- PUBLICATION
- 論文
小型生物トラッカー
-
Triazole Fungicides Inhibit Zebrafish Hatching by Blocking the Secretory Function of Hatching Gland Cells
Javiera F. De la Paz, Natalia Beiza, Susana Paredes-Zúñiga, Misque S. Hoare, and Miguel L. Allende
(System: WMicrotracker ONE Model organism: Haemonchus contortus)
Abstract: In animals, hatching represents the transition point from a developing embryo to a free-living individual, the larva. This process is finely regulated by many endogenous and environmental factors and has been shown to be sensitive to a variety of chemical agents. It is commonly evaluated in bioassays in order to establish the effects of different agents on early development and reproductive capabilities in fish and other aquatic animals. In fish, the breakdown of the chorion is achieved by the secretion of choriolysin by hatching gland cells (HGCs) into the perivitelline space (PVS), coupled with spontaneous movements of the developing larva. In this work, we used zebrafish to assay the effects of a family of widely used agrochemicals—triazoles Triadimefon (FON), Triadimenol (NOL) and free triazole (1,2,4-T)—on hatching success.
-
Use of Caenorhabditis elegans based assays as an alternative strategy in toxicity testing
Author(s)Gregori Balaguer, María Tutor(s)Sanmartín Santos, Isaias Salvador
(System: WMicrotracker ONE Model organism: Haemonchus contortus)
Abstract: In vivo models are an essential working tool in the biomedicine laboratory. Specially, they are important for obtaining complete biological responses to certain stimuli or substances, which needs to be evaluated as a whole complete organism. Toxicology particularly is a demanding field of new models, which needs to administer investigators, reliable results in a short period of time. The use in investigations of the nematode Caenorhabditis elegans (C. elegans) was first boosted and developed by Sydney Brenner. It is a small invertebrate animal (round worm), used extensively in basic biological investigations, and reaching a great popularity as an in vivo model.
-
High-Throughput Phenotypic Assay to Screen for Anthelmintic Activity on Haemonchus contortus
Aya C. Taki ,Joseph J. Byrne ,Tao Wang ,Brad E. Sleebs,Nghi Nguyen ,Ross S. Hall,Pasi K. Korhonen,Bill C.H. Chang,Paul Jackson,Abdul Jabbarand Robin B. Gasser
(System: WMicrotracker ONE Model organism: Haemonchus contortus)
Abstract: 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.
-
Chemical characterization and in vitro anthelmintic activity of Citrus bergamia Risso and Citrus X paradisii Macfad essential oil against Haemonchus contortus Kirby isolate
Vivien PatriciaGarbin, BeatrizMunguía, Jenny CarolinSaldaña, Cícero Deschamps, Roger Raupp Cipriano, Marcelo Beltrão Molentoa
(System: WMicrotracker ONE Model organism: Haemonchus contortus)
Abstract
Haemonchus contortus, a blood-sucking parasite of small ruminants, produces very important economic losses in the productive sector. This abomasum parasite has become resistant to most commercial drugs worldwide, and alternatives to fight this problem are urgently needed. Essential oils (EO) are a complex mixture of volatile secondary metabolites, composed mainly by terpenoids and phenolic compounds, from plants that have several pharmacological properties, including anthelmintic activity. Particularly, citrus peel is a source of cold-pressed EO, where limonene is its major component, and can be used as an additional food component for ruminants. -
Sensitivity of Haemonchus contortus to anthelmintics using different in vitro screening assays: a comparative study
Beatriz Munguía, Jenny Saldaña, Magdalena Nieves, María Elisa Melian, Manuela Ferrer, Ramiro Teixeira, Williams Porcal, Eduardo Manta & Laura Domínguez
(System: WMicrotracker ONE Model organism: Haemonchus contortus)
Abstract
Background
Helminthiasis and resistance to commercial anthelmintic compounds are major causes of economic losses for livestock producers, resulting in an urgent need for new drugs and reliable in vitro screening tests capable of detecting potentially active products. Considering this, a series of novel benzimidazole derivatives (5-methylbenzimidazole 1,2-disubstituted, 5-carboxybenzimidazole, 5-methylbenzimidazole 2-one) was screened on exsheathed L3 (xL3) and on the adult stage of Haemonchus contortus (Kirby anthelmintic-susceptible McMaster isolate).
