Variaciones del índice de cono en suelo movilizado posterior al tráfico de un tractor equipado con dos tipos de neumáticos y a cuatro velocidades de desplazamiento

The tractor is one of the machines that more traffics over the soil during the processes involving agricultural production. The interaction tractor/soil is made by the tires which, in most of the cases, are pneumatic. The tire type and the tractor travel speed, interfere directly on the pressure over the soil. One of the techniques employed to evaluate the alterations that tractor traffic causes in the soil is to measure its Cone Index. The aim of this research was to evaluate the same Cone Index alterations caused by an agricultural tractor equipped with both radial tires and bias ply tires, trafficking mobilized soil in four different travel speeds. The experiment was performed in a LATOSSOLO VERMELHO, located 22°51' S, 48°25'W and 770 m of altitude, in Botucatu-SP, Brazil. The soil mobilization was performed with a chisel plow and a disc arrow. The traction was accomplished with a John Deere tractor, model 6600, with 88 kW of power and 6,723 kg. Equipment requiring a force of 25kN was traced by the tractor draw bar. The experimental design was in randomized blocks with 4 × 2 factorial arrangements, with two distinct treatments corresponding to the types of tires (bias and radial) and the four travel speeds, with six replications. There were selected the following speeds: 3.5, 3.9, 5.1 and 5.9 km h-1. To determine the soil resistance, there was utilized MSSU - Mobile Soil Sampling Unit, with which the Cone Index was obtained in layers from 0-100, 100-200, 200-300, 300-400, 400-500 and over 500 mm deep. The Cone Index where evaluated in areas with non contact between tire and soil (ICn) and in the tire footprint track (ICp). There were calculated the Cone Index increments caused by the tractor tire (AIC) and the results showed that as the tractor travel speed increased, there were observed decrements in the medium values of cone index. The radial tire provided smaller values of the Cone Index in the superficial layer of the soil (0 to 100 mm) in relation to the bias ply tire, when the speed was approximately 6 km h-1. The increment in the Cone Index, promoted by the tractor, was more intense in the first 200 mm depth, but it also reached the layer from 200 to 300 mm

Improvement of DNA minicircle production by optimization of the secondary structure of the 5′-UTR of ParA resolvase

The use of minicircles in gene therapy applications is dependent on the availability of high-producer cell systems. In order to improve the performance of minicircle production in Escherichia coli by ParA resolvase-mediated in vivo recombination, we focus on the 5′ untranslated region (5′-UTR) of parA messenger RNA (mRNA). The arabinose-inducible P[subscript BAD]/araC promoter controls ParA expression and strains with improved arabinose uptake are used. The 27-nucleotide-long 5′-UTR of parA mRNA was optimized using a predictive thermodynamic model. An analysis of original and optimized mRNA subsequences predicted a decrease of 8.6–14.9 kcal/mol in the change in Gibbs free energy upon assembly of the 30S ribosome complex with the mRNA subsequences, indicating a more stable mRNA-rRNA complex and enabling a higher (48–817-fold) translation initiation rate. No effect of the 5′-UTR was detected when ParA was expressed from a low-copy number plasmid (∼14 copies/cell), with full recombination obtained within 2 h. However, when the parA gene was inserted in the bacterial chromosome, a faster and more effective recombination was obtained with the optimized 5′-UTR. Interestingly, the amount of this transcript was 2.6–3-fold higher when compared with the transcript generated from the original sequence, highlighting that 5′-UTR affects the level of the transcript. A Western blot analysis confirmed that E. coli synthesized higher amounts of ParA with the new 5′-UTR (∼1.8 ± 0.7-fold). Overall, these results show that the improvements made in the 5′-UTR can lead to a more efficient translation and hence to faster and more efficient minicircle generation.MIT-Portugal ProgramFundação para a Ciência e a Tecnologia (PhD grant SFRH/BD/33786/2009

Evidence that the insertion events of IS2 transposition are biased towards abrupt compositional shifts in target DNA and modulated by a diverse set of culture parameters

Insertion specificity of mobile genetic elements is a rather complex aspect of DNA transposition, which, despite much progress towards its elucidation, still remains incompletely understood. We report here the results of a meta-analysis of IS2 target sites from genomic, phage, and plasmid DNA and find that newly acquired IS2 elements are consistently inserted around abrupt DNA compositional shifts, particularly in the form of switch sites of GC skew. The results presented in this study not only corroborate our previous observations that both the insertion sequence (IS) minicircle junction and target region adopt intrinsically bent conformations in IS2, but most interestingly, extend this requirement to other families of IS elements. Using this information, we were able to pinpoint regions with high propensity for transposition and to predict and detect, de novo, a novel IS2 insertion event in the 3′ region of the gfp gene of a reporter plasmid. We also found that during amplification of this plasmid, process parameters such as scale, culture growth phase, and medium composition exacerbate IS2 transposition, leading to contamination levels with potentially detrimental clinical effects. Overall, our findings provide new insights into the role of target DNA structure in the mechanism of transposition of IS elements and extend our understanding of how culture conditions are a relevant factor in the induction of genetic instability.Fundação para a Ciência e a Tecnologia (PTDC/EBB-EBI/113650/2009)MIT-Portugal Progra

