scholarly journals Unidirectional Single-File Transport of Full-Length Proteins Through a Nanopore

2021 ◽  
Author(s):  
Luning Yu ◽  
Xinqi Kang ◽  
Fanjun Li ◽  
Behzad Mehrafrooz ◽  
Amr Makhamreh ◽  
...  
Keyword(s):  
High Speed ◽  
Protein Transport ◽  
Ionic Current ◽  
Single Nucleotide ◽  
Osmotic Effect ◽  
Single File ◽  
Protein Fingerprinting ◽  
High Concentrations ◽  
Single File Transport ◽  
Dynamics Simulations

AbstractNanopore technology offers long, accurate sequencing of an DNA or RNA strand via enzymatic ratcheting of the strand through a nanopore in single nucleotide steps, producing stepwise modulations of the nanopore ion current. In contrast to nucleic acids, their daughter molecules, proteins, have neutral peptide backbones and side chains of varying charges. Further, proteins have stable secondary and higher order structures that obstruct protein linearization required for single file nanopore transport. Here, we describe a general approach for realizing unidirectional transport of proteins through a nanopore that neither requires the protein to be uniformly charged nor a pull from a biological enzyme. At high concentrations of guanidinium chloride, we find fulllength proteins to translocate unidirectionally through an a-hemolysin nanopore in a polymer-based membrane, provided that one of the protein ends is decorated with a short anionic peptide. Molecular dynamics simulations show that such surprisingly steady protein transport is driven by a giant electro-osmotic effect caused by binding of guanidinium cations to the inner surface of the nanopore. We show that ionic current signals produced by protein passage can be used to distinguish two biological proteins and the global orientation of the same protein (N-to-C vs. C-to-N terminus) during the nanopore transport. With the average transport rate of one amino acid per 10 μs, our method may enable direct enzyme-free protein fingerprinting or perhaps even sequencing when combined with a high-speed nanopore reader instrument.

scholarly journals Aerodynamic analysis of uphill drafting in cycling

2021 ◽  
Vol 24 (1) ◽  
Author(s):  
T. van Druenen ◽  
B. Blocken
Keyword(s):  
Fluid Dynamics ◽  
Computational Fluid Dynamics ◽  
High Speed ◽  
Scientific Literature ◽  
Sensitivity Analyses ◽  
Air Resistance ◽  
Wind Tunnel Measurements ◽  
Aerodynamic Effect ◽  
Computational Fluid Dynamics Simulations ◽  
Dynamics Simulations

AbstractSome teams aiming for victory in a mountain stage in cycling take control in the uphill sections of the stage. While drafting, the team imposes a high speed at the front of the peloton defending their team leader from opponent’s attacks. Drafting is a well-known strategy on flat or descending sections and has been studied before in this context. However, there are no systematic and extensive studies in the scientific literature on the aerodynamic effect of uphill drafting. Some studies even suggested that for gradients above 7.2% the speeds drop to 17 km/h and the air resistance can be neglected. In this paper, uphill drafting is analyzed and quantified by means of drag reductions and power reductions obtained by computational fluid dynamics simulations validated with wind tunnel measurements. It is shown that even for gradients above 7.2%, drafting can yield substantial benefits. Drafting allows cyclists to save over 7% of power on a slope of 7.5% at a speed of 6 m/s. At a speed of 8 m/s, this reduction can exceed 16%. Sensitivity analyses indicate that significant power savings can be achieved, also with varying bicycle, cyclist, road and environmental characteristics.

scholarly journals Colchicine-binding protein of the liver. Its characterization and relation to microtubules.

1975 ◽  
Vol 66 (3) ◽  
pp. 609-620 ◽  
Author(s):  
C Patzelt ◽  
A Singh ◽  
Y L Marchand ◽  
L Orci ◽  
B Jeanrenaud
Keyword(s):  
High Speed ◽  
Specific Binding ◽  
Electrophoretic Analysis ◽  
Binding Activity ◽  
Low Concentrations ◽  
Electrophoretic Behavior ◽  
High Affinity Binding Site ◽  
High Concentrations