Link -
Screening of a drug repurposing library with a nematode motility assay identifies promising anthelmintic hits against Cooperia oncophora and other ruminant parasites
Maoxuan Liu, Bart Landuyt, bHugo Klaassen, Peter Geldhof, Walter Luyten
(System: WMicrotracker ONE Model organism: Nematode)
Parasitic nematodes continue to cause significant economic losses in livestock globally. Given the limited number of anthelmintic drugs on the market and the currently increasing drug resistance, there is an urgent need for novel anthelmintics. Most motility assays of anthelmintic activity for parasitic nematodes are laborious and low throughput, and therefore not suitable for screening large compound libraries. Cooperia oncophora accounts for a large proportion of reports on the drug-resistance development of parasites globally.
-
A validated high-throughput method for assaying rat lungworm (Angiostrongylus cantonensis) motility when challenged with potentially anthelmintic natural products from Hawaiian fungi
Randi L. Rollins, Mallique Qader, William L. Gosnell, Cong Wang, Shugeng Cao and Robert H. Cowie
(System: WMicrotracker ONE Model organism: A.cantonensis)
Parasitic nematodes devastate human and animal health. The limited number of anthelmintics available is concerning, especially because of increasing drug resistance. Anthelmintics are commonly derived from natural products, e.g. fungi and plants. This investigation aimed to develop a high-throughput whole organism screening method based on a motility assay using the wMicroTracker system. Anthelmintic activity of extracts from Hawaiian fungi was screened against third-stage larvae of the parasitic nematode Angiostrongylus cantonensis, categorized according to the degree of motility reduction. Of the 108 crude samples and fractionated products, 48 showed some level of activity, with 13 reducing motility to 0–25% of the maximum exhibited, including two pure compounds, emethacin B and epicoccin E, neither previously known to exhibit anthelmintic properties.
-
In vitro screening methods for parasites: the wMicroTracker & the WormAssay
Emma Gunderson, Christina Bulman, Mona Luo, and Judy Sakanari
(System: WMicrotracker ONE Model organism: Parasites)
Description: The purpose of this study was to determine the utility of the wMicroTracker as a screening platform to assess the motility of various parasites. We tested three species of parasites: the adult and larval stages of the filarial nematode Brugia pahangi, the schistosomula stage of the trematode Schistosoma mansoni, and the epimastigote stage of the protozoan parasite Trypanosoma cruzi.We optimized the assay for the number of parasites per well, plate type and media volume using the wMicroTracker and compared those readouts to readouts from the WormAssay (Marcellino et al. 2012) when possible. The WormAssay has been used in phenotypic drug screens to identify new compounds for the treatment of lymphatic filariasis, onchocerciasis and schistosomiasis (Storey et al. 2014; Bulman et al. 2015; Weeks et al. 2018; Tyagi et al. 2019). The original WormAssay was developed by Marcellino et al. 2012 and was subsequently modified to the “Worminator” by Storey et al. 2014 to observe smaller worms with an inverted microscope. wMicroTracker (InVivo Biosystems) protocols optimized for C. elegans adults (1 mm in length by 80 µm in width, highly motile) were used to optimize parasite assays based on the size and motility of each of the parasite species as compared to C. elegans adults.
-
Disruption of genes associated with Charcot-Marie-Tooth type 2 lead to common behavioural, cellular and molecular defects in Caenorhabditis elegans
Ming S. Soh,Xinran Cheng,Tarika Vijayaraghavan,Arwen Vernon,Jie Liu,Brent Neumann
(System: WMicrotracker ONE Model organism: C.elegans)
Abstract: Charcot-Marie-Tooth (CMT) disease is an inherited peripheral motor and sensory neuropathy. The disease is divided into demyelinating (CMT1) and axonal (CMT2) neuropathies, and although we have gained molecular information into the details of CMT1 pathology, much less is known about CMT2. Due to its clinical and genetic heterogeneity, coupled with a lack of animal models, common underlying mechanisms remain elusive. In order to gain an understanding of the normal function of genes associated with CMT2, and to draw direct comparisons between them, we have studied the behavioural, cellular and molecular consequences of mutating nine different genes in the nematode Caenorhabditis elegans (lin-41/TRIM2, dyn-1/DNM2, unc-116/KIF5A, fzo-1/MFN2, osm-9/TRPV4, cua-1/ATP7A, hsp-25/HSPB1, hint-1/HINT1, nep-2/MME).
-
A single copy transgenic mutant FUS strain reproduces age-dependent ALS phenotypes in C. elegans
Audrey Labarre,Gilles Tossing,Claudia Maios,ames J Doyle,and J Alex Parker
(System: WMicrotracker ONE Model organism: C.elegans)
Abstract: Mutations in the human DNA/RNA binding protein FUS are associated with amyotrophic lateral sclerosis and frontotemporal lobar degeneration, including some aggressive and juvenile onset forms. Cytoplasmic inclusions of human FUS proteins are observed in various neurodegenerative disorders, such as Huntington’s disease or spinocerebellar ataxia, suggesting that FUS proteinopathy may be a key player in neurodegeneration. To better understand the pathogenic mechanisms of FUS, we created single copy transgenic Caenorhabditis elegans strains expressing full-length, untagged human FUS in the worm’s GABAergic neurons.
