The Development and Evaluation of Brain and Heart Cell Lines from a Marine Fish for Use in Xenobiotic-Induced Cytotoxicity Testing

Two cell lines derived from the brain and heart of a Pacific white snook specimen ( Centropomus viridis) were developed and evaluated in terms of their responsiveness to glyphosate-induced cytotoxicity. The cells were grown in Leibovitz-15 (L-15) medium supplemented with 10% fetal bovine serum (FBS) and were passaged 36 times. Growth was tested at different concentrations of FBS (5, 10 and 20%) at 27°C. The cell lines were cryopreserved at different passages and were successfully thawed, with a survival rate greater than 80% without detectable contamination. At passage 36, the cells were used to assess the deleterious effects of glyphosate, and cell proliferation was measured by direct counting and with the MTT assay. Similar LC50 values were obtained with both methods. Although the principles behind these two assessment methods differ, our results show that both are suitable for evaluating glyphosate toxicity. In addition, heart- and brain-derived cells showed similar sensitivity, suggesting that the same mode of action might be responsible for the toxicity of glyphosate at the cellular level. The newly developed Pacific white snook brain and heart cell lines could be useful to investigate cellular and molecular mechanisms of toxicity, satisfying the need to reduce the use of animals in experiments. Glyphosate-related toxicological data obtained in the present study will allow us to continue investigating the effects of this herbicide directly on brain and heart fish cells since similar studies have only been carried out on either live organisms or on human cell lines such as neuroblastoma, which are immortalised by oncogenes or similar.

New Models to Predict the Acute and Chronic Toxicities of Representative Species of the Main Trophic Levels of Aquatic Environments

To assess the impact of chemicals on an aquatic environment, toxicological data for three trophic levels are needed to address the chronic and acute toxicities. The use of non-testing methods, such as predictive computational models, was proposed to avoid or reduce the need for animal models and speed up the process when there are many substances to be tested. We developed predictive models for Raphidocelis subcapitata, Daphnia magna, and fish for acute and chronic toxicities. The random forest machine learning approach gave the best results. The models gave good statistical quality for all endpoints. These models are freely available for use as individual models in the VEGA platform and for prioritization in JANUS software.

Preparation of a Nonclinical Overview Based on Scientific Literature

Current requirements for the registration dossier include submission of a preclinical (nonclinical) overview, including scientific literature data on preclinical studies and actual preclinical data on the medicinal product. For some groups of medicines, scientific literature data may be used instead of actual preclinical data, which may be redundant. One of the important functions of the scientific literature review is the analysis of updated preclinical information on the medicinal product, which reflects the product’s characteristics, supports conclusions on its efficacy or safety, and may affect the results of the benefit/risk assessment. The aim of the study was to determine the optimal format for presenting scientific literature data in a nonclinical overview that would reflect the methodological aspects of preclinical pharmacology and toxicology studies of medicines. The authors analysed the regulations of the Russian Federation and the Eurasian Economic Union containing requirements for the scientific literature review submitted instead of actual preclinical data as part of the registration dossier for a medicinal product. The authors also considered potential difficulties in preparing a nonclinical overview based on scientific literature. In order to systematise scientific literature data, it is recommended to provide pharmacodynamic, pharmacokinetic, and toxicological data using a format consistent with the common technical document. The proposed recommendations help to harmonise the process of preparation and design of a nonclinical overview which should contain data and facts enabling a reasoned assessment of the benefit/risk ratio. The standardised format of literature data presentation will help the developer prepare an adequate nonclinical overview and will speed up assessment of clinical trial or marketing authorisation applications.

