Good Laboratory Practice Research Articles

Role of model organisms and nanocompounds in human health risk assessment.

Safeguarding the environment is one of the most serious modern challenges, as increasing amounts of chemical compounds are produced and released into the environment, causing a serious threat to the future health of the Earth as well as organisms and humans on a global scale. Ecotoxicology is an integrative science involving different physical, chemical, biological, and social aspects concerned with the study of toxic effects caused by natural or synthetic pollutants on any constituents of ecosystems, including animals (including humans), plants, or microorganisms, in an integral context. In recent decades, this science has undergone considerable development by addressing environmental risk assessments through the biomonitoring of indicator species using biomarkers, model organisms, and nanocompounds in toxicological assays. Since a single taxon cannot be representative of complex ecotoxicological effects and mechanisms of action of a chemical, the use of test batteries is widely accepted in ecotoxicology. Test batteries include properly chosen organisms that are easy to breed, adapt easily to laboratory conditions, and are representative of the environmental compartment under consideration. One of the main issues of toxicological and ecotoxicological research is to gain a deeper understanding of how data should be obtained through laboratory and field approaches using experimental models and how they could be extrapolated to humans. There is a tendency to replace animal tests with in vitro systems and to perform them according to standardized analytical methods and the rules of the so-called good laboratory practice(GLP). This paper aims to review this topic to stimulate both efforts to understand the toxicological and ecotoxicological properties of natural and synthetic chemicals and the possible use of such data for application to humans.

Accurate Quantification of AAV Vector Genomes by Quantitative PCR.

The ability to accurately determine the dose of an adeno-associated viral (AAV) therapeutic vector is critical to the gene therapy process. Quantitative PCR (qPCR) is one of the common methods to quantify the AAV vector titre, but different variables can lead to inconsistent results. The aim of this study was to analyze the influence of the conformation of the DNA used as the standard control, and the enzymatic digestion was performed to release the viral genome from the protein capsid on the physical genome titration of a clinically relevant AAV8.RPGR vector, made to good laboratory practice standards in an academic setting. The results of this study showed that the conformation of the DNA used as standard has a significant impact on the accuracy of absolute quantification by qPCR. The use of supercoiled undigested plasmid DNA template generated a higher apparent titer, as compared to the use of linearized plasmid as the standard. In contrast to previous studies, the pre-treatment of the samples with Proteinase K, in addition to the high temperature step used after DNase I digestion, resulted in a reduction on AAV titers. Ideally, all AAV documentation should state which form of reference plasmid and which pre-treatment of the samples have been used to calculate titers, so that appropriate comparisons relating to dose toxicity and transduction efficacy can be made in the clinical scenario.

Ripple effects of research capacity strengthening: a study of the effects of a project to support test facilities in three African countries towards Good Laboratory Practice certification

Background: Strengthening capacity for public health research is essential to the generation of high-quality, reliable scientific data. This study focuses on a research capacity strengthening project supporting seven test facilities in Africa conducting studies on mosquito vector control products towards Good Laboratory Practice (GLP) certification. It captures the primary effects of the project on each facility’s research capacity, the secondary effects at the individual and institutional level, and the ripple effects that extend beyond the research system. The relationships between effects at different levels are identified and compared to an existing framework for the evaluation of research capacity strengthening initiatives.Methods: To capture the views of individuals engaged in the project at all levels within each facility, a maximum-variation purposive sampling strategy was used. This allowed triangulation between different data sources. Semi-structured interviews were conducted with individuals in three facilities and a combination of email and remote video-call interviews were conducted with individuals at two further facilities.Results: We found that, despite a focus of the GLP certification project at the institutional level, the project had effects also at individual (including enhanced motivation, furtherment of careers) and national/international levels (including development of regional expertise). In addition, we detected ripple effects of the project which extended beyond the research system.Conclusion: This study shows that research capacity strengthening interventions that are focussed on institutional level goals require actions also at individual and national/international levels. The effects of engagement at all three levels can be amplified by collaborative actions at the national/international level. These findings show that research capacity strengthening projects must develop plans that address and evaluate impact at all three levels. Capturing the ripple effects of investment in research capacity strengthening should also be planned for from the beginning of projects to support further engagement of all stakeholders.

