scholarly journals A Portable Device for Simple Exosome Separation from Biological Samples

Micromachines ◽  
2021 ◽  
Vol 12 (10) ◽  
pp. 1182
Author(s):  
Wenwen Chen ◽  
Yingying Xie ◽  
Yuang Chang ◽  
Yuhai Xu ◽  
Mengqian Zhao ◽  
...  

Exosomes are membrane-bound nanovesicles secreted by most types of cells, which contain a series of biologically important molecules, such as miRNAs, proteins, and lipids, etc. Emerging evidence show that exosomes can affect the physiological status of cells and are involved in various pathological processes. However, due to their small size and density close to body fluids, it is challenging to separate exosomes from a small volume of biological samples in a simple manner. Herein, we propose a new strategy for isolating circulating exosomes from biological samples in a portable device. This method synergistically integrates chitosan electrostatic-adsorption, scaffold substrates, and shuttle flow to enable the highly effective capture of circulating exosomes with a recovery rate of over 80% within 20 minutes, which is much better than the performance of traditional ultracentrifugation (5–25%, 3 h). Besides, the isolated exosomes from samples could be lysed in situ and further subjected to RNA concentration detection and protein analysis. In particular, all the necessary procedures for exosome separation could be integrated into a single device without the need for bulky equipment. This established device is portable and easy to operate, which provides a promising platform for the study of exosome biology and clinical diagnosis.

Author(s):  
P.R. Swann ◽  
A.E. Lloyd

Figure 1 shows the design of a specimen stage used for the in situ observation of phase transformations in the temperature range between ambient and −160°C. The design has the following features a high degree of specimen stability during tilting linear tilt actuation about two orthogonal axes for accurate control of tilt angle read-out high angle tilt range for stereo work and habit plane determination simple, robust construction temperature control of better than ±0.5°C minimum thermal drift and transmission of vibration from the cooling system.


1964 ◽  
Vol 45 (4) ◽  
pp. 535-559 ◽  
Author(s):  
E. Bolté ◽  
S. Mancuso ◽  
G. Eriksson ◽  
N. Wiqvist ◽  
E. Diczfalusy

ABSTRACT In 15 cases of therapeutic abortion by laparotomy the placenta was disconnected from the foetus and perfused in situ with tracer amounts of radioactive dehydroepiandrosterone (DHA), dehydroepiandrosterone sulphate (DHAS), androst-4-ene-3,17-dione (A), testosterone (T) and 17β-oestradiol (OE2). Analysis of the placentas, perfusates and urine samples revealed an extensive aromatisation of DHA, A and T; more than 70% of the radioactive material recovered was phenolic, and at least 80 % of this phenolic material was identified as oestrone (OE1), 17β-oestradiol (OE2) and oestriol (OE3), the latter being detected only in the urine. Comparative studies indicated that A and T were aromatised somewhat better than DHA and that all three unconjugated steroids were aromatised to a much greater extent than DHAS. Radioactive OE1 and OE2 were isolated and identified in the placentas and perfusates, but no OE3, epimeric oestriols, or ring D ketols could be detected in these sources, not even when human chorionic gonadotrophin (HCG) was added to the blood prior to perfusion. Lack of placental 16-hydroxylation was also apparent when OE2 was perfused. Regardless of the precursor perfused, there was three times more OE2 than OE1 in the placenta and three times more OE1 than OE2 in the perfusate. This was also the case following perfusion with OE2. The results are interpreted as suggesting the existence in the pregnant human of a placental »barrier« limiting the passage of circulating androgen. The barrier consists of a) limited ability to transfer directly DHAS and b) an enzymic mechanism resulting in the rapid and extensive aromatisation of the important androgens DHA, A and T.


2017 ◽  
Author(s):  
Amanda H. Schmidt ◽  
◽  
Paul R. Bierman ◽  
Veronica Sosa-Gonzalez ◽  
Thomas B. Neilson ◽  
...  
Keyword(s):  

2019 ◽  
Vol 219 ◽  
pp. 08003
Author(s):  
Maja Verstraeten

The SoLid Collaboration is currently operating a 1.6 ton neutrino detector near the Belgian BR2 reactor. Its main goal is the observation of the oscillation of electron antineutrinos to previously undetected flavour states. The highly segmented SoLid detector employs a compound scintillation technology based on PVT scintillator in combination with LiF-ZnS(Ag) screens containing the 6Li isotope. The experiment has demonstrated a channel-to-channel response that can be controlled to the level of a few percent, an energy resolution of better than 14% at 1 MeV, and a determination of the interaction vertex with a precision of 5 cm. This contribution highlights the major outcomes of the R&D program, the quality control during component manufacture and integration, the current performance and stability of the full-scale system, as well as the in-situ calibration of the detector with various radioactive sources.


2014 ◽  
Vol 81 (3) ◽  
pp. 1024-1031 ◽  
Author(s):  
Bhagyalakshmi Kalidass ◽  
Muhammad Farhan Ul-Haque ◽  
Bipin S. Baral ◽  
Alan A. DiSpirito ◽  
Jeremy D. Semrau

ABSTRACTIt is well known that copper is a key factor regulating expression of the two forms of methane monooxygenase found in proteobacterial methanotrophs. Of these forms, the cytoplasmic, or soluble, methane monooxygenase (sMMO) is expressed only at low copper concentrations. The membrane-bound, or particulate, methane monooxygenase (pMMO) is constitutively expressed with respect to copper, and such expression increases with increasing copper. Recent findings have shown that copper uptake is mediated by a modified polypeptide, or chalkophore, termed methanobactin. Although methanobactin has high specificity for copper, it can bind other metals, e.g., gold. Here we show that inMethylosinus trichosporiumOB3b, sMMO is expressed and active in the presence of copper if gold is also simultaneously present. Such expression appears to be due to gold binding to methanobactin produced byM. trichosporiumOB3b, thereby limiting copper uptake. Such expression and activity, however, was significantly reduced if methanobactin preloaded with copper was also added. Further, quantitative reverse transcriptase PCR (RT-qPCR) of transcripts of genes encoding polypeptides of both forms of MMO and SDS-PAGE results indicate that both sMMO and pMMO can be expressed when copper and gold are present, as gold effectively competes with copper for binding to methanobactin. Such findings suggest that under certain geochemical conditions, both forms of MMO may be expressed and activein situ. Finally, these findings also suggest strategies whereby field sites can be manipulated to enhance sMMO expression, i.e., through the addition of a metal that can compete with copper for binding to methanobactin.


