scholarly journals A Screening Approach for Identifying Gliadin Neutralizing Antibodies on Epithelial Intestinal Caco-2 Cells

2017 ◽  
Vol 22 (8) ◽  
pp. 1035-1043
Author(s):  
Harald Hundsberger ◽  
Anita Koppensteiner ◽  
Elisabeth Hofmann ◽  
Doris Ripper ◽  
Maren Pflüger ◽  
...  
Keyword(s):  
High Throughput Screening ◽  
Neutralizing Antibodies ◽  
Inflammatory Condition ◽  
Gluten Free ◽  
Label Free ◽  
Industrialized Countries ◽  
Therapeutic Approaches ◽  
Impedance Sensing ◽  
Cell Substrate ◽  
Chronic Inflammatory Condition

Celiac disease (CD) is a chronic inflammatory condition caused by the ingestion of gliadin-containing food in genetically susceptible individuals. Undigested peptides of gliadin exert various effects, including increased intestinal permeability and inflammation in the small intestine. Although many therapeutic approaches are in development, a gluten-free diet is the only effective treatment for CD. Affecting at least 1% of the population in industrialized countries, it is important to generate therapeutic options against CD. Here, we describe the establishment of a high-throughput screening (HTS) platform based on AlphaLISA and electrical cell–substrate impedance sensing (ECIS) technology for the identification of anti-inflammatory and barrier-protective compounds in human enterocytes after pepsin-trypsin-digested gliadin (PT-gliadin) treatment. Our results show that the combination of these HTS technologies enables fast, reliable, simple, and label-free screening of IgY antibodies against PT-gliadin. Using this platform, we have identified a new chicken anti-PT-gliadin IgY antibody as a potential anti-CD agent.

Electric cell-substrate impedance sensing in kidney research

2019 ◽  
Author(s):  
Takamasa Iwakura ◽  
Julian A Marschner ◽  
Zhi Bo Zhao ◽  
Monika Katarzyna Świderska ◽  
Hans-Joachim Anders
Keyword(s):  
Continuous Monitoring ◽  
Adherent Cells ◽  
Label Free ◽  
Impedance Sensing ◽  
Barrier Integrity ◽  
Practical Applications ◽  
Non Invasive ◽  
Glomerular Epithelial Cells ◽  
Cell Substrate ◽  
Wide Range

Abstract Electric cell-substrate impedance sensing (ECIS) is a quantitative, label-free, non-invasive analytical method allowing continuous monitoring of the behaviour of adherent cells by online recording of transcellular impedance. ECIS offers a wide range of practical applications to study cell proliferation, migration, differentiation, toxicity and monolayer barrier integrity. All of these applications are relevant for basic kidney research, e.g. on endothelial cells, tubular and glomerular epithelial cells. This review gives an overview on the fundamental principles of the ECIS technology. We name strengths and remaining hurdles for practical applications, present an ECIS array reuse protocol, and review its past, present and potential future contributions to preclinical kidney research.

scholarly journals Comparison of Leading Biosensor Technologies to Measure Endothelial Adhesion, Barrier Properties, and Responses to Cytokines in Real-Time

Proceedings ◽  
2020 ◽  
Vol 60 (1) ◽  
pp. 62
Author(s):  
James J. W. Hucklesby ◽  
Akshata Anchan ◽  
Simon J. O’Carroll ◽  
Catherine E. Angel ◽  
E. Scott Graham
Keyword(s):  
Real Time ◽  
Barrier Properties ◽  
Microvascular Endothelial Cell ◽  
Label Free ◽  
Adhesion Barrier ◽  
Impedance Sensing ◽  
Non Invasive ◽  
Reduced Frequency ◽  
Cell Substrate ◽  
Higher Sensitivity

Electric Cell-substrate Impedance Sensing (ECIS), xCELLigence and cellZscope are commercially available instruments which are able to measure the impedance of cellular monolayers continuously and with high precision. The small currents used allow the label-free, real-time monitoring of the cells in a non-invasive manner. Despite the widespread use of these systems individually, direct comparisons between the systems have not been published. In order to compare the sensitivity of the instruments, the responses of the brain microvascular endothelial cell line hCMVEC to the inflammatory cytokines TNFα and IL1β were measured on all three instruments simultaneously. All three instruments showed transient decreases, followed by prolonged increases in impedance. Although xCELLigence could detect these changes, it was unable to determine which component of the barrier was affected. In contrast, ECIS and cellZscope were both able to attribute responses to particular barrier components, and ECIS had a higher sensitivity than cellZscope. Finally, as cellZscope uses Transwells, it allows access to the basolateral compartment, an important advantage of this technology. Furthermore, although xCELLigence readings are equivalent to ECIS, the reduced frequency range greatly limits interpretation. This work demonstrates that instruments must be carefully selected in order to ensure that they are appropriate for the experimental questions being asked.