Are carboxyhaemoglobin levels and distance of the corpse from the site of explosion linked? Results from the retrospective analysis of a terroristic attack

We present a retrospective analysis of a terrorist attack with incendiary grenades. We tried to analyse the correlation between carboxyhaemoglobin levels and distance of the corpse from the explosion site to check whether there is a direct relationship between them. In most fatal cases caused by grenade explosions reported in literature, death results from disruptive injuries or following multiorgan complications of the trauma, but here the terrorists used incendiary grenades causing serious burns to the victims with flames at the site of explosion. As a result, the external injuries were not fatal for most victims, especially those further from the explosion site where the flames and carbon monoxide from the fire played a significant role in causing death. The convergence of circumstantial data, autopsy results, and toxicological data led us to conclude that a direct link between the carboxyhaemoglobin level and distance from the site of explosion could not be made. Understanding the limitation of the value of linking such data is important both for police and forensic pathologists as this can reduce potential errors in reconstructing the dynamics of the event.

Pharmacogenetics and Forensic Toxicology: A New Step towards a Multidisciplinary Approach

Pharmacogenetics analyzes the individual behavior of DNA genes after the administration of a drug. Pharmacogenetic research has been implemented in recent years thanks to the improvement in genome sequencing techniques and molecular genetics. In addition to medical purposes, pharmacogenetics can constitute an important tool for clarifying the interpretation of toxicological data in post-mortem examinations, sometimes crucial for determining the cause and modality of death. The purpose of this systematic literature review is not only to raise awareness among the forensic community concerning pharmacogenetics, but also to provide a workflow for forensic toxicologists to follow in cases of unknown causes of death related to drug use/abuse. The scientific community is called on to work hard in order to supply evidence in forensic practice, demonstrating that this investigation could become an essential tool both in civil and forensic contexts. The following keywords were used for the search engine: (pharmacogenetics) AND (forensic toxicology); (pharmacogenetics) AND (post-mortem); (pharmacogenetics) AND (forensic science); and (pharmacogenetics) AND (autopsy). A total of 125 articles were collected. Of these, 29 articles were included in this systematic review. A total of 75% of the included studies were original articles (n = 21) and 25% were case reports (n = 7). A total of 78% (n = 22) of the studies involved deceased people for whom a complete autopsy was performed, while 22% (n = 6) involved people in good health who were given a drug with a subsequent pharmacogenetic study. The most studied drugs were opioids (codeine, morphine, and methadone), followed by antidepressants (tricyclic antidepressants and venlafaxine). Furthermore, all studies highlighted the importance of a pharmacogenetics study in drug-related deaths, especially in cases of non-overdose of drugs of abuse. This study highlights the importance of forensic pharmacogenetics, a field of toxicology still not fully understood, which is of great help in cases of sudden death, deaths from overdose, deaths after the administration of a drug, and also in cases of complaint of medical malpractice.

Guidance Level for Brevetoxins in French Shellfish

Brevetoxins (BTXs) are marine biotoxins responsible for neurotoxic shellfish poisoning (NSP) after ingestion of contaminated shellfish. NSP is characterized by neurological, gastrointestinal and/or cardiovascular symptoms. The main known producer of BTXs is the dinoflagellate Karenia brevis, but other microalgae are also suspected to synthesize BTX-like compounds. BTXs are currently not regulated in France and in Europe. In November 2018, they have been detected for the first time in France in mussels from a lagoon in the Corsica Island (Mediterranean Sea), as part of the network for monitoring the emergence of marine biotoxins in shellfish. To prevent health risks associated with the consumption of shellfish contaminated with BTXs in France, a working group was set up by the French Agency for Food, Environmental and Occupational Health & Safety (Anses). One of the aims of this working group was to propose a guidance level for the presence of BTXs in shellfish. Toxicological data were too limited to derive an acute oral reference dose (ARfD). Based on human case reports, we identified two lowest-observed-adverse-effect levels (LOAELs). A guidance level of 180 µg BTX-3 eq./kg shellfish meat is proposed, considering a protective default portion size of 400 g shellfish meat.

