High throughput screening methods for freeze-drying of fungi and mammalian cells

Mosquito-borne arthropod-borne viruses (arboviruses) such as the dengue virus (DENV), Zika virus (ZIKV), and chikungunya virus (CHIKV) are important human pathogens that are responsible for significant global morbidity and mortality. The recent emergence and re-emergence of mosquito-borne viral diseases (MBVDs) highlight the urgent need for safe and effective vaccines, therapeutics, and vector-control approaches to prevent MBVD outbreaks. In nature, arboviruses circulate between vertebrate hosts and arthropod vectors; therefore, disrupting the virus lifecycle in mosquitoes is a major approach for combating MBVDs. Several strategies were proposed to render mosquitoes that are refractory to arboviral infection, for example, those involving the generation of genetically modified mosquitoes or infection with the symbiotic bacterium Wolbachia. Due to the recent development of high-throughput screening methods, an increasing number of drugs with inhibitory effects on mosquito-borne arboviruses in mammalian cells were identified. These antivirals are useful resources that can impede the circulation of arboviruses between arthropods and humans by either rendering viruses more vulnerable in humans or suppressing viral infection by reducing the expression of host factors in mosquitoes. In this review, we summarize recent advances in small-molecule antiarboviral drugs in mammalian and mosquito cells, and discuss how to use these antivirals to block the transmission of MBVDs.

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Identification of Anti-malarial Drug Candidate Inhibitors using Ultra-high Throughput Screening Methods and Subsequent Mechanism of Action Studies Aimed at the Treatment and Prevention of Plasmodium falciparum

10.14264/uql.2019.548 ◽

2019 ◽

Author(s):

Timothy Patrick Spicer

Keyword(s):

Plasmodium Falciparum ◽

High Throughput ◽

Mechanism Of Action ◽

High Throughput Screening ◽

Drug Candidate ◽

Screening Methods ◽

Treatment And Prevention

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High throughput method development and optimised production of leaf protein concentrates with potential to support the agri-industry

Journal of Food Measurement & Characterization ◽

10.1007/s11694-021-01136-w ◽

2021 ◽

Author(s):

Ajay Iyer ◽

Lisa Guerrier ◽

Salomé Leveque ◽

Charles S. Bestwick ◽

Sylvia H. Duncan ◽

...

Keyword(s):

Amino Acid ◽

High Throughput ◽

High Throughput Screening ◽

Protein Extraction ◽

Leaf Protein ◽

Protein Recovery ◽

Screening Methods ◽

Extraction Technology ◽

Protein Concentrates ◽

Ethanol Precipitation

AbstractInvasive plants offer an interesting and unconventional source of protein and the considerable investment made towards their eradication can potentially be salvaged through their revalorisation. To identify viable sources, effective and high-throughput screening methods are required, as well as efficient procedures to isolate these components. Rigorous assessment of low-cost, high-throughput screening assays for total sugar, phenolics and protein was performed, and ninhydrin, Lever and Fast Blue assays were found to be most suitable owing to high reliability scores and false positive errors less than 1%. These assays were used to characterise invasive Scottish plants such as Gorse (Ulex europeans), Broom (Cystisus scoparius) and Fireweed (Chamaenerion angustifolium). Protein extraction (alkali-, heat- and enzyme assisted) were tested on these plants, and further purification (acid and ethanol precipitation, as well as ultrafiltration) procedures were tested on Gorse, based on protein recovery values. Cellulase treatment and ethanol precipitation gave the highest protein recovery (64.0 ± 0.5%) and purity (96.8 ± 0.1%) with Gorse. The amino acid profile of the purified protein revealed high levels of essential amino acids (34.8 ± 0.0%). Comparison of results with preceding literature revealed a strong association between amino acid profiles and overall protein recovery with the extraction method employed. The final purity of the protein concentrates was closely associated to the protein content of the initial plant mass. Leaf protein extraction technology can effectively raise crop harvest indices, revalorise underutilised plants and waste streams.