-
Caenorhabditis elegans Infrared-Based Motility Assay Identified New Hits for Nematicide Drug Development
Gastón Risi,Elena Aguilera,Enrique Ladós,Gonzalo Suárez,Inés Carrera,4ORCID,Guzmán Álvarez,and Gustavo Salinas
(System: WMicrotracker ONE Model organism: C.elegans)
Abstract: Nematode parasites have a profound impact on humankind, infecting nearly one-quarter of the world’s population, as well as livestock. There is a pressing need for discovering nematicides due to the spread of resistance to currently used drugs. The free-living nematode Caenorhabditis elegans is a formidable experimentally tractable model organism that offers key advantages in accelerating nematicide discovery. We report the screening of drug-like libraries using an overnight high-throughput C. elegans assay, based on an automated infrared motility reader. As a proof of concept, we screened the “Pathogen Box” library.
-
Biochemical characterization, cytotoxic, antimutagenic, anticancer and molecular docking studies on Tecomella undulata
SanaRiaz,Muhammad ArslanJaved, IqraNawaz, TariqJaved,
(System: WMicrotracker ONE Model organism: C.elegans)
Abstract: In this study bioassay-guided screening of Tecomella undulate was performed for its cytotoxic, antimutagenic and anticancer potential. The ariel parts were extracted on a polarity basis (methanol, dichloromethane and hexane). The in vivo toxicity was assessed on Caenorhabditis elegans, and its locomotion was affected by Tecomella undulata hexane (TUAH) the most. Ames test for antimutagenicity showed Tecomella undulata methanol (TUAM) exhibited against mutagen 2AA showed inhibition of 71.03% and 26.32% 2AA in TA98 while in in vitro MTT assay on carcinoma cell lines TUAM showed 68.1% cytotoxicity.
-
Small Molecule Rescue of ATXN3 Toxicity in C. elegans via TFEB/HLH-30
Yasmin Fardghassemi, Claudia Maios & J. Alex Parker
(System: WMicrotracker ONE Model organism: C.elegans)
Abstract: Spinocerebellar ataxia type 3 (SCA3), also known as Machado-Joseph disease (MJD), is a polyglutamine expansion disease arising from a trinucleotide CAG repeat expansion in exon 10 of the gene ATXN3. There are no effective pharmacological treatments for MJD, thus the identification of new pathogenic mechanisms, and the development of novel therapeutics is urgently needed. In this study, we performed a comprehensive, blind drug screen of 3942 compounds (many FDA approved) and identified small molecules that rescued the motor-deficient phenotype in transgenic ATXN3 Caenorhabditis elegans strain.
-
Assessment of the effects of organic vs. inorganic arsenic and mercury in Caenorhabditis elegans
JessicaCamachoaAlinede ContibIgor P.PogribnybRobert L.SprandoaPiper ReidHunta
(System: WMicrotracker ONE Model organism: C.elegans)
Abstract: Exposures to mercury and arsenic are known to pose significant threats to human health. Effects specific to organic vs. inorganic forms of these toxic elements are less understood however, especially for organic dimethylarsinic acid (DMA), which has recently been detected in pups of rodent dams orally exposed to inorganic sodium (meta)arsenite (NaAsO2). Caenorhabditis elegans is a small animal alternative toxicity model. To fill data gaps on the effects of DMA relative to NaAsO2, C. elegans were exposed to these two compounds alongside more thoroughly researched inorganic mercury chloride (HgCl2) and organic methylmercury chloride (meHgCl). For timing of developmental milestone acquisition in C. elegans, meHgCl was 2 to 4-fold more toxic than HgCl2, and NaAsO2 was 20-fold more toxic than DMA, ranking the four compounds meHgCl > HgCl2 > NaAsO2 ≫ DMA for developmental toxicity.
-
Disruption of mitochondrial dynamics affects behaviour and lifespan in Caenorhabditis elegans
Joseph J. Byrne, Ming S. Soh, Gursimran Chandhok, Tarika Vijayaraghavan, Jean-Sébastien Teoh, Simon Crawford, Ansa E. Cobham, Nethmi M. B. Yapa, Christen K. Mirth & Brent Neumann
(System: WMicrotracker ONE Model organism: C.elegans)
Abstract: Mitochondria are essential components of eukaryotic cells, carrying out critical physiological processes that include energy production and calcium buffering. Consequently, mitochondrial dysfunction is associated with a range of human diseases. Fundamental to their function is the ability to transition through fission and fusion states, which is regulated by several GTPases. Here, we have developed new methods for the non-subjective quantification of mitochondrial morphology in muscle and neuronal cells of Caenorhabditis elegans. Using these techniques, we uncover surprising tissue-specific differences in mitochondrial morphology when fusion or fission proteins are absent.
