| 1. |
- Chang, Keke, et al.
(författare)
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Considerations on Circuit Design and Data Acquisition of a Portable Surface Plasmon Resonance Biosensing System
- 2015
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Ingår i: Sensors. - : MDPI AG. - 1424-8220. ; 15:8, s. 20511-20523
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Tidskriftsartikel (refereegranskat)abstract
- The aim of this study was to develop a circuit for an inexpensive portable biosensing system based on surface plasmon resonance spectroscopy. This portable biosensing system designed for field use is characterized by a special structure which consists of a microfluidic cell incorporating a right angle prism functionalized with a biomolecular identification membrane, a laser line generator and a data acquisition circuit board. The data structure, data memory capacity and a line charge-coupled device (CCD) array with a driving circuit for collecting the photoelectric signals are intensively focused on and the high performance analog-to-digital (A/D) converter is comprehensively evaluated. The interface circuit and the photoelectric signal amplifier circuit are first studied to obtain the weak signals from the line CCD array in this experiment. Quantitative measurements for validating the sensitivity of the biosensing system were implemented using ethanol solutions of various concentrations indicated by volume fractions of 5%, 8%, 15%, 20%, 25%, and 30%, respectively, without a biomembrane immobilized on the surface of the SPR sensor. The experiments demonstrated that it is possible to detect a change in the refractive index of an ethanol solution with a sensitivity of 4.99838 × 10(5) ΔRU/RI in terms of the changes in delta response unit with refractive index using this SPR biosensing system, whereby the theoretical limit of detection of 3.3537 × 10(-5) refractive index unit (RIU) and a high linearity at the correlation coefficient of 0.98065. The results obtained from a series of tests confirmed the practicality of this cost-effective portable SPR biosensing system.
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| 2. |
- Aref, Mohaddeseh, et al.
(författare)
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Potentiometric pH Nanosensor for Intracellular Measurements: Real-Time and Continuous Assessment of Local Gradients
- 2021
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Ingår i: Analytical Chemistry. - : American Chemical Society (ACS). - 0003-2700 .- 1520-6882. ; 93:47, s. 15744-15751
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Tidskriftsartikel (refereegranskat)abstract
- We present a pH nanosensor conceived for single intracellular measurements. The sensing architecture consisted of a two-electrode system evaluated in the potentiometric mode. We used solid-contact carbon nanopipette electrodes tailored to produce both the indicator (pH nanosensor) and reference electrodes. The indicator electrode was a membrane-based ion-selective electrode containing a receptor for hydrogen ions that provided a favorable selectivity for intracellular measurements. The analytical features of the pH nanosensor revealed a Nernstian response (slope of -59.5 mV/pH unit) with appropriate repeatability and reproducibility (variation coefficients of <2% for the calibration parameters), a fast response time (<5 s), adequate medium-term drift (0.7 mV h(-)(1)), and a linear range of response including physiological and abnormal cell pH levels (6.0-8.5). In addition, the position and configuration of the reference electrode were investigated in cell-based experiments to provide unbiased pH measurements, in which both the indicator and reference electrodes were located inside the same cell, each of them inside two neighboring cells, or the indicator electrode inside the cell and the reference electrode outside of (but nearby) the studied cell. Finally, the pH nanosensor was applied to two cases: (i) the tracing of the pH gradient from extra-to intracellular media over insertion into a single PC12 cell and (ii) the monitoring of variations in intracellular pH in response to exogenous administration of pharmaceuticals. It is anticipated that the developed pH nanosensor, which is a label-free analytical tool, has high potential to aid in the investigation of pathological states that manifest in cell pH misregulation, with no restriction in the type of targeted cells.
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| 3. |
- Da Silva Lima, Alex, et al.
(författare)
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Electrochemistry at and in single cells
- 2020
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Ingår i: Electrochemistry for Bioanalysis. Bhavik Patel (red.). - : Elsevier. - 9780128212035 ; , s. 125-160
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Bokkapitel (övrigt vetenskapligt/konstnärligt)abstract
- This chapter introduces the reader to electrochemical methods for analysis of extracellular events and applications of electrodes for measurements in single cell cytoplasm and organelles. Applications include measurements of exocytosis, the process by which chemical species are released by cells, and excretion of reactive oxygen and nitrogen species as well as cholesterol in cell membranes. In addition, new methods have been devised to measure the electroactive contents of single organelles both removed from cells and in living cells. A large part of the success of these methods has come from the development of new micro and nano electrodes. © 2020 Elsevier Inc. All rights reserved.
