Accumbal dopamine function in postpartum rats that were raised without their mothers

Postpartum rats that had been previously raised in an artificial rearing (AR) apparatus, without their mothers or siblings during the preweaning period, show altered maternal responses towards their own offspring in adulthood. In mother-reared (MR) rats, nucleus accumbens (NAC) dopamine (DA) responses to pups evoke a robust sustained rise during the postpartum period and following treatment with estrogen/progesterone parturient-like hormones (Afonso et al., 2009). These MR females had siblings that received AR rearing with varying amounts of preweaning tactile stimulation (ARmin; ARmax). The present study examined NACshell DA responses to pup and food stimuli in these AR rats, and statistically compared them to their MR siblings. Microdialysis samples were collected from adult (90 days postnatal) AR females in different parity states (cycling vs. postpartum, Exp. 1), or after ovariectomy with different hormone treatments (sham vs. hormone, Exp. 2. After basal sample collection, pup and then food stimuli were individually presented to the females in the dialysis chamber. As with their MR siblings, basal DA concentrations were lower and pup-evoked DA responses greater in hormonally-primed AR females than in non-primed AR controls. Compared to their postpartum MR sisters (Exp. 1), AR rats had increased basal DA levels, reduced pup related DA elevations, and disrupted maternal behavior. The postpartum AR impairment in pup-evoked DA was reversed by additional pre-weaning tactile stimulation. Exogenous hormones (Exp. 2) eliminated AR impairments on pup-evoked DA responses. Although MR and AR siblings had comparable DA responses to food stimuli, upon reanalyzing MR data it was found that only postpartum dams had DA responses to pups greater than to food. These data suggest that that the hormonally induced suppression of basal DA levels may reflect saliency of pups which was greater in MR than in AR dams. Preweaning tactile stimulation could partially reverse these effects only in naturally cycling or parturient animals.     

Effect of duration of storage at ambient temperature on fertilizing ability and mucus penetration ability of fresh bovine sperm

Although the use of fresh semen in the Irish dairy AI industry only accounts for 5% of total AI usage, this may peak to over 25% during the spring breeding season due to the increased demand for Irish proven sires of high genetic merit. The aim of this study was to examine the effect of storage of fresh semen for up to 7 d at ambient temperature on fertilization and embryo development in vitro, and on the ability of sperm to penetrate artificial mucus in vitro. In vitro matured bovine oocytes were inseminated with fresh semen stored in a caprogen-based diluent, with or without prior Percoll separation. Irrespective of sire, storage of fresh semen at ambient temperature for up to 7 d post collection had no effect on cleavage rate or blastocyst development after IVF. In addition, blastocyst quality, as assessed by the proportion of blastocysts hatching from the zona, was not affected by semen storage. Higher numbers of fresh sperm migrated through artificial mucus on Day 0 (day of semen collection) compared with frozen-thawed sperm. On Day 1 and 2 postcollection there was no difference in the number of sperm migrating through the mucus, but storage of sperm at ambient temperature for longer than 2 d resulted in a significant decline in their ability to penetrate mucus compared with frozen sperm from the same ejaculate. In conclusion, bovine sperm retain the ability to fertilize oocytes in vitro for up to 7 d following storage at ambient temperature. However, the ability of sperm to migrate through artificial mucus in vitro is severely depressed after 2 d storage which may have significant implications for the ability of these sperm to reach the site of fertilization in vivo after AI.     

栎树繁殖生物技术进展

栎树是重要的森林树种.由于常规营养繁殖困难、种子繁育开花结实迟等原因,栎树的繁育问题一直未得到很好的解决.综述栎树繁殖生物技术的历史与进展;分析栎树的体细胞胚胎从诱导到萌发的过程及机理;阐述欧洲栓皮栎等树种的小孢子胚胎发生技术、双单倍体培养技术、体胚遗传转化技术、人工种子技术以及相关的遗传稳定性与变异性检测技术.     

A chromogenic substrate for solid-phase detection of phospholipase A2

A method for solid-phase detection of phospholipase A₂ (PLA₂) was developed. The method uses 1-octanoyloxynaphthalene-3-sulfonic acid, which was found to be a good substrate of PLA₂. The substrate is hydrolyzed by PLA₂ into 1-naphthol-3-sulfonic acid, which is spontaneously coupled with coexisting diazonium salt to form a red-purple azo dye. Streptomyces and bovine pancreatic PLA₂ spotted on a nitrocellulose membrane could be detected by this method with considerable sensitivity. In addition, colonies of recombinant Escherichia coli producing bacterial PLA₂ were distinguishable from those producing an inactive mutant PLA₂, facilitating high-throughput screening in directed evolution of the enzyme.     

