The potential use of Drosophila as an in vivo model organism for COVID-19-related research: A review

The term cosmetics refers to a product applied to the body for the purpose of beautifying, cleansing or improving appearance and enhancing attractive features. The natural cosmetics market has grown since the consumer took consciousness of the concept of natural-based ingredients. A great number of cosmetics have noxious and chemically-potent substances and have an ecological impact on the environment. A study performed by the Danish Council THINK Chemicalsfound that in total 65 chemicals of concern were found in 39 products. This means consumers are exposed to these chemicals, perhaps in a daily basis. They also found that three products contained illegal ingredients in the European Union. Thus, the use of natural and organic cosmetics becomes increasingly important. This requires a strong investigation into the benefits that fruits and plants can bring to health. The PhD project will focus on four natural ingredients common in the Trás-os-Montes area: almond (Prunus dulcis), elderberry (Sambucus nigra), olive (Olea europaea) and grapes (Vitis vinifera). The general purpose of this PhD project is to evaluate the cosmetic properties of the natural ingredients towards the DNA integrity promotion. Additionally, it is intended to evaluate genoprotection, longevity and prolificacy of the natural ingredients in Drosophila melanogaster. The short life cycle, the distinct developmental stages, the availability of various tools and reagents, known genome sequence and the physiological similarity of Drosophila with humans make them an excellent in vivo model organism to rapidly test toxicity in whole organism and elucidate the molecular mechanisms underlying the toxicity. The natural product with the best result will be used to evaluate genoprotection in human lymphocytes. These are used as a surrogate tissue, as they are easily obtained, in large numbers, do not require cell culture, are diploids and are almost all in the same phase of the cell cycle. This project is in an initial phase and lacks results, which will be available along this year.

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Thrombin Generation in Zebrafish

Blood ◽

10.1182/blood.v124.21.4218.4218 ◽

2014 ◽

Vol 124 (21) ◽

pp. 4218-4218

Author(s):

Evelien Schurgers ◽

Martijn Moorlag ◽

Hilde Kelchtermans ◽

Coenraad Hemker ◽

Bas De Laat

Keyword(s):

Whole Blood ◽

Thrombin Generation ◽

Conflicts Of Interest ◽

Genetic Research ◽

Model Organism ◽

Lag Time ◽

In Vivo Model ◽

Time To Peak ◽

Paper Disk

Abstract Introduction Recently, the whole blood calibrated automated thrombogram (CAT) was miniaturized enabling the measurement of thrombin generation (TG) in a limited volume (5 µl) of whole blood. Consequently, this approach may be used to determine TG in small lab animals. Zebrafish are readily available test animals for genetic research. However, their small size has been a hurdle in thrombosis and hemostasis research since most assays require large amounts of plasma. In this study we verified the possibility to measure TG in zebrafish using our newly developed miniaturized whole blood assay. Methods For TG, 5 µl of whole blood was mixed with 5 µl of buffer containing a rhodamin-based thrombin-sensitive P2Rho substrate (final concentration (fc) 300 µM). 5 µl of this mixture was put on a paper disk and covered with mineral oil to prevent evaporation. Calibration was done as described previously (Ninivaggi et al. Clin Chem 2012) by adding 5 µl of whole blood to 5 µl of a mixture containing P2Rho (fc 300 µM), a2M-thrombin calibrator (fc 100 nM) and citrate (fc 9,8 mM). Fluorescence was detected with a fluorometer (485/538 nm). Results Due to their limited blood volume, it is impossible to perform both a TG and calibrator measurement on the same fish. Since calibrator measurements performed on blood from different fish demonstrated acceptable variation (CV calibrator slopes < 15%), the average calibrator slope was used for calculations. The calculated TG parameters from 2 independent experiments are depicted in Table 1. TG measured in individual fish showed the same amount of inter-individual variation as in humans. Striking differences with human TG parameters were the short lag time, high peak and high velocity index. Moreover a further analysis of the fibrin network of the clot, by means of scanning electron microscopy (SEM), showed a much denser network composed of thinner fibers compared to humans. Table 1. Thrombin generation parameters and their coefficient of variance (CV) in zebrafish in two independent experiments. Experiment 1(n=4) Experiment 2(n=9) Mean CV Mean CV Peak ETP (nM.min) 575.68 19.69 689.71 24.91 Peak (nM) 1573.16 29.58 1668.05 14.46 Lag time (min) 0.27 0.00 0.49 10.92 Time to peak (min) 1.08 5.30 1.37 11.49 Velocity (nM/min) 4002.33 38.21 3826.35 17.04 Conclusion These results demonstrate the feasibility of measuring TG in whole blood collected from zebrafish. Consequently, zebrafish may be used as a in vivo model to test the effect of (novel) anticoagulant therapeutics on thrombin generation and serve as a model organism for mechanistical research in thrombosis and haemostasis. Disclosures No relevant conflicts of interest to declare.

