scholarly journals An automated high-throughput system for phenotypic screening of chemical libraries onC. elegansand parasitic nematodes

2017 ◽  
Author(s):  
Frederick A. Partridge ◽  
Anwen E. Brown ◽  
Steven D. Buckingham ◽  
Nicky J. Willis ◽  
Graham M. Wynne ◽  
...  
Keyword(s):  
High Throughput ◽  
Crop Damage ◽  
Parasitic Nematodes ◽  
Chemical Library ◽  
Trichuris Muris ◽  
Chemical Screening ◽  
Automated Phenotyping ◽  
Global Threat ◽  
Therapeutic Compounds ◽  
Parasitic Worms

AbstractParasitic nematodes infect hundreds of millions of people and farmed livestock. Further, plant parasitic nematodes result in major crop damage. The pipeline of therapeutic compounds is limited and parasite resistance to the existing anthelmintic compounds is a global threat. We have developed an INVertebrate Automated Phenotyping Platform (INVAPP) for high-throughput, plate-based chemical screening, and an algorithm (Paragon) which allows screening for compounds that have an effect on motility and development of parasitic worms. We have validated its utility by determining the efficacy of a panel of known anthelmintics against model and parasitic nematodes:Caenorhabditis elegans, Haemonchus contortus, Teladorsagia circumcincta, andTrichuris muris. We then applied the system to screen the Pathogen Box chemical library in a blinded fashion and identified known anthelmintics, including tolfenpyrad, auranofin, and mebendazole and 14 compounds previously undescribed as anthelmintics, including benzoxaborole and isoxazole chemotypes. This system offers an effective, high-throughput system for the discovery of novel anthelmintics.

scholarly journals Automated phenotyping of mosquito larvae enables high-throughput screening for novel larvicides and smartphone-based detection of larval insecticide resistance

2020 ◽  
Author(s):  
Steven D. Buckingham ◽  
Frederick A. Partridge ◽  
Beth C. Poulton ◽  
Ben Miller ◽  
Rachel A. McKendry ◽  
...  
Keyword(s):  
Insecticide Resistance ◽  
High Throughput ◽  
High Throughput Screening ◽  
Pyrethroid Resistance ◽  
Standard Test ◽  
Mosquito Larvae ◽  
Mosquito Vector ◽  
Chemical Screening ◽  
Field Assay ◽  
Automated Phenotyping

AbstractPyrethroid-impregnated nets have contributed significantly to halving the burden of malaria but resistance threatens their future efficacy and the pipeline of new insecticides is short. Here we report that an invertebrate automated phenotyping platform (INVAPP), combined with the algorithm Paragon, provides a robust system for measuring larval motility in Anopheles gambiae (and An. coluzzi) as well as Aedes aegypti with the capacity for high-throughput screening for new larvicides. By this means, we reliably quantified both time- and concentration-dependent actions of chemical insecticides faster than using the WHO standard larval assay. We illustrate the effectiveness of the system using an established larvicide (temephos) and demonstrate its capacity for library-scale chemical screening using the Medicines for Malaria (MFP) Pathogen-Box library. As a proof-of-principle, this library screen identified a compound, subsequently confirmed to be tolfenpyrad, as an effective larvicide. We have also used the INVAPP / Paragon system to enable detection of resistance to deltamethrin. We show how this approach to monitoring larval susceptibility to insecticides can be adapted for use with a smartphone camera application and therefore has potential for further development as a simple portable field-assay for insecticide resistance with associated real-time, geo-located information to identify hotspots.Author summaryWe have developed an automated platform for recording the motility of mosquito larvae and applied it to larvae of a mosquito vector of malaria and a mosquito vector of dengue, Zika, yellow fever and other human diseases. The platform facilitates high-throughput, chemical screening for new compounds to control mosquito larvae and also allows detection of larval resistance to the pyrethroid insecticide deltamethrin. Pyrethroid-impregnated bednets have helped to halve the deaths from malaria in recent years but pyrethroid resistance is an important threat to this progress. Our approach assays insecticide actions faster than the current WHO standard test and we show that it can be adapted for use with a smartphone, which offers the prospect of a future field assay for resistance with the added benefit of precise satellite-based location.

