scholarly journals 709 Development of a high-throughput, cell-based phenotypic assay to identify novel anti-fibrotic medicines to prevent scar formation after burn injury

2019 ◽  
Vol 139 (5) ◽  
pp. S122
Author(s):  
A.R. Lapthorn ◽  
M.M. Ilg ◽  
J.V. Sullivan ◽  
P. Dziewulski ◽  
S. Cellek
Keyword(s):  
High Throughput ◽  
Burn Injury ◽  
Scar Formation ◽  
Phenotypic Assay

scholarly journals Breeding for boron tolerance in lentil (Lens culinaris Medik.) using a high-throughput phenotypic assay and molecular markers

2018 ◽  
Vol 137 (4) ◽  
pp. 492-501 ◽  
Author(s):  
Matthew S. Rodda ◽  
Shimna Sudheesh ◽  
Muhammad Javid ◽  
Dianne Noy ◽  
Annathurai Gnanasambandam ◽  
...  
Keyword(s):  
Molecular Markers ◽  
High Throughput ◽  
Lens Culinaris ◽  
Boron Tolerance ◽  
Phenotypic Assay

scholarly journals High Content and High Throughout Phenotypic Assay for the Hourly Resolution of the Malaria Parasite Erythrocytic Cycle

2021 ◽  
Author(s):  
Donald Bell ◽  
Sophie Ridewood ◽  
Asha P. Patel ◽  
Sun Hyeok Lee ◽  
Young-Tae Chang ◽  
...  
Keyword(s):  
Drug Resistance ◽  
High Throughput ◽  
Population Level ◽  
Drug Resistant ◽  
Malaria Research ◽  
Phenotypic Assay ◽  
Rbc Membrane ◽  
Erythrocytic Cycle ◽  
Technical Advances ◽  
Drug Resistant Strains

AbstractOver the last 20 years increased funding for malaria research has resulted in very significant technical advances to study the biology of Plasmodium species. High throughput phenotypic assays have been developed to screen millions of compounds and identify small molecules with antiparasitic activity. At the same time, advances in malaria genetic have greatly facilitated the generation of genetically modified parasites, and whole genome genetic screens are now feasible in Plasmodium species. Finally, there has been an increased interest to study malaria parasites at the population level, in particular in the area of drug resistance. Drug resistant field isolates have been collected around the world, and drug resistant strains are routinely generated in the lab to study the mechanisms of drug resistance. As a result, one of the current bottlenecks in malaria research is our ability to quickly characterize the phenotype associated with compound treatment or genetic modification, or to quickly compare differences in intracellular development between strains. Here, we present a high content/high throughput phenotypic assay that combines highly selective RNA, DNA, and RBC membrane dyes to provide hourly resolution of the full erythrocytic cycle for both P. falciparum and P. knowlesi. A flow cytometry assay allows the analysis of samples in a 384-well format and a quick way to determine the parasite developmental stage. On the other hand, the fluorescence microscopy format allows for a detailed visualization of parasite morphology. Finally, using open source software we have developed protocols for the automated cluster analysis of microscopy images. This assay can be applied to any Plasmodium species, requires very little amount of sample, is performed with fixed cells, and is easily scalable. Overall, we believe this assay will be a great tool for the malaria community to study Plasmodium species.

scholarly journals A Microscopic Phenotypic Assay for the Quantification of Intracellular Mycobacteria Adapted for High-throughput/High-content Screening

10.3791/51114 ◽  
2014 ◽  
Author(s):  
Christophe. J Queval ◽  
Ok-Ryul Song ◽  
Vincent Delorme ◽  
Raffaella Iantomasi ◽  
Romain Veyron-Churlet ◽  
...  
Keyword(s):  
High Throughput ◽  
High Content Screening ◽  
Phenotypic Assay

scholarly journals High-Throughput Phenotypic Assay for Compounds That Influence Mitochondrial Health Using iPSC-Derived Human Neurons

2021 ◽  
pp. 247255522110006
Author(s):  
Courtney MacMullen ◽  
Ronald L. Davis
Keyword(s):  
Neurodegenerative Diseases ◽  
High Throughput ◽  
High Throughput Screening ◽  
Mitochondrial Dynamics ◽  
Live Cell ◽  
Phenotypic Assay ◽  
Microtiter Plates ◽  
Human Neurons ◽  
Induced Pluripotent ◽  
And Function