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Danio Rerio as Model Organism for Adenoviral Vector Evaluation

Genes ◽

10.3390/genes10121053 ◽

2019 ◽

Vol 10 (12) ◽

pp. 1053

Author(s):

Paola Gulías ◽

Jorge Guerra-Varela ◽

Manuela Gonzalez-Aparicio ◽

Ana Ricobaraza ◽

Africa Vales ◽

...

Keyword(s):

Danio Rerio ◽

Viral Vector ◽

Early Stage ◽

High Capacity ◽

Model Organism ◽

Adenoviral Vectors ◽

Regulatory Agencies ◽

In Vivo Model ◽

Efficient Alternative

Viral vector use is wide-spread in the field of gene therapy, with new clinical trials starting every year for different human pathologies and a growing number of agents being approved by regulatory agencies. However, preclinical testing is long and expensive, especially during the early stages of development. Nowadays, the model organism par excellence is the mouse (Mus musculus), and there are few investigations in which alternative models are used. Here, we assess the possibility of using zebrafish (Danio rerio) as an in vivo model for adenoviral vectors. We describe how E1/E3-deleted adenoviral vectors achieve efficient transduction when they are administered to zebrafish embryos via intracranial injection. In addition, helper-dependent (high-capacity) adenoviral vectors allow sustained transgene expression in this organism. Taking into account the wide repertoire of genetically modified zebrafish lines, the ethical aspects, and the affordability of this model, we conclude that zebrafish could be an efficient alternative for the early-stage preclinical evaluation of adenoviral vectors.

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Panel Discussion II: Usefulness of in vitro and in vivo Model Systems for Predicting the Adverse Public Health Outcome for Antimicrobial Drug Residues in Food

Microbial Ecology in Health and Disease ◽

10.1080/08910600050216174 ◽

2000 ◽

Vol 12 (3) ◽

pp. 45-53

Author(s):

Andrew Beaulieu ◽

Jacques Boisseau ◽

Carl Cerniglia ◽

Denis Corpet ◽

A. Haydée Fernández ◽

...

Keyword(s):

Public Health ◽

Health Outcome ◽

Panel Discussion ◽

Antimicrobial Drug ◽

Model Systems ◽

In Vivo Model ◽

Drug Residues ◽

Public Health Outcome

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P1735 Arginine supplementation and oxidative processes: in vivo model

European Heart Journal ◽

10.1016/s0195-668x(03)94873-1 ◽

2003 ◽

Vol 24 (5) ◽

pp. 330

Author(s):

A KOPFF

Keyword(s):

In Vivo Model ◽

Oxidative Processes ◽

Arginine Supplementation

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Establishment of an in vivo model for KCNJ5 dependent hyperaldosteronism

Experimental and Clinical Endocrinology & Diabetes ◽

10.1055/s-0035-1547718 ◽

2015 ◽

Vol 122 (03) ◽

Cited By ~ 2

Author(s):

U Lichtenauer ◽

PL Schmid ◽

A Oßwald ◽

I Renner-Müller ◽

M Reincke ◽

...

Keyword(s):

In Vivo Model

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An in vivo Model for the Assessment of Acute Fibrinolytic Capacity of the Endothelium

Thrombosis and Haemostasis ◽

10.1055/s-0038-1657722 ◽

1997 ◽

Vol 78 (04) ◽

pp. 1242-1248 ◽

Cited By ~ 68

Author(s):

David E Newby ◽

Robert A Wright ◽

Christopher A Ludlam ◽

Keith A A Fox ◽

Nicholas A Boon ◽

...

Keyword(s):

Blood Flow ◽

Substance P ◽

Sodium Nitroprusside ◽

Plasma Concentrations ◽

In Vivo Model ◽

Forearm Circulation ◽

Von Willebrand ◽

Local Release ◽

Willebrand Factor

SummaryThe effects on blood flow and plasma fibrinolytic and coagulation parameters of intraarterial substance P, an endothelium dependent vasodilator, and sodium nitroprusside, a control endothelium independent vasodilator, were studied in the human forearm circulation. At subsystemic locally active doses, both substance P (2-8 pmol/min) and sodium nitroprusside (2-8 μg/min) caused dose-dependent vasodilatation (p <0.001 for both) without affecting plasma concentrations of PAI-1, von Willebrand factor antigen or factor VIII:C activity. Substance P caused local increases in t-PA antigen and activity (p <0.001) in the infused arm while sodium nitroprusside did not. At higher doses, substance P increased blood flow and t-PA concentrations in the noninfused arm. We conclude that brief, locally active and subsystemic infusions of intraarterial substance P cause a rapid and substantial local release of t-PA which appear to act via a flow and nitric oxide independent mechanism. This model should provide a useful and selective method of assessing the in vivo capacity of the forearm endothelium to release t-PA acutely.

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