Purification of rectangle DNA origami by rate-zonal centrifugation

DNA origami technique, a breakthrough in DNA nanotechnology, has been widely used to prepare complex DNA nanostructures with nanoscale addressability. However, the purity and yield are generally the bottleneck to application of DNA nanostructures, and current methods for purifying DNA origami nanostructures in large quantities are time-consuming and laborious. This study aims to develop a scalable, cost-effective and contamination-free method of purifying DNA origami nanostructures. We employ an effective and convenient purification approach to purify planar rectangle DNA origami structures through rate-zonal centrifugation. By subjecting DNA origami samples to high centrifugal force in a density gradient media of glycerol, well-folded nanostructures and by-products are separated successfully, which are confirmed by agarose gel electrophoresis and atomic force microscopy(AFM). This method will aid the production of pure rectangle DNA origami nanostructures in large quantity.

Investigation of the block toppling evolution of a layered model slope by centrifuge test and discrete element modeling

Primary toppling usually occurs in layered rock slopes with large anti-dip angles.In this paper,the block toppling evolution was explored using a large-scale centrifuge system.Each block column in the layered model slope was made of cement mortar.Some artificial cracks perpendicular to the block column were prefabricated.Strain gages,displacement gages,and high-speed camera measurements were employed to monitor the deformation and failure processes of the model slope.The centrifuge test results show that the block toppling evolution can be divided into seven stages,i.e.layer compression,formation of major tensile crack,reverse bending of the block column,closure of major tensile crack,strong bending of the block column,formation of failure zone,and complete failure.Block toppling is characterized by sudden large deformation and occurs in stages.The wedge-shaped cracks in the model incline towards the slope.Experimental observations show that block toppling is mainly caused by bending failure rather than by shear failure.The tensile strength also plays a key factor in the evolution of block toppling.The simulation results from discrete element method(DEM)is in line with the testing results.Tensile stress exists at the backside of rock column during toppling deformation.Stress concentration results in the fragmented rock column and its degree is the most significant at the slope toe.

Experimental Analysis of Radial Centrifugal Pump Shutdown

Centrifugal pumps are widely used in the metallurgy,coal,and building sectors.In order to study the hydraulic characteristics of a closed impeller centrifugal pump during its shutdown in the so-called power frequency and frequency conversion modes,experiments were carried to determine the characteristic evolution of parameters such as speed,inlet and outlet pressure,head,flow rate and shaft power.A quasi-steady-state method was also used to further investigate these transient behaviors.The results show that,compared to the power frequency input,the performance parameter curves for the frequency conversion input are less volatile and smoother.The characteristic time is longer and the response to shutdown is slower.The quasi-steady-state theoretical head-flow curves match the experimental head-flow curves more closely at low flow rates when the frequency conversion input is considered.Moreover,in this case,the similarity law predicts the hydraulic performance more accurately.

Blade Wrap Angle Impact on Centrifugal Pump Performance:Entropy Generation and Fluid-Structure Interaction Analysis

The centrifugal pump is a prevalent power equipment widely used in different engineering patterns,and the impeller blade wrap angle significantly impacts its performance.A numerical investigation was conducted to analyze the influence of the blade wrap angle on flow characteristics and energy distribution of a centrifugal pump evaluated as a low specific speed with a value of 69.This study investigates six impellermodels that possess varying blade wrap angles(95°,105°,115°,125°,135°,and 145°)that were created while maintaining the same volute and other geometrical characteristics.The investigation of energy loss was conducted to evaluate the values of total and entropy generation rates(TEG,EGR).The fluid-structure interaction was considered numerically using the software tools ANSYS Fluent and ANSYSWorkbench.The elastic structural dynamic equation was used to estimate the structural response,while the shear stress transport k–ωturbulence model was utilized for the fluid domain modeling.The findings suggest that the blade wrap angle has a significant influence on the efficiency of the pump.The impeller featuring a blade wrap angle of 145°exhibits higher efficiency,with a notable increase of 3.76%relative to the original model.Variations in the blade wrap angle impact the energy loss,shaft power,and pump head.The model with a 145°angle exhibited a maximum equivalent stress of 14.8MPa and a total deformation of 0.084 mm.The results provide valuable insights into the intricate flow mechanism of the centrifugal pump,particularly when considering various blade wrap angles.