There is a critical need to develop high-throughput assays to identify compounds that offer therapy for individuals suffering from neurodegenerative diseases. Most brain disorders, including neurodegenerative diseases, share the common neuropathology of mitochondria dysfunction, which can lead to apoptosis of neurons, overproduction of reactive oxygen species (ROS), and other cellular neuropathologies characteristic of these diseases. Human induced pluripotent stem cells (iPSCs) with a stable genomic insertion of the neurogenin-2 transcription factor under the control of the TetOn promoter can be differentiated into excitatory human neurons (i3Neurons) within 3 days of exposure to doxycycline. These neurons have been used to develop and validate a live-cell assay for parameters of mitochondrial dynamics and function using two compounds known to promote mitochondrial elongation in mouse neurons, 4-hydroxychalcone and 2,4-dihyrdroxychalcone. The assay involves plating the neurons in 384-well microtiter plates, treating them with known or unknown substances, and then capturing morphological information for the neuronal mitochondria using a lentivirus vector to express a mitochondrial-targeted fluorescence reporter. The i3Neuron cultures exposed to these two compounds for 24 h exhibit significantly decreased circularity and significantly increased length compared to controls, two morphological parameters correlated with increased mitochondrial health. The assay is rapid, with results obtained after a one-week-long i3Neuron culture or one month if neurons are co-cultured with astrocytes. This live-cell, mitochondrial phenotypic assay can be used for high-throughput screening or as an orthogonal assay for compounds obtained via other high-throughput screening campaigns.

The Use of Dermal Substitute in Deep Burns of Functional/Mobile Anatomic Areas at Acute Phase After Early Excision and Subsequent Skin Autografting: Dermal Substitute Prevents Functional Limitations

2020 ◽  
Vol 41 (5) ◽  
pp. 1079-1083
Author(s):  
Kayhan Gurbuz ◽  
Mete Demir ◽  
Koray Das
Keyword(s):  
Acute Phase ◽  
Skin Graft ◽  
Burn Injury ◽  
Functional Limitations ◽  
Graft Loss ◽  
Scar Formation ◽  
Functional Evaluation ◽  
Dermal Substitute ◽  
Major Burn ◽  
Skin Autografting

Abstract We aimed to evaluate the results of dermal substitute implantation after early excision in the acute phase of major burn cases within the scope of efforts to reduce contractions and scar formation in functional anatomic areas (face, neck, axilla, elbow, popliteal). Twelve patients with major burn who were treated in the burn center between September 2017 and September 2018 were included in the study. In these patients, Nevelia® dermal substitute was implanted into 24 functional areas with deep partial or full-thickness burns after surgical debridement of the wound. Autologous split-thickness skin graft was applied to these areas after 14 to 21 days. The patients were followed for 4 to 14 months (mean 6 months). Postoperative scar formation was assessed by the Vancouver Scar Scale at the end of the follow-up period. A simple qualitative staging system was used for aesthetic and functional evaluation. The time from burn injury to dermal substitute implantation was 3 to 21 days. Skin graft take was complete in 22 of 24 regions and partial in one of them, while graft loss developed in one region. In the implantation sites, the Vancouver Scar Scale ranged from 1 to 7. The aesthetic and functional evaluation showed excellent/good results in 21 of 24 anatomic regions, moderate results in 2 regions, and poor results in 1 region. The use of dermal substitute in deep burns of functional/mobile anatomic areas at the acute phase after early excision and subsequent skin autografting has opened a new alternative area in the burn surgery arena to prevent contractures and functional limitations.

Clodronate liposomes reduce excessive scar formation in a mouse model of burn injury by reducing collagen deposition and TGF-β1 expression

2014 ◽  
Vol 41 (4) ◽  
pp. 2143-2149 ◽  
Author(s):  
Shao-wei Lu ◽  
Xing-mei Zhang ◽  
Hong-min Luo ◽  
Yu-cai Fu ◽  
Ming-yan Xu ◽  
...  
Keyword(s):  
Mouse Model ◽  
Burn Injury ◽  
Scar Formation ◽  
Collagen Deposition ◽  
Clodronate Liposomes ◽  
Tgf Β1

Silicone Gel and Hypertrophic Scar Formation: A Literature Review

1992 ◽  
Vol 59 (2) ◽  
pp. 78-86 ◽  
Author(s):  
Kathy Farquhar
Keyword(s):  
Hypertrophic Scar ◽  
Burn Injury ◽  
Statistical Significance ◽  
Treatment Protocol ◽  
Scar Formation ◽  
Treatment Technique ◽  
Hypertrophic Scarring ◽  
Duration Of Treatment ◽  
Silicone Gel ◽  
Treatment Techniques

Since 1982 a number of studies have investigated the use of silicone gel sheets in the prevention and reduction of hypertrophic scar formation. The statistical significance of several of these studies has been reduced by poor standardization of research methods, and the lack of a reliable and valid method of assessing hypertrophic scarring. When these studies are considered together, however, several clinically significant trends become apparent. The application of silicone gel appears to normalize the texture, colour, evaluation, and occurrence of subjective complaints (i.e., pain and pruritis) typically associated with hypertrophic scarring. These results are independent of the patient's age, method of attachment of the gel, or the location, age, or cause of the scar. The most appropriate treatment protocol for the application of silicone gel has yet to be determined. Current literature suggests that daily wearing tolerance be developed over the course of one to two weeks, the gel and scar be cleaned twice daily, and skin monitored regularly for signs of irritation. The optimal duration of treatment has not been established. Further research using standardized procedures and valid, reliable methods of measurement are required before this treatment can be ethically used to replace more established treatment techniques. However, the literature currently available would indicate that silicone gel may be an effective treatment technique for the prevention and reduction of hypertrophic scar formation following burn injury.

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