scholarly journals Assessment of Cytotoxicity of Magnesium Oxide and Magnesium Hydroxide Nanoparticles using the Electric Cell-Substrate Impedance Sensing

2020 ◽  
Vol 10 (6) ◽  
pp. 2114
Author(s):  
Manishi Pallavi ◽  
Jenora Waterman ◽  
Youngmi Koo ◽  
Jagannathan Sankar ◽  
Yeoheung Yun
Keyword(s):  
Magnesium Oxide ◽  
Magnesium Hydroxide ◽  
Bone Repair ◽  
Corrosion Products ◽  
Label Free ◽  
Animal Testing ◽  
Impedance Sensing ◽  
Cell Substrate

Magnesium (Mg)-based alloys have the potential for bone repair due to their properties of biodegradation, biocompatibility, and structural stability, which can eliminate the requirement for a second surgery for the removal of the implant. Nevertheless, uncontrolled degradation rate and possible cytotoxicity of the corrosion products at the implant sites are known current challenges for clinical applications. In this study, we assessed in vitro cytotoxicity of different concentrations (0 to 50 mM) of possible corrosion products in the form of magnesium oxide (MgO) and magnesium hydroxide (Mg(OH)2) nanoparticles (NPs) in human fetal osteoblast (hFOB) 1.19 cells. We measured cell proliferation, adhesion, migration, and cytotoxicity using a real-time, label-free, non-invasive electric cell-substrate impedance sensing (ECIS) system. Our results suggest that 1 mM concentrations of MgO/Mg(OH)2 NPs are tolerable in hFOB 1.19 cells. Based on our findings, we propose the development of innovative biodegradable Mg-based alloys for further in vivo animal testing and clinical trials in orthopedics.

scholarly journals The Measurement and Analysis of Impedance Response of HeLa Cells to Distinct Chemotherapy Drugs

Micromachines ◽  
2021 ◽  
Vol 12 (2) ◽  
pp. 202
Author(s):  
Xiangbin Du ◽  
Jinlong Kong ◽  
Yang Liu ◽  
Qianmin Xu ◽  
Kaiqun Wang ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Hela Cells ◽  
Electrode System ◽  
Evaluation Methodology ◽  
Label Free ◽  
Impedance Sensing ◽  
Chemotherapy Drugs ◽  
Substrate Adhesion ◽  
Cell Substrate ◽  
Cell Electrode

Electric cell–substrate impedance sensing exhibits a real-time and label-free feature to monitor the response of cells stimulated by various biochemical and mechanical signals. Alterations in the currents passing through the cell–electrode system characterize the impedance variations of cells. The impedance responses of HeLa cells under distinct chemotherapy drugs combine the effects of cell proliferation and cell–substrate adhesion. Optimal interdigitated electrodes were selected to explore the impedance responses of HeLa cells. Measurements of impedance of cells in response to three widely used chemotherapy drugs in clinical practice, namely cisplatin, doxorubicin, 5-fluorouracil, were performed. The results demonstrated that distinct impedance responses of HeLa cells to drugs were exhibited and a decrease in measured impedance was observed after drug treatment, accompanied by alterations in the distribution and intensity of the adhesion-related protein vinculin and the rate of cell proliferation. The link between the impedance profiles of HeLa cells and their biological functions was developed based on the circuit model. This study demonstrated the weights of cell proliferation and adhesion of HeLa cells under the treatments of DDP, DOX, and 5-FU, resulted in distinct impedance responses of cells, providing an impedance-based evaluation methodology for cervical cancer treatment.

scholarly journals Electric Cell-Substrate Impedance Sensing of Cellular Effects under Hypoxic Conditions and Carbonic Anhydrase Inhibition

2017 ◽  
Vol 2017 ◽  
pp. 1-10
Author(s):  
Luciana Stanica ◽  
Mihnea Rosu-Hamzescu ◽  
Mihaela Gheorghiu ◽  
Miruna Stan ◽  
Loredana Antonescu ◽  
...  
Keyword(s):  
Carbonic Anhydrase ◽  
Real Time ◽  
Ph Regulation ◽  
Electrical Impedance Spectroscopy ◽  
High Temporal Resolution ◽  
Label Free ◽  
Human Colon Cancer ◽  
Impedance Sensing ◽  
Hypoxic Conditions ◽  
Cell Substrate