Application of a probabilistic model for analysing the abortion of seeds and pods in winter oilseed rape (Brassica napus)

In winter oilseed rape (WOSR), only a subset of ovules can develop into seeds in the majority of pods. Any difficulty during the process of seed production may result in seed or pod abortion. This study aimed to reproduce the process of seed development in WOSR based on a limited number of parameters. As a result of the complexity of the developmental patterns of WOSR, it is challenging to identify the roles of various factors that influence seed production using an experimental approach. Here, we present a stochastic probabilistic model of seed development. The generalised least squares method was implemented to estimate the model parameters using the experimental data. Experiments were done in Grignon (France) in 2008 and 2009. The variations in the parameters were analysed according to the following four factors: year, pod rank, inflorescence position and ramification‐clipping treatment. The year had no effect on the number of ovules per ovary (μ) and the probability of seed viability (p). The proportion of effective pollen grains (k) significantly decreased with pod rank at the end of the main stem. Inflorescence position influenced the number of ovules per ovary (μ: 30.8–33.8 from top to bottom) and the parameter k. The mean number of seeds per pod on the main stem and the bottom ramification along the stem was larger than the other ramifications within one plant. Ramification‐clipping treatment increased the number of ovules per ovary (μ: 31 for control plants and 32 for clipped plants), the parameter k and the number of seeds per pod (p). This effect could be due to the competition for assimilates between the pods and seeds. Furthermore, the distribution parameters of the pollen number per stigma (m) remained stable, and the probability of pod survival (Bo) varied with different factors, including the year, pod rank and inflorescence position. Our results indicate that pollen germination is a factor that determines final seed number. This model can identify the impact of each of the factors that lead to the abortion of seeds and pods in WOSR, which include the position effect, assimilate competition and pollination limitation. However, further studies on the pollination process in WOSR should help to refine this model.     

Effects of altering the dose and timing of triptorelin when given as an intravaginal gel for advancing and synchronizing ovulation in weaned sows

Previous studies have shown that triptorelin gel (TG) given intravaginally in gel form is effective for advancing the time of ovulation in weaned sows. Three experiments were performed to determine the effects of altering the dose and timing of administration of intravaginal TG for advancing and synchronizing ovulation in weaned sows. In all experiments, estrus was detected twice or three times daily and ultrasound was performed to determine ovulation at 8-hour intervals. In experiment 1, sows (n = 131) received intravaginal gel containing 0 (Placebo), 25, 100, or 200 μg of TG at 96 hours after weaning and sows were inseminated on each day of standing estrus. Wean-to-estrus interval and duration of estrus were correlated (P < 0.0001) with estrus duration longer in TG (P < 0.05) compared with Placebo. More sows ovulated (P < 0.001) by 48 hours after treatment with 200 (81%), 100 (64%), and 25 μg (63%) of TG compared with Placebo (42%). The farrowing rate and total pigs born did not differ (P > 0.10). In experiment 2, sows (n = 126) received 200 μg of TG at 72, 84, or 96 hours after weaning or were untreated (Control-96). Sows receiving TG were inseminated once 24 to 28 hours after treatment. Control-96 sows were inseminated on each day of standing estrus. Wean-to-estrus interval was not affected by treatment, but wean-to-ovulation interval was reduced (P < 0.05) by TG-72 and TG-84 compared with TG-96 and Control-96. More sows ovulated 40 hours after treatment (P < 0.001) with TG-72 (56.5%) and TG-84 (32.2%) compared with TG-96 and Control-96 (13%) and for all TG treatments 48 hours after treatment (64%) compared with Control-96 (34%, P < 0.05). The farrowing rate was lower (P < 0.05) for sows assigned to TG-72 and TG-84 compared with TG-96 and Control-96, whereas the number of liveborn pigs did not differ (P > 0.10). In experiment 3, sows (n = 113) were assigned to receive no treatment (Control), intravaginal gel alone (Placebo), or 200 μg of TG given intravaginally (OvuGel) at 96 hours after weaning. Wean-to-estrus interval did not differ, but the duration of estrus tended (P < 0.10) to be reduced with OvuGel compared with the other treatments. More sows ovulated (P < 0.001) by 48 hours after OvuGel treatment (79.1%) compared with Control (46.4%) and Placebo (37.9%) and by 56 hours (P < 0.05). The farrowing rate and the number of liveborn pigs did not differ among treatments. The results of these studies indicate that 200 μg of TG given intravaginally at 96 hours after weaning (OvuGel) synchronizes ovulation and results in fertility similar to Controls.     