-
Bioassay-guided isolation of three anthelmintic compounds from Warburgia ugandensis Sprague subspecies ugandensis, and the mechanism of action of polygodial
MaoxuanLiu PurityKipanga Anh HungMai InekeDhondt Bart P.Braeckman Wim De Borggraeve
WalterLuyten(System: WMicrotracker ONE Model organism: C.elegans)
Abstract: Parasitic helminths continue to pose problems in human and veterinary medicine, as well as in agriculture. Resistance to current anthelmintics has prompted the search for new drugs. Anthelmintic metabolites from medicinal plants could be good anthelmintic drug candidates. However, the compounds active against nematodes have not been identified in most medicinal plants with anthelmintic activity. In this study, we aimed to identify the active compounds against helminths in Warburgia ugandensis Sprague subspecies ugandensis (Canellaceae) and study the underlying mechanism of action.
-
C. elegans Development and Activity Test detects mammalian developmental neurotoxins
Piper ReidHunt NicholasOlejnik Keenan D.BaileyCory A.VaughtRobert L.Sprando
(System: WMicrotracker ONE Model organism: C.elegans)
Abstract: Due to the high cost and long duration of traditional testing methods for developmental neurotoxicity (DNT), only a small fraction of chemicals that humans are exposed to have been assessed for DNT activity. In order to ensure public safety, human-predictive methods for DNT detection that are faster and less resource intensive are urgently required. Using Caenorhabditis elegans, a novel worm Development and Activity test (wDAT) has been designed that uses a relatively inexpensive small-animal activity tracker and takes less than 4 days to complete.
-
Antibacterial, Antifungal, Antiviral, and Anthelmintic Activities of Medicinal Plants of Nepal Selected Based on Ethnobotanical Evidence
Bishnu Joshi ,Sujogya Kumar Panda ,Ramin Saleh Jouneghani,Maoxuan Liu,1 Niranjan Parajuli ,Pieter Leyssen,Johan Neyts,and Walter Luyten
(System: WMicrotracker ONE Model organism: C.elegans)
Abstract: Background. Infections by microbes (viruses, bacteria, and fungi) and parasites can cause serious diseases in both humans and animals. Heavy use of antimicrobials has created selective pressure and caused resistance to currently available antibiotics, hence the need for finding new and better antibiotics. Natural products, especially from plants, are known for their medicinal properties, including antimicrobial and anthelmintic activities. Geoclimatic variation, together with diversity in ethnomedicinal traditions, has made the Himalayas of Nepal an invaluable repository of traditional medicinal plants.
-
Increased sensitivity of an infrared motility assay for nematicide discovery
Franco Vairoletti,Antonela Baron, Cecilia Saiz,Graciela Mahler,and Gustavo Salinas
(System: WMicrotracker ONE Model organism: C.elegans)
Abstract: Parasitic nematodes constitute a health problem for humans, livestock and crops, and cause huge economic losses to developing-country economies. Due to the spread of nematicide resistance, there is an urgent need for new drugs. C. elegans is now recognized as a cost-effective alternative for the screening of compound libraries with potential nematicidal activity, as parasitic organisms are hard to maintain under laboratory conditions.
-
Practical High-Throughput Method to Screen Compounds for Anthelmintic Activity against Caenorhabditis elegans
Aya C. Taki Joseph J. Byrne Peter R. Boag 2Abdul Jabbar and Robin B. Gasser
(System: WMicrotracker ONE Model organism: C.elegans)
Abstract: In the present study, we established a practical and cost-effective high throughput screening assay, which relies on the measurement of the motility of Caenorhabditis elegans by infrared lightinterference. Using this assay, we screened 14,400 small molecules from the “HitFinder” library (Maybridge), achieving a hit rate of 0.3%. We identified small molecules that reproducibly inhibited the motility of C. elegans (young adults) and assessed dose relationships for a subset of compounds.