Comparison of Apoptotic Cells Between Cryopreserved Ejaculated Sperm and Epididymal Sperm in Stallions

AbstractThe development of a reliable technique to freeze epididymal semen would provide a unique opportunity to preserve valuable genetic material from unexpectedly lost stallions. The aim of this study was to compare the apoptotic indices of sperm obtained from ejaculate, sperm recently recovered from the epididymides (EP), and sperm recovered from epididymides stored at 5°C for 24 hours (EP-stored). For the first category, two ejaculates from seven stallions were collected and then submitted to cryopreservation using an egg yolk-based extender. One week after the last semen collection, the stallions were submitted to bilateral orchiectomy, and sperm from one of the cauda epididymis was harvested immediately after castration (EP). The remaining testicle was stored in a passive refrigeration container at 5°C for 24 hours before the cauda epididymal sperm was harvested (EP-stored). Sperm harvesting from the epididymis for EP and EP-stored was performed by retrograde flushing of the caudal portion of the epididymis using a skim milk-based extender. The recovered sperm was then cryopreserved using the egg yolk-based extender. Sperm motility parameters were studied by computer-assisted semen analysis, and apoptosis was estimated by measuring caspase activity and membrane phospholipid translocation using epifluorescence microscopy. The samples were evaluated immediately (0 hour) and 8 hours after thawing. At 0 hour, no differences in sperm parameters were observed among the groups, but after 8 hours, significant statistical differences were observed in sperm motility parameters and plasma membrane integrity among the treatment groups. In addition, viable cells with no apoptotic signs were more prevalent in EP and EP-stored, suggesting that epididymal sperm is less sensitive to the cold shock caused by sperm cryopreservation

Design of an intrinsically safe series-series compensation WPT system for automotive LiDAR

The earliest and simplest impedance compensation technique used in inductive wireless power transfer (WPT) design is the series-series (SS) compensation circuit, which uses capacitors in series with both primary and secondary coils of an air-gapped transformer. Despite of its simplicity at the resonant condition, this configuration exhibits a major sensitivity to variations of the load attached to the secondary, especially when higher coupling coefficients are used in the design. In the extreme situation that the secondary coil is left at open circuit, the current at the primary coil may increase above the safety limits for either the power converter driving the primary coil or the components in the primary circuit, including the coil itself. An approach often used to minimize this problem is detuning, but this also reduces the electrical efficiency of the power transfer. In low power, fixed-distance stationary WPT, a fair trade-off between efficiency and safety must be verified. This paper aims to consolidate a simple design procedure for such a SS-compensation, exemplifying its use in the prototype of a WPT system for automotive light detection and ranging (LiDAR) equipment. The guidelines herein provided should equally apply to other low power applications.This work has been supported by national funds through FCT—Fundação para a Ciência e Tecnologia within the Project Scope: UID/CEC/00319/2019, and also European Structural and Investment Funds in the FEDER component, through the Operational Competitiveness and Internationalization Programme (COMPETE 2020) [Project nº 037902; Funding Reference: POCI-01-0247-FEDER-037902]

Protein-DNA interactions define the mechanistic aspects of circle formation and insertion reactions in IS2 transposition