Colchicine-binding activity of mouse liver high-speed supernate has been investigated. It has been found to be time and temperature dependent. Two binding activities with different affinities for colchicine seem to be present in this high-speed supernate, of which only the high-affinity binding site (half maximal binding at 5 x 10(-6) M colchicine) can be attributed to microtubular protein by comparison with purified tubulin. Vinblastine interacted with this binding activity by precipitating it when used at high concentrations (2 x 10(-3) M), and by stabilizing it at low concentrations (10(-5) M). Lumicolchicine was found not to compete with colchicine. The colchicine-binding activity was purified from liver and compared with that of microtubular protein from brain. The specific binding activity of the resulting preparation, its electrophoretic behavior, and the electron microscope appearance of the paracrystals obtained upon its precipitation with vinblastine permitted its identification as microtubular protein (tubulin). Electrophoretic analysis of the proteins from liver supernate that were precipitated by vinblastine indicated that this drug was not specific for liver tubulin. Preincubation of liver supernate with 5 mM EGTA resulted in a time-dependent decrease of colchicine-binding activity, which was partly reversed by the addition of Ca++. However, an in vitro formation of microtubules upon lowering the Ca++ concentration could not be detected. Finally, a method was developed enabling that portion of microtubular protein which was present as free tubulin to be measured and to be compared with the total amount of this protein in the tissue. This procedure permitted demonstration of the fact that, under normal conditions, only about 40% of the tubulin of the liver was assemled as microtubules. It is suggested that, in the liver, rapid polymerization and depolymerization of microtubules occur and may be an important facet of the functional role of the microtubular system.

Development of a hybrid ozonation biofilm-membrane filatration process for the production of drinking water

2005 ◽  
Vol 51 (6-7) ◽  
pp. 241-248 ◽  
Author(s):  
T. Leiknes ◽  
M. Lazarova ◽  
H. Ødegaard
Keyword(s):  
Drinking Water ◽  
Organic Matter ◽  
High Speed ◽  
Membrane Surface ◽  
Drinking Water Treatment ◽  
Rotating Disk ◽  
Operating Conditions ◽  
Operating Modes ◽  
High Concentrations ◽  
Biomass Separation

Drinking water sources in Norway are characterized by high concentrations of natural organic matter (NOM), low alkalinity and low turbidity. The removal of NOM is therefore a general requirement in producing potable water. Drinking water treatment plants are commonly designed with coagulation direct filtration or NF spiral wound membrane processes. This study has investigated the feasibility and potential of a hybrid process combining ozonation and biofiltration with a rotating disk membrane for treating drinking water with high NOM concentrations. Ozonation will oxidize the NOM content removing colour and form biodegradable organic compounds, which can be removed in biological filters. A constructed water was used in this study which is representative of ozonated NOM-containing water. A rotating membrane disk bioreactor downstream the ozonation process was used to carry out both the biodegradation as well as biomass separation in the same reactor. Maintenance of biodegradation of the organic matter while controlling biofouling of the membrane and acceptable water production rates was the focus in the study. Three operating modes were investigated. Removal of the biodegradable organics was consistent throughout the study indicating that sufficient biomass was maintained in the reactor for all operating conditions tested. Biofouling control was not achieved through shear-induced cleaning by periodically rotating the membrane disks at high speed. By adding a small amount of sponges in the membrane chamber the biofouling could be controlled by mechanical cleaning of the membrane surface during disk rotation. The overall results indicate that the system can favorably be used in an ozonation/biofiltration process by carrying out both biodegradation as well as biomass separation in the same reactor.

Measurements of Periodic Reynolds Stress Oscillations in a Forced Turbulent Premixed Swirling Flame

2018 ◽  
Author(s):  
Christopher Douglas ◽  
Jamie Lim ◽  
Travis Smith ◽  
Benjamin Emerson ◽  
Timothy Lieuwen ◽  
...  
Keyword(s):  
Reynolds Stress ◽  
High Speed ◽  
Large Scale ◽  
Turbulent Transport ◽  
Stereoscopic Particle Image Velocimetry ◽  
Thermoacoustic Instability ◽  
Image Velocimetry ◽  
Planar Laser ◽  
Swirling Flame ◽  
Dynamics Simulations

This work is motivated by the thermoacoustic instability challenges associated with ultra-low emissions gas turbine combustors. It demonstrates the first use of high-speed dual-plane orthogonally-polarized stereoscopic-particle image velocimetry and synchronized OH planar laser-induced fluorescence in a premixed swirling flame. We use this technique to explore the effects of combustion and longitudinal acoustic forcing on the time- and phase-averaged flow field — particularly focusing on the behavior of the Reynolds stress in the presence of harmonic forcing. We observe significant differences between ensemble averaged and time averaged Reynolds stress. This implies that the large-scale motions are non-ergodic, due to coherent oscillations in Reynolds stress associated with the convection of periodic vortical structures. This result has important implications on hydrodynamic stability models and reduced order computational fluid dynamics simulations, which do show the importance of turbulent transport on the problem, but do not capture these coherent oscillations in their models.

