Principles Of Good Laboratory Practice Research Articles

Application of chitosan-based wound dressings in local treatment of parenchymal organ wounds: a preclinical experimental study

Background. Injuries of various origins with damage to parenchymal organs remain one of the urgent issues of emergency surgery, despite all the achievements of medical science of the first quarter of the XXI century. One way to improve the quality of treatment for patients with this pathology consists in developing new wound dressings or modifying the properties of existing ones. Hemostatic efficacy is claimed to be one of the main requirements for such dressings.Objective. To perform a comparative analysis of the efficacy of the developed biodegradable wound dressings based on chitosan in the treatment of experimental wounds of parenchymal organs.Methods. The study involved laboratory animals (male rabbits weighing 2900 ± 150 g) to form experimental wounds of parenchymatous organs. The methodology involved wounding the liver via internal method, with the creation of a tissue defect of a given volume, or forming a linear incised wound with fixed parameters followed by application of a developed wound dressing. In the control group of animals, a collagen hemostatic sponge was used as a wound dressing. The animals used in the experiment were kept in accordance with GOST 33044-2014 “Principles of good laboratory practice”. The experiment involved studying the properties of two samples of wound dressings developed on the basis of modified Chitosan Soft (Ch-S) and Chitosan Hard (Ch-H) chitosan, having different properties due to their different structural parameters. The applied ultrasound control of the experimental wound area, with visualization of the samples of the investigated material in the process of biodegradation, enabled the features of the wound process to be studied in more detail. Statistical processing of the study results was carried out by methods of variational statistics, including the calculation of mean values (M), standard errors of mean values (± m). The criterion of the level of statistical significance considered a value of p <0.05. Histomorphological examination of samples was carried out using classical methods.Results. An experimental study conducted on laboratory animals in the treatment of liver wounds, followed by a comprehensive assessment of the results obtained, revealed high efficacy in the developed wound dressings in comparison with the control. The following parameters predicted in the development of wound dressing samples were confirmed: stable hemostatic and scaffold functions, biodegradability, adhesiveness and stimulating effect on bioreparative processes in the wound area. In addition, the ultrasound control algorithm in vivo was adapted in animals to assess the dynamics of the wound process and biodegradation of the studied wound dressings in wounds of parenchymal organs in the experiment, previously used in studying the dynamics of the wound process in soft tissues.Conclusion. The developed samples of chitosan-based wound dressings proved their efficacy in an experimental model of the wound process in parenchymal organs. The applied ultrasonic method for monitoring the biodegradation of the studied wound dressing, as well as the dynamics of the wound process in the liver tissue, revealed the correspondence between the visualized image and the histomorphological picture of the studied tissues, the structure and properties of the wound dressing.

Preclinical Safety Assessment of Gadopiclenol: A High-Relaxivity Macrocyclic Gadolinium-Based MRI Contrast Agent.

Gadopiclenol is a new high-relaxivity macrocyclic gadolinium-based contrast agent for magnetic resonance imaging of the central nervous system and other body regions. The product has been approved by US Food and Drug Administration and is currently being evaluated by European Medicines Agency. For risk assessment of the single diagnostic use in humans, the safety profile of gadopiclenol was evaluated with a series of preclinical studies. With exception of dose-ranging studies, all safety pharmacology and toxicology studies were performed in compliance with Good Laboratory Practice principles. Safety pharmacology studies were conducted to assess potential effects on cardiovascular (in vitro and in dogs), respiratory (in rats and guinea pigs), neurological (in rats), and renal endpoints (in rats). Toxicology studies were also performed to investigate acute toxicity (in rats and mice), extended single-dose (in rats and dogs) and repeated-dose toxicity (in rats and dogs), reproductive (in rats), developmental (in rats and rabbits) and juvenile toxicity (in rats), as well as genotoxicity (in vitro and in rats), local tolerance (in rabbits), potential immediate hypersensitivity (in guinea pigs), and potential tissue retention of gadolinium (in rats). Safety pharmacology studies conducted at high intravenous (IV) doses showed a satisfactory tolerance of gadopiclenol in the main body systems. After either single or repeated IV dosing (14 and 28 days) in rats and dogs, gadopiclenol was well tolerated even at high doses. The no-observed-adverse-effect level values (ie, the highest experimental dose without adverse effects) representing between 8 times in rats and 44 times in dogs (based on the exposure), the exposure achieved in humans at the intended diagnostic dose, provide a high safety margin. No or only minor and reversible effects on body weight, food consumption, clinical signs, clinical pathology parameters, or histology were observed at the highest doses. The main histological finding consists in renal tubular vacuolations (exacerbated after repeated exposure), which supports a well-known finding for this class of compounds that has no physiological consequence on kidney function. Reproductive toxicity studies showed no evidence of effects on reproductive performance, fertility, perinatal and postnatal development in rats, or reproductive development in rats or rabbits. The safety profile of gadopiclenol in juvenile rats was satisfactory like in adults. Gadopiclenol was not genotoxic in vitro in the Ames test, a mouse lymphoma assay, and a rat in vivo micronucleus test. There were no signs of local intolerance at the injection site after IV and intra-arterial administration in rabbits. However, because of minor signs of intolerance after perivenous administration, misadministration must be avoided. Gadopiclenol exhibited no signs of potential to induce immediate hypersensitivity in guinea pigs. High safety margins were observed between the single diagnostic dose of 0.05 mmol/kg in humans and the doses showing effects in animal studies. Gadopiclenol is, therefore, well tolerated in various species (mice, rats, dogs, rabbits, and guinea pigs). All observed preclinical data support the clinical approval.

