scholarly journals Cellular effects of gliotoxin: Evaluation of a proteomic, isotope-based method to detect reactive cysteines

2021 ◽  
Author(s):  
◽  
Sven Sondhauss
Keyword(s):  
Large Scale ◽  
Thiol Group ◽  
Disulfide Bridge ◽  
Reversed Phase ◽  
Cellular Damage ◽  
Full Potential ◽  
Label Free ◽  
Cysteine Modification ◽  
Affinity Tag ◽  
Cell Vitality

<p>Cysteinyl residues in proteins are important for many cellular processes and unregulated modification of the cysteine thiol group can have negative effects on cell vitality and viability. In this thesis, the potential for use of the isotope coded affinity tag (ICAT) method for detection of cysteine modification has been investigated. ICAT reagents label free cysteine thiols. The aim of this study was to use HL-60 cells treated with gliotoxin, a fungal metabolite with a reactive disulfide bridge, as a system to evaluate the performance of ICAT for identification of cysteine modification in a whole cell proteome. Gliotoxin has antimicrobial, antitumor, immunosuppressive and cytotoxic properties that have been related to cysteine modification in proteins. Cellular assays including viability using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, cell cycle analysis, and measurement of reactive oxygen species using dichlorofluorescin diacetate were used to establish conditions for measuring the effects of gliotoxin on HL-60 cells prior to large-scale cellular damage. Cells exposed to gliotoxin and control cells were then labeled with ICAT reagents and analysed by offline reversed phase liquid chromatography followed by matrix-assisted laser desorption/ionization tandem mass spectrometry. The pilot results identified tubulin, glyceraldehyde-3-phosphate dehydrogenase and peptidyl-prolyl cis-trans isomerase as putative targets of gliotoxin. Additionally, this study showed that ICAT can be used to detect modified cysteines from a highly complex sample, but further optimization is needed to unlock the full potential for detection of cysteine modification in complex samples.</p>

scholarly journals Cellular effects of gliotoxin: Evaluation of a proteomic, isotope-based method to detect reactive cysteines

2021 ◽  
Author(s):  
◽  
Sven Sondhauss
Keyword(s):  
Large Scale ◽  
Thiol Group ◽  
Disulfide Bridge ◽  
Reversed Phase ◽  
Cellular Damage ◽  
Full Potential ◽  
Label Free ◽  
Cysteine Modification ◽  
Affinity Tag ◽  
Cell Vitality

<p>Cysteinyl residues in proteins are important for many cellular processes and unregulated modification of the cysteine thiol group can have negative effects on cell vitality and viability. In this thesis, the potential for use of the isotope coded affinity tag (ICAT) method for detection of cysteine modification has been investigated. ICAT reagents label free cysteine thiols. The aim of this study was to use HL-60 cells treated with gliotoxin, a fungal metabolite with a reactive disulfide bridge, as a system to evaluate the performance of ICAT for identification of cysteine modification in a whole cell proteome. Gliotoxin has antimicrobial, antitumor, immunosuppressive and cytotoxic properties that have been related to cysteine modification in proteins. Cellular assays including viability using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, cell cycle analysis, and measurement of reactive oxygen species using dichlorofluorescin diacetate were used to establish conditions for measuring the effects of gliotoxin on HL-60 cells prior to large-scale cellular damage. Cells exposed to gliotoxin and control cells were then labeled with ICAT reagents and analysed by offline reversed phase liquid chromatography followed by matrix-assisted laser desorption/ionization tandem mass spectrometry. The pilot results identified tubulin, glyceraldehyde-3-phosphate dehydrogenase and peptidyl-prolyl cis-trans isomerase as putative targets of gliotoxin. Additionally, this study showed that ICAT can be used to detect modified cysteines from a highly complex sample, but further optimization is needed to unlock the full potential for detection of cysteine modification in complex samples.</p>

A microfluidic device with integrated impedance detection for λ-DNA

2003 ◽  
Vol 773 ◽  
Author(s):  
Myung-Il Park ◽  
Jonging Hong ◽  
Dae Sung Yoon ◽  
Chong-Ook Park ◽  
Geunbae Im
Keyword(s):  
Microfluidic Device ◽  
Electrochemical Impedance ◽  
Direct Detection ◽  
Full Potential ◽  
Label Free ◽  
Buffer Solution ◽  
Detection Systems ◽  
Significant Difference ◽  
Detection Of Biomolecules ◽  
Impedance Magnitude