Multiresidue Method for Determination of 67 Pesticides in Water Samples Using Solid-Phase Extraction with Centrifugation and Gas Chromatography-Mass Spectrometry

A new multi-residue method based on solid-phase extraction (SPE) with centrifugation was developed for determination and quantitation of 67 pesticides in water samples. Two SPE cartridges were tested: Chromabond C18 and Oasis HLB. Parameters that influence the extraction efficiency such as the eluent volume, the sample loading volume, the addition of organic solvent to water sample, sorbent drying and elute concentration were optimized. The innovation of this work was the examination of the use of a centrifugation technique in both the drying and elution steps. When combined with centrifugation, the volume of the elution solvent was reduced to 2 mL and the time for sorbent drying decreased also to 10 min under vacuum. Under the optimized conditions, this method showed good recoveries higher than 65% - 68% for the 67 analyzed pesticides using the C18 and HLB cartridges with relative standard deviations lower than 9.7% - 12.3%. Limits of quantification were between 2 and 20 ng.L–1. The simplicity of the described method, use of less of organic solvent, short procedure time, and good recoveries demonstrate the advantages of this environmentally friendly approach for routine analysis of numerous samples.

Centrifugation: an important pre-analytic procedure that influences plasma microRNA quantification during blood processing

Circulating microRNAs are robustly present in plasma or serum and have become a research focus as biomarkers for tumor diagnosis and prognosis.Centrifugation is a necessary procedure for obtaining highquality blood supernatant.Herein,we investigated one-step and two-step centrifugations,two centrifugal methods routinely used in microRNA study,to explore their effects on plasma microRNA quantification.The microRNAs obtained from one-step and two-step centrifugations were quantified by microarray and TaqMan-based real-time quantitative polymerase chain reaction(Q-PCR).Dynamic light scattering was performed to explore the difference underlying the two centrifugal methods.The results from the microarray containing 1,347 microRNAs showed that the signal detection rate was greatly decreased in the plasma sample prepared by two-step centrifugation.More importantly,the microRNAs missing in this plasma sample could be recovered and detected in the precipitate generated from the second centrifugation.Consistent with the results from microarray,a marked decrease of three representative microRNAs in twostep centrifugal plasma was validated by Q-PCR.According to the size distribution of all nanoparticles in plasma,there were fewer nanoparticles with size>1,000 nm in two-step centrifugal plasma.Our experiments directly demonstrated that different centrifugation methods produced distinct quantities of plasma microRNAs.Thus,exosomes or protein complexes containing microRNAs may be involved in large nanoparticle formation and may be precipitated after two-step centrifugation.Our results remind us that sample processing methods should be first considered in conducting research.

Purification of specularite by centrifugation instead of flotation to produce iron oxide red pigment

This study used specularite, a high-gradient magnetic separation concentrate, as a raw material in reverse flotation.An iron concentrate with a grade of 65.1 wt% and a recovery rate of 75.31% were obtained.A centrifugal concentrator served as the deep purification equipment for the preparation of iron oxide red pigments, and its optimal rotating drum speed, feed concentration, and other conditions were determined.Under optimal conditions, a high-purity iron oxide concentrate with a grade of 69.38 wt% and a recovery rate of 80.89% were obtained and used as a raw material for preparing iron oxide red pigment.Calcining with sulfuric acid produced iron red pigments with different hues.Simultaneously, middlings with a grade of 60.20 wt% and a recovery rate of 17.51% were obtained and could be used in blast furnace ironmaking.High-value utilization of specularite beneficiation products was thus achieved.

Different Re-centrifugation Conditions for Infectious Disease Screening Using the Abbott 12000

We investigated whether an ordinary centrifuge can achieve the standard centrifugal effect required according to specifications for infectious disease screening using the Abbott i2000.Samples were collected and centrifuged following a standard operating procedure(SOP).They were then divided into three groups according to the results of the initial screening tests:a negative group,weak-positive group,and positive group.Twenty negative samples and all weak-positive and positive samples were re-analyzed.Two tubes for each re-analyzed sample were centrifuged simultaneously,one for 10 min at 10000x g,per recommendations,and one for 10 min at 2750^g.No significant difference was found between the groups using different centrifugal forces.There was a strong correlation in the quantitative values between the two conditions of centrifugation.Consistency analysis showed a Cronbach's alpha>0.8 for detection of Treponema pallidum,human immunodeficiency virus(HIV),hepatitis C virus(HCV),and hepatitis B surface antigen in the three groups(negative group,weak-positive group,and positive group)under different centrifugation conditions.Strong consistency was found under different centrifugal conditions,regardless of the initial testing results.In conclusion,we conducted centrifugation steps in duplicate,according to infectious disease screening protocols.Our study showed that all samples should be centrifuged using a recommended relative centrifugal force after a proper clotting time,as in the standard operating procedure of our laboratory.In this way,we were able to obtain the same results using an ordinary centrifuge as those obtained using a high-speed centrifuge,such as the Abbott i2000.

