Production of monoclonal antibody against recombinant bovine sex-determining region Y (SRY) and their preferential binding to Y chromosome-bearing sperm

Separation of X and Y chromosome-bearing sperm is an appropriate method for the selection of desired sex of offspring to increase the profit in livestock industries. The purpose of this study was the production of a monoclonal antibody against recombinant bovine sex-determining region Y protein for separation Y sperm. The hybridoma cells from splenocytes of immunized female's balb/C mice and Sp2/0 cells were made. The binding affinity of our monoclonal antibody (mAbSRY2) was compared with mouse monoclonal SRY-15. The Western blot method indicated that mAbSRY2 successfully detected the rbSRY protein. The specificity and sensitivity of mAbSRY2 is comparable to SRY-15 commercially ones. The SRY gene in 100% of bull semen contains the Y chromosome that had the strongest binding affinity to mAbSRY2 was synthesized. In other words, the binding affinity of semen contains the X sperms near the negative control. In general, this immunological method can help to separate X from Y sperms. However, the mAbSRY2 is bind to Y-bearing sexed sperm, but in the future; the sexed sperms need to apply in farms.     

Accumulation of Nutrients in Above and Below Ground Biomass in Response to Ammonium Sulphate Addition in a Norway Spruce Stand in Southwest Sweden

The effects of ammonium sulphate (NS) on the accumulation of nutrients in above and below ground biomass and soil were studied in a Norway spruce stand in south-west Sweden during 1988–1993. Ammonium sulphate addition resulted in nitrogen accumulation with 326 and 16 kg ha^?1 in above and below ground biomass, respectively. Corresponding figures for the control plots (C) were 34 and 3 kg ha^?1. Nitrogen accumulation in forest floor of NS was 266 kg ha^?1 and 47 kg ha^?1 in mineral soil. About 70% of added sulphate by fertiliser was retained in NS plots (482 kg S ha^?1) of which 274 kg ha^?1 was adsorbed in the mineral soil. The sulphate addition resulted in increased leaching of nitrogen, magnesium, calcium and sulphur. It is suggested that the spruce stand at the study site has a high capacity to accumulate nitrogen with a high above ground production. The high input of ammonium sulphate may in the long run result in increased losses of cations to ground water.     

Bayesian methods for predicting LAI and soil water content

LAI of winter wheat (Triticum aestivum L.) and soil water content of the topsoil (200 mm) and of the subsoil (500 mm) were considered as state variables of a dynamic soil-crop system. This system was assumed to progress according to a Bayesian probabilistic state space model, in which real values of LAI and soil water content were daily introduced in order to correct the model trajectory and reach better future evolution. The chosen crop model was mini STICS which can reduce the computing and execution times while ensuring the robustness of data processing and estimation. To predict simultaneously state variables and model parameters in this non-linear environment, three techniques were used: extended Kalman filtering (EKF), particle filtering (PF), and variational filtering (VF). The significantly improved performance of the VF method when compared to EKF and PF is demonstrated. The variational filter has a low computational complexity and the convergence speed of states and parameters estimation can be adjusted independently. Detailed case studies demonstrated that the root mean square error of the three estimated states (LAI and soil water content of two soil layers) was smaller and that the convergence of all considered parameters was ensured when using VF. Assimilating measurements in a crop model allows accurate prediction of LAI and soil water content at a local scale. As these biophysical properties are key parameters in the crop-plant system characterization, the system has the potential to be used in precision farming to aid farmers and decision makers in developing strategies for site-specific management of inputs, such as fertilizers and water irrigation.     

Evaluation of genetic diversity of Aegilops tauschii accessions using morphological characters

A collection of sixty-three accessions of Ae. tauschii belonging to different eco-geographical regions were used to evaluate its genetic diversity by 15 morphological characters. The data recorded were analyzed on all accessions using multivariate analyses. Principal component analysis (PCA) indicated that first six principal components with eigenvalues more than 1 explained 77.2% of the variability amongst accessions. Through cluster analysis according to Euclidian distance and UPGMA method, accessions divided into three groups. Knowledge of genetic diversity in Aegilops species provides different levels of information which is important in the management of germplasm resources.     