Assessment of Safety Profile of Activated Curcumin C3 Complex (AC3®), Enriched Extract of Bisdemethoxycurcumin from the Rhizomes of Curcuma longa.

The present work was carried out to investigate the toxic effects of Activated Curcumin C3 Complex (AC3®) through the methods of acute, subacute, subchronic, reproductive/developmental toxicity, and genotoxicity when administered orally in experimental rodents. The studies were carried out in line with OECD principles of good laboratory practice. A single-dose acute oral toxicity study was conducted on female Wistar rats that produced no toxic effects after 14 days (the observation period) of treatment. Subacute, subchronic, and reproductive/developmental studies were conducted in Wistar rats, divided equally into vehicle control, 125, 250, and 500 mg/kg dose groups along with recovery groups for vehicle control and high dose. In all the studies, there were no abnormal clinical signs/behavioral changes, reproductive and developmental parameters, or gross and histopathological changes. Likewise, no alteration was found in the body weight, hematology, and other biochemical parameters. Also, it did not show mutagenicity in the in vitro AMES test or clastogenicity and aneugenicity in the in vivo micronucleus test, indicating that AC3® did not induce any genotoxic effects. This revealed that oral administration of AC3® is safe in rodents, nonmutagenic, and had no observed adverse effects under experimental conditions.

Bezpieczeństwo chemiczne i gwarancje jego zapewnienia w Unii Europejskiej

This article analyses the guarantees of chemical safety defined by the provisions of European Union law. It defines the essence of chemical safety; the author presents the importance of the principles of Good Laboratory Practice, which are a manifestation of the uniformity of standards for dealing with chemical substances. Next, it discusses EU Bezpieczeństwo chemiczne i gwarancje jego zapewnienia w Unii Europejskiej RAP 2023 (9) legal solutions to the registration and marketing authorization of chemical substances (REACH regulation). It also analyses the issue of the export and import into the EU of substances classified as dangerous. It argues that the conditions of European integration have enabled the unification of regulations and standardization of procedures for dealing with chemical substances. Therefore, the author is convinced that the legal regulations of the EU constitute an important guarantee of chemical safety.

The safety of BCG vaccination in cattle: results from good laboratory practice safety studies in calves and lactating cows

Bovine tuberculosis (bTB) is a global disease of livestock that has damaging economic, animal health and public health consequences. Conventional bTB disease control strategies, based around the testing and slaughter of cattle infected with bTB, are typically used to help limit or reduce the transmission of this disease but in many low- and middle-income countries such strategies may often be economically unviable, culturally unacceptable or logistically impracticable. The use of vaccination to protect cattle against bTB could provide a potentially more affordable, ethically acceptable and practical additional disease control measure. The protective efficacy of the commercially produced and readily available human vaccine against tuberculosis (Mycobacterium bovis Bacille Calmette-Guérin; BCG) in cattle has been demonstrated in many experimental laboratory and field studies. However, Good Laboratory Practice (GLP) studies assessing the safety of BCG vaccination in cattle have not previously been reported. We describe here the results of two GLP safety studies in which calves and lactating cows were vaccinated with BCG (Danish 1331 strain). From an animal health and welfare perspective, the results of these studies indicate that BCG vaccine is well tolerated in these categories of cattle with only transient and minor local or systemic reactions. Furthermore, there was no evidence that BCG was shed in raw milk, saliva or faeces collected from vaccinates and vaccination did not have a detrimental effect on milk yields in lactating cattle. These data, underpinned by GLP principles, further support the existing data on the safety of BCG vaccine in cattle and complement the abundant available cattle efficacy data for this potential cattle bTB vaccine.

