Quality characteristics and fertilizing ability of ram sperm subpopulations separated by partition in an aqueous two-phase system

Centrifugal countercurrent distribution (CCCD) in an aqueous two-phase system (TPS) is a resolute technique revealing sperm heterogeneity and for the estimation of the fertilizing potential of a given semen sample. However, separated sperm subpopulations have never been tested for their fertilizing ability yet. Here, we have compared sperm quality parameters and the fertilizing ability of sperm subpopulations separated by the CCCD process from ram semen samples maintained at 20 degrees C or cooled down to 5 degrees C. Total and progressive sperm motility was evaluated by computer-assisted analysis using a CASA system and membrane integrity was evaluated by flow cytometry by staining with CFDA/Pl. The capacitation state, staining with chlortetracycline, and apoptosis-related markers, such as phosphatidylserine (PS) translocation detected with Annexin V. and DNA damage detected by the TUNEL assay, were determined by fluorescence microscopy. Additionally, the fertilizing ability of the fractionated subpopulations was comparative assessed by zona binding assay (ZBA). CCCD analysis revealed that the number of spermatozoa displaying membrane and DNA alterations was higher in samples chilled at 5 degrees C than at 20 degrees C. which can be reflected in the displacement to the left of the CCCD profiles. The spermatozoa located in the central and right chambers (more hydrophobic) presented higher values (P<0.01) of membrane integrity, lower PS translocation (P<0.05) and DNA damage (P<0.001) than those in the left part of the profile, where apoptotic markers were significantly increased and the proportion of viable non-capacitated sperm was reduced. We have developed a new protocol to recover spermatozoa from the CCCD fractions and we proved that these differences were related with the fertilizing ability determined by ZBA, because we found that the number of spermatozoa attached per oocyte was significantly higher for spermatozoa recovered from the central and right chambers, in both types of samples. This is the first time, to our knowledge that sperm recovered from a two-phase partition procedure are used for fertilization assays. These results open up new possibilities for using specific subpopulations of sperm for artificial insemination or in vitro fertilization, not only regarding better sperm quality but also certain characteristics such as subpopulations enriched in spermatozoa bearing X or Y chromosome that we have already isolated or any other feature. (C) 2011 Elsevier B.V. All rights reserved.

Cromatografia em contra-corrente de gotejamento na separação de substâncias fenólicas de Neuropogon aurantiaco-ater

This work reports the separation of phenolic compounds present in the metabolic extract of the lichen Neuropogon aurantiaco-ater (Usniaceae). The crude extract was fractionated by droplet counter-current chromatography using a solvent mixture of high polarity (chloroform:methanol:water at 43:37:20, in ascending mode). The separation resulted in the isolation of usnic and protocetratic acid, which were identified by TLC, HPLC, and NMR tests.

Quick preparative separation of natural naphthopyranones with antioxidant activity by high-speed counter-current chromatography

The natural naphthopyranones paepalantine (1), paepalantine-9O-β-D-glucopyranoside (2) and paepalantine-9-O-β-D-allopyranosyl-(1→6)-O-β-D-glucopyranoside (3) were separated in a preparative scale from the ethanolic extract of the capitula of Paepalanthus bromelioides by high-speed counter-current chromatography (HSCCC). The solvent system used was composed of water-ethanol-ethyl acetate-hexane (10:4:10:4, v/v/v/v). This technique led to the separation of the three different naphthopyranone glycosides in pure form in approximately 7 hours. Paepalantine showed a good antioxidant activity when assayed by the DPPH radical spectrophotometric assay.