Future work will critically evaluate the potential of some of these hits as candidates for subsequentoptimisation or repurposing as nematocides or nematostats. -
Comparison of electrophysiological and motility assays to study anthelmintic effects in Caenorhabditis elegans
Steffen R.Hahnela1William M.RobertsbIringHeisleraDanielKulkea2Janis C.Weeksb
(System: WMicrotracker ONE Model organism: C.elegans)Abstract:Currently, only a few chemical drug classes are available to control the global burden of nematode infections in humans and animals. Most of these drugs exert their anthelmintic activity by interacting with proteins such as ion channels, and the nematode neuromuscular system remains a promising target for novel intervention strategies. Many commonly-used phenotypic readouts such as motility provide only indirect insight into neuromuscular function and the site(s) of action of chemical compounds. Electrophysiological recordings provide more specific information but are typically technically challenging and lack high throughput for drug discovery. Because drug discovery relies strongly on the evaluation and ranking of drug candidates, including closely related chemical derivatives, precise assays and assay combinations are needed for capturing and distinguishing subtle drug effects.
-
The nervous and prenervous roles of serotonin in Echinococcus spp
F Camicia , M Herz, L C Prada, L Kamenetzky, S H Simonetta, M A Cucher, J I Bianchi, C Fernández, K Brehm, M C Rosenzvit
(System: ONE, Model organism: Echinococcus spp)
PMID: 23639266 DOI: 10.1016/j.ijpara.2013.03.006
2013 Jul;43(8):647-59. doi: 10.1016/j.ijpara.2013.03.006. Epub 2013 Apr 29.ABSTRACT:Serotonin (5-hydroxytryptamine, 5-HT) is an important neuroactive and morphogenetic molecule in several metazoan phyla, including flatworms. Serotoninergic nervous system studies are incomplete and 5-HT function/s are unknown in Echinococcus spp.the flatworm parasites that cause hydatid disease. In the present work, we searched for genes of the serotoninergic pathway and performed immunocytochemical and functional analyses of 5-HT in Echinococcus spp. Bioinformatic analysis using the recently available Echinococcus multilocularis and Echinococcus granulosus genomes suggests the presence of genes encoding enzymes, receptors and transporters participating in 5-HT synthesis, sensing and transport in these parasites.
-
An automated tracking system for Caenorhabditis elegans locomotor behavior and circadian studies application
ergio H Simonetta, Diego A Golombek
(System: ONE, Model organism: C. Elegans)
PMID: 17207862 DOI: 10.1016/j.jneumeth.2006.11.015
2007 Apr 15;161(2):273-80. doi: 10.1016/j.jneumeth.2006.11.015.ABSTRACT:Automation of simple behavioral patterns, such as locomotor activity, is fundamental for pharmacological and genetic screening studies. Recently, circadian behaviors in locomotor activity and stress responses were reported in the nematode Caenorhabditis elegans, a well-known model in genetics and developmental studies. Here we present a new method for long-term recordings of C. elegans (as well as other similar-sized animals) locomotor activity based on an infrared microbeam scattering. Individual nematodes were cultured in a 96-well microtiter plate; we tested L15, CeMM and E. coli liquid cultures in long-term activity tracking experiments, and found CeMM to be the optimal medium. Treatment with 0.2% azide caused an immediate decrease in locomotor activity as recorded with our system. In addition to the validation of the method (including hardware and software details), we report its application in chronobiological studies. Circadian rhythms in animals entrained to light-dark and constant dark conditions (n=48 and 96 worms, respectively) at 16 degrees C, were analyzed by LS periodograms. We obtained a 24.2+/-0.44 h period (52% of significantly rhythmic animals) in LD, and a 23.1+/-0.40 h period (37.5% of significantly rhythmic animals) under DD. The system is automateable using microcontrollers, of low-cost construction and highly reproducible.
-
Neurodegeneration in C. elegans models of ALS requires TIR-1/Sarm1 immune pathway activation in neurons
Julie Vérièpe, Lucresse Fossouo, J Alex Parker
(System: ONE, Model organism: C. Elegans)
PMID: 26059317 DOI: 10.1038/ncomms8319 Free article
2015 Jun 10;6:7319. doi: 10.1038/ncomms8319.ABSTRACT:Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease thought to employ cell non-autonomous mechanisms where neuronal injury engages immune responses to influence disease progression. Here we show that the expression of mutant proteins causative for ALS in Caenorhabditis elegans motor neurons induces an innate immune response via TIR-1/Sarm1. Loss of function mutations in tir-1, associated downstream kinases, and the transcription factor atf-7 all suppress motor neuron degeneration. The neurosecretory proteins UNC-13 and UNC-31 are required for induction of the immune response as well as the degeneration of motor neurons. The human orthologue of UNC-13, UNC13A, has been identified as a genetic modifier of survival in ALS, and we provide functional evidence of UNC-13/UNC13A in regulating motor neuron degeneration. We propose that the innate immune system reacts to the presence of mutant proteins as a contagion, recruiting a pathogen resistance response that is ultimately harmful and drives progressive neurodegeneration.