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Evaluation of Methods To Assess the Biofilm-Forming Ability of Listeria monocytogenes

Journal of Food Protection ◽

10.4315/0362-028x.jfp-11-464 ◽

2012 ◽

Vol 75 (8) ◽

pp. 1411-1417 ◽

Cited By ~ 25

Author(s):

ANTÓNIO LOURENÇO ◽

FRANCISCO REGO ◽

LUISA BRITO ◽

JOSEPH F. FRANK

Keyword(s):

Stainless Steel ◽

Listeria Monocytogenes ◽

High Throughput ◽

Biofilm Formation ◽

High Throughput Screening ◽

Screening Methods ◽

Genetic Lineage ◽

Production Lines ◽

Genetic Characteristics

The contamination of ready-to-eat products with Listeria monocytogenes has been related to the presence of biofilms in production lines, as biofilms protect cells from chemical sanitizers. The ability of L. monocytogenes to produce biofilms is often evaluated using in vitro methodologies. This work aims to compare the most frequently used methodologies, including high-throughput screening methods based on microplates (crystal violet and the Calgary Biofilm Device) and methods based on CFU enumeration and microscopy after growth on stainless steel. Thirty isolates with diverse origins and genetic characteristics were evaluated. No (or low) correlations between methods were observed. The only significant correlation was found between the methods using stainless steel. No statistically significant correlation (P > 0.05) was detected among genetic lineage, serovar, and biofilm-forming ability. Because results indicate that biofilm formation is influenced by the surface material, the extrapolation of results from high-throughput methods using microplates to more industrially relevant surfaces should be undertaken with caution.

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High-throughput Screening Methods Developed for Oxidoreductases

Enzyme Assays ◽

10.1002/3527607846.ch3 ◽

2006 ◽

pp. 77-93 ◽

Cited By ~ 6

Author(s):

Tyler W. Johannes ◽

Ryan D. Woodyer ◽

Huimin Zhao

Keyword(s):

High Throughput ◽

High Throughput Screening ◽

Screening Methods

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Paper-Based SERS Platform for One-Step Screening of Tetracycline in Milk

Scientific Reports ◽

10.1038/s41598-019-54380-y ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 4

Author(s):

Ana Marques ◽

Bruno Veigas ◽

Andreia Araújo ◽

Beatriz Pagará ◽

Pedro Viana Baptista ◽

...

Keyword(s):

High Throughput ◽

High Throughput Screening ◽

Point Of Care ◽

Dairy Industry ◽

Principal Component ◽

Surface Enhanced Raman Spectroscopy ◽

Economic Losses ◽

Screening Methods ◽

Sers Substrate ◽

Molecular Signatures

AbstractThroughout the last decade, the expansion of food testing has been gradually moving towards ordinary high throughput screening methods performed on-site. The demand for point-of-care testing, able to distinguish molecular signatures with high accuracy, sensitivity and specificity has been significantly increasing. This new requirement relies on the on-site detection and monitorization of molecular signatures suitable for the surveillance of food production and processing. The widespread use of antibiotics has contributed to disease control of livestock but has also created problems for the dairy industry and consumers. Its therapeutic and subtherapeutic use has increased the risk of contamination in milk in enough concentrations to cause economic losses to the dairy industry and have a health impact in highly sensitive individuals. This study focuses on the development of a simple Surface-Enhanced Raman Spectroscopy (SERS) method for fast high throughput screening of tetracycline (TET) in milk. For this, we integrate a paper-based low-cost, fully recyclable and highly stable SERS platform, with a minimal sample preparation protocol. A two-microliter sample of milk solutions spiked with TET (from 0.01 to 1000 ppm) is dried on a silver nanoparticle coated cardboard substrate and measured via a Raman spectrophotometer. The SERS substrate showed to be extremely stable with a shelf life of several months. A global spectrum principal component analysis approach was used to test all the detected vibrational modes and their correlation with TET concentration. Peak intensity ratios (455 cm−1/1280 cm−1 and 874 cm−1/1397 cm−1) were found to be correlated with TET concentrations in milk, achieving a sensitivity as low as 0.1 ppm. Results indicate that this SERS method combined with portable Raman spectrometer is a potential tool that can be used on-site for the monitoring of TET residues and other antibiotics.

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