Centrifugal processing of cell debris and inclusion bodies from recombinant Escherichia coli

The settling characteristics of cell debris and inclusion bodies prior to, and following, fractionation in a disc-stack centrifuge were measured using Cumulative Sedimentation Analysis (CSA) and Centrifugal Disc photosedimentation (CDS). The impact of centrifuge feedrate and repeated homogenisation on both cell debris and inclusion body collection efficiency was investigated. Increasing the normalised centrifuge feedrate (Q/Sigma) from 1.32 x 10(-9) m s(-1) to 3.97 x 10(-9) m s(-1) leads to a 36% increase in inclusion body paste purity. Purity may also be improved by repeated homogenisation. Increasing the number of homogeniser passes results in smaller cell debris size whilst leaves inclusion body size unaltered. At a normalised centrifuge feedrate of 2.65 x 10(-9) m s(-1), increasing the number of homogeniser passes from two (2) to ten (10) improved overall inclusion body paste purity by 58%. Grade-efficiency curves for both the cell debris and inclusion bodies have also been generated in this study. The data are described using an equation developed by Mannweiler (1989) with parameters of k = 0.15-0.26 and n = 2.5-2.6 for inclusion bodies, and k = 0.12-0.14 and n = 2.0-2.2 for cell debris. This is the first accurate experimentally-determined grade efficiency curve for cell debris. Previous studies have simply estimated debris grade efficiency curves using an approximate debris size distribution and grade efficiency curves determined with 'ideal particles' (e.g. spherical PVA particles). The findings of this study may be used to simulate and optimise the centrifugal fractionation of inclusion bodies from cell debris.

Computational and Experimental Analysis of Flow Field in the Diffusers of Centrifugal Compressors

Centrifugal compressors are widely used for example in process industry, oil and gas industry, in small gas turbines and turbochargers. In order to achieve lower consumption of energy and operation costs the efficiency of the compressor needs to be improve. In the present work different pinches and low solidity vaned diffusers were utilized in order to improve the efficiency of a medium size centrifugal compressor. In this study, pinch means the decrement of the diffuser flow passage height. First different geometries were analyzed using computational fluid dynamics. The flow solver Finflo was used to solve the flow field. Finflo is a Navier-Stokes solver. The solver is capable to solve compressible, incompressible, steady and unsteady flow fields. Chien's k-e turbulence model was used. One of the numerically investigated pinched diffuser and one low solidity vaned diffuser were studied experimentally. The overall performance of the compressor and the static pressure distribution before and after the diffuser were measured. The flow entering and leaving the diffuser was measured using a three-hole Cobra-probe and Kiel-probes. The pinch and the low solidity vaned diffuser increased the efficiency of the compressor. Highest isentropic efficiency increment obtained was 3\% of the design isentropic efficiency of the original geometry. It was noticed in the numerical results that the pinch made to the hub and the shroud wall was most beneficial to the operation of the compressor. Also the pinch made to the hub was better than the pinchmade to the shroud. The pinch did not affect the operation range of the compressor, but the low solidity vaned diffuser slightly decreased the operation range.The unsteady phenomena in the vaneless diffuser were studied experimentally andnumerically. The unsteady static pressure was measured at the diffuser inlet and outlet, and time-accurate numerical simulation was conducted. The unsteady static pressure showed that most of the pressure variations lay at the passing frequency of every second blade. The pressure variations did not vanish in the diffuser and were visible at the diffuser outlet. However, the amplitude of the pressure variations decreased in the diffuser. The time-accurate calculations showed quite a good agreement with the measured data. Agreement was very good at the design operation point, even though the computational grid was not dense enough inthe volute and in the exit cone. The time-accurate calculation over-predicted the amplitude of the pressure variations at high flow.

Computational Analysis and Optimization of Real Gas Flow In Small Centrifugal Compressors