-
Reliable Screening of Dye Phototoxicity by Using a Caenorhabditis elegans Fast Bioassay
Javier Ignacio Bianchi, Juan Carlos Stockert, Lucila Ines Buzzi, Alfonso Blázquez-Castro, Sergio Hernán Simonetta
(System: ONE, Model organism: C. Elegans)
PMID: 26039060 PMCID: PMC4454604 DOI: 10.1371/journal.pone.0128898
2015 Jun 3;10(6):e0128898. doi: 10.1371/journal.pone.0128898. eCollection 2015.ABSTRACT: Phototoxicity consists in the capability of certain innocuous molecules to become toxic when subjected to suitable illumination. In order to discover new photoactive drugs or characterize phototoxic pollutants, it would be advantageous to use simple biological tests of phototoxicy. In this work, we present a pilot screening of 37 dyes to test for phototoxic effects in the roundworm Caenorhabditis elegans. Populations of this nematode were treated with different dyes, and subsequently exposed to 30 min of white light. Behavioral outcomes were quantified by recording the global motility using an infrared tracking device (WMicrotracker). Of the tested compounds, 17 dyes were classified as photoactive, being phloxine B, primuline, eosin Y, acridine orange and rose Bengal the most phototoxic. To assess photoactivity after uptake, compounds were retested after washing them out of the medium before light irradiation. Dye uptake into the worms was also analyzed by staining or fluorescence. All the positive drugs were incorporated by animals and produced phototoxic effects after washing. We also tested the stress response being triggered by the treatments through reporter strains. Endoplasmic reticulum stress response (hsp-4::GFP strain) was activated by 22% of phototoxic dyes, and mitochondrial stress response (hsp-6::GFP strain) was induced by 16% of phototoxic dyes. These results point to a phototoxic perturbation of the protein functionality and an oxidative stress similar to that reported in cell cultures. Our work shows for the first time the feasibility of C. elegans for running phototoxic screenings and underscores its application on photoactive drugs and environmental pollutants assessment.
-
Bioactivity of nanosilver in Caenorhabditis elegans: Effects of size, coat, and shape
Piper ReidHuntaZacharyKeltneraXiugongGaoaSteven J.OldenburgbPriyankaBushanaaNicholasOlejnikaRobert L.Sprandoa
(System: ONE, Model organism: C. Elegans)
ABSTRACT: The in vivo toxicity to eukaryotes of nanosilver (AgNP) spheres and plates in two sizes each was assessed using the simple model organism Caenorhabditis elegans. For each shape, smaller AgNP size correlated with higher toxicity, as indicated by reduced larval growth. Smaller size also correlated with significant increases in silver uptake for silver nanospheres. Citrate coated silver spheres of 20 nm diameter induced an innate immune response that increased or held steady over 24 h, while regulation of genes involved in metal metabolism peaked at 4 h and subsequently decreased. For AgNP spheres, coating altered bioactivity, with a toxicity ranking of polyethylene glycol (PEG) > polyvinylpyrrolidone (PVP) ≅ branched polyethyleneimine (BPEI) > citrate, but silver uptake ranking of PEG > PVP > citrate > BPEI. Our findings in C. elegans correlate well with findings in rodents for AgNP size vs. uptake and toxicity, as well as for induction of immune effectors, while using methods that are faster and far less expensive, supporting the use of C. elegans as an alternative model for early toxicity screening.
-
Development of Nutraceutical Emulsions as Risperidone Delivery Systems: Characterization and Toxicological Studies
Daniela Edith Igartúa, María Natalia Calienni, Daniela Agustina Feas, Nadia Silvia Chiaramoni, Silvia Del Valle Alonso, María Jimena Prieto
(System: ONE, Model organism: Zebrafish)
PMID: 26359783 DOI: 10.1002/jps.24636
2015 Dec;104(12):4142-4152. doi: 10.1002/jps.24636. Epub 2015 Sep 11.
ABSTRACT:Emulsions are gaining increasing interest to be applied as drug delivery systems. The main goal of this work was the formulation of an oil/water nutraceutical emulsion (NE) for oral administration, enriched in omega 3 (ω3) and omega 6 (ω6), and able to encapsulate risperidone (RISP), an antipsychotic drug widely used in the treatment of autism spectrum disorders (ASD). RISP has low solubility in aqueous medium and poor bioavailability because of its metabolism and high protein binding. Coadministration of ω3, ω3, and vitamin E complexed with RISP might increase its bioavailability and induce a synergistic effect on the treatment of ASD. Here, we developed an easy and quick method to obtain NEs and then optimized them. The best formulation was chosen after characterization by particle size, defects of the oil-in-water interface, zeta potential (ZP), and in vitro drug release. The formulation selected was stable over time, with a particle size of around 3 μm, a ZP lower than -20 mV and controlled drug release. -
Reliable Screening of Dye Phototoxicity by Using a Caenorhabditis elegans Fast Bioassay.