scholarly journals 3D Printed Biomodels for Flow Visualization in Stenotic Vessels: An Experimental and Numerical Study

Micromachines ◽  
2020 ◽  
Vol 11 (6) ◽  
pp. 549
Author(s):  
Violeta Carvalho ◽  
Nelson Rodrigues ◽  
Ricardo Ribeiro ◽  
Pedro F. Costa ◽  
Rui A. Lima ◽  
...  
Keyword(s):  
High Speed ◽  
Complex Disease ◽  
Numerical Study ◽  
Numerical Data ◽  
In Vivo Studies ◽  
Physiological Variables ◽  
Microscopy Technique ◽  
Dynamics Simulations

Atherosclerosis is one of the most serious and common forms of cardiovascular disease and a major cause of death and disability worldwide. It is a multifactorial and complex disease that promoted several hemodynamic studies. Although in vivo studies more accurately represent the physiological conditions, in vitro experiments more reliably control several physiological variables and most adequately validate numerical flow studies. Here, a hemodynamic study in idealized stenotic and healthy coronary arteries is presented by applying both numerical and in vitro approaches through computational fluid dynamics simulations and a high-speed video microscopy technique, respectively. By means of stereolithography 3D printing technology, biomodels with three different resolutions were used to perform experimental flow studies. The results showed that the biomodel printed with a resolution of 50 μm was able to most accurately visualize flow due to its lowest roughness values (Ra = 1.8 μm). The flow experimental results showed a qualitatively good agreement with the blood flow numerical data, providing a clear observation of recirculation regions when the diameter reduction reached 60%.

scholarly journals Comparison of Tetrodotoxin and Procaine in Internally Perfused Squid Giant Axons

1967 ◽  
Vol 50 (5) ◽  
pp. 1413-1428 ◽  
Author(s):  
Toshio Narahashi ◽  
Nels C. Anderson ◽  
John W. Moore
Keyword(s):  
Steady State ◽  
Action Potential ◽  
Ionic Current ◽  
Transient Current ◽  
Resting Potential ◽  
Giant Axons ◽  
Time To Peak ◽  
Sucrose Gap ◽  
High Concentrations ◽  
Very High

Squid giant axons were internally perfused with tetrodotoxin and procaine, and excitability and electrical properties were studied by means of current-clamp and sucrose-gap voltage-clamp methods. Internally perfused tetrodotoxin was virtually without effect on the resting potential, the action potential, the early transient membrane ionic current, and the late steady-state membrane ionic current even at very high concentrations (1,000–10,000 nM) for a long period of time (up to 36 min). Externally applied tetrodotoxin at a concentration of 100 nM blocked the action potential and the early transient current in 2–3 min. Internally perfused procaine at concentrations of 1–10 mM reversibly depressed or blocked the action potential with an accompanying hyperpolarization of 2–4 mv, and inhibited both the early transient and late steady-state currents to the same extent. The time to peak early transient current was increased. The present results and the insolubility of tetrodotoxin in lipids have led to the conclusion that the gate controlling the flow of sodium ions through channels is located on the outer surface of the nerve membrane.

scholarly journals Design and Fabrication of a Micropump With Mixing Function for LOC

2008 ◽  
Vol 2 (2) ◽  
Author(s):  
Ho-jin Kang ◽  
Bum-kyoo Choi
Keyword(s):  
Lorentz Force ◽  
High Speed ◽  
Low Voltage ◽  
Flow Direction ◽  
Ionic Current ◽  
Digital Camera ◽  
Working Medium ◽  
Electrode Length ◽  
Portable System ◽  
Phosphate Buffered Saline

A micropump, which includes a mixing function, has been fabricated. For the application to LOC (Lab On a chip), the micropump utilized PBS (Phosphate Buffered Saline) solution as the working medium. The solution is commonly used in biochemistry and cell culturing. The portable system and low energy consumption are important to realize the LOC device. In spite of a low voltage of 4V, the flow rate of the micropump was 0.02466ml∕min. The new micropump shows more enhanced performance than existing micropumps. The micropump uses Lorentz force actuation. The Lorentz force acting onto the ionic current in the PBS solution generates the fluid flow in the micropump. For the accurate prediction on flow direction, a computer simulation has been made using commercial CFD code. The results of simulation showing circulation direction were verified by experiment. The fluid circulation from each electrode combined and acted as the mixer in the micropump. The micropump was fabricated 20.2mm in length, 1mm in width and 400 μm in electrode length. To measure accurately, a high speed digital camera was used.

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