Small centrifugal compressors are more and more widely used in many industrialsystems because of their higher efficiency and better off-design performance comparing to piston and scroll compressors as while as higher work coefficient perstage than in axial compressors. Higher efficiency is always the aim of the designer of compressors. In the present work, the influence of four partsof a small centrifugal compressor that compresses heavy molecular weight real gas has been investigated in order to achieve higher efficiency. Two parts concern the impeller: tip clearance and the circumferential position of the splitter blade. The other two parts concern the diffuser: the pinch shape and vane shape. Computational fluid dynamics is applied in this study. The Reynolds averaged Navier-Stokes flow solver Finflo is used. The quasi-steady approach is utilized. Chien's k-e turbulence model is used to model the turbulence. A new practical real gas model is presented in this study. The real gas model is easily generated, accuracy controllable and fairly fast. The numerical results and measurements show good agreement. The influence of tip clearance on the performance of a small compressor is obvious. The pressure ratio and efficiency are decreased as the size of tip clearance is increased, while the total enthalpy rise keeps almost constant. The decrement of the pressure ratio and efficiency is larger at higher mass flow rates and smaller at lower mass flow rates. The flow angles at the inlet and outlet of the impeller are increased as the size of tip clearance is increased. The results of the detailed flow field show that leakingflow is the main reason for the performance drop. The secondary flow region becomes larger as the size of tip clearance is increased and the area of the main flow is compressed. The flow uniformity is then decreased. A detailed study shows that the leaking flow rate is higher near the exit of the impeller than that near the inlet of the impeller. Based on this phenomenon, a new partiallyshrouded impeller is used. The impeller is shrouded near the exit of the impeller. The results show that the flow field near the exit of the impeller is greatly changed by the partially shrouded impeller, and better performance is achievedthan with the unshrouded impeller. The loading distribution on the impeller blade and the flow fields in the impeller is changed by moving the splitter of the impeller in circumferential direction. Moving the splitter slightly to the suction side of the long blade can improve the performance of the compressor. The total enthalpy rise is reduced if only the leading edge of the splitter ismoved to the suction side of the long blade. The performance of the compressor is decreased if the blade is bended from the radius direction at the leading edge of the splitter. The total pressure rise and the enthalpy rise of thecompressor are increased if pinch is used at the diffuser inlet. Among the fivedifferent pinch shape configurations, at design and lower mass flow rates the efficiency of a straight line pinch is the highest, while at higher mass flow rate, the efficiency of a concave pinch is the highest. The sharp corner of the pinch is the main reason for the decrease of efficiency and should be avoided. The variation of the flow angles entering the diffuser in spanwise direction is decreased if pinch is applied. A three-dimensional low solidity twisted vaned diffuser is designed to match the flow angles entering the diffuser. The numerical results show that the pressure recovery in the twisted diffuser is higher than in a conventional low solidity vaned diffuser, which also leads to higher efficiency of the twisted diffuser. Investigation of the detailed flow fields shows that the separation at lower mass flow rate in the twisted diffuser is later than in the conventional low solidity vaned diffuser, which leads to a possible wider flow range of the twisted diffuser.

Experimental and Numerical Analysis of Different Volutes in a Centrifugal Compressor

In a centrifugal compressor the flow around the diffuser is collected and led to the pipe system by a spiral-shaped volute. In this study a single-stage centrifugal compressor with three different volutes is investigated. The compressorwas first equipped with the original volute, the cross-section of which was a combination of a rectangle and semi-circle. Next a new volute with a fully circular cross-section was designed and manufactured. Finally, the circular volute wasmodified by rounding the tongue and smoothing the tongue area. The overall performance of the compressor as well as the static pressure distribution after the impeller and on the volute surface were measured. The flow entering the volute was measured using a three-hole Cobra-probe, and flow visualisations were carriedout in the exit cone of the volute. In addition, the radial force acting on theimpeller was measured using magnetic bearings. The complete compressor with thecircular volute (inlet pipe, full impeller, diffuser, volute and outlet pipe) was also modelled using computational fluid dynamics (CFD). A fully 3-D viscous flow was solved using a Navier-Stokes solver, Finflo, developed at Helsinki University of Technology. Chien's k-e model was used to take account of the turbulence. The differences observed in the performance of the different volutes were quite small. The biggest differences were at low speeds and high volume flows,i.e. when the flow entered the volute most radially. In this operating regime the efficiency of the compressor with the modified circular volute was about two percentage points higher than with the other volutes. Also, according to the Cobra-probe measurements and flow visualisations, the modified circular volute performed better than the other volutes in this operating area. The circumferential static pressure distribution in the volute showed increases at low flow, constant distribution at the design flow and decrease at high flow. The non-uniform static pressure distribution of the volute was transmitted backwards across the vaneless diffuser and observed at the impeller exit. At low volume flow a strong two-wave pattern developed into the static pressure distribution at the impeller exit due to the response of the impeller to the non-uniformity of pressure. The radial force of the impeller was the greatest at the choke limit, the smallest atthe design flow, and moderate at low flow. At low flow the force increase was quite mild, whereas the increase at high flow was rapid. Thus, the non-uniformityof pressure and the force related to it are strong especially at high flow. Theforce caused by the modified circular volute was weaker at choke and more symmetric as a function of the volume flow than the force caused by the other volutes.