AbstractThe large optical detection systems that are typically utilized at present may not be able to reach their full potential as portable analysis tools. Accurate, early, and fast diagnosis for many diseases requires the direct detection of biomolecules such as DNA, proteins, and cells. In this research, a glass microchip with integrated microelectrodes has been fabricated, and the performance of electrochemical impedance detection was investigated for the biomolecules. We have used label-free λ-DNA as a sample biomolecule. By changing the distance between microelectrodes, the significant difference between DW and the TE buffer solution is obtained from the impedance-frequency measurements. In addition, the comparison for the impedance magnitude of DW, the TE buffer, and λ-DNA at the same distance was analyzed.

scholarly journals Graphene-Based Artificial Synapses with Tunable Plasticity

2021 ◽  
Vol 17 (4) ◽  
pp. 1-21
Author(s):  
He Wang ◽  
Nicoleta Cucu Laurenciu ◽  
Yande Jiang ◽  
Sorin Cotofana
Keyword(s):  
Synaptic Plasticity ◽  
Weight Change ◽  
Time Scale ◽  
Large Scale ◽  
Synaptic Weight ◽  
Low Voltage ◽  
Logic Gates ◽  
Full Potential ◽  
Back Gate ◽  
Input Spike

Design and implementation of artificial neuromorphic systems able to provide brain akin computation and/or bio-compatible interfacing ability are crucial for understanding the human brain’s complex functionality and unleashing brain-inspired computation’s full potential. To this end, the realization of energy-efficient, low-area, and bio-compatible artificial synapses, which sustain the signal transmission between neurons, is of particular interest for any large-scale neuromorphic system. Graphene is a prime candidate material with excellent electronic properties, atomic dimensions, and low-energy envelope perspectives, which was already proven effective for logic gates implementations. Furthermore, distinct from any other materials used in current artificial synapse implementations, graphene is biocompatible, which offers perspectives for neural interfaces. In view of this, we investigate the feasibility of graphene-based synapses to emulate various synaptic plasticity behaviors and look into their potential area and energy consumption for large-scale implementations. In this article, we propose a generic graphene-based synapse structure, which can emulate the fundamental synaptic functionalities, i.e., Spike-Timing-Dependent Plasticity (STDP) and Long-Term Plasticity . Additionally, the graphene synapse is programable by means of back-gate bias voltage and can exhibit both excitatory or inhibitory behavior. We investigate its capability to obtain different potentiation/depression time scale for STDP with identical synaptic weight change amplitude when the input spike duration varies. Our simulation results, for various synaptic plasticities, indicate that a maximum 30% synaptic weight change and potentiation/depression time scale range from [-1.5 ms, 1.1 ms to [-32.2 ms, 24.1 ms] are achievable. We further explore the effect of our proposal at the Spiking Neural Network (SNN) level by performing NEST-based simulations of a small SNN implemented with 5 leaky-integrate-and-fire neurons connected via graphene-based synapses. Our experiments indicate that the number of SNN firing events exhibits a strong connection with the synaptic plasticity type, and monotonously varies with respect to the input spike frequency. Moreover, for graphene-based Hebbian STDP and spike duration of 20ms we obtain an SNN behavior relatively similar with the one provided by the same SNN with biological STDP. The proposed graphene-based synapse requires a small area (max. 30 nm 2 ), operates at low voltage (200 mV), and can emulate various plasticity types, which makes it an outstanding candidate for implementing large-scale brain-inspired computation systems.

scholarly journals Label-free fluorescence predictions from large-scale correlative light and electron microscopy data

2021 ◽  
Vol 27 (S1) ◽  
pp. 94-95
Author(s):  
Ryan Lane ◽  
Luuk Balkenende ◽  
Simon van Staalduine ◽  
Anouk Wolters ◽  
Ben Giepmans ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Large Scale ◽  
Light And Electron Microscopy ◽  
Label Free ◽  
Electron Microscopy Data ◽  
Microscopy Data

Structure Controlled Synthesis of Vertically Aligned Carbon Nanotubes Using Thermal Chemical Vapor Deposition Process

2010 ◽  
Vol 133 (3) ◽  
Author(s):  
Myung Gwan Hahm ◽  
Young-Kyun Kwon ◽  
Ahmed Busnaina ◽  
Yung Joon Jung
Keyword(s):  
Carbon Nanotubes ◽  
Chemical Vapor Deposition ◽  
Vapor Deposition ◽  
Large Scale ◽  
Chemical Vapor ◽  
Controlled Synthesis ◽  
Full Potential ◽  
Thermal Chemical Vapor Deposition ◽  
Vertically Aligned ◽  
Walled Carbon Nanotubes