MYTOXDATA MYTOXDATA: A TOXICOLOGICAL DATA MANAGEMENT FOR NATIONAL POISON CENTRE

The aim of the present paper is to evaluate the Malaysian Toxicological Database (MyToxData), a newly developed system for toxicological data management. MyToxData is a local system for collection of case reporting and toxicity statistics as well as its database of chemical products and toxicity information. An online questionnaire using JotForm was distributed among healthcare professionals in Malaysian hospitals. The participants were asked to perform tasks on MyToxData system before rating their satisfaction experience. The questionnaire was adopted from previous literature and comprised 12 items, each having a five-point scale that ranged from Strongly Disagree to Strongly Agree. The collected data were then analysed using the SPSS software package. Based on the results, the satisfaction testing showed a high satisfaction score for the login at 95.7% and case registration and toxicological/poisoning case notification at 65.2%. The poisoning substance identification component received a 78.2% agreement on the usefulness with a very high acceptance of 91.3% on the treatment and clinical information retrieval made available on the system. One sample t-test found that all constructs for the satisfaction testing of MyToxData, namely, accuracy, content, ease of use, format and timeliness, were significant. The higher acceptance by healthcare professionals on the satisfaction testing of MyToxData has identified the effectiveness of the development and subsequent usage of a dedicated system for toxicological data management.

Gas chromatography/tandem mass spectrometry with long-term alkaline transesterification for the determination of esters of monochloropropanediols and glycidol with fatty acids in edible vegetable oils and fats

Introduction. The article discusses the method of determination of 2-, 3-monochloropropanediol (2-MCPDE, 3-MCPDE) and glycidyl (GE) fatty acids esters in edible vegetable oils and fats by slow alkaline transesterification and GC-MS/MS. Many toxicological data showed these substances to have the potential for human health risks if consumed; in addition, existing determination methods have various limitations. So, there is a need to expand the variety of 2-MCPDE, 3-MCPDE and GE determination methods. Materials and methods. 2- and 3-MCPD esters, and glycidyl esters in their corresponding free forms were measured by slow alkaline transesterification with GC-MS/MS. Results. Multiple reactions monitoring mode parameters were optimized. Compared to SIM operating mode, MRM mode improved the selectivity and increased the signal-to-noise ratio. The method was linear in the range of 0.3 mg/kg - 30.0 mg/kg for each analyte. Recovery, tested in this range, was found to be 87.5%-113.6 %. Conclusion. The method discussed here was based on the ISO 18363-2: 2018 standard. In contrast to the last one, we used tandem chromatography-mass spectrometry to detect analytes and a method of absolute calibration with internal standards to quantitate them. In addition, it is proposed to analyze a spiked sample during the sequence of unknown samples to improve the quality of results.

Pharmacokinetics, Biodistribution, and Biosafety of PEGylated Gold Nanoparticles In Vivo

Despite the obvious advantages of gold nanoparticles for biomedical applications, controversial and incomplete toxicological data hamper their widespread use. Here, we present the results from an in vivo toxicity study using gold nanoparticles coated with polyethylene glycol (PEG-AuNPs). The pharmacokinetics and biodistribution of PEG-AuNPs were examined in the rat’s liver, lung, spleen, and kidney after a single i.v. injection (0.7 mg/kg) at different time intervals. PEG-AuNPs had a relatively long blood circulation time and accumulated primarily in the liver and spleen, where they remained for up to 28 days after administration. Increased cytoplasmic vacuolation in hepatocytes 24 h and 7 days after PEG-AuNPs exposure and apoptotic-like cells in white splenic pulp 24 h after administration has been detected, however, 28 days post-exposure were no longer observed. In contrast, at this time point, we identified significant changes in lipid metabolism, altered levels of liver injury markers, and elevated monocyte count, but without marked biological relevance. In blood cells, no DNA damage was present in any of the studied time intervals, with the exception of DNA breakage transiently detected in primary kidney cells 4 h post-injection. Our results indicate that the tissue accumulation of PEG-AuNPs might result in late toxic effects.