Numerical simulation of rotor-stator interaction and tip clearance flow in centrifugal compressors

The effect of the tip clearance and vaneless diffuser width on the stage performance and flow fields of a centrifugal compressor were studied numerically and results were compared to the experimental measurements. The diffuser width was changed by moving the shroud side of the diffuser axially and six tip clearances size from 0.5 to 3 mm were studied. Moreover, the effects of rotor-stator interaction on the diffuser and impeller flow fields and performance were studied. Also transient simulations were carried out in order to investigate the influence of the interaction on the impeller and diffuser performance parameters. It was seen that pinch could improve the performance and it help to get more uniform flow at exit and less back flow from diffuser to the impeller.

TRANSVERSAL DISTRIBUTION COMPARISON OF SOLID FERTILIZERS APPLIED OVER ANNUAL CROPS

Centrifugal spreaders dominate the application of solid materials in agriculture offering expressive operational field capacity and extended range of applied rates. Field tests for characterization of theirperformance are conducted without any physical obstacles (such as the presence of plants) during the parabolic trajectory of the falling particles of fertilizer to the soil. The purpose of this study was to comparatively evaluate the transverse distribution of solid fertilizers applied on cropped corn, soybeans and cotton. Evaluations of the spreaders were designed according to ASAE S341.3/99 Standard. Tests consisted in aligning side by side collectors in-between the cropped rows and weighting the material deposited. The results showed that transverse distribution of solid fertilizers applied over the cotton and corn crops is affected by the crop height, interfering directly on the effective width of the spreader application, which was not observedin the soybean crop, once the fertilizer application is done when the crop was still below the collector's height. The results suggest that evaluation of effective width of the spreaders application need to be done under real crop environment.

Renal insufficiency is independently associated with a distal distribution pattern of symptomatic lower-limb atherosclerosis

Objectives The purpose of this study was to assess the impact of renal insufficiency (RI) on the distribution pattern of peripheral arterial disease (PAD). We hypothesised that RI is associated with a distally accentuated involvement of the peripheral arterial tree. Design This is a retrospective analysis. Materials and Methods Analysis was based on a consecutive series of 2709 patients with chronic PAD of atherosclerotic origin undergoing primary endovascular treatment of lower-extremity arteries. Atherosclerotic pattern was grouped into femoropopliteal (n = 2085) and infragenicular (n = 892) disease according to target lesions treated while using iliac disease (n = 1133) as reference. Univariable and multivariable multinomial regression analyses were performed to assess relation with RI. Results are shown as relative risk ratio (RRRs) with 95% confidence intervals (95% CIs). A p < 0.05 was considered statistically significant. RI was defined as glomerular filtration rate (GFR) < 60 ml min−1 1.73 m−2. Results Presence of RI was an independent risk factor for a centrifugal lesion pattern (RRR 1.48, 95% CI: 1.17–1.86, p = 0.001). Moreover, a decrease in GFR by 10 ml min−1 1.73 m−2 was associated with an RRR of 1.08 for below-the-knee arterial disease (95% CI: 1.03–1.13, p = 0.003). Conclusion Presence and severity of RI are independent predictors of a distal obstructive pattern in patients with symptomatic PAD.