Managing the bacterial contamination risk in an axenic mice animal facility.

A gap exists between good laboratory practices with axenic animals and the procedures applied. This work examined the efficacy of sodium dichloroisocyanurate (MB-10) and potassium peroxymonosulfate (Virkon™) disinfectants, as well as the appropriate soaking time for materials used with the ISOcage Biosafety Station™. We also compared the microbial load in cage systems hosting mice over 2 weeks in axenic rooms (ARs) and in typical specific-pathogen-free (SPF) non-axenic rooms (NARs) to identify resistant microorganisms, targeted for longer soaking disinfection, and evaluated the necessary procedures for reducing the microbial load in AR. Staphylococcus was the most frequently isolated genus (in both ARs and NARs). An average of three spore-forming microorganisms per cage were counted from AR. The disinfection time to reach 1 log reduction for Bacillus atrophaeus spores varied from 138 s (100 ppm MB-10) to 290 (Virkon™) to <20 s for S. epidermidis (100 ppm MB-10). AR management protocols lead to a microbial load that is 1000 times lower than that found in NARs. Data comparing the microbial load in SPF and axenic facilities can be used to improve the effectiveness of their microbial control procedures.

Seroprevalencia y seguimiento de anticuerpos IgG contra SARS-CoV-2 en personal del Laboratorio Estatal de Salud Pública de Guanajuato LaESaP

El sector salud, al frente de la actual pandemia, está altamente expuesto al contagio de COVID- 19. Dada la variada sintomatología que presenta la enfermedad y ante la escasez de estudios de seroprevalencia en unidades de diagnóstico por Laboratorio, surge la necesidad de estudiar la presencia y comportamiento de los anticuerpos anti SARS-CoV-2 en el personal del Laboratorio Estatal de Salud Pública de Guanajuato (LaESaP), lugar donde se efectúa el diagnóstico molecular de COVID-19 en Guanajuato, desde marzo de 2020. Este estudio pretende estimar la seroprevalencia de anticuerpos IgG anti SARS-CoV-2 en sus trabajadores, así como su permanencia tras un periodo de 5 meses. Método: Se recolectaron las muestras sanguíneas de 100 trabajadores pertenecientes al LaESaP, que representan la totalidad de los trabajadores que mantuvieron sus actividades laborales desde marzo de 2020 y se analizaron mediante inmunoanálisis quimioluminiscente de micropartículas (CMIA), para la detección cualitativa de anticuerpos IgG frente al virus del SARS-CoV-2. Resultados: Se encontró una seroprevalencia del 6%, sin embargo, solo el 67% conservó un título de anticuerpos positivo después de 5 meses. No se establecieron correlaciones en los títulos de anticuerpos por género o sintomatología. Los 4 trabajadores con antecedentes de prueba rRT-PCR positiva presentaron anticuerpos al inicio del estudio, así como 2 trabajadores sin antecedente de prueba rRT-PCR, considerados como asintomáticos. Discusión: Se observó una seroprevalencia baja en el personal de laboratorio en comparación con otros estudios realizados a personal de salud. Lo anterior puede deberse al adecuado uso de equipo de protección personal, buenas prácticas de laboratorio e implementación de medidas de contención dentro de la institución.

Industrial Development of Standardized Fetal Progenitor Cell Therapy for Tendon Regenerative Medicine: Preliminary Safety in Xenogeneic Transplantation.

Tendon defects require multimodal therapeutic management over extensive periods and incur high collateral burden with frequent functional losses. Specific cell therapies have recently been developed in parallel to surgical techniques for managing acute and degenerative tendon tissue affections, to optimally stimulate resurgence of structure and function. Cultured primary human fetal progenitor tenocytes (hFPT) have been preliminarily considered for allogeneic homologous cell therapies, and have been characterized as stable, consistent, and sustainable cell sources in vitro. Herein, optimized therapeutic cell sourcing from a single organ donation, industrial transposition of multi-tiered progenitor cell banking, and preliminary preclinical safety of an established hFPT cell source (i.e., FE002-Ten cell type) were investigated. Results underlined high robustness of FE002-Ten hFPTs and suitability for sustainable manufacturing upscaling within optimized biobanking workflows. Absence of toxicity or tumorigenicity of hFPTs was demonstrated in ovo and in vitro, respectively. Furthermore, a 6-week pilot good laboratory practice (GLP) safety study using a rabbit patellar tendon partial-thickness defect model preliminarily confirmed preclinical safety of hFPT-based standardized transplants, wherein no immune reactions, product rejection, or tumour formation were observed. Such results strengthen the rationale of the multimodal Swiss fetal progenitor cell transplantation program and prompt further investigation around such cell sources in preclinical and clinical settings for musculoskeletal regenerative medicine.