An Effective Method for Depleting MatureT Lymphocytes from Bone Marrow Cells──Two－step Percoll Centrifugation

In the present paper we have observed the effect of discontinuous gradient two-step Percoll centrifugation on depleting mature T lymphocytes from normal bone marrow. The pre-/post-Percoll percentage of CD34+, and Leu4+ cells in MNC was counted by using APAAP technique. As the two-step Percoll centrifugation is simple,and time saving, and decreases the incidence of contamination of cultured cells as well,This technique may be of value in serum-free culture of hematopoietic cells and immunological studies.

Single and double layer centrifugation improve the quality of cryopreserved bovine sperm from poor quality ejaculates

Background： Density gradient centrifugation was reported as a technique of semen preparation in assisted reproductive techniques in humans and animals. This technique was found to be efficient in improving semen quality after harmful techniques such as cryopreservation. Recently a modified technique, single layer centrifugation,was proposed as a technique providing a large amount of high quality spermatozoa, and this treatment was performed before conservation. Single layer centrifugation has been studied prevalently in stallions and in boars,but limited data were available for bulls. Occasionally bulls are known to experience a transient reduction in semen quality, thus techniques that allow improvement in semen quality could be applied in this context. The aim of this study was the evaluation of single layer and double layer centrifugation by the use of iodixanol, compared with conventional centrifugation and non-centrifuged semen, on the sperm characteristics during the cryopreservation process in bulls with normal and poor semen quality.Results： Single layer centrifugation and double layer centrifugation both significantly increased the percentage of normal spermatozoa and decreased the percentage of non-sperm cells in poor quality samples, while both were ineffective in those of normal quality. Sperm characteristics in poor quality samples increased after single layer centrifugation and double layer centrifugation, reaching values similar to those recorded in normal samples, and this trend is maintained after equilibration and after cryopreservation. On the other hand, SLC and DLC resulted in a consistent reduction in the spermatozoa recovered, and this resulted in a reduction of the absolute amount of spermatozoa cryopreserved in the normal samples, without a clear improvement in sperm characteristics in this type of sample.Conclusions： These data suggested that both SLC and DLC could be performed in practice, but their application should be limited to the cases in which the quality of the spermatozoa recovered is more important than the total amount of spermatozoa.

Preliminary Study of Effects of Centrifugation of Stallion Semen on Motility and Pro-AKAP4 Concentration over 72 Hours Post-collection

The study aimed to investigate the effects of centrifugation of stallion semen on progressive sperm motility over a 72-hour period;and determine if the protein marker,precursor of A-kinase anchor protein 4(pro-AKAP4),may be maintained at a higher level over time when compared to non-centrifuged samples.Pro-AKAP4 has been studied for its function in stabilizing the structure of spermatozoa and its use as a marker for fertilizing capacity of spermatozoa.Semen collections were obtained from four fertile,commercially used Standardbred stallions collected three times each.Semen samples were diluted to a final concentration of 50 million total cells per milliliter.One extended aliquot of semen from each collection was used for the non-centrifuged treatment,and one extended aliquot of semen was centrifuged at 750×g for 10 min.The supernatant from the centrifuged sample was then removed with a 10 mL plastic transfer pipette,and the spermatozoa cell pellet was suspended in fresh extender to a final concentration of 50 million total cells per milliliter.Both centrifuged and non-centrifuged samples from each collection were stored in Equine Express II stallion semen shipping containers for 72 h and fresh ice packs were added to each container every 24 h.Both progressive sperm motility and levels of pro-AKAP4 were determined at times 0,12,24,48 and 72 h after collection.Differences were found between stallions,times and treatments with the main finding being that pro-AKAP4 concentration in centrifuged samples was maintained at a much higher level over the 72-hour period studied.