scholarly journals Automated phenotyping of mosquito larvae enables high-throughput screening for novel larvicides and offers potential for smartphone-based detection of larval insecticide resistance

2021 ◽  
Vol 15 (6) ◽  
pp. e0008639
Author(s):  
Steven D. Buckingham ◽  
Frederick A. Partridge ◽  
Beth C. Poulton ◽  
Benjamin S. Miller ◽  
Rachel A. McKendry ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Mosquito Larvae ◽  
Chemical Screening ◽  
Field Assay ◽  
Library Screen ◽  
Automated Phenotyping ◽  
Robust System ◽  
Further Development ◽  
Proof Of Principle

Pyrethroid-impregnated nets have contributed significantly to halving the burden of malaria but resistance threatens their future efficacy and the pipeline of new insecticides is short. Here we report that an invertebrate automated phenotyping platform (INVAPP), combined with the algorithm Paragon, provides a robust system for measuring larval motility in Anopheles gambiae (and An. coluzzi) as well as Aedes aegypti with the capacity for high-throughput screening for new larvicides. By this means, we reliably quantified both time- and concentration-dependent actions of chemical insecticides faster than using the WHO standard larval assay. We illustrate the effectiveness of the system using an established larvicide (temephos) and demonstrate its capacity for library-scale chemical screening using the Medicines for Malaria Venture (MMV) Pathogen Box library. As a proof-of-principle, this library screen identified a compound, subsequently confirmed to be tolfenpyrad, as an effective larvicide. We have also used the INVAPP / Paragon system to compare responses in larvae derived from WHO classified deltamethrin resistant and sensitive mosquitoes. We show how this approach to monitoring larval response to insecticides can be adapted for use with a smartphone camera application and therefore has potential for further development as a simple portable field-assay with associated real-time, geo-located information to identify hotspots.

scholarly journals High-Throughput Phenotypic Assay to Screen for Anthelmintic Activity on Haemonchus contortus

2021 ◽  
Vol 14 (7) ◽  
pp. 616
Author(s):  
Aya C. Taki ◽  
Joseph J. Byrne ◽  
Tao Wang ◽  
Brad E. Sleebs ◽  
Nghi Nguyen ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Small Molecules ◽  
High Throughput ◽  
Haemonchus Contortus ◽  
Well Being ◽  
World Health ◽  
Infrared Light ◽  
Parasitic Nematodes ◽  
Parasitic Worms ◽  
Anthelmintic Drugs

Parasitic worms cause very significant diseases in animals and humans worldwide, and their control is critical to enhance health, well-being and productivity. Due to widespread drug resistance in many parasitic worms of animals globally, there is a major, continuing demand for the discovery and development of anthelmintic drugs for use to control these worms. Here, we established a practical, cost-effective and semi-automated high throughput screening (HTS) assay, which relies on the measurement of motility of larvae of the barber’s pole worm (Haemonchus contortus) using infrared light-interference. Using this assay, we screened 80,500 small molecules and achieved a hit rate of 0.05%. We identified three small molecules that reproducibly inhibited larval motility and/or development (IC50 values of ~4 to 41 µM). Future work will critically assess the potential of selected hits as candidates for subsequent optimisation or repurposing against parasitic nematodes. This HTS assay has a major advantage over most previous assays in that it achieves a ≥ 10-times higher throughput (i.e., 10,000 compounds per week), and is thus suited to the screening of libraries of tens of thousands to hundreds of thousands of compounds for subsequent hit-to-lead optimisation or effective repurposing and development. The current assay should be adaptable to many socioeconomically important parasitic nematodes, including those that cause neglected tropical diseases (NTDs). This aspect is of relevance, given the goals of the World Health Organization (WHO) Roadmap for NTDs 2021–2030, to develop more effective drugs and drug combinations to improve patient outcomes and circumvent the ineffectiveness of some current anthelmintic drugs and possible drug resistance.