ИЗУЧЕНИЕ ОБЩЕТОКСИЧЕСКОГО ДЕЙСТВИЯ ТРАВЫ КИПРЕЯ УЗКОЛИСТНОГО

The results of the toxicological experiment on extracts from Epilobium angustifolium L. herb are presented. Absence of reliable negative changes, structural and functional disorders was established in accordance with the criteria for assessing the harmfulness and danger of toxic effect of the raw materials studied in Good Laboratory Practice (GLP). Significant differences in some hematological parameters in the experimental group of animals where the dose of Epilobium angustifolium extract making 800 mg/kg of animal weight was chosen for study compared to the control one are not diagnostically significant and are within the fluctuations of physiological norms for the animals species given. It was found that the extracts in doses of 200, 500 and 800 mg/kg (by dry residue) in repeated oral administration for 28 days do not cause detectable toxic effects.

Comprehensive review of genotoxicity data for diclofenac

Diclofenac is a non-steroidal anti-inflammatory drug discovered several decades ago, which has since been used by an estimated one billion patients and has demonstrated an acceptable safety profile. In support of its marketing approval, a comprehensive set of genotoxicity studies were conducted in vitro and in vivo. Despite the fact that these studies preceded both Good Laboratory Practice (GLP) requirements and International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH) guidelines on genotoxicity testing, they were conducted using the best scientific principles and are considered appropriate by contemporary standards. In addition to bacterial mutagenicity and mammalian in vitro assays, repeat-dose somatic, germ cell and dominant lethal assays were conducted. These data are made available for the first time to offer researchers an opportunity to review the existing data set that unequivocally demonstrates that diclofenac sodium is not genotoxic. This is further substantiated by long-term bioassay data demonstrating that diclofenac sodium has no carcinogenic potential in rodents. However, more recently, new studies have been published showing a genotoxic potential for diclofenac in novel or modified in vitro test systems. These new publications are discussed in the context of the existing comprehensive data package.

Safety and chronic lesion characterization of pulsed field ablation in a Porcine model.

BackgroundPulsed field ablation (PFA) has been identified as an alternative to thermal‐based ablation systems for treatment of atrial fibrillation patients. The objective of this Good Laboratory Practice (GLP) study was to characterize the chronic effects and safety of overlapping lesions created by a PFA system at intracardiac locations in a porcine model.MethodsA circular catheter with nine gold electrodes was used for overlapping low‐ or high‐dose PFA deliveries in the superior vena cava (SVC), right atrial appendage (RAA), and right superior pulmonary vein (RSPV) in six pigs. Electrical isolation was evaluated acutely and chronic lesions were assessed via necropsy and histopathology after 4‐week survival. Acute and chronic safety data were recorded peri‐ and post‐procedurally.ResultsNo animal experienced ventricular arrhythmia during PFA delivery, and there was no evidence of periprocedural PFA‐related adverse events. Lesions created in all anatomies resulted in electrical isolation postprocedure. Lesions were circumferential, contiguous, and transmural, with all converting into consistent lines of chronic replacement fibrosis, regardless of trabeculated or smooth endocardial surface structure. Ablations were non‐thermally generated with only minimal post‐delivery temperature rises recorded at the electrodes. There was no evidence of extracardiac damage, stenosis, aneurysms, endocardial disruption, or thrombus.ConclusionPFA deliveries to the SVC, RAA, and RSPV resulted in complete circumferential replacement fibrosis at 4‐week postablation with an excellent chronic myocardial and collateral tissue safety profile. This GLP study evaluated the safety and efficacy of a dosage range in preparation for a clinical trial and characterized the non‐thermal nature of PFA.