Tumor hypoxia provides a dynamic environment for the cancer cells to thrive and metastasize. Evaluation of cell growth, cell-cell, and cell surface interactions in hypoxic conditions is therefore highly needed in the establishment of treatment options. Electric cell-substrate impedance sensing (ECIS) has been traditionally used in the evaluation of cellular platforms as a real-time, label-free impedance-based method to study the activities of cells grown in tissue cultures, but its application for hypoxic environments is seldom reported. We present real-time evaluation of hypoxia-induced bioeffects with a focus on hypoxic pH regulation of tumor environment. To this end, multiparametric real-time bioanalytical platform using electrical impedance spectroscopy (EIS) and human colon cancer HT-29 cells is advanced. A time series of EIS data enables monitoring with high temporal resolution the alterations occurring within the cell layer, especially at the cell-substrate level. We reveal the dynamic changes of cellular processes during hypoxic conditions and in response to application of acetazolamide (AZA), a carbonic anhydrase inhibitor. Optical evaluation and pH assessment complemented the electrical analysis towards establishing a pattern of cellular changes. The proposed bioanalytical platform indicates wide applicability towards evaluation of bioeffects of hypoxia at cellular level.

scholarly journals Can ECIS Biosensor Technology Be Used to Measure the Cellular Responses of Glioblastoma Stem Cells?

Biosensors ◽  
2021 ◽  
Vol 11 (12) ◽  
pp. 498
Author(s):  
Laverne Diana Robilliard ◽  
Jane Yu ◽  
Sung-Min Jun ◽  
Akshata Anchan ◽  
Graeme Finlay ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cellular Responses ◽  
Cellular Adhesion ◽  
Label Free ◽  
Migratory Behaviour ◽  
Glioblastoma Cells ◽  
Glioblastoma Stem Cells ◽  
Impedance Sensing ◽  
Cell Substrate ◽  
Cellular Behaviour

Glioblastoma is considered the most aggressive and lethal form of brain cancer. Glioblastoma tumours are complex, comprising a spectrum of oncogenically transformed cells displaying distinct phenotypes. These can be generated in culture and are called differentiated-glioblastoma cells and glioblastoma stem cells. These cells are phenotypically and functionally distinct, where the stem-like glioblastoma cells give rise to and perpetuate the tumour. Electric cell-substrate impedance sensing (ECIS) is a real-time, label-free, impedance-based method for the analysis of cellular behaviour, based on cellular adhesion. Therefore, we asked the question of whether ECIS was suitable for, and capable of measuring the adhesion of glioblastoma cells. The goal was to identify whether ECIS was capable of measuring glioblastoma cell adhesion, with a particular focus on the glioblastoma stem cells. We reveal that ECIS reliably measures adhesion of the differentiated glioblastoma cells on various array types. We also demonstrate the ability of ECIS to measure the migratory behaviour of differentiated glioblastoma cells onto ECIS electrodes post-ablation. Although the glioblastoma stem cells are adherent, ECIS is substantially less capable at reliably measuring their adhesion, compared with the differentiated counterparts. This means that ECIS has applicability for some glioblastoma cultures but much less utility for weakly adherent stem cell counterparts.

scholarly journals Development of a Robust High-Throughput Screening Platform for Inhibitors of the Striatal-Enriched Tyrosine Phosphatase (STEP)

2021 ◽  
Vol 22 (9) ◽  
pp. 4417
Author(s):  
Lester J Lambert ◽  
Stefan Grotegut ◽  
Maria Celeridad ◽  
Palak Gosalia ◽  
Laurent JS De Backer ◽  
...  
Keyword(s):  
Drug Discovery ◽  
High Throughput ◽  
Tyrosine Kinases ◽  
High Throughput Screening ◽  
Kinase Inhibitors ◽  
Tyrosine Phosphatase ◽  
Protein Tyrosine Phosphatases ◽  
Synaptic Function ◽  
Label Free ◽  
Protein Tyrosine

Many human diseases are the result of abnormal expression or activation of protein tyrosine kinases (PTKs) and protein tyrosine phosphatases (PTPs). Not surprisingly, more than 30 tyrosine kinase inhibitors (TKIs) are currently in clinical use and provide unique treatment options for many patients. PTPs on the other hand have long been regarded as “undruggable” and only recently have gained increased attention in drug discovery. Striatal-enriched tyrosine phosphatase (STEP) is a neuron-specific PTP that is overactive in Alzheimer’s disease (AD) and other neurodegenerative and neuropsychiatric disorders, including Parkinson’s disease, schizophrenia, and fragile X syndrome. An emergent model suggests that the increase in STEP activity interferes with synaptic function and contributes to the characteristic cognitive and behavioral deficits present in these diseases. Prior efforts to generate STEP inhibitors with properties that warrant clinical development have largely failed. To identify novel STEP inhibitor scaffolds, we developed a biophysical, label-free high-throughput screening (HTS) platform based on the protein thermal shift (PTS) technology. In contrast to conventional HTS using STEP enzymatic assays, we found the PTS platform highly robust and capable of identifying true hits with confirmed STEP inhibitory activity and selectivity. This new platform promises to greatly advance STEP drug discovery and should be applicable to other PTP targets.