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| 4. |
- Fei, Keke, et al.
(författare)
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CpxR regulates the Rcs phosphorelay system in controlling the Ysc-Yop type III secretion system in Yersinia pseudotuberculosis
- 2021
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Ingår i: Microbiology. - : Microbiology Society. - 1350-0872 .- 1465-2080. ; 167:1
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Tidskriftsartikel (refereegranskat)abstract
- The CpxRA two-component regulatory system and the Rcs phosphorelay system are both employed by the Enterobacteriaceae family to preserve bacterial envelope integrity and function when growing under stress. Although both systems regulate several overlapping physiological processes, evidence demonstrating a molecular connection between Cpx and Rcs signalling outputs is scarce. Here, we show that CpxR negatively regulates the transcription of the rcsB gene in the Rcs phosphorelay system in Yersinia pseudotuberculosis. Interestingly, transcription of rcsB is under the control of three promoters, which were all repressed by CpxR. Critically, synthetic activation of Cpx signalling through mislocalization of the NlpE lipoprotein to the inner membrane resulted in an active form of CpxR that repressed activity of rcsB promoters. On the other hand, a site-directed mutation of the phosphorylation site at residue 51 in CpxR generated an inactive non-phosphorylated variant that was unable to regulate output from these rcsB promoters. Importantly, CpxR-mediated inhibition of rcsB transcription in turn restricted activation of the Ysc-Yop type III secretion system (T3SS). Moreover, active CpxR blocks zinc-mediated activation of Rcs signalling and the subsequent activation of lcrF transcription. Our results demonstrate a novel regulatory cascade linking CpxR-RcsB-LcrF to control production of the Ysc-Yop T3SS.
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| 5. |
- Fei, Keke, et al.
(författare)
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LcrQ coordinates with the YopD-LcrH complex to repress lcrF expression and control type III secretion by Yersinia pseudotuberculosis
- 2021
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Ingår i: mBio. - : American Society for Microbiology (ASM). - 2161-2129 .- 2150-7511. ; 12:3
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Tidskriftsartikel (refereegranskat)abstract
- Human-pathogenic Yersinia species employ a plasmid-encoded type III secretion system (T3SS) to negate immune cell function during infection. A critical element in this process is the coordinated regulation of T3SS gene expression, which involves both transcriptional and posttranscriptional mechanisms. LcrQ is one of the earliest identified negative regulators of Yersinia T3SS, but its regulatory mechanism is still unclear. In a previous study, we showed that LcrQ antagonizes the activation role played by the master transcriptional regulator LcrF. In this study, we confirm that LcrQ directly interacts with LcrH, the chaperone of YopD, to facilitate the negative regulatory role of the YopD-LcrH complex in repressing lcrF expression at the posttranscriptional level. Negative regulation is strictly dependent on the YopD-LcrH complex, more so than on LcrQ. The YopD-LcrH complex helps to retain cytoplasmic levels of LcrQ to facilitate the negative regulatory effect. Interestingly, RNase E and its associated protein RhlB participate in this negative regulatory loop through a direct interaction with LcrH and LcrQ. Hence, we present a negative regulatory loop that physically connects LcrQ to the posttranscriptional regulation of LcrF, and this mechanism incorporates RNase E involved in mRNA decay.
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| 6. |
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| 7. |
- Hu, Keke, et al.
(författare)
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Correlating Molecule Count and Release Kinetics with Vesicular Size Using Open Carbon Nanopipettes
- 2020
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Ingår i: Journal of the American Chemical Society. - : American Chemical Society (ACS). - 0002-7863 .- 1520-5126. ; 142:40, s. 16910-16914
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Tidskriftsartikel (refereegranskat)abstract
- In this work, open carbon nanopipettes (CNPs) with radius between 50 and 600 nm were used to control translocation of different-sized vesicles through the pipette orifice followed by nanoelectrochemical analysis. Vesicle impact electrochemical cytometry (VIEC) was used to determine the number of catecholamine molecules expelled from single vesicles onto an inner-wall carbon surface, where the duration of transmitter release was quantified and correlated to the vesicle size all in the same nanotip. This in turn allowed us to both size and count molecules for vesicles in a living cell. Here, small and sharp open CNPs were employed to carry out intracellular VIEC with minimal invasion and high sensitivity. Our findings with VIEC reveal that the vesicular content increases with vesicle size. The release kinetics of vesicular transmitters and dense core size have the same relation with the vesicle size, implying that the vesicular dense core size determines the speed of each release event. This direct correlation unravels one of the complexities of exocytosis.