End-grain butt joints obtained by friction welding of high density eucalyptus wood

End-grain-to-end-grain welding gave butt joints of good strength with three high density Australian eucalyptus woods, namely Sidney blue gum (Eucalyptus saligna), Spotted gum (Eucalyptus maculata, Corymbia maculata spp.) and Black butt (Eucalyptus pilularis). The absence or limited tendency to defibration in end-grain-to-end-grain welding for wood densities as high as these, around 800–900 kg/m³, indicates that end-grain-to-end-grain welding is possible and yields good joint strengths, but with different appearance of the welded interphase. At anatomical level, three features were noticeable: (1) the paintbrush-like appearance of the cell tips bent so by the frictional movement during welding. This leaves this cell tips free to intertwine with the same one from the surface of the opposite wood piece. (2) The absence of any great amount of molten material between the straight shafts of the parallel bundles of cells, showing that the greater part of the intercellular material has molten and has flowed away from the cells towards the interphase. (3) The amount of molten intercellular material found on the upper surface of the cell tips, giving the joint the appearance of an anatomical fingerjoint.     

Inotropic and chronotropic effects of 6-hydroxy-4-methylquinolin-2(1H)-one derivatives in isolated rat atria

Background: Selective phosphodiesterase (PDE3) inhibitors improve cardiac contractility and may use in congestive heart failure. However, their proarrhythmic potential is the most important side effect. Methods: In this research, we evaluated the potential cardiotonic activity of six new synthesized selective PDE3 inhibitors (6-hydroxy-4-methylquinolin-2(1H)-one derivatives) using the spontaneously beating atria model. In each experiment, atrium of reserpine-treated rat was isolated and the contractile and chronotropic effects of a synthesized compound were assessed. The 3-isobutyl-1-methylxanthine, a non-selective PDE inhibitor, was used for comparison. Results: The results showed that, among new compounds, the best pharmacological profile was obtained with the compound 6-[4-(4-methylpiperazine-1-yl)-4-oxobutoxy]-4-methylquinolin-2(1H)-one, C6, which displayed selectivity for increasing the force of contraction (168 +/- 5% change over the control) rather than the frequency rate (138 +/- 5% change over the control) at 300 muM. However, C6 at concentrations of 10 and 100 muM produced left and upward shift in the positive inotropic concentration-response curve of isoprenaline. The -log EC50 of isoprenaline was 8.843 +/- 0.171 in the absence, 9.448 +/- 0.138 and 9.456 +/- 0.107 in the presence of 10, 100 muM of C6, respectively (P<0.001, n = 6). Also, amrinone, a selective PDE3 inhibitor, shifted the isoprenaline concentration-response curve to the left and upward. The concentration of 10 and 100 muM amrinone decreased -log EC50 of isoprenaline to 9.527 +/- 0.287 and 9.423 +/- 0.243, respectively (P<0.001, n = 6). Moreover, the positive chronotropic effect of isoprenaline was not affected by amrinone or C6. Conclusion: This study provides functional evidence for the positive inotropic effect of C6. Considering the augmentation of isoprenaline positive inotropic concentration-response with C6 and amrinone, we conclude that C6 produces its effect via potentiation of cAMP-dependent signaling system and possibly by inhibition of PDE3 activity.     

Effects of slash retention and wood ash addition on fine root biomass and production and fungal mycelium in a Norway spruce stand in SW Sweden

In the study reported here we examined the short-term effects (1–3 years) of slash retention (SR) and the long-term effects (13–15 years) of wood-ash application (A) on fine roots and mycorrhizae in a 40-year-old Norway spruce forest in southwest Sweden. Soil cores were used to obtain estimates of the biomass (g m⁻²) of roots in three diameter classes (<0.5, 0.5–1 and 1–2 mm), root length density (RLD), specific root length (SRL) and mycorrhizal root tip density (RTD). Fine root (<1 mm) length production and mortality, and mycelium production, were estimated using minirhizotron and mesh bag techniques, respectively. Compared with the control plots (C), the biomass of fine roots in diameter classes <0.5 mm and 0.5–1 mm was significantly higher in A plots, but lower in SR plots. In addition, RLD was significantly lower in the humus layer of SR plots than in the humus layers of C and A plots, but not in the other layers. None of the treatments affected the SRL. In all soil layers, the SR treatment resulted in significant reductions in the number of ectomycorrhizal root tips, and the mycelia production of fungi in mesh bags, relative to the C treatment, but the C and A treatments induced no significant changes in these variables. Fine root length production in the C, A and SR plots amounted to 94, 87 and 70 mm tube⁻¹ during the 2003 growing season, respectively. Fine root mortality in treated plots did not change over the course of the study. We suggest that leaving logging residues on fertile sites may result in nitrogen mineralisation, which may in turn induce reductions in root biomass, and both root and mycelium production, and consequently affect nutrient uptake and the accumulation of organic carbon in soil derived from roots and mycorrhizae.     