Distribution and biomass of seston in the East Atlantic

Biomass of seston in the surface layers of coastal waters off Namibia reaches 1 g/m**2 and decreases with distance from the shore. Two regions of high seston biomass, one northern and one southern, are distinguished. A subsurface maximum of seston biomass, presumably coinciding with the stream of compensating countercurrent, is identified in the 200-500 m layer. Similar vertical distribution of plankton is known in upwelling areas of the eastern shores of the Atlantic and Pacific Oceans and in several other ocean areas, such as the area of the Kuril-Kamchatka Trench. This fact probably indicates that life cycles of pelagic animal forms of various taxonomic groups that inhabit them and phases of their ontogenic migrations are similar.

Vertical distribution of suspended aggregates in the equatorial Pacific

Marine snow (MS) distribution from the surface to 1000 m depth was determined in the equatorial Pacific using the underwater video profiler during the Etude du Broutage en Zone Equatoriale cruise in fall 1996. The latitudinal transect was carried out at 17 stations along the 180° meridian from 8°S to 8°N during a cold phase of El Niño-Southern Oscillation. Higher MS concentrations were found below the equatorial zone than poleward. At the equator the estimated integrated MS carbon/m**2 in the upper kilometer was 5.7 g/m**2, while both southward and northward (between 1° and 8°) the mean integrated MS carbon was about 2.7 g/m**2. In the upper 50 m the MS carbon was twofold lower than the combined carbon of autotrophic and heterotrophic protists and four times lower than the mesozooplankton carbon biomass, both measured concurrently during the cruise. Different water bodies had different MS content. The highest concentrations were found in the South Equatorial Current, the South Equatorial Counter Current, and the North Equatorial Countercurrent. Tropical waters at the south in the South Subsurface Countercurrents and the warm northern superficial waters had the lowest MS biomass. Mechanistically, a latitudinal "conveyor belt", a poleward divergence of upwelled waters that return to the equator after being downwelled at north and south convergent zones, may partially explain the vertical distribution of particulate matter observed during the studied period.

Brief Announcement: Node Sampling Using Centrifugal Random Walks.

We propose distributed algorithms for sampling networks based on a new class of random walks that we call Centrifugal Random Walks (CRW). A CRW is a random walk that starts at a source and always moves away from it. We propose CRW algorithms for connected networks with arbitrary probability distributions, and for grids and networks with regular concentric connectivity with distance based distributions. All CRW sampling algorithms select a node with the exact probability distribution, do not need warm-up, and end in a number of hops bounded by the network diameter.

Node sampling using random centrifugal walks

Sampling a network with a given probability distribution has been identified as a useful operation. In this paper we propose distributed algorithms for sampling networks, so that nodes are selected by a special node, called the source, with a given probability distribution. All these algorithms are based on a new class of random walks, that we call Random Centrifugal Walks (RCW). A RCW is a random walk that starts at the source and always moves away from it. Firstly, an algorithm to sample any connected network using RCW is proposed. The algorithm assumes that each node has a weight, so that the sampling process must select a node with a probability proportional to its weight. This algorithm requires a preprocessing phase before the sampling of nodes. In particular, a minimum diameter spanning tree (MDST) is created in the network, and then nodes weights are efficiently aggregated using the tree. The good news are that the preprocessing is done only once, regardless of the number of sources and the number of samples taken from the network. After that, every sample is done with a RCW whose length is bounded by the network diameter. Secondly, RCW algorithms that do not require preprocessing are proposed for grids and networks with regular concentric connectivity, for the case when the probability of selecting a node is a function of its distance to the source. The key features of the RCW algorithms (unlike previous Markovian approaches) are that (1) they do not need to warm-up (stabilize), (2) the sampling always finishes in a number of hops bounded by the network diameter, and (3) it selects a node with the exact probability distribution.

Physical and operational features of a pulsed continuous countercurrent liquid-solids contactor /

"National Reactor Testing Station"--Cover.