Evaluation of oral toxicity and genotoxicity of Achyranthis Radix extract

Ethnopharmacological relevanceThe root of Achyranthes bidentata Blume, Achyranthis Radix (AR), is used as a traditional medicine ingredient in East Asia. It has anti-inflammatory, anti-oxidative, and anti-diabetic activities. Aim of the studyIn the present study, we aimed to evaluate the oral toxicity and genotoxicity of single-dose and 4-week repeated-doses of AR hot water extract (ARE), under the good laboratory practice principles. Materials and methodsFor oral toxicity studies, SD rats (n = 5 per sex and group) were administered ARE at concentrations of 500, 1000, and 2000 mg/kg/day once (single dose) or once per day for 4 weeks (repeated dose). The non-clinical genotoxicity study consisted of bacterial reverse mutation using Escherichia coli (WP2 uvrA) and Salmonella typhimurium (TA98, TA100, TA1535, and TA1537), in vitro chromosomal aberration test with Chinese hamster lung cells (CHL/IU), and in vivo mouse bone marrow micronucleus test using bone marrow cells collected from male ICR mice (n = 5) that were orally administered ARE. ResultsIn the single-dose oral toxicity study, mortality and treatment-related changes in body weight were not observed throughout the study, and the lethal dose was estimated to be > 2000 mg/kg in rats. In the 4-week repeated-dose oral toxicity study, ARE did not induce significant changes in body weight, organ weight, food intake, or hematological and serum biochemical parameters in any group. In the bacterial reverse mutation test, ARE did not induce gene mutations in any tested strain. In the chromosomal aberration test, ARE did not cause chromosomal aberrations. The micronucleus test showed no significant increase in the number of micronucleated polychromatic erythrocytes or the mean ratio of polychromatic to total erythrocytes. ConclusionsThese results showed that ARE does not induce oral toxicity and genotoxicity in the in vivo and in vitro test systems.

Detection of Barrett's Neoplasia with Near-Infrared Fluorescent Heterodimeric Peptide: Feasibility Results from a Phase 1 Study

Background: Esophageal adenocarcinoma is a molecularly heterogeneous disease that is rising rapidly in incidence and has a poor prognosis. Current methods of endoscopic surveillance in patients with Barrett’s esophagus (BE) using white light illumination are not sensitive to pre-malignant lesions that are flat in appearance and patchy in distribution. We aim to demonstrate specific binding by a peptide heterodimer to Barrett’s neoplasia in human subjects. Methods: Peptide monomers specific for EGFR and ErbB2 were arranged in a heterodimer configuration and labeled with IRDye800. The near-infrared (NIR)-labeled heterodimer was synthesized using current good manufacturing practices (cGMP) methods and performed pharmacology/toxicology study in animals using good laboratory practices (GLP). The NIR contrast agent was topically administered in the distal esophagus of patients with history of neoplastic BE undergoing endoscopic therapy or surveillance. Fluorescence images were collected using a flexible fiber accessory passed through the instrument channel of an upper endoscope. Fluorescence images were collected from n = 31 BE patients, and a deep learning model was used to segment the target (T) and background (B) regions to calculate the T/B ratio. Findings: Between August 2018 and November 2020, 25 human subjects were enrolled for safety study and 31 patients for either evaluation or therapy of Barrett’s neoplasia were recruited for imaging study. No adverse events attributed to the heterodimer were found. The mean T/B ratio in patients for HGD and EAC were significantly greater than that for BE with low grade dysplasia, without dysplasia, or squamous mucosa. At a T/B ratio of 1.5, 94.1% sensitivity and 92.6% specificity for detection of Barrett’s neoplasia was achieved with an AUC = 0.95. The presence of Barrett’s neoplasia was validated by pathological assessment of resected specimens. Interpretation: This “first-in-human” clinical study demonstrates feasibility to detect early Barrett’s neoplasia using a NIR-labeled peptide heterodimer. Trial Registration: This study was registered online at ClinicalTrials.gov (NCT03643068) Funding National Institutes of Health. Declaration of Interest: Authors (JC, TDW) are inventors on patents filed by the University of Michigan on the peptide heterodimer used in this study. Author (EJS) is an inventor on patents filed by the University of Washington on the multimodal scanning fiber endoscope (mmSFE) used in this study. The other authors disclose no conflicts. Ethical Approval: All study protocols were reviewed and approved by the Institutional Review Board (IRB) at Michigan Medicine