2009 ◽  
Vol 390 (2) ◽  
pp. 137-144 ◽  
Author(s):  
Yingmiao Liu ◽  
Chien-Tsun Kuan ◽  
Jing Mi ◽  
Xiuwu Zhang ◽  
Bryan M. Clary ◽  
...  

Abstract Epidermal growth factor receptor variant III (EGFRvIII) is a glycoprotein uniquely expressed in glioblastoma, but not in normal brain tissues. To develop targeted therapies for brain tumors, we selected RNA aptamers against the histidine-tagged EGFRvIII ectodomain, using an Escherichia coli system for protein expression and purification. Representative aptamer E21 has a dissociation constant (Kd) of 33×10-9 m, and exhibits high affinity and specificity for EGFRvIII in ELISA and surface plasmon resonance assays. However, selected aptamers cannot bind the same protein expressed from eukaryotic cells because glycosylation, a post-translational modification present only in eukaryotic systems, significantly alters the structure of the target protein. By transfecting EGFRvIII aptamers into cells, we find that membrane-bound, glycosylated EGFRvIII is reduced and the percentage of cells undergoing apoptosis is increased. We postulate that transfected aptamers can interact with newly synthesized EGFRvIII, disrupt proper glycosylation, and reduce the amount of mature EGFRvIII reaching the cell surface. Our work establishes the feasibility of disrupting protein post-translational modifications in situ with aptamers. This finding is useful for elucidating the function of proteins of interest with various modifications, as well as dissecting signal transduction pathways.


2010 ◽  
Vol 97-101 ◽  
pp. 814-817 ◽  
Author(s):  
Jun Deng

One of the greatest drawbacks to predicting the behaviour of bonded joints has been the lack of reliable data on the mechanical properties of adhesives. In this study, methods for determining mechanical properties of structural adhesive were discussed. The Young’s modulus, Poisson’s ratio and tensile strength of the adhesive were tested by dogbone specimens (bulk form) and butt joint specimens (in situ form). The shear modulus and shear strength were test by V-notched specimens (bulk form) and thick adherend lap-shear (TALS) joint specimens (in situ form). The test results show that the elastic modulus provided by the manufacturer is too low, the dogbone specimen is better than the butt joint specimen to test the tensile strength and elastic modulus and the TALS joint specimen is better than the V-notched specimen to test the shear strength.


Author(s):  
RUBING HUANG ◽  
XIAODONG XIE ◽  
DAVE TOWEY ◽  
TSONG YUEH CHEN ◽  
YANSHENG LU ◽  
...  

Combinatorial interaction testing is a well-recognized testing method, and has been widely applied in practice, often with the assumption that all test cases in a combinatorial test suite have the same fault detection capability. However, when testing resources are limited, an alternative assumption may be that some test cases are more likely to reveal failure, thus making the order of executing the test cases critical. To improve testing cost-effectiveness, prioritization of combinatorial test cases is employed. The most popular approach is based on interaction coverage, which prioritizes combinatorial test cases by repeatedly choosing an unexecuted test case that covers the largest number of uncovered parameter value combinations of a given strength (level of interaction among parameters). However, this approach suffers from some drawbacks. Based on previous observations that the majority of faults in practical systems can usually be triggered with parameter interactions of small strengths, we propose a new strategy of prioritizing combinatorial test cases by incrementally adjusting the strength values. Experimental results show that our method performs better than the random prioritization technique and the technique of prioritizing combinatorial test suites according to test case generation order, and has better performance than the interaction-coverage-based test prioritization technique in most cases.


2018 ◽  
Vol 22 (10) ◽  
pp. 5341-5356 ◽  
Author(s):  
Seyed Hamed Alemohammad ◽  
Jana Kolassa ◽  
Catherine Prigent ◽  
Filipe Aires ◽  
Pierre Gentine

Abstract. Characterizing soil moisture at spatiotemporal scales relevant to land surface processes (i.e., of the order of 1 km) is necessary in order to quantify its role in regional feedbacks between the land surface and the atmospheric boundary layer. Moreover, several applications such as agricultural management can benefit from soil moisture information at fine spatial scales. Soil moisture estimates from current satellite missions have a reasonably good temporal revisit over the globe (2–3-day repeat time); however, their finest spatial resolution is 9 km. NASA's Soil Moisture Active Passive (SMAP) satellite has estimated soil moisture at two different spatial scales of 36 and 9 km since April 2015. In this study, we develop a neural-network-based downscaling algorithm using SMAP observations and disaggregate soil moisture to 2.25 km spatial resolution. Our approach uses the mean monthly Normalized Differenced Vegetation Index (NDVI) as ancillary data to quantify the subpixel heterogeneity of soil moisture. Evaluation of the downscaled soil moisture estimates against in situ observations shows that their accuracy is better than or equal to the SMAP 9 km soil moisture estimates.


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