Cell-based models for mycotoxin screening and toxicity evaluation: an update

2014 ◽  
Vol 7 (2) ◽  
pp. 153-166 ◽  
Author(s):  
F. Cheli ◽  
E. Fusi ◽  
A. Baldi

This review presents the applications of cell-based models in mycotoxin research, with a focus on models for mycotoxin screening and cytotoxicity evaluation. Various cell-based models, cell and cell culture condition related factors, toxicity endpoints and culture systems as well as predictive value of cell-based bioassays are reviewed. Advantages, drawbacks and technical problems regarding set up and validation of consistent, robust, reproducible and high-throughput cell-based models are discussed. Various cell-based models have been developed and used as screening tests for mycotoxins but the data obtained are difficult to compare. However, the results highlight the potential of cell-based models as promising in vitro platforms for the initial screening and cytotoxicity evaluation of mycotoxins and as a significant analytical approach in mycotoxin research before any animal or human clinical studies. To develop cell-based models as powerful high-throughput laboratory platforms for the analysis of large numbers of samples, there are mainly two fundamental requirements that should be met, i.e. the availability of easy-to-use and, if possible, automated cell platforms and the possibility to obtain reproducible results that are comparable between laboratories. The transition from a research model to a test model still needs optimisation, standardisation, and validation of analytical protocols. The validation of a cell-based bioassay is a complex process, as several critical points, such as the choice of the cellular model, the assay procedures, and the appropriate use and interpretation of the results, must be strictly defined to ensure more consistency in the results. The development of cell-based models exploring the third dimension together with automation and miniaturisation will bring cellular platforms to a level appropriate for cost-effective and large-scale analysis in the field of mycotoxin research.

Sensors ◽  
2021 ◽  
Vol 21 (3) ◽  
pp. 736
Author(s):  
Kamila Malecka ◽  
Edyta Mikuła ◽  
Elena E. Ferapontova

Improved outcomes for many types of cancer achieved during recent years is due, among other factors, to the earlier detection of tumours and the greater availability of screening tests. With this, non-invasive, fast and accurate diagnostic devices for cancer diagnosis strongly improve the quality of healthcare by delivering screening results in the most cost-effective and safe way. Biosensors for cancer diagnostics exploiting aptamers offer several important advantages over traditional antibodies-based assays, such as the in-vitro aptamer production, their inexpensive and easy chemical synthesis and modification, and excellent thermal stability. On the other hand, electrochemical biosensing approaches allow sensitive, accurate and inexpensive way of sensing, due to the rapid detection with lower costs, smaller equipment size and lower power requirements. This review presents an up-to-date assessment of the recent design strategies and analytical performance of the electrochemical aptamer-based biosensors for cancer diagnosis and their future perspectives in cancer diagnostics.


Reproduction ◽  
2021 ◽  
Author(s):  
Zoe Claire Johnston ◽  
Franz S Gruber ◽  
Sean Brown ◽  
Neil R Norcross ◽  
Jason R Swedlow ◽  
...  

Despite recent advances in male reproductive health research, there remain many elements of male (in)fertility where our understanding is incomplete. Consequently, diagnostic tools and treatments for men with sperm dysfunction, other than medically assisted reproduction, are limited. On the other hand, the gaps in our knowledge of the mechanisms which underpin sperm function have hampered the development of male non-hormonal contraceptives. The study of mature spermatozoa is inherently difficult. They are a unique and highly specialised cell type which does not actively transcribe or translate proteins and cannot be cultured for long periods of time or matured in vitro. One, large scale, approach to both increasing understanding of sperm function, and the discovery and development of compounds that can modulate sperm function, is to directly observe responses to compounds with phenotypic screening techniques. These target agnostic approaches can be developed into high-throughput screening platforms with the potential to drastically increase advances in the field. Here we discuss the rationale and development of high-throughput phenotypic screening platforms for mature human spermatozoa, and the multiple potential applications these present, as well as the current limitations and leaps in our understanding and capabilities needed to overcome them. Further development and use of these technologies could lead to the identification of compounds which positively or negatively affect sperm cell motility or function, or novel platforms for toxicology or environmental chemical testing among other applications. Ultimately, each of these potential applications is also likely to increase understanding within the field of sperm biology.


2018 ◽  
Vol 46 (08) ◽  
pp. 1825-1840 ◽  
Author(s):  
Taiyi Wang ◽  
Xiaonan Chen ◽  
Jiahui Yu ◽  
Qunqun Du ◽  
Jie Zhu ◽  
...  