Study the Effect of Naturally Low Temperature in Animals Experiment

In modern conditions of Arctic development, the problem of hypothermia of humans and animals is becoming fundamental and applicable nature. Studying deep hypothermia effect and body recovery after it on experimental models of animals is relevant for world science. The article presents some aspects of the naturally low temperatures affect (-40° C and below) on the domestic pig’s body under the conditions of Yakutia (experimental research). To perform this task, we simulated the conditions of natural deep hypothermia, in which we studied the mechanisms of entering an anabiosis state and got the data of experimental animals’ brain and heart bioelectrical activity. The experiment was conducted according to the ethical standards of the European Convention for the Protection of Vertebrate Animals used for Experimental and Other Scientific Purposes No. 123 of 18 March 1986, Strasbourg, and the order of the Ministry of Health of the Russian Federation No. 199n of 01.04.2016 “on approval of the regulations of good laboratory practice”. Permission of the bioethical commission is available.

Features of the macro and microscopic structure of the lungs of a mature rabbit – Oryctolagus cuniculus L. 1758

The macro- and microscopic structure of the lungs of mature rabbits – Oryctolagus cuniculus L., 1758 – European rabbit was studied. This work was performed using comprehensive research methods: anatomical, organometric, histological and histometric. The research was carried out in the conditions of the laboratory of pathomorphology in the Department of Anatomy and Histology. The scientific work was performed on the subject of research work of the Department of Anatomy and Histology of Polissya National University on the theme: “Development, morphology and histochemistry of animal organs in normal and pathological”, according to the state registration number – № 0113V000900. The morphological studies followed the basic rules of Good Laboratory Practice GLP (1981), the provisions of the “General Ethical Principles of Animal Experiments”, adopted by the First National Congress of Bioethics (Kyiv, 2001) and the requirements for the “Rules for carrying out work with experimental animals”, approved by the order of the Ministry of Health № 281 of November 1, 2000 “On measures to further improve the organizational forms of work with the use of experimental animals”. The material for histomorphological studies were pieces of lungs selected from clinically healthy adult rabbits (n = 5). Slices were made from paraffin blocks on a sled microtome MS–2. The thickness of histological sections did not exceed 8–12 μm, which were then stained with hematoxylin and eosin and Van Gizon to study the morphofunctional activity of cells and tissues of the respiratory tract. Histometric studies of histological specimens were performed using a microscope “Micros” with a digital camera at a magnification of × 100–1000. Studies have shown that macroscopically the lungs of mature rabbits have a reflection of the shape of the thoracic cavity, and gradually expands ventrally. The right and left lungs are surrounded by pleural right and left sacs. The pleural cavities of both lungs in the test animals are not interconnected. Organometry showed that the absolute lung mass of rabbits is 18.05 ± 1.32 g, relative to 0.624 ± 0.013 %. The histoarchitectonics of the lungs is formed by the lung lobes. The respiratory department of the lungs is represented by a set of acinuses. The latter are represented by respiratory bronchioles, alveolar passages, alveolar sacs and alveoli that form the alveolar tree. The results of morphometric studies showed that the respiratory (respiratory) part of the lungs of rabbits occupies 39.6 ± 0.62 %, and the connective tissue base, respectively, 58.5 ± 1.27 %. The average volume of alveoli, which are divided into small, medium and large, is 42.3 ± 4.35 thousand μm3. Thus, studies of the macro- and microscopic structure of the lungs of experimental animals are of great general biological importance, as they are an important criterion for objective assessment for the differential diagnosis of diseases of various origins.

Critical review of the publications on the genotoxicology of aluminium salts: 1990-2018.

