal subgraph topology and dynamics can predict brain regions that generate seizures, and (iii) subgraphs undergo slower and more coordinated fluctuations during ictal epochs compared to interictal epochs. Our observations suggest that the epileptic network drives dysfunction by controlling dynamics of functional interactions between brain regions that generate seizures and those that underlie normal function.
Upgrading short-read animal genome assemblies to chromosome level using comparative genomics and a universal probe set
Authors: Damas Joana , O'Connor Rebecca , Farré Marta , Lenis Vasileios Panagiotis E. , Martell Henry J. , Mandawala Anjali , Fowler Katie , Joseph Sunitha , Swain Martin T. , Griffin Darren K. , Larkin Denis M.
Published: December 2016

Most recent initiatives to sequence and assemble new species’ genomes de novo fail to achieve the ultimate endpoint to produce contigs, each representing one whole chromosome. Even the best-assembled genomes (using contemporary technologies) consist of subchromosomal-sized scaffolds. To circumvent this problem, we developed a novel approach that combines computational algorithms to merge scaffolds into chromosomal fragments, PCR-based scaffold verification, and physical mapping to chromosomes. Multigenome-alignment-guided probe selection led to the development of a set of universal avian BAC clones that permit rapid anchoring of multiple scaffolds to chromosomes on all avian genomes. As proof of principle, we assembled genomes of the pigeon (Columbia livia) and peregrine falcon (Falco peregrinus) to chromosome levels comparable, in continuity, to avian reference genomes. Both species are of interest for breeding, cultural, food, and/or environmental reasons. Pigeon has a typical avian karyotype (2n = 80), while falcon (2n = 50) is highly rearranged compared to the avian ancestor. By using chromosome breakpoint data, we established that avian interchromosomal breakpoints appear in the regions of low density of conserved noncoding elements (CNEs) and that the chromosomal fission sites are further limited to long CNE “deserts.” This corresponds with fission being the rarest type of rearrangement in avian genome evolution. High-throughput multiple hybridization and rapid capture strategies using the current BAC set provide the basis for assembling numerous avian (and possibly other reptilian) species, while the overall strategy for scaffold assembly and mapping provides the basis for an approach that (provided metaphases can be generated) could be applied to any animal genome.
Performing Yeast One-Hybrid Library Screens
Authors: Fuxman Bass Juan I. , Reece-Hoyes John S. , Walhout Albertha J.M.
Published: December 2016

Yeast one-hybrid (Y1H) assays are used to identify which transcription factor (TF) “prey” molecules can bind a DNA fragment of interest that is used as “bait”. Y1H assays involve introducing plasmids that encode TFs into a yeast “bait strain” in which the DNA fragment of interest is integrated upstream of one or more reporters, and activation of these reporters indicates that a TF–DNA interaction has occurred. These plasmids express each TF as a hybrid protein (hence the “one-hybrid” name) fused to the activation domain (AD) of the yeast TF Gal4. The AD moiety activates reporter expression even if the TF to which it is fused typically functions as a repressor. Here, we describe how to perform a Y1H screen of a library of cDNA fragments cloned into a pPC86 plasmid expressing the protein encoded by the cDNA as an AD fusion. The method assumes availability of either commercially available libraries or libraries generated in house using mRNA extracted from a tissue of interest. We also assume that users have access to a yeast bait strain that possesses the DNA fragment of interest integrated upstream of two different reporters—HIS3, an auxotrophic marker, and LacZ, a colorimetric marker that changes colorless X-gal into a blue compound. Briefly, the screen involves transforming the AD-cDNA library into the yeast bait strain, identifying colonies that show activation of both reporters, retesting the interaction in a freshly grown bait strain, and sequencing the cDNA insert to identify the interacting TF.
Preparation of Intracellular Metabolite Extracts from Liquid Schizosaccharomyces pombe Cultures
Authors: Pluskal Tomáš , Nakamura Takahiro , Yanagida Mitsuhiro
Published: December 2016

The success of metabolomic analysis relies heavily on the sample preparation protocol. Here we present a protocol for intracellular metabolite extraction from liquid fission yeast cultures based on rapid quenching in pure methanol at −40°C, bead-beating in 50% methanol for cell disruption, and 10 kDa cutoff ultrafiltration for removal of proteins. Samples are concentrated by vacuum evaporation and resuspended in 50% acetonitrile for mass spectrometric analysis. This protocol is optimal for extraction of polar metabolites such as amino acids, organic acids, nucleotides, sugars, or sugar-phosphates. Its implementation requires <6 h and allows preparation of multiple samples in parallel.