Although the efficacy and the health care advantages of Chinese herbal medicine (CHM) have become increasingly recognized worldwide, the potential side effects and toxicity still restrict its broader application. This study established and applied an integrated platform anchored on automatic patch clamp system to screen and evaluate a collection of CHM extracts, compositions and monomeric compounds for in vitro cardiac toxicity. Of 1036 CHM samples screened, 2.79% significantly inhibited hERG channel activity. Among them, Strychnine was identified for the first time as a potent hERG inhibitor with an IC[Formula: see text] of [Formula: see text]M in comparison to that of Dofetilide at [Formula: see text]M and Quinidine at [Formula: see text]M. Langendorff-perfusion experiments confirmed that strychnine increased QT interphase from [Formula: see text][Formula: see text]ms to [Formula: see text][Formula: see text]ms and decreased heart rates from [Formula: see text][Formula: see text]bmp to [Formula: see text][Formula: see text]bmp in isolated rat hearts. The cardiac toxicity effect of strychnine appears to be specific to hERG channel since an in vitro multiplex imaging analysis showed that it did not affect cellular phenotypes such as cell vitality, nucleus area, mitochondria mass and function, nor intracellular calcium in rat primary myocytes. This integrated high-throughput hERG patch clamp and high-content multi-parameter imaging cardiac toxicity screen approach should be useful for large-scale preclinical evaluation of complex Chinese herbal medicine.


2019 ◽  
Vol 48 (1) ◽  
pp. 1-18 ◽  
Author(s):  
Celia Blanco ◽  
Evan Janzen ◽  
Abe Pressman ◽  
Ranajay Saha ◽  
Irene A. Chen

The function of fitness (or molecular activity) in the space of all possible sequences is known as the fitness landscape. Evolution is a random walk on the fitness landscape, with a bias toward climbing hills. Mapping the topography of real fitness landscapes is fundamental to understanding evolution, but previous efforts were hampered by the difficulty of obtaining large, quantitative data sets. The accessibility of high-throughput sequencing (HTS) has transformed this study, enabling large-scale enumeration of fitness for many mutants and even complete sequence spaces in some cases. We review the progress of high-throughput studies in mapping molecular fitness landscapes, both in vitro and in vivo, as well as opportunities for future research. Such studies are rapidly growing in number. HTS is expected to have a profound effect on the understanding of real molecular fitness landscapes.


2021 ◽  
Vol 9 (11) ◽  
pp. 2373
Author(s):  
Rima Jeske ◽  
Larissa Dangel ◽  
Leander Sauerbrey ◽  
Dimitrios Frangoulidis ◽  
Lauren R. Teras ◽  
...  

The causative agent of Q fever, the bacterium Coxiella burnetii (C. burnetii), has gained increasing interest due to outbreak events and reports about it being a potential risk factor for the development of lymphomas. In order to conduct large-scale studies for population monitoring and to investigate possible associations more closely, accurate and cost-effective high-throughput assays are highly desired. To address this need, nine C. burnetii proteins were expressed as recombinant antigens for multiplex serology. This technique enables the quantitative high-throughput detection of antibodies to multiple antigens simultaneously in a single reaction. Based on a reference group of 76 seropositive and 91 seronegative sera, three antigens were able to detect C. burnetii infections. Com1, GroEL, and DnaK achieved specificities of 93%, 69%, and 77% and sensitivities of 64%, 72%, and 47%, respectively. Double positivity to Com1 and GroEL led to a combined specificity of 90% and a sensitivity of 71%. In a subgroup of seropositives with an increased risk for chronic Q fever, the double positivity to these markers reached a specificity of 90% and a sensitivity of 86%. Multiplex serology enables the detection of antibodies against C. burnetii and appears well-suited to investigate associations between C. burnetii infections and the clinical manifestations in large-scale studies.


2020 ◽  
Author(s):  
Namrata Singh ◽  
Komal Patel ◽  
Ambuja Navalkar ◽  
Pradeep Kadu ◽  
Debalina Datta ◽  
...  

AbstractBiomaterials mimicking extracellular matrices (ECM) for three-dimensional (3D) cultures have gained immense interest in tumor modeling and in vitro organ development. Here, we introduce versatile, thixotropic amyloid hydrogels as a bio-mimetic ECM scaffold for 3D cell culture as well as high-throughput tumor spheroid formation using a drop cast method. The unique cross-β-sheet structure, sticky surface, and thixotropicity of amyloid hydrogels allow robust cell adhesion, survival, proliferation, and migration, which are essential for 3D tumor modeling with various cancer cell types. The spheroids formed show overexpression of the signature cancer biomarkers and confer higher drug resistance compared to two-dimensional (2D) monolayer cultures. Using breast tumor tissue from mouse xenograft, we showed that these hydrogels support the formation of tumor spheroids with a well-defined necrotic core, cancer-associated gene expression, higher drug resistance, and tumor heterogeneity reminiscent of the original tumor. Altogether, we have developed a rapid and cost-effective platform for generating in vitro cancer models for the screening of anti-cancer therapeutics and developing personalized medicines.