scholarly journals Electric Cell-Substrate Impedance Sensing To Monitor Viral Growth and Study Cellular Responses to Infection with Alphaherpesviruses in Real Time

mSphere ◽  
2017 ◽  
Vol 2 (2) ◽  
Author(s):  
Matthew R. Pennington ◽  
Gerlinde R. Van de Walle
Keyword(s):  
Real Time ◽  
Cellular Response ◽  
Cellular Responses ◽  
Cellular Damage ◽  
Impedance Sensing ◽  
Novel Technologies ◽  
Antiviral Therapies ◽  
Mucosal Surfaces ◽  
Cell Substrate ◽  
Hsv 1

ABSTRACT Alphaherpesviruses, including those that commonly infect humans, such as HSV-1 and HSV-2, typically infect and cause cellular damage to epithelial cells at mucosal surfaces, leading to disease. The development of novel technologies to study the cellular responses to infection may allow a more complete understanding of virus replication and the creation of novel antiviral therapies. This study demonstrates the use of ECIS to study various aspects of herpesvirus biology, with a specific focus on changes in cellular morphology as a result of infection. We conclude that ECIS represents a valuable new tool with which to study alphaherpesvirus infections in real time and in an objective and reproducible manner. Electric cell-substrate impedance sensing (ECIS) measures changes in an electrical circuit formed in a culture dish. As cells grow over a gold electrode, they block the flow of electricity and this is read as an increase in electrical impedance in the circuit. ECIS has previously been used in a variety of applications to study cell growth, migration, and behavior in response to stimuli in real time and without the need for cellular labels. Here, we demonstrate that ECIS is also a valuable tool with which to study infection by alphaherpesviruses. To this end, we used ECIS to study the kinetics of cells infected with felid herpesvirus type 1 (FHV-1), a close relative of the human alphaherpesviruses herpes simplex virus 1 (HSV-1) and HSV-2, and compared the results to those obtained with conventional infectivity assays. First, we demonstrated that ECIS can easily distinguish between wells of cells infected with different amounts of FHV-1 and provides information about the cellular response to infection. Second, we found ECIS useful in identifying differences between the replication kinetics of recombinant DsRed Express2-labeled FHV-1, created via CRISPR/Cas9 genome engineering, and wild-type FHV-1. Finally, we demonstrated that ECIS can accurately determine the half-maximal effective concentration of antivirals. Collectively, our data show that ECIS, in conjunction with current methodologies, is a powerful tool that can be used to monitor viral growth and study the cellular response to alphaherpesvirus infection. IMPORTANCE Alphaherpesviruses, including those that commonly infect humans, such as HSV-1 and HSV-2, typically infect and cause cellular damage to epithelial cells at mucosal surfaces, leading to disease. The development of novel technologies to study the cellular responses to infection may allow a more complete understanding of virus replication and the creation of novel antiviral therapies. This study demonstrates the use of ECIS to study various aspects of herpesvirus biology, with a specific focus on changes in cellular morphology as a result of infection. We conclude that ECIS represents a valuable new tool with which to study alphaherpesvirus infections in real time and in an objective and reproducible manner.

scholarly journals Pilonidal Cyst of the Penis Mimicking Carcinoma

2013 ◽  
Vol 2013 ◽  
pp. 1-2
Author(s):  
Luigi Cormio ◽  
Francesca Sanguedolce ◽  
Paolo Massenio ◽  
Giuseppe Di Fino ◽  
Giuseppe Carrieri
Keyword(s):  
Differential Diagnosis ◽  
Pilonidal Sinus ◽  
Rare Case ◽  
Subcutaneous Tissue ◽  
Sinus Tract ◽  
Penile Carcinoma ◽  
Inflammatory Condition ◽  
Pilonidal Cyst ◽  
Chronic Inflammatory Condition

Pilonidal sinus is a long-standing chronic inflammatory condition consisting of a sinus tract from the skin-lined orifice extending into subcutaneous tissue, with hairs attached to the wall of the tract and projecting outside of the opening. Penile location is rare, and differential diagnosis with severe balanoposthitis, epidermal cysts, and neoplasms can be difficult. We report a rare case of pilonidal cyst located between coronal sulcus and prepuce which, due to its ulcerated aspect and absence of a tract with projecting hairs, simulated a penile carcinoma.

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