Since the mid-1970s, there have been many reports that purport to implicate aluminium in the aetiology of neurodegenerative disease. After several decades of research, the role of aluminium in such disease remains controversial and is not the subject of this review. However, if aluminium is implicated in such disease then it follows that there must be a toxicological mechanism or mode of action, and many researchers have investigated various potential mechanisms including the involvement of oxidative damage, cytotoxicity and genotoxicity. This paper reviews many of the publications of studies using various salts of aluminium and various genotoxicity end points, both in vitro and in vivo, with a focus on oxidative damage. The conclusion of this review is that the majority, if not all, of the publications that report positive results have serious technical flaws and/or implausible findings and consequently should contribute little or no weight to a weight of evidence (WoE) argument. There are many high-quality, Good Laboratory Practice (GLP)-compliant genotoxicity studies, that follow relevant OECD test guidelines and the European Chemicals Agency (ECHA) integrated mutagenicity testing strategy, on several salts of aluminium; all demonstrate clear negative results for both in vitro and in vivo genotoxicity. In addition, the claim for an oxidative mode of action for aluminium can be shown to be spurious. This review concludes that there are no reliable studies that demonstrate a potential for genotoxicity, or oxidative mode of action, for aluminium.

Managing COVID-19 Lockdown Impacts: Sustaining GLP Compliance and Man Material Medium (MMM) Strategy for Augmenting Prevention of Workplace Infections.

Good Laboratory Practices (GLP) is a well-established global system that encompass a set of principles or a framework for defining how laboratory studies are planned, performed, monitored, recorded, reported, and stored for future reference. It is important that compliance with the principles of GLP continues to be maintained. Coronavirus disease 2019 (COVID-19) pandemic lockdowns in various countries, including India, have been sudden and over an extended duration. Although every GLP laboratory has Standard Operating Procedure for disaster management, the sudden lockdown due to COVID-19 created specific emergency procedures related to this situation such as travel bans, safe distancing, and work from home notifications. Good Laboratory Practice compliances in the context of animal experimentation during and post lockdown period need effective managerial responses that are not just flexible and innovative but can ensure they are well-calibrated to the challenges of business continuity and maintenance of health directives. On-the-ground realities suggest there may still be practical challenges to compliance, and guidelines may not always be complied with. This article discusses the issues that may be encountered due to COVID-19 that could potentially impact the GLP status of a study and suggests ways to manage them so as to minimize or prevent infection with COVID-19. We propose an MMM (Man, Material, and Medium) strategy to ensure compliance with health directives and guidelines that will help staff to keep themselves and others safe in the workplace while endeavoring to comply with GLP requirements.

Good laboratory practice-compliant bio-efficacy testing of repellent products against Aedes aegypti

The Medical Entomology Unit of the Institute for Medical Research, Kuala Lumpur, is a laboratory that was certified in 2019 to conduct evaluation of vector control products in compliance to the Organisation for Economic Co-operation and Development principles of good laboratory practice (GLP) laboratory. To ensure the product evaluation practice complies with the GLP principles and corrections are made for improving test procedures, internal inspections by the Quality Assurance team play a crucial role. Before commencing GLP-compliant studies, study plans are developed according to the study objectives and critically examined to ensure they are performed in accordance with the GLP principles. We described the quality of test system, test item and test method used for evaluation of repellent effect of topically applied repellent products in accordance to the GLP test requirements.

Acute and Subchronic Toxicities and Safety Pharmacology Studies of a Bacillus Subtilisin in Dogs.

Subtilisin NAT, a Bacillus subtilisin, is widely applied as a functional food and considered to be one of the most exploitable potential oral thrombolytic agents. Subtilisin QK, another Bacillus subtilisin, is a serine protease fermented by Bacillus subtilis 02 and has a better thrombolytic effect. Therefore, subtilisin QK is typically used for evaluating the safety of Bacillus subtilisins. Here, we conduct several good laboratory practice (GLP)-compliant studies in non-rodent animal, i.e., in Beagle dogs, including acute toxicity, subchronic toxicity, and safety pharmacology studies. No adverse effects were evident in the acute and 28-d subchronic toxicity studies at doses up to 40000 FU/kg and 16000 FU/kg/d, respectively. In evaluating the pharmacological safety of up to 2000FU/kg subtilisin QK, we found no significant differences between the electrocardiograms, blood pressures, and respiration of beagle dogs. These findings suggest the safety of Bacillus subtilisin, providing reliable pharmacological and toxicological data for its development and popularization as a functional food and drug.