Javier Ignacio Bianchi, Juan Carlos Stockert, Lucila Ines Buzzi, Alfonso Blázquez-Castro, Sergio Hernán Simonetta
(System: ONE ,Model organism: C. Elegans)
PMID: 26039060 PMCID: PMC4454604 DOI: 10.1371/journal.pone.0128898 Free PMC article
2015 Jun 3;10(6):e0128898. doi: 10.1371/journal.pone.0128898. eCollection 2015.ABSTRACT:Phototoxicity consists in the capability of certain innocuous molecules to become toxic when subjected to suitable illumination. In order to discover new photoactive drugs or characterize phototoxic pollutants, it would be advantageous to use simple biological tests of phototoxicy. In this work, we present a pilot screening of 37 dyes to test for phototoxic effects in the roundworm Caenorhabditis elegans. Populations of this nematode were treated with different dyes, and subsequently exposed to 30 min of white light. Behavioral outcomes were quantified by recording the global motility using an infrared tracking device (WMicrotracker). Of the tested compounds, 17 dyes were classified as photoactive, being phloxine B, primuline, eosin Y, acridine orange and rose Bengal the most phototoxic. To assess photoactivity after uptake, compounds were retested after washing them out of the medium before light irradiation. Dye uptake into the worms was also analyzed by staining or fluorescence. All the positive drugs were incorporated by animals and produced phototoxic effects after washing. We also tested the stress response being triggered by the treatments through reporter strains. Endoplasmic reticulum stress response (hsp-4::GFP strain) was activated by 22% of phototoxic dyes, and mitochondrial stress response (hsp-6::GFP strain) was induced by 16% of phototoxic dyes. These results point to a phototoxic perturbation of the protein functionality and an oxidative stress similar to that reported in cell cultures. Our work shows for the first time the feasibility of C. elegans for running phototoxic screenings and underscores its application on photoactive drugs and environmental pollutants assessment.
-
Optimization of a locomotion-based zebrafish seizure model
Philip Anthony Gilbert Shaw, Sujogya Kumar Panda, Alexandru Stanca, Walter Luyten
(System: ONE, Model organism: Zebrafish)
ABSTRACT:Background: Locomotor assays in zebrafish have emerged as a screening test in early drug discovery for antiseizure compounds. However, parameters differ considerably between published studies, which may explain some discrepant results with (candidate) antiseizure medications. New method: We optimized a locomotor-based seizure assay in zebrafish with pentylenetetrazol (PTZ) as the pharmacological proconvulsant to generate a therapeutic window in which proconvulsant-treated zebrafish larvae could be discriminated from a non-treated control. To generate a reliable control, exposure time and concentration of valproate (VPA, anticonvulsant) was optimized.
Results: Wells with one or three larvae show a similar PTZ dose-dependent increase in locomotion with less variability in motility for the latter. Zebrafish immersed in 10 mM PTZ showed a significant increase in movement with a sustained effect, without any indication of toxicity. Animals treated with 3 mM VPA showed the strongest reduction of PTZ-induced movement without toxicity. The decrease in PTZ-induced locomotion was greater after 18 h versus 2 h.
Comparison with existing method(s): For the larval zebrafish PTZ-induced seizure model, varying experimental parameters have been reported in literature. Our results show that PTZ is often used at toxic concentrations and we provide instead reliable conditions to quantify convulsant behaviour using an infrared-beam motility assay.
Conclusions: We recommend using three zebrafish larvae per well to quantify locomotion in 96-multiwell plates.