centrifugal用热控装置设计

目的评估飞行器产品在飞行过程对热–离心综合环境的适应性,研制一套热–离心综合环境试验用热控装置。方法从热–离心试验需求出发,完成综合试验系统总体研制目标及系统方案设计。针对高g值离心环境(离心加速度值≥90g)下的热载荷加载需求,开展基于热传导的热载荷加载方法研究,并完成适用于高g值环境的热控装置硬件布局优化设计。提出基于分段包络思想的产品响应温度梯度分布效应模拟方法及实现,减少过试验考核。结果利用该热控装置完成了热–离心综合环境功能验证试验,设置最大加速度为92g,保载时间为3 min,最高加热温度为150℃,最大温升速率为3℃/min,3温区控制,升温过程中,温度误差优于±1.5℃。结论该热–离心综合环境试验用热控装置可用于飞行器产品热–离心综合环境试验的考核。

Experimental and Numerical Investigation on High-Pressure Centrifugal Pumps:Ultimate Pressure Formulation,Fatigue Life Assessment and Topological Optimization of Discharge Section

A high percentage of failure in pump elements originates from fatigue.This study focuses on the discharge section behavior,made of ductile iron,under dynamic load.An experimental protocol is established to collect the strain under pressurization and depressurization tests at specific locations.These experimental results are used to formulate the ultimate pressure expression function of the strain and the lateral surface of the discharge section and to validate finite element modeling.Fe-Safe is then used to assess the fatigue life cycle using different types of fatigue criteria(Coffin-Manson,Morrow,Goodman,and Soderberg).When the pressure is under 3000 PSI,pumps have an unlimited service life of 107 cycles,regardless of the criterion.However,for a pressure of 3555 PSI,only the Morrow criterion denotes a significant decrease in fatigue life cycles,as it considers the average stress.The topological optimization is then applied to the most critical pump model(with the lowest fatigue life cycle)to increase its fatigue life.Using the solid isotropic material with a penalization approach,the Abaqus Topology OptimizationModule is employed.The goal is to reduce the strain energy density while keeping the volume within bounds.According to the findings,a 5%volume reduction causes the strain energy density to decrease from 1.06 to 0.66106 J/m^(3).According to Morrow,the fatigue life cycle at 3,555 PSI is 782,425 longer than the initial 309,742 cycles.

Discrete Optimization on Unsteady Pressure Fluctuation of a Centrifugal Pump Using ANN and Modified GA

Pressure fluctuation due to rotor-stator interaction in turbomachinery is unavoidable,inducing strong vibration in the equipment and shortening its lifecycle.The investigation of optimization methods for an industrial centrifugal pump was carried out to reduce the intensity of pressure fluctuation to extend the lifecycle of these devices.Considering the time-consuming transient simulation of unsteady pressure,a novel optimization strategy was proposed by discretizing design variables and genetic algorithm.Four highly related design parameters were chosen,and 40 transient sample cases were generated and simulated using an automatic program.70%of them were used for training the surrogate model,and the others were for verifying the accuracy of the surrogate model.Furthermore,a modified discrete genetic algorithm(MDGA)was proposed to reduce the optimization cost owing to transient numerical simulation.For the benchmark test,the proposed MDGA showed a great advantage over the original genetic algorithm regarding searching speed and effectively dealt with the discrete variables by dramatically increasing the convergence rate.After optimization,the performance and stability of the inline pump were improved.The efficiency increased by more than 2.2%,and the pressure fluctuation intensity decreased by more than 20%under design condition.This research proposed an optimization method for reducing discrete transient characteristics in centrifugal pumps.

A modified generalized scaling law for the similitude of dynamic strain in centrifuge modeling

Soil strain is the key parameter to control the elasto-plastic deformation and even the failure processes.To overcome the defect that the strain of the model soil is always smaller than that of the prototype in Iai′s generalized scaling law(GSL),a modified scaling law was proposed based on Iai′s GSL to secure the same dynamic shear strain between the centrifuge model and the prototype by modulating the amplitude and frequency of the input motion at the base.A suite of dynamic centrifuge model tests of dry sand level ground was conducted with the same overall scaling factor(λ=200)under different centrifugal accelerations by using the technique of“modeling of models”to validate the modified GSL.The test results show that the modified GSL could achieve the same dynamic strain in model as that of the prototype,leading to better modeling for geotechnical problems where dynamic strain dominates the response or failure of soils.Finally,the applicability of the proposed scaling law and possible constraints on geometry scaling due to the capability limits of existing centrifuge shaking tables are discussed.

