Effect of Oil Overlay on Inhibition Potential of Roscovitine in Sheep Cumulus‐Oocyte Complexes

Inhibitors of cyclin‐dependent kinases, as roscovitine, have been used to prevent the spontaneous resumption of meiosis in vitro and to improve the oocyte developmental competence. In this study, the interference of oil overlay on the reversible arrest capacity of roscovitine in sheep oocytes as well as its effects on cumulus expansion was evaluated. For this, cumulus‐oocyte complexes (COCs) were cultured for 20 h in TCM 199 with 10% foetal bovine serum (Control) containing 75 μm roscovitine (Rosco). Subsequently, they were in vitro matured (IVM) for further 18 h in inhibitor‐free medium with LH and FSH. The culture was performed in Petri dishes under mineral oil (+) or in 96 well plates without oil overlay (?) at 38.5°C and 5% CO₂. At 20 and 38 h, the cumulus expansion and nuclear maturation were evaluated under stereomicroscope and by Hoechst 33342 staining, respectively. No group presented cumulus expansion at 20 h. After additional culture with gonadotrophins, a significant rate of COCs from both Control groups (+/?) exhibited total expansion while in both Rosco groups (+/?) the partial expansion prevailed. Among the oocytes treated with roscovitine, 65.2% were kept at GV in the absence of oil overlay while 40.6% of them reached MII under oil cover (p < 0.05). This meiotic arrest was reversible, and proper meiosis progression also occurred in the Control groups (+/?). So, the culture system without oil overlay improved the meiotic inhibition promoted by roscovitine without affecting the cumulus expansion rate or the subsequent meiosis progression.     

Effect of pH on rheotaxis of bull sperm using microfluidics

The aim of the present research is to study the effect of pH values on the sperm rheotaxis properties. Semen collected from bulls was diluted with SOF medium (1:10). pH of the medium was adjusted using a digital pH meter to the following pH values: 6.0, 6.2, 6.4, 6.4, 6.8, 7.0. All kinetic parameters of sperm (n = 3,385) were determined through a computer‐assisted sperm analysis (CASA) system using microfluidic devices with controlled flow velocity. The following parameters were determined: total motility (TM%), positive rheotaxis (PR%), straightline velocity (VSL, μm/s), average path velocity (VAP, μm/s), linearity (LIN, as VSL/VCL, %), beat cross‐frequency (BCF, Hz) and curvilinear velocity (VCL, μm/s). Nitric oxide, calcium and potassium were estimated in semen at different pH values. To confirm the effect of nitric oxide and K⁺, we used sodium nitroprusside (an NO donor) and KCL as (a K⁺ donor) to see their effect on sperm PR%. The results showed no difference in TM% at pH (6–7). The PR% was the lowest at pH 6 and 7. The best parameters for the PR% were at pH 6.4–6.6. The concentration of Ca⁺² did not change at different pH values. The mean NO values decreased with the increase of pH; however, the mean values of K⁺ increased with the increase of pH. Addition of high concentration of NO and K⁺ to the semen media at fixed pH level had a negative effect on TM% and PR%. In conclusion, the bull sperm had the best rheotaxis properties at pH 6.4–6.6 and sensitive to the change of seminal NO and K⁺.     

Antagonism of detomidine with tolazoline in isoflurane-anaesthetised ponies

The effects of tolazoline (4.0 mg/kg iv) antagonism of detomidine (0.02 mg/kg iv) were evaluated in isoflurane-anaesthetised, ventilated ponies. Each of 6 ponies received both tolazoline and saline treatment during separate anaesthetic episodes only (no surgery was performed). Detomidine administration produced an increase in blood pressure, decrease in heart rate and decrease in P_aO₂ Tolazoline treatment transiently increased heart rate while blood pressure returned to baseline after both treatments. Arterial oxygenation decreased further after tolazoline treatment while oxgenation recovered towards baseline with saline treatment. No other cardiopulmonary effects were detected. Recovery from anaesthesia tended to be more rapid when detomidine was antagonized. The potential benefit of antagonizing detomidine-induced bradycardia with tolazoline, during isoflurane anaesthesia should be weighed against the potential to produce a decrease in arterial oxygenation. The mechanism for this effect is not clear.     

Simrank: Rapid and sensitive general-purpose k-mer search tool

Background  

Terabyte-scale collections of string-encoded data are expected from consortia efforts such as the Human Microbiome Project . Intra- and inter-project data similarity searches are enabled by rapid k-mer matching strategies. Software applications for sequence database partitioning, guide tree estimation, molecular classification and alignment acceleration have benefited from embedded k-mer searches as sub-routines. However, a rapid, general-purpose, open-source, flexible, stand-alone k-mer tool has not been available.     

Tunneling into a single magnetic atom: spectroscopic evidence of the kondo resonance

The Kondo effect arises from the quantum mechanical interplay between the electrons of a host metal and a magnetic impurity and is predicted to result in local charge and spin variations around the magnetic impurity. A cryogenic scanning tunneling microscope was used to spatially resolve the electronic properties of individual magnetic atoms displaying the Kondo effect. Spectroscopic measurements performed on individual cobalt atoms on the surface of gold show an energetically narrow feature that is identified as the Kondo resonance-the predicted response of a Kondo impurity. Unexpected structure in the Kondo resonance is shown to arise from quantum mechanical interference between the d orbital and conduction electron channels for an electron tunneling into a magnetic atom in a metallic host.     