Modern Approach to Assessing Quality of Laboratory Animals Used for Experimental Purposes

Introduction: The main task of conducting a biomedical research is to obtain statistically significant experimental results. The studies with laboratory animals used as test systems are conducted in research organizations. Owing to the introduction of good laboratory practice (GLP) principles into the world practice, stricter requirements have been imposed on the quality of test systems. The purpose of our study was to analyze the regulatory, methodological, and legislative framework governing laboratory research using biomodels in order to update a number of regulatory documents that are mandatory for implementation. Methods: We searched electronic databases of references and abstracts (Web of Science, PubMed, eLibrary) for appropriate keywords and, having reviewed search results, selected 22 literary sources on the topic. Results: We established that, based on existing regulations, specialized institutions develop internal instructions to fulfil their goals and objectives. The presence of a program of standard operating procedures allows experiments to be performed with appropriate quality. The quality of laboratory animals can be affected by unfavorable conditions of keeping, lack of balanced feeding, and violation of sanitary and hygienic standards of care. For different categories of laboratory animals, different requirements are imposed on their housing in accordance with the microbiological status. Institutions are implementing a specially developed program for health monitoring in laboratory animals. Healthy-looking animals are carriers of a number of infectious agents that can distort results of an experiment. Veterinarians specializing in laboratory animals should have knowledge of biomodeling and effects of drugs on physiological and pathological processes in animals and should perform adequate anesthesia during painful manipulations. Only employees with high qualifications and skills in working with animals are authorized to handle biomodels. Conclusions: Our findings demonstrate that the use of high-quality laboratory animals and standardization of all stages of an experiment ensures a quality research compliant with international standards.

Development and validation of a simple and sensitive LC-MS/MS method for quantification of ampicillin and sulbactam in human plasma and its application to a clinical pharmacokinetic study

Ampicillin-sulbactam is a broad-spectrum combination antibiotic used for a variety of clinical applications, including as a prophylactic agent to reduce the risk of surgical site infection. The pharmacokinetics of ampicillin-sulbactam after redosing during prolonged surgeries remains incompletely understood. In anticipation of further studying the intra-operative pharmacokinetics of this drug, we have developed a novel liquid chromatography-tandem mass spectrometry (LC–MS/MS) method for the quantification of ampicillin and sulbactam. The plasma samples were prepared using a simple protein precipitation method. Gradient chromatographic elution was used to separate analytes, and MS/MS analysis was performed in negative ionization mode for both analytes via multiple reaction monitoring (MRM). All validation parameters were evaluated under a good laboratory practice (GLP) environment. For both ampicillin and sulbactam, the lower limit of quantitation (LLOQ) was established as 0.25 μg/mL. The calibration curve ranged from 0.25 to 200 μg/mL for ampicillin and 0.25–100 μg/mL for sulbactam. Inter- and intra-day precisions for both analytes were ≤11.5 % for quality controls and ≤17.4 % for LLOQ; accuracies ranged from -11.5 to 12.5% for 3 quality control levels and -18.1–18.7% for LLOQ. In addition to sensitivity, accuracy and precision, 13 other parameters were also validated for both analytes, and the results met the acceptance criteria. Our method was successfully applied to quantify ampicillin and sulbactam concentrations in patients undergoing surgery.

Compounded conservatism in European re-entry worker risk assessment of pesticides

We review the risk parameters and drivers in the current European Union (EU) worker risk assessment for pesticides, for example considering crop maintenance, crop inspection or harvesting activities, and show that the current approach is very conservative due to multiple worst-case default assumptions.As a case study, we compare generic exposure model estimates with measured worker re-entry exposure values which shows that external cumulative exposure is overpredicted by about 50-fold on average. For this exercise, data from 16 good laboratory practice (GLP)-compliant worker exposure studies in 6 crops were evaluated with a total number of 184 workers.As generic overprediction does not allow efficient risk management or realistic risk communication, we investigate how external exposure can be better predicted within the generic model, and outline options for possible improvements in the current methodology. We show that simply using averages achieves more meaningful exposure estimates, while still being conservative, with an average exposure overprediction of about 9-fold.Overall, EU risk assessment includes several numerically unaccounted “hidden safety factors”, which means that workers are well protected; but simultaneously risk assessments are biased towards failing due to compounded conservatism. This should be considered for further global or regional guidance developments and performing more exposure-relevant risk assessment.