Stand age and fine root biomass, distribution and morphology in a Norway spruce chronosequence in southeast Norway

We assessed the influence of stand age on fine root biomass and morphology of trees and understory vegetation in 10-, 30-, 60- and 120-year-old Norway spruce stands growing in sandy soil in southeast Norway. Fine root (< 1, 1-2 and 2-5 mm in diameter) biomass of trees and understory vegetation (< 2 mm in diameter) was sampled by soil coring to a depth of 60 cm. Fine root morphological characteristics, such as specific root length (SRL), root length density (RLD), root surface area (RSA), root tip number and branching frequency (per unit root length or mass), were determined based on digitized root data. Fine root biomass and morphological characteristics related to biomass (RLD and RSA) followed the same tendency with chronosequence and were significantly higher in the 30-year-old stand and lower in the 10-year-old stand than in the other stands. Among stands, mean fine root (< 2 mm) biomass ranged from 49 to 398 g m(-2), SLR from 13.4 to 19.8 m g(-1), RLD from 980 to 11,650 m m(-3) and RSA from 2.4 to 35.4 m(2) m(-3). Most fine root biomass of trees was concentrated in the upper 20 cm of the mineral soil and in the humus layer (0-5 cm) in all stands. Understory fine roots accounted for 67 and 25% of total fine root biomass in the 10- and 120-year-old stands, respectively. Stand age had no affect on root tip number or branching frequency, but both parameters changed with soil depth, with increasing number of root tips and decreasing branching frequency with increasing soil depth for root fractions < 2 mm in diameter. Specific (mass based) root tip number and branching density were highest for the finest roots (< 1 mm) in the humus layer. Season (spring or fall) had no effect on tree fine root biomass, but there was a small and significant increase in understory fine root biomass in fall relative to spring. All morphological characteristics showed strong seasonal variation, especially the finest root fraction, with consistently and significantly higher values in spring than in fall. We conclude that fine root biomass, especially in the finest fraction (< 1 mm in diameter), is strongly dependent on stand age. Among stands, carbon concentration in fine root biomass was highest in the 30-year-old stand, and appeared to be associated with the high tree and canopy density during the early stage of stand development. Values of RLD and RSA, morphological features indicative of stand nutrient-uptake efficiency, were higher in the 30-year-old stand than in the other stands.     

The Relationship Between Developmental Accumulation of Leaf Soluble Proteins and Vernalization Response of Wheat （Triticum aestivum L.em. Thell）

The relationship between vernalization requirement and quantitative and qualitative changes in total leaf soluble proteins were determined in one spring （cv. Kohdasht） and two winter （cvs. Sardari and Norstar） cultivars of wheat （Triticum aestivum L.） exposed to 4℃. Plants were sampled on days 2, 14, 21 and 35 of exposure to 4℃. The final leaf number （FLN） was determined throughout the vernalization periods （0, 7, 14, 24, and 35 d） at 4℃. The final leaf number decreased until days 24 and 35 in Sardari and Norstar eultivars, respectively, indicating the vernalization saturation at these times. No clear changes were detected in the final leaf number of Kohdash cultivar, verifying no vernalization requirement for this spring wheat cultivar. Comparing with control, clear cold-induced 2-fold increases in proteins quantity occurred after 48 h following the 4℃-treatment in the leaves of the both winter wheat cultivars but, such response was not detected in the spring cultivar. However, the electrophoretic protein patterns showed between-cultivar and between-temperature treatment differences. With increasing exposure time to 4℃, the winter cultivars tended to produce more HMW polypeptides than the spring cultivar. Similar proteins were induced in both Sardari and Norstar winter wheat cultivars, however, the long vernalization requirement in Norstar resulted in high level and longer duration of expression of cold-induced proteins compared to Sardari with a short vernalization requirement. These observations indicate that vernalization response regulates the expression of low temperature （LT） tolerance proteins and determines the duration of expression of LT- induced proteins.