Larvae should preferably be exposed to 10 mM PTZ for 1 h, consisting of 30 min acclimation and 30 min subsequent recording. As positive control for anticonvulsant activity, we recommend exposure to 3 mM VPA for 18 h before administration of PTZ. -
Nutraceutical emulsion containing valproic acid (NE-VPA): a drug delivery system for reversion of seizures in zebrafish larvae epilepsy model
Daniela Agustina Feas· Daniela Edith Igartúa· María Natalia Calienni·
Carolina Soledad Martinez· Marina Pifano· Nadia Silvia Chiaramoni·
Silvia del Valle Alonso · María Jimena Prieto(System: ONE, Model organism: Zebrafish)
Published: 27 February 2017
ABSTRACT: Valproic acid (VPA) is an antiepileptic drug which is currently used in neurodegenerative diseases. However a high dose is required to obtain a therapeutic effect. Long-chain polyunsaturated fatty acids (PUFAs) such as omega 3 and omega 6, are efficient complements in treatments for neurological diseases. Previous studies have reported that a dietary supplement containing PUFAs together with the administration of antiepileptic drugs significantlyreduces the frequency of seizures. Based on this the main goal of this work was to obtain a complex based on VPA encapsulation in an oil/water (o/w) nutraceutical emulsion (NE) enriched with PUFAs for oral administration.Besides, encapsulation of VPA might reduce its dose and increase its therapeutic effect. In order to study its effect, we used a zebrafish larvae model of induced epileptiform behavior with the proconvulsant drug pentylenetetrazol (PTZ). Results have shown that when 100 μM VPA and fatty acids were combined in the NE (NE-VPA), the epileptiform behavior of PTZ-treated zebrafish larvae decreased significantly. Additionally, morphological changes hepatotoxicity,lethality and heart rate were studied. Despite the fact that a high dose of VPA exerted a cardiotoxic effect this was no longer detected after addition of this drug in the NE. This treatment exerted a significant antiepileptic effectand did not result in highly toxic or lethal effects. In order
-
Set-up of an infrared fast behavioral assay using zebrafish (Danio rerio) larvae, and its application in compound biotoxicity screening
Darío Bicharaa,b, Nora B. Calcaterrab, Silvia Arranzb, Pablo Armasb and
Sergio H. Simonettaa(System: ONE, Model organism: Zebrafish)
PMID: 23401233 DOI: 10.1002/jat.2856
2014 Feb;34(2):214-9. doi: 10.1002/jat.2856. Epub 2013 Feb 11.ABSTRACT:Zebrafish (Danio rerio) is increasingly employed for evaluating toxicity and drug discovery assays. Commonly experimental approaches for biotoxicity assessment are based on visual inspection or video recording. However, these techniques are limited for large-scale assays, as they demand either a time-consuming detailed inspection of the animals or intensive computing resources in order to analyze a considerable amount of screenshots. Recently, we have developed a simple methodology for tracking the locomotor activity of small animals cultured in microtiter plates. In this work, we implemented this automatic methodology, based on infrared (IR) microbeam scattering, for measuring behavioral activity in zebrafish larvae. We determined the appropriate culture conditions, number of animals and stage of development to get robust results.
-
Developmental toxicity assessment of 4-MBC in Danio rerio embryo-larval stages
Ved Prakash, Veena Jain, Shweta Singh Chauhan, Ramakrishnan Parthasarathi, Somendu K. Roy, Sadasivam Anbumani
(System: ARENA, Model organism: Zebrafish)
PMID: 34509837 DOI: 10.1016/j.scitotenv.2021.149920
2022 Jan 15;804:149920. doi: 10.1016/j.scitotenv.2021.149920. Epub 2021 Sep 2.ABSTRACT:Enormous production of cosmetic products and its indiscriminate use tends to discharge into the aquatic environment and might threaten non-target organisms inhabiting aquatic ecosystems. In the present study, developmental toxicity of 4-methylbenzylidene camphor (4-MBC), a widely used organic UV filter in personal care
products has been evaluated using zebrafishembryo-larval stages.Waterborne exposure induced developmental toxicity and deduced 2.71 mg/L as 96 h LC50 whereas embryos exposed to sub-lethal concentrations (50 and 500 μg/L) caused a significant delay in hatching rate, heart rate, reduced larval length, and restricted hatchlings motility besides the axial curvature. Chronic exposure to 10 dpf resulted in significant decrease in SOD activity at 500 μg/L with no changes in CAT level besides a significant increase in GST enzyme at 5 μg/L concentration in 5 dpf sampled larvae. However, all the three enzymes were significantly elevated in 10 dpf larvae indicating differential oxidative stress during the stages of development. Similar trend is noticed for acetylcholine esterase enzymeactivity.A concentration dependent increase inmalondialdehyde contentwas noted in larvae sampled at 5 and 10 dpf. In addition, multixenobiotic resistance (MXR) activity inhibition, and elevated oxidative tissue damage were noticed at 5 dpf with no significant changes in 10 dpf larvae. Furthermore, immunoblot analysis confirms 4-MBC induced apoptosis in zebrafish larvae with promoted cleaved Caspase-3, Bax and inhibited
Bcl-2 proteins expression. Subsequently, docking studies revealed the binding potential of 4-MBC to zebrafishAbcb4 and CYP450 8A1 proteinswith the binding energy of−8.1 and−8.5 kcal/mol representing target proteins