Spatiotemporal Mapping of the Contracting Gravid Uterus of the Rabbit Shows Contrary Changes With Increasing Gestation and Dosage With Oxytocin.

Spontaneous and oxytocin induced contractile activity was quantified in the bicornuate uteri of pregnant rabbits maintained in situ, using data from two- and uni- dimensional video spatiotemporal maps (VSTM) of linear and area strain rate and compared statistically. Spontaneous contractions occurred over a range of frequencies between 0.1 and 10 cpm, in gravid animals at 18-21 and at 28 days of gestation, and propagated both radially and longitudinally over the uterine wall overlying each fetus. Patches of contractions were randomly distributed over the entire surface of the cornua and were pleomorphic in shape. No spatial coordination was evident between longitudinal and circular muscle layers nor temporal coordination that could indicate the activity of a localized pacemaker. The density and duration of contractions decreased, and their frequency increased with the length of gestation in the non-laboring uterus. Increasing intravenous doses of oxytocin had no effect on the mean frequencies, or the mean durations of contractions in rabbits of 18-21 days gestation, but caused frequencies to decrease and durations to increase in rabbits of 28 days gestation, from greater spatial and temporal clustering of individual contractions. This was accompanied by an increase in the distance of propagation, the mean size of the patches of contraction, the area of the largest patch of contraction and the overall density of patches. Together these results suggest that progressive smooth muscle hypertrophy and displacement with increasing gestation is accompanied by a decrease in smooth muscle connectivity causing an increase in wall compliance and that oxytocin restores connectivity and decreases compliance, promoting volumetric expulsion rather than direct propulsion of the fetus by peristalsis. The latter effects were reversed by the ß2 adrenergic receptor agonist salbutamol thus reducing area of contraction, and the duration and distance of propagation.

Evaluation of the use of the polyubiquitin genes, Ubi4 and Ubi10 as reference genes for expression studies in Brachypodium distachyon.

BACKGROUND: Brachypodium distachyon is emerging as the model plant for temperate grass research and the genome of the community line Bd21 has been sequenced. Additionally, techniques have been developed for Agrobacterium-mediated transformation for the generation of T-DNA insertional lines. Recently, it was reported that expression of the polyubiquitin genes, Ubi4 and Ubi10 are stable in different tissues and growth hormone-treated plant samples, leading to the conclusion that both Ubi4 and Ubi10 are good reference genes for normalization of gene expression data using real-time, quantitative PCR (qPCR). PRINCIPAL FINDINGS: Mining of the Joint Genome Institute (JGI) 8X Brachypodium distachyon genome assembly showed that Ubi4 and Ubi10 share a high level of sequence identity (89%), and in silico analyses of the sequences of Ubi4 (Bradi3g04730) and Ubi10 (Bradi1g32860) showed that the primers used previously exhibit multiple binding sites within the coding sequences arising from the presence of tandem repeats of the coding regions. This can potentially result in over-estimation of steady-state levels of Ubi4 and Ubi10. Additionally, due to the high level of sequence identity between both genes, primers used previously for amplification of Ubi4 can bind to Ubi10 and vice versa, resulting in the formation of non-specific amplification products. CONCLUSIONS: The results from this study indicate that the primers used previously were not sufficiently robust and specific. Additionally, their use would result in over-estimation of the steady-state expression levels of Ubi4. Our results question the validity of using the previously proposed primer sets for qPCR amplification of Ubi4 and Ubi10. We demonstrate that primers designed to target the 3'-UTRs of Ubi4 and Ubi10 are better suited for real-time normalization of steady-state expression levels in Brachypodium distachyon.

Electroencephalographic responses of tramadol, parecoxib and morphine to acute noxious electrical stimulation in anaesthetised dogs.

This study compared the efficacy of different classes of analgesics in preventing the changes in electroencephalographic (EEG) indices of nociception in anaesthetised dogs, subjected to a standard electrical stimulus. In a crossover study, eight dogs received morphine (0.5mg/kg) or tramadol (3mg/kg) or parecoxib (1mg/kg) or 0.9% saline subcutaneously (SC) at the time of pre-anaesthetic medication. After induction with intravenous propofol, anaesthesia was maintained with halothane at a stable concentration between 0.85% and 0.95%. EEG was recorded in a three electrode montage, using SC needle electrodes, before and after electrical stimulation of dogs during anaesthesia. Post-stimulation median frequency (a reliable indicator of nociception) of the EEG increased significantly in tramadol, parecoxib and saline groups compared to that of morphine. Total EEG power decreased in all treatment groups following stimulation. These results indicate that the changes in EEG responses to noxious stimulation can be used for evaluating anti-nociceptive efficacy of analgesics.

An inducible, modular system for spatio-temporal control of gene expression in stomatal guard cells.

Stomata, flanked by pairs of guard cells, are small pores on the leaf surfaces of plants and they function to control gas exchange between plants and the atmosphere. Stomata will open when water is available to allow for the uptake of carbon dioxide for photosynthesis. During periods of drought, stomata will close to reduce desiccation stress. As such, optimal functioning of stomata will impact on water use efficiency by plants. The development of an inducible, modular system for robust and targeted gene expression in stomatal guard cells is reported here. It is shown that application of ethanol vapour to activate the gene expression system did not affect the ability of stomata to respond to ABA in bioassays to determine the promotion of stomatal closure and the inhibition of stomatal opening. The system that has been developed allows for robust spatio-temporal control of gene expression in all cells of the stomatal lineage, thereby enabling molecular engineering of stomatal function as well as studies on stomatal development.