Due to their unique one-dimensional nanostructure along with excellent mechanical, electrical, and optical properties, carbon nanotubes (CNTs) become a promising material for diverse nanotechnology applications. However, large-scale and structure controlled synthesis of CNTs still have many difficulties due to the lack of understanding of the fundamental growth mechanism of CNTs, as well as the difficulty of controlling atomic-scale physical and chemical reactions during the nanotube growth process. Especially, controlling the number of graphene wall, diameter, and chirality of CNTs are the most important issues that need to be solved to harness the full potential of CNTs. Here we report the large-scale selective synthesis of vertically aligned single walled carbon nanotubes (SWNTs) and double walled carbon nanotubes (DWNTs) by controlling the size of catalyst nanoparticles in the highly effective oxygen assisted thermal chemical vapor deposition (CVD) process. We also demonstrate a simple but powerful strategy for synthesizing ultrahigh density and diameter selected vertically aligned SWNTs through the precise control of carbon flow during a thermal CVD process.

scholarly journals Technology Acceptance in Healthcare: A Systematic Review

2021 ◽  
Vol 11 (22) ◽  
pp. 10537
Author(s):  
Adi A. AlQudah ◽  
Mostafa Al-Emran ◽  
Khaled Shaalan
Keyword(s):  
Systematic Review ◽  
Technology Acceptance ◽  
Large Scale ◽  
Empirical Studies ◽  
Influential Factors ◽  
Full Potential ◽  
Eligibility Criteria ◽  
Factors Affecting ◽  
The Usa ◽  
Healthcare Technologies

Understanding the factors affecting the use of healthcare technologies is a crucial topic that has been extensively studied, specifically during the last decade. These factors were studied using different technology acceptance models and theories. However, a systematic review that offers extensive understanding into what affects healthcare technologies and services and covers distinctive trends in large-scale research remains lacking. Therefore, this review aims to systematically review the articles published on technology acceptance in healthcare. From a yield of 1768 studies collected, 142 empirical studies have met the eligibility criteria and were extensively analyzed. The key findings confirmed that TAM and UTAUT are the most prevailing models in explaining what affects the acceptance of various healthcare technologies through different user groups, settings, and countries. Apart from the core constructs of TAM and UTAUT, the results showed that anxiety, computer self-efficacy, innovativeness, and trust are the most influential factors affecting various healthcare technologies. The results also revealed that Taiwan and the USA are leading the research of technology acceptance in healthcare, with a remarkable increase in studies focusing on telemedicine and electronic medical records solutions. This review is believed to enhance our understanding through a number of theoretical contributions and practical implications by unveiling the full potential of technology acceptance in healthcare and opening the door for further research opportunities.

scholarly journals Endogenous formaldehyde scavenges cellular glutathione resulting in cytotoxic redox disruption

2020 ◽  
Author(s):  
Carla Umansky ◽  
Agustín Morellato ◽  
Marco Scheidegger ◽  
Matthias Rieckher ◽  
Manuela R. Martinefski ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Dna Repair ◽  
Fanconi Anemia ◽  
Redox Homeostasis ◽  
Thiol Group ◽  
Cellular Damage ◽  
Repair Pathway ◽  
Cellular Redox ◽  
Redox Active ◽  
Development And Aging

AbstractFormaldehyde (FA) is a ubiquitous endogenous and environmental metabolite that is thought to exert cytotoxicity through DNA and DNA-protein crosslinking. We show here that FA can cause cellular damage beyond genotoxicity by triggering oxidative stress, which is prevented by the enzyme alcohol dehydrogenase 5 (ADH5/GSNOR). Mechanistically, we determine that endogenous FA reacts with the redox-active thiol group of glutathione (GSH) forming S-hydroxymethyl-GSH, which is metabolized by ADH5 yielding reduced GSH thus preventing redox disruption. We identify the ADH5-ortholog gene in Caenorhabditis elegans and show that oxidative stress also underlies FA toxicity in nematodes. Moreover, we show that endogenous GSH can protect cells lacking the Fanconi Anemia DNA repair pathway from FA, which might have broad implications for Fanconi Anemia patients and for healthy BRCA2-mutation carriers. We thus establish a highly conserved mechanism through which endogenous FA disrupts the GSH-regulated cellular redox homeostasis that is critical during development and aging.

scholarly journals Widespread multi-targeted therapy resistance via drug-induced secretome fucosylation

2021 ◽  
Author(s):  
Mark Borris D. Aldonza ◽  
Junghwa Cha ◽  
Insung Yong ◽  
Jayoung Ku ◽  
Dabin Lee ◽  
...  
Keyword(s):  
Targeted Therapies ◽  
Large Scale ◽  
Kinase Inhibitors ◽  
Tumor Regression ◽  
Therapy Resistance ◽  
Critical Component ◽  
Label Free ◽  
Drug Induced ◽  
Pan Cancer ◽  
Core Fucosylation