scholarly journals Identification of a juvenile-hormone signaling inhibitor via high-throughput screening of a chemical library

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Takumi Kayukawa ◽  
Kenjiro Furuta ◽  
Keisuke Nagamine ◽  
Tetsuro Shinoda ◽  
Kiyoaki Yonesu ◽  
...  
Keyword(s):  
Juvenile Hormone ◽  
Cell Line ◽  
Signaling Pathway ◽  
High Throughput ◽  
High Throughput Screening ◽  
Large Scale ◽  
Ex Vivo ◽  
Agricultural Field ◽  
Chemical Library ◽  
Field Development

Abstract Insecticide resistance has recently become a serious problem in the agricultural field. Development of insecticides with new mechanisms of action is essential to overcome this limitation. Juvenile hormone (JH) is an insect-specific hormone that plays key roles in maintaining the larval stage of insects. Hence, JH signaling pathway is considered a suitable target in the development of novel insecticides; however, only a few JH signaling inhibitors (JHSIs) have been reported, and no practical JHSIs have been developed. Here, we established a high-throughput screening (HTS) system for exploration of novel JHSIs using a Bombyx mori cell line (BmN_JF&AR cells) and carried out a large-scale screening in this cell line using a chemical library. The four-step HTS yielded 69 compounds as candidate JHSIs. Topical application of JHSI48 to B. mori larvae caused precocious metamorphosis. In ex vivo culture of the epidermis, JHSI48 suppressed the expression of the Krüppel homolog 1 gene, which is directly activated by JH-liganded receptor. Moreover, JHSI48 caused a parallel rightward shift in the JH response curve, suggesting that JHSI48 possesses a competitive antagonist-like activity. Thus, large-scale HTS using chemical libraries may have applications in development of future insecticides targeting the JH signaling pathway.

MP44-19 HIGH-THROUGHPUT CHEMICAL SCREENING FOR SENSITIZATION OF BLADDER CANCER TO GEMCITABINE AND CISPLATIN CHEMOTHERAPY

2017 ◽  
Vol 197 (4S) ◽  
Author(s):  
Yuki Kita ◽  
Takashi Kobayashi ◽  
Atsuro Sawada ◽  
Ryouichi Saito ◽  
Toshinari Yamasaki ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
High Throughput ◽  
Chemical Screening ◽  
Gemcitabine And Cisplatin

scholarly journals Prospects of Using High-Throughput Proteomics to Underpin the Discovery of Animal Host–Nematode Interactions

Pathogens ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 825
Author(s):  
Tao Wang ◽  
Robin Gasser
Keyword(s):  
Mass Spectrometry ◽  
High Throughput ◽  
Anthelmintic Resistance ◽  
Nematode Species ◽  
Economic Losses ◽  
Future Research ◽  
Parasitic Nematodes ◽  
Public Health Burden ◽  
Host Parasite Interactions ◽  
Host Parasite

Parasitic nematodes impose a significant public health burden, and cause major economic losses to agriculture worldwide. Due to the widespread of anthelmintic resistance and lack of effective vaccines for most nematode species, there is an urgent need to discover novel therapeutic and vaccine targets, informed through an understanding of host–parasite interactions. Proteomics, underpinned by genomics, enables the global characterisation proteins expressed in a particular cell type, tissue and organism, and provides a key to insights at the host–parasite interface using advanced high-throughput mass spectrometry-based proteomic technologies. Here, we (i) review current mass-spectrometry-based proteomic methods, with an emphasis on a high-throughput ‘bottom-up’ approach; (ii) summarise recent progress in the proteomics of parasitic nematodes of animals, with a focus on molecules inferred to be involved in host–parasite interactions; and (iii) discuss future research directions that could enhance our knowledge and understanding of the molecular interplay between nematodes and host animals, in order to work toward new, improved methods for the treatment, diagnosis and control of nematodiases.