Size fractions of organic matter pools influence their stability: Application of the Rock-Eval® analysis to beech forest soils

Soil organic matter (SOM) is a complex heterogeneous mixture formed through decomposition and organo-mineral interactions, and characterization of its composition and biogeochemical stability is challenging. From this perspective, Rock-Eval^® is a rapid and efficient thermal analytical method that combines the quantitative and qualitative information of SOM, including several parameters related to thermal stability. This approach has already been used to monitor changes in organic matter (OM) properties at the landscape, cropland, and soil profile scales. This study was aimed to assess the stability of SOM pools by characterizing the grain size fractions from forest litters and topsoils using Rock-Eval^® thermal analysis. Litter (organic) and topsoil samples were collected from a beech forest in Normandy (France), whose management in the last 200 years has been documented. Fractionation by wet sieving was used to separate large debris (> 2 000 μm) and coarse (200-2 000 μm) and fine particulate OM (POM) (50-200 μm) in the organic samples as well as coarse (200-2 000 μm), medium (50-200 μm), and fine (< 50 μm) fractions of the topsoil samples. Rock-Eval^® was able to provide thermal parameters sensitive enough to study fine-scale soil processes. In the organic layers, quantitative and qualitative changes were explained by the progressive decomposition of labile organic compounds from plant debris to the finest organic particles. Meanwhile, the grain size fractions of topsoils presented different characteristics. The coarse organo-mineral fractions showed higher C contents, albeit with a different composition, higher thermal stability, and greater decomposition degree than the plant debris forming the organic layer. These results are consistent with those of previous studies that microbial activity is more effective in this fraction. The finest fractions of topsoils showed low C contents, the highest thermal stability, and low decomposition degree, which can be explained by the stronger interactions with the mineral matrix. Therefore, it is suggested that the dynamics of OM in the different size fractions be interpreted in the light of a plant-microbe-soil continuum. Finally, three distinct thermostable C pools were highlighted through the grain size heterogeneity of SOM:free coarse OM (large debris and coarse and fine particles), weakly protected OM in (bio)aggregates (coarse fraction of topsoil), and stabilized OM in the fine fractions of topsoil, which resulted from the interactions within organo-mineral complexes. Therefore, Rock-Eval^® thermal parameters can be used to empirically illustrate the conceptual models emphasizing the roles of drivers played by the gradual decomposition and protection of the most thermally labile organic constituents.     

High Embryonic Recovery Rates with In vivo and Ex vivo Techniques in the Bitch

The embryonic collection techniques in dogs present a vast methodological variation and low recovery rates. The objectives were to compare and describe two techniques as to the recovery of canine embryos, on the 12th day after the first mating or artificial insemination. Embryos were recovered through uterine horn flushing in vivo, before performing the ovariohysterectomy (OHE) (Group 1; n = 9) or ex vivo, immediately after the OHE (Group 2; n = 9). In total, 43 and 47 embryonic structures were recovered in Groups 1 and 2, respectively. There was no significant difference (p > 0.05) between groups on recovery rates (72.8% and 81.0%, respectively). We inferred that both in vivo and ex vivo techniques allow a high rate of embryonic recovery; in the collection technique prior to the OHE, it is essential to carefully handle the reproductive system during the trans‐surgical period and that the 12th day (D12) after the first mating/artificial insemination is an efficient option for the high recovery rate of morulae and blastocysts.     

Population pharmacokinetic modelling and simulation of single and multiple dose administration of meloxicam in cats

Lehr, T., Narbe, R., Jöns, O., Kloft, C., Staab, A. Population pharmacokinetic modelling and simulation of single and multiple dose administration of meloxicam in cats. J. vet. Pharmacol. Therap. 33 , 277–286. The objectives of these investigations were: first, to describe the pharmacokinetic properties of meloxicam in cats following single and multiple oral administration and secondly, to simulate different oral dosage regimes for meloxicam in cats after multiple dose administration to illustrate and evaluate those dosage regimes for the alleviation of inflammation and pain in cats. Six healthy domestic short hair cats were treated orally with various dosage regimes (0.05–0.2 mg/kg/day). Plasma samples were collected at predefined times and quantitatively analysed using liquid/liquid extraction followed by reverse phase HPLC with UV‐detection. Meloxicam plasma concentration data were analysed using the population pharmacokinetic approach (software: NONMEM). The final model was used to simulate different dosage regimes. The plasma concentration–time profiles of meloxicam in cats after oral single and multiple dose administration were best described by an open one‐compartment model with first‐order absorption and first‐order elimination. Pharmacokinetic parameters were estimated to be 0.00656 L/h/kg for the total apparent body clearance (CL/F), 0.245 L/kg for the apparent volume of distribution (V/F), 1.26 1/h for the absorption constant (K_A) and 25.7 h for the mean plasma terminal half‐life. Simulations showed that the median trough steady‐state concentrations of 228 ng/mL were reached after five, one or 6 days following a single initial dose of 0.05, 0.1 and 0.2 mg/kg each followed by 0.05 mg/kg/day.