<p>Abstract</p> <p>Background</p> <p>Transposition in IS<it>3</it>, IS<it>30</it>, IS<it>21 </it>and IS<it>256 </it>insertion sequence (IS) families utilizes an unconventional two-step pathway. A figure-of-eight intermediate in Step I, from asymmetric single-strand cleavage and joining reactions, is converted into a double-stranded minicircle whose junction (the abutted left and right ends) is the substrate for symmetrical transesterification attacks on target DNA in Step II, suggesting intrinsically different synaptic complexes (SC) for each step. Transposases of these ISs bind poorly to cognate DNA and comparative biophysical analyses of SC I and SC II have proven elusive. We have prepared a native, soluble, active, GFP-tagged fusion derivative of the IS<it>2 </it>transposase that creates fully formed complexes with single-end and minicircle junction (MCJ) substrates and used these successfully in hydroxyl radical footprinting experiments.</p> <p>Results</p> <p>In IS2, Step I reactions are physically and chemically asymmetric; the left imperfect, inverted repeat (IRL), the exclusive recipient end, lacks donor function. In SC I, different protection patterns of the cleavage domains (CDs) of the right imperfect inverted repeat (IRR; extensive <it>in cis</it>) and IRL (selective <it>in trans</it>) at the single active cognate IRR catalytic center (CC) are related to their donor and recipient functions. In SC II, extensive binding of the IRL CD <it>in trans </it>and of the abutted IRR CD <it>in cis </it>at this CC represents the first phase of the complex. An MCJ substrate precleaved at the 3' end of IRR revealed a temporary transition state with the IRL CD disengaged from the protein. We propose that in SC II, sequential 3' cleavages at the bound abutted CDs trigger a conformational change, allowing the IRL CD to complex to its cognate CC, producing the second phase. Corroborating data from enhanced residues and curvature propensity plots suggest that CD to CD interactions in SC I and SC II require IRL to assume a bent structure, to facilitate binding <it>in trans</it>.</p> <p>Conclusions</p> <p>Different transpososomes are assembled in each step of the IS<it>2 </it>transposition pathway. Recipient versus donor end functions of the IRL CD in SC I and SC II and the conformational change in SC II that produces the phase needed for symmetrical IRL and IRR donor attacks on target DNA highlight the differences.</p

Probing the properties of convective cores through g modes: high-order g modes in SPB and gamma Doradus stars

In main sequence stars the periods of high-order gravity modes are sensitive probes of stellar cores and, in particular, of the chemical composition gradient that develops near the outer edge of the convective core. We present an analytical approximation of high-order g modes that takes into account the effect of the mu gradient near the core. We show that in main-sequence models, similarly to the case of white dwarfs, the periods of high-order gravity modes are accurately described by a uniform period spacing superposed to an oscillatory component. The periodicity and amplitude of such component are related, respectively, to the location and sharpness of the mu gradient. We investigate the properties of high-order gravity modes for stellar models in a mass domain between 1 and 10 Msun, and the effects of the stellar mass, evolutionary state, and extra-mixing processes on period spacing features. In particular, we show that for models of a typical SPB star, a chemical mixing that could likely be induced by the slow rotation observed in these stars, is able to significantly change the g-mode spectra of the equilibrium model. Prospects and challenges for the asteroseismology of gamma Doradus and SPB stars are also discussed.Comment: 18 pages, 29 figures, accepted for publication in MNRA

Sensitivity of the g-mode frequencies to pulsation codes and their parameters

From the recent work of the Evolution and Seismic Tools Activity (ESTA, Lebreton et al. 2006; Monteiro et al. 2008), whose Task 2 is devoted to compare pulsational frequencies computed using most of the pulsational codes available in the asteroseismic community, the dependence of the theoretical frequencies with non-physical choices is now quite well fixed. To ensure that the accuracy of the computed frequencies is of the same order of magnitude or better than the observational errors, some requirements in the equilibrium models and the numerical resolutions of the pulsational equations must be followed. In particular, we have verified the numerical accuracy obtained with the Saclay seismic model, which is used to study the solar g-mode region (60 to 140$\mu$Hz). We have compared the results coming from the Aarhus adiabatic pulsation code (ADIPLS), with the frequencies computed with the Granada Code (GraCo) taking into account several possible choices. We have concluded that the present equilibrium models and the use of the Richardson extrapolation ensure an accuracy of the order of $0.01 \mu Hz$ in the determination of the frequencies, which is quite enough for our purposes.Comment: 10 pages, 5 figures, accepted in Solar Physic

Fluorometric determination of ethidium bromide efflux kinetics in Escherichia coli

This is an Open Access article distributed under the terms of the Creative Commons Attribution Licens

Control algorithms based on the active and non-active currents for a UPQC without series transformers

This study presents control algorithms for a new unified power quality conditioner (UPQC) without the series transformers that are frequently used to make the insertion of the series converter of the UPQC between the power supply and the load. The behaviour of the proposed UPQC is evaluated in presence of voltage imbalances, as well as under non-sinusoidal voltage-and current conditions. The presented algorithms derive from the concepts involving the active and non-active currents, together with a phase-locked-loop circuit. Based on these real-time algorithms, and considering the proposed hardware topology, the UPQC is able to compensate the harmonic components of the voltages and currents, correct the power factor, and keep the load voltages regulated, all of this in a dynamic way, responding instantaneously to changes in the loads or in the electrical power system. The control algorithms were distributed in a two-DSP digital control architecture, without any communication between them. Consequently, can be increased the sampling frequency of the acquired voltages and currents and improve the UPQC performance. Furthermore, some constraints of the proposed UPQC are evidenced, particularly when the main voltages are imbalanced. Simulation and experimental results are presented to verify the UPQC performance under transient and steady state conditions.info:eu-repo/semantics/publishedVersio