First field results on the use of stallion sex-sorted semen in a large-scale embryo transfer program

Flow cytometry sex-sorting technology was developed in 1989. However, it is only the bovine species in which offspring of the desired sex are obtained at a commercial level. The aim of the present work was to evaluate efficiency parameters when using fresh sexed semen in a large-scale equine commercial embryo transfer program. During the 2009, 2010 and 2011 breeding seasons, 938 synchronized cycles were artificially inseminated. One hundred (10.6%) mares failed to ovulate, and for the remaining 838 useable cycles, 887 doses of sexed semen were used, representing 1.06 doses per cycle. In general, 435 (51.9%) out of 838 flushing performed resulted in the recovery of at least one embryo and 496 (59.1%) embryos were recovered, including twins and triplets. Pregnancy rate at 25 days achieved 81.5% (one embryo transferred per recipient). Embryo recovery rate was not statistically different either between preovulatory and postovulatory artificially inseminated mares or when increased quantities of sexed sperm per dose were used (15–45 million) (P > 0.05). A broad variation in embryo recovery rate was observed between the different stallions used in this study. Sex accuracy of the sex sorting assessed by ultrasound fetal sex determination was 90.3%. Finally, overall efficiency (female embryo pregnancies per useable cycles) was 39% (325/838), meaning that to obtain a female pregnancy of at least 75 days it was necessary to perform 2.5 flushing.     

Importance of a traditional irrigation system in amphibian conservation in the Cinque Terre National Park (NW Italy)

In Mediterranean agricultural landscapes the impact of irrigation on biodiversity, and in particular on wildlife, remains controversial. This study investigated the role of traditional irrigation in the conservation of amphibians in the smallest and most densely populated national park in Italy, the Cinque Terre National Park (CTNP). The coastline in this area is so steep that dry stone walls were built to create arable land surfaces, irrigated by water stored in small open tanks. An almost complete census of these tanks and a survey of amphibian populations in natural and artificial habitats were undertaken from 2009 to 2012. A total of 58 water tanks were censused, 12 of them being empty, damaged or abandoned. All the tanks containing water were built in concrete and had small volume capacities (mean = 3.6, range 0.3–12.4 m³) but, overall, hosted all the seven aquatic amphibian species still present in the Park's natural freshwater habitats. Amphibians bred in 66% of the tanks still used for irrigation; large tanks were occupied more often than small tanks and hosted a different, more species-rich amphibian community. These results have management implications and confirm that, in the CTNP, extensive agriculture and biodiversity are compatible, and that maintaining traditional irrigation systems will benefit amphibian populations, especially in the case of drier climate scenarios.     

Human Rights and Broken Cisterns: Counterpublic Christianity and Rights-based Discourse in Contemporary England

Although human rights are often framed as the result of centuries of Western Christian thought, many English evangelicals are wary of the U.K.’s recent embrace of rights-based law. Yet this wariness does not preclude their use of human rights instruments in the courts. Drawing upon fieldwork with Christian lobbyists and lawyers in London, I argue that evangelical activists instrumentalise rights-based law so as to undermine the universalist claims on which they rest. By constructing themselves as a marginalised counterpublic whose rights are frequently ‘trumped’ by the competing claims of others, they hope to convince their fellow Britons that a society built upon the logic of equal rights cannot hope to deliver the human flourishing it promises. Given the salience of contemporary political conservatism, I call for further ethnographic research into counterpublic movements, and offer my interlocutors’ instrumentalisation of human rights as a critique of the inconsistencies of secular law.     

In silico,in vitro,and in vivo machine learning in synthetic biology and metabolic engineering

Among the main learning methods reviewed in this study and used in synthetic biology and metabolic engineering are supervised learning, reinforcement and active learning, and in vitro or in vivo learning.In the context of biosynthesis, supervised machine learning is being exploited to predict biological sequence activities, predict structures and engineer sequences, and optimize culture conditions.Active and reinforcement learning methods use training sets acquired through an iterative process generally involving experimental measurements. They are applied to design, engineer, and optimize metabolic pathways and bioprocesses.The nascent but promising developments with in vitro and in vivo learning comprise molecular circuits performing simple tasks such as pattern recognition and classification.