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| 8. |
- Hu, Keke, et al.
(författare)
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Electrochemical Measurements Reveal Reactive Oxygen Species in Stress Granules**
- 2021
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Ingår i: Angewandte Chemie - International Edition. - : Wiley. - 1433-7851 .- 1521-3773. ; 60:28, s. 15302-15306
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Tidskriftsartikel (refereegranskat)abstract
- Stress granules (SGs) are membrane-less organelles that assemble in the cytoplasm to organize cellular contents and promote rapid adaptation during stress. To understand how SGs contribute to physiological functions, we used electrochemical measurements to detect electroactive species in SGs. With amperometry, we discovered that reactive oxygen species (ROS) are encapsulated inside arsenite-induced SGs, and H2O2 is the main species. The release kinetics of H2O2 from single SGs and the number of H2O2 molecules were quantified. The discovery that SGs contain ROS implicates them as communicators of the cellular stresses rather than a simple endpoint. This may explain how SGs regulate cellular metabolism and stress responses. This may also help better understand their cytoprotective functions in pathological conditions associated with SGs such as neurodegenerative diseases (NDs), cancers and viral infections. © 2021 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH.
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| 9. |
- Hu, Keke, et al.
(författare)
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Quantifying Intracellular Single Vesicular Catecholamine Concentration with Open Carbon Nanopipettes to Unveil the Effect of L-DOPA on Vesicular Structure
- 2022
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Ingår i: Angewandte Chemie-International Edition. - : Wiley. - 1433-7851 .- 1521-3773. ; 61:1
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Tidskriftsartikel (refereegranskat)abstract
- Understanding the regulatory mechanisms of exocytosis is essential for uncovering the pathologies of neuronal disorders and developing related pharmaceuticals. In this work intracellular vesicle impact electrochemical cytometry (IVIEC) measurements with different-sized (50-500 nm radius) open carbon nanopipettes (CNPs) were performed to quantify the vesicular content and release kinetics of specific vesicle populations grouped by orifice sizes. Intracellular vesicles with radius below 100 nm were captured and narrowed between 50 and 100 nm. On the basis of this, single vesicular catecholamine concentrations in the intracellular environment were quantified as 0.23-1.1 M. Our results with L-3,4-dihydroxyphenylalanine (L-DOPA)-exposure indicate that L-DOPA regulates exocytosis by increasing the dense core size and vesicular content while catecholamine concentrations did not show obvious alterations. These were all achieved simultaneously and relatively noninvasively with open CNPs.
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| 10. |
- Zhan, Chengcheng, et al.
(författare)
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Mechanical property enhancement of NbTiZr refractory medium-entropy alloys due to Si-induced crystalline-to-amorphous transitions
- 2022
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Ingår i: Surface & Coatings Technology. - : Elsevier. - 0257-8972 .- 1879-3347. ; 433
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Tidskriftsartikel (refereegranskat)abstract
- Crystalline-to-amorphous transitions induced by chemical modulation have been attracting great research interests and an in-depth understanding of such transitions is always demanding. Here we design the Nb-Ti-Zr-(Si) alloy systems employing the empirical criteria and the related phase diagrams for enhancing mechanical properties accompanied by Si-induced microstructure evolutions. (NbTiZr)100-xSix (x = 0, 3.0, 10.2, 22.2, and 25.0 at.%) refractory medium-entropy alloy (RMEA) coatings are synthesized by magnetron co-sputtering. With increasing Si content, the pristine body-centered cubic (bcc) single phase transforms into a nanocomposite structure consisting of bcc nanocrystals embedded in an amorphous matrix and eventually into an entirely amorphous structure. This is well rationalized with a thermodynamic database of the Nb-Ti-Zr-Si system constructed using the CALPHAD (CALculation of PHAse Diagrams) approach, which suggests the bcc structure to be thermodynamically stable at low Si contents and the formation of amorphous RMEA to be preferred at higher Si contents. The superior mechanical property of the amorphous (NbTiZr)75.0Si25.0 (at.%) coating compared to the Si-free counterpart is achieved, i.e., the hardness (H) increases by 115% and the elastic modulus (E) increases by 70%. The Si-induced crystalline-to-amorphous transition in RMEA which leads to a consistently impressive strengthening effect was rarely found in other alloys or coatings.
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