Unlocking the Potential of High-Throughput Experimentation for Electrochemistry with a Standardized Microscale Reactor

2021 ◽  
Author(s):  
Jonas Rein ◽  
James R. Annand ◽  
Michael K. Wismer ◽  
Jiantao Fu ◽  
Juno C. Siu ◽  
...  
Keyword(s):  
High Throughput ◽  
Electrochemical Reaction ◽  
Electrochemical Reactor ◽  
Constant Current ◽  
Library Synthesis ◽  
Chemical Library ◽  
Reaction Discovery ◽  
Reaction Optimization ◽  
High Throughput Experimentation ◽  
Broad Array

Organic electrochemistry has emerged as an enabling and sustainable technology in modern organic synthesis. Despite the recent renaissance of electrosynthesis, the broad adoption of electrochemistry in the synthetic community and, especially in industrial settings, has been hindered by the dearth of general, standardized platforms for high-throughput experimentation (HTE). Herein, we disclose the design of the HT<i>e<sup>-</sup></i>Chem, a high-throughput microscale electrochemical reactor that is compatible with existing HTE infrastructure, and enables rapid evaluation of a broad array of electrochemical reaction parameters. Utilizing the HT<i>e<sup>-</sup></i>Chem to accelerate reaction optimization, reaction discovery, and chemical library synthesis is illustrated using a suite of oxidative and reductive transformations under constant current, constant voltage, and electrophotochemical conditions.

scholarly journals Advances in the Use of Microfluidics to Study Crystallization Fundamentals

2019 ◽  
Vol 10 (1) ◽  
pp. 59-83 ◽  
Author(s):  
Nadine Candoni ◽  
Romain Grossier ◽  
Mehdi Lagaize ◽  
Stéphane Veesler
Keyword(s):  
Phase Diagram ◽  
Diffraction Analysis ◽  
High Throughput ◽  
Concentration Measurement ◽  
Ultraviolet Spectroscopy ◽  
X Ray Diffraction ◽  
Chemical Library ◽  
Microfluidic Platform ◽  
X Ray ◽  
Phase Diagram Determination

This review compares droplet-based microfluidic systems used to study crystallization fundamentals in chemistry and biology. An original high-throughput droplet-based microfluidic platform is presented. It uses nanoliter droplets, generates a chemical library, and directly solubilizes powder, thus economizing both material and time. It is compatible with all solvents without the need for surfactant. Its flexibility permits phase diagram determination and crystallization studies (screening and optimizing experiments) and makes it easy to use for nonspecialists in microfluidics. Moreover, it allows concentration measurement via ultraviolet spectroscopy and solid characterization via X-ray diffraction analysis.

scholarly journals Using Weighted Entropy to Rank Chemicals in Quantitative High-Throughput Screening Experiments

2013 ◽  
Vol 19 (3) ◽  
pp. 344-353 ◽  
Author(s):  
Keith R. Shockley
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Nonlinear Models ◽  
Hill Equation ◽  
Parameter Estimates ◽  
Chemical Library ◽  
Data Set ◽  
Screening Experiments ◽  
The Hill ◽  
Weighted Entropy

Quantitative high-throughput screening (qHTS) experiments can simultaneously produce concentration-response profiles for thousands of chemicals. In a typical qHTS study, a large chemical library is subjected to a primary screen to identify candidate hits for secondary screening, validation studies, or prediction modeling. Different algorithms, usually based on the Hill equation logistic model, have been used to classify compounds as active or inactive (or inconclusive). However, observed concentration-response activity relationships may not adequately fit a sigmoidal curve. Furthermore, it is unclear how to prioritize chemicals for follow-up studies given the large uncertainties that often accompany parameter estimates from nonlinear models. Weighted Shannon entropy can address these concerns by ranking compounds according to profile-specific statistics derived from estimates of the probability mass distribution of response at the tested concentration levels. This strategy can be used to rank all tested chemicals in the absence of a prespecified model structure, or the approach can complement existing activity call algorithms by ranking the returned candidate hits. The weighted entropy approach was evaluated here using data simulated from the Hill equation model. The procedure was then applied to a chemical genomics profiling data set interrogating compounds for androgen receptor agonist activity.

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