2013 ◽  
Vol 35 (2) ◽  
pp. 270-270 ◽  
Author(s):  
Hongzhi Cao ◽  
Yu Wang ◽  
Wei Zhang ◽  
Xianghua Chai ◽  
Xiandong Zhang ◽  
...  

1996 ◽  
Vol 59 (6) ◽  
pp. 631-641 ◽  
Author(s):  
ANTONIO-JAVIER RAMOS ◽  
JOHANA FINK-GREMMELS ◽  
ENRIQUE HERNÁNDEZ

Mycotoxins comprise a family of fungal toxins, many of which have been implicated as chemical progenitors of toxicity in man and animals. The most thoroughly studied are the aflatoxins. A variety of physical, chemical, and biological methods to counteract the mycotoxin problem have been reported, but large-scale, practical, and cost-effective methods for detoxifying mycotoxin-containing feedstuffs are currently not available. The most recent approach to the problem has been the addition to the animal's diet of nonnutritive sorbents that sequester mycotoxins and reduce their gastrointestinal absorption, avoiding their toxic effects on livestock and toxin carryover into animal products. This review comments on the in vitro efficacy of several of the adsorbents assayed, and their in vivo applications in a range of animals will be discussed. The sorbents reviewed are activated charcoal, bentonite, zeolite, hydrated sodium calcium aluminosilicate (HSCAS) and a wide variety of clays and synthetic ion-exchange resins.


2020 ◽  
Vol 25 (9) ◽  
pp. 1038-1046
Author(s):  
Eun Jeong Cho ◽  
Ashwini K. Devkota ◽  
Gabriel Stancu ◽  
Ramakrishna Edupunganti ◽  
Ginamarie Debevec ◽  
...  

Hypoxic solid tumors induce the stabilization of hypoxia-inducible factor 1 alpha (HIF1α), which stimulates the expression of many glycolytic enzymes and hypoxia-responsive genes. A high rate of glycolysis supports the energetic and material needs for tumors to grow. Fructose-1,6-bisphosphate aldolase A (ALDOA) is an enzyme in the glycolytic pathway that promotes the expression of HIF1α. Therefore, inhibition of ALDOA activity represents a potential therapeutic approach for a range of cancers by blocking two critical cancer survival mechanisms. Here, we present a luminescence-based strategy to determine ALDOA activity. The assay platform was developed by integrating a previously established ALDOA activity assay with a commercial NAD/NADH detection kit, resulting in a significant (>12-fold) improvement in signal/background (S/B) compared with previous assay platforms. A screening campaign using a mixture-based compound library exhibited excellent statistical parameters of Z′ (>0.8) and S/B (~20), confirming its robustness and readiness for high-throughput screening (HTS) application. This assay platform provides a cost-effective method for identifying ALDOA inhibitors using a large-scale HTS campaign.


1994 ◽  
Vol 22 (4) ◽  
pp. 243-253
Author(s):  
Boris Isomaa ◽  
Henrik Lilius ◽  
Christina Råbergh

There is an urgent need for effective in vitro tests in aquatic toxicology, because only a very small proportion of the chemicals in common use have been adequately tested for their toxicity to aquatic organisms and aquatic ecosystems. Toxicity tests with higher animals, besides being time-consuming and expensive, are ethically questionable, which further increases the importance of developing efficient in vitro toxicity tests. In developing in vitro tests for toxicity assessments, aquatic toxicology lags behind mammalian toxicology. Aqueous environmental chemistry is complex, and the sensitivity of the organisms living in a particular aquatic environment may vary considerably. The predictive value of single-species or cell culture tests is therefore generally considered to be low. Nevertheless, single-species tests, utilising bacteria, algae, protozoans and invertebrates, have frequently been used in in vitro toxicity studies of aquatic pollutants (mainly as screening tests). Attempts at large-scale validations are few. Such attempts seem to be hampered by the complexity of the aquatic ecosystem. Although cells from aquatic organisms have been isolated and cultured for many years, the use of isolated or cultured cells in aquatic toxicology has been limited. However, during the last few years, interest in the use of fish cells in toxicity testing has grown rapidly. For aquatic in vitro toxicology to develop further, a more comparative and mechanistic approach needs to be adopted.


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