Performance Prediction of an Optimized Centrifugal Pump with High Efficiency

The main structural parameters of the IR100-80-100A type chemical centrifugal pump have been optimized by means of an orthogonal test approach.The centrifugal pump has been modeled using the CFturbo software,and 16 sets of orthogonal-test schemes have been defined on the basis of 4 parameters,namely,the blade number,blade outlet angle,impeller outlet diameter,and impeller outlet width.Such analysis has been used to determine the influence of each index parameter on the pump working efficiency and identify a set of optimal combinations of such parameters.The internalflowfield in the centrifugal pump has been simulated by using the PumLinx software.These numerical results have shown that,compared with the prototype pump,the outlet pressure and shaft power of the optimized pump can be significantly reduced,and the pump working efficiency can be improved by 5.59%.In the present study,some arguments are also provided to demonstrate that,with respect to other optimization methods,the orthogonal test approach is more convenient,and requires less test times.

Optimal Structural Parameters for a Plastic Centrifugal Pump Inducer

The aim of the study is to determine the optimal structural parameters for a plastic centrifugal pump inducer within the framework of an orthogonal experimental method.For this purpose,a numerical study of the related flow field is performed using CFX.The shaft power and the head of the pump are taken as the evaluation indicators.Accordingly,the examined variables are the thickness(S),the blade cascade degree(t),the blade rim angle(β1),the blade hub angle(β2)and the hub length(L).The impact of each structural parameter on each evaluation index is examined and special attention is paid to the following combinations:S2 mm,t 2,β1235°,β2360°and L 140 mm(corresponding to a maximum head of 98.15 m);S 5 mm,t 1.6,β1252°,β2350°and L 140 mm(corresponding to a minimum shaft power of 63.06 KW).Moreover,using least squares and fish swarm algorithms,the pump shaft power and head are further optimized,yielding the following optimal combination:S 5 mm,t 1.9,β1252°,β2360°and L 145 mm(corresponding to the maximum head of 91.90 m,and a minimum shaft power of 64.83 KW).

Review:Measurement-Based Monitoring and Fault Identification in Centrifugal Pumps

Condition based maintenance(CBM) is one of the solutions to machinery maintenance requirements. Latest approaches to CBM aim at reducing human engagement in the real-time fault detection and decision making. Machine learning techniques like fuzzy-logic-based systems, neural networks, and support vector machines help to reduce human involvement. Most of these techniques provide fault information with 100% confidence. It is undeniably apparent that this area has a vast application scope. To facilitate future exploration, this review is presented describing the centrifugal pump faults, the signals they generate, their CBM based diagnostic schemes, and case studies for blockage and cavitation fault detection in centrifugal pump(CP) by performing the experiment on test rig. The classification accuracy is above 98% for fault detection. This review gives a head-start to new researchers in this field and identifies the un-touched areas pertaining to CP fault diagnosis.

Single Layer Centrifugation with Androcoll-E^(TM) improved progressive motility and percentage of live spermatozoa with intact acrosome of chilled stallion semen but did not have an effect on DNA integrity

A significant number of stallions produce low quality ejaculates with high sensibility to chilling. Single Layer Centrifugation (SLC) with Andro-coll-ETM has been presented as an efficient method of selecting good quality spermatozoa. In the current study, changes in sperm quality (motility, viability, acrosome integrity and DNA damage) occurring during storage at 5?C for a maximum of 72 h, were investigated. For that, one ejaculate from 12 stallions was split in two aliquots: control and SLC-selected. Both aliquots were chilled and stored at 5?C and spermatozoa were evaluated for motility, viability and acrosome integrity at 24, 48 and 72 h post collection. DNA damage was evaluated at 48 h post collection using the comet assay. In the SLC-selected aliquots, there was a significant improvement in terms of progressive motility (0 h: P = 0.005;24 h: P 0.05). SLC with Androcoll-ETM improved semen quality prolonging sperm longevity of chilled semen (P = 0.012). This positive effect was more evident in ejaculates most sensitive to chilling that had a sharp decrease in motility in the first 24 h of refrigeration and for all ejaculates at 72 h post-chilling. Therefore, this method reveals to be a useful technique for selecting spermatozoa and maintain sperm quality during storage.