AbstractCancer secretome is a reservoir for aberrant glycosylation. How therapies alter this post-translational cancer hallmark and the consequences thereof remain elusive. Here we show that an elevated secretome fucosylation is a pan-cancer signature of both response and resistance to multiple targeted therapies. Large-scale pharmacogenomics revealed that fucosylation genes display widespread association with resistance to these therapies. In both cancer cell cultures and patients, targeted kinase inhibitors distinctively induced core fucosylation of secreted proteins less than 60 kDa. Label-free proteomics of N-glycoproteomes revealed that fucosylation of the antioxidant PON1 is a critical component of the therapy-induced secretome. Core fucosylation in the Golgi impacts PON1 stability and folding prior to secretion, promoting a more degradation-resistant PON1. Non-specific and PON1-specific secretome de-N-glycosylation both limited the expansion of resistant clones in a tumor regression model. Our findings demonstrate that core fucosylation is a common modification indirectly induced by targeted therapies that paradoxically promotes resistance.

scholarly journals The Co-Construction of Energy Provision and Everyday Practice: Integrating Heat Pumps in Social Housing in England

2015 ◽  
Vol 28 (3) ◽  
pp. 26-53 ◽  
Author(s):  
Ellis P Judson ◽  
Sandra Bell ◽  
Harriet Bulkeley ◽  
Gareth Powells ◽  
Stephen Lyon
Keyword(s):  
Large Scale ◽  
New Technologies ◽  
Heat Pumps ◽  
Consumer Information ◽  
Hot Water ◽  
Everyday Practice ◽  
Full Potential ◽  
Low Carbon ◽  
Energy Services ◽  
The North

Challenges of energy security, low carbon transitions, and electricity network constraints have led to a shift to new, efficient technologies for household energy services. Studies of such technological innovations usually focus on consumer information and changes in behaviour to realise their full potential. We suggest that regarding such technologies in existing energy provision systems opens up questions concerning how and why such interventions are delivered. We argue that we must understand the ways by which energy systems are co-constituted through the habits and expectations of households, their technologies and appliances, alongside arrangements associated with large-scale socio-technical infrastructures. Drawing on research with air-source-to-water heat pumps (ASWHP), installed as part of a large trans-disciplinary, utility-led research and demonstration project in the north of England, we investigate how energy services provision and everyday practice shapes new technologies uptake, and how such technologies mediate and reconfigure relations between users, providers and infrastructure networks. While the installation of ASWHP has led to role differentiation through which energy services are provided, the space for new forms of co-provision to emerge is limited by existing commitments to delivering energy services. Simultaneously, new forms of interdependency emerge between users, providers and intermediaries through sites of installation, instruction, repair and feedback. We find that although new technologies do lead to the rearrangement of practices, this is often disrupted by obduracy in the conventions and habits around domestic heating and hot water practices that have been established in relation to existing systems of provision. Rather being simply a matter of increasing levels of knowledge in order to ensure that such technologies are adopted effi ciently and effectively, our paper demonstrates how systemic arrangements of energy provision and everyday practice are co-implicated in socio-technical innovation by changing the nature of energy supply and use.

scholarly journals Proteomics and metabolomics in cow fertility: a systematic review

Reproduction ◽  
2020 ◽  
Vol 160 (5) ◽  
pp. 639-658
Author(s):  
Nicolas Aranciaga ◽  
James D Morton ◽  
Debra K Berg ◽  
Jessica L Gathercole
Keyword(s):  
Oxidative Stress ◽  
Systematic Review ◽  
Large Scale ◽  
Reproductive Tract ◽  
Early Embryo ◽  
Female Reproductive Tract ◽  
Full Potential ◽  
Technical Standards ◽  
The Impact

Cow subfertility is a multi-factorial problem in many countries which is only starting to be unravelled. Molecular biology can provide a substantial source of insight into its causes and potential solutions, particularly through large scale, untargeted omics approaches. In this systematic review, we set out to compile, assess and integrate the latest proteomic and metabolomic research on cow reproduction, specifically that on the female reproductive tract and early embryo. We herein report a general improvement in technical standards throughout the temporal span examined; however, significant methodological limitations are also identified. We propose easily actionable avenues for ameliorating these shortcomings and enhancing the reach of this field. Text mining and pathway analysis corroborate the relevance of proteins and metabolites related to the triad oxidative stress-inflammation-disease on reproductive function. We envisage a breakthrough in cattle reproductive molecular research within the next few years as in vivo sample techniques are improved, omics analysis equipment becomes more affordable and widespread, and software tools for single- and multi-omics data processing are further developed. Additional investigation of the impact of local oxidative stress and inflammation on fertility, both at the local and systemic levels, is key towards realising the full potential of this field.

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