Сучасні методи дослідження гострої токсичності полютантів для представників водної і наземної екосистем та впровадження їх у практику наукового центру превентивної та регуляторної токсикології

The paper provides a brief historical background on the formation and development of ecotoxicology as a science, reflects the main modern methods and principles for studying the acute toxicity of xenobiotics for representatives of aquatic and terrestrial ecosystems recommended by the OECD (Organization for Economic Cooperation and Development). The classification of acute ecotoxicity of chemicals for land and water test systems is given. Further perspective development of the unit, which will be aimed at expanding ecotoxicological research in accordance with international OECD methods and GLP requirements, has been outlined. Aim. Revealing the main methods and principles recommended by the OECD for determining acute toxicity for biological objects of aquatic and terrestrial ecosystems and putting these methods and principles into practice of the Scientific Centre. Materials and Methods. The work was based on the implementation of international OECD standards and GLP principles. The OECD public electronic library of methods, particularly section 2 of the Instructions for Testing Chemicals, was used and analysed. Results and Conclusions. International methods for determining the acute toxicity of pollutants for non-target environmental objects were put into practice in the Scientific Centre, such as OECD 201 (Freshwater Alga and Cyanobacteria, Growth Inhibition Test); OECD 202 (Daphnia sp. Acute Immobilisation Test); OECD 203 (Fish, Acute Toxicity Test); OECD 207 (Earthworm, Acute Toxicity Tests); OECD 213 (Honeybees, Acute Oral Toxicity Test); OECD 216 (Soil Microorganisms: Nitrogen Transformation Test); OECD 217 (Soil Microorganisms: Carbon Transformation Test); OECD 223 (Avian Acute Oral Toxicity Test). Systems for the quality and control of ecotoxicological studies have been developed, which made it possible to obtain national and international accreditation for the conduct of the above studies. Further activities of the department will be aimed at expanding ecotoxicological research using international OECD methods and compliance with GLP requirements, which will contribute to the development of ecotoxicological research both in the Scientific Centre and in Ukraine as a whole. Key Words: ecotoxicology, pesticides and agrochemicals, biological test systems, OECD methods, good laboratory practice (GLP).

Implementing OECD GLP principles for the evaluation of novel vector control tools: a case study with two novel LLINs, SafeNet® and SafeNet NF®

BackgroundTo sustain high universal Long-Lasting Insecticidal Nets (LLINs) coverage, affordable nets that provide equivalent or better protection than standard LLINs, are required. Test facilities evaluating new LLINs require compliance to Good Laboratory Practice (GLP) standards to ensure the quality and integrity of test data. Following GLP principles allows for the reconstruction of activities during the conduct of a study and minimizes duplication of efficacy testing. This case study evaluated the efficacy of two LLINs: SafeNet NF® and SafeNet® LLIN.MethodsThe study was conducted according to GLP principles and followed World Health Organization guidelines for evaluating LLINs. The LLINs were assessed in experimental huts against wild, pyrethroid-resistant Anopheles arabiensis mosquitoes. Nets were either unwashed or washed 20 times and artificially holed to simulate a used torn net. Blood-feeding inhibition and mortality were compared with a positive control (Interceptor® LLIN) and an untreated net.ResultsMosquito entry in the huts was reduced compared to negative control for the unwashed SafeNet NF, washed Safenet LLIN and the positive control arms. Similar exiting rates were found for all the treatment arms. Significant blood-feeding inhibition was only found for the positive control, both when washed and unwashed. All insecticide treatments induced significantly higher mortality compared to an untreated net. Compared to the positive control, the washed and unwashed SafeNet NF® resulted in similar mortality. For the SafeNet® LLINs the unwashed net had an equivalent performance, but the mortality for the washed net was significantly lower than the positive control.Internal audits of the study confirmed that all critical phases complied with Standard Operating Procedures (SOPs) and the study plan. The external audit confirmed that the study complied with GLP standards.ConclusionsSafeNet NF® and SafeNet® LLIN offered equivalent protection to the positive control (Interceptor® LLIN). However, further research is needed to investigate the durability, acceptability, and residual efficacy of these nets in the community. This study demonstrated that GLP-compliant evaluation of LLINs can be successfully conducted by African research institutions.

Study of novel 1,4-dihydropyridine derivatives as prospective anti-inflammatory remedies: a randomised controlled trial

Background. Over the past decades, scientific community is motivated on finding new anti-inflammatory agents with a safe and high-effective profile to manage pathology.Objectives. A study of the anti-inflammatory action of novel compounds, 1,4-dihydropyridine derivatives, in a classical formalin-induced paw oedema test in white rats.Methods. Originally synthesised 1,4-dihydropyridine derivatives were preliminarily subjected to virtual screening using the SwissTargetPrediction toolkit. White laboratory rats (130 animals) were divided into a control (formalin oedema) and intact group, 4 comparison (meloxicam, sodium metamizole, sodium diclofenac and indomethacin) and 7 experiment groups by the number of 1,4-dihydropyridine derivatives studied. The samples anti-inflammatory efficacy was evaluated in acute formalin-induced paw oedema model simulated by right hind limb subplantar injection of 0.1 mL 2% formalin. The studied substances were administered intragastrically at 5 mg/kg 1.5 h prior to oedema induction. Oncometry was assessed quantitatively by limb circumference. Animals were managed in compliance with GOST 33044–2014 “Principles of Good Laboratory Practice” at all experiment steps. Experimental data were analysed statistically to describe quantitative variability with variance σ2, mean limb girth a and standard deviation σ. Data homogeneity and reliability were estimated by variation coefficient V and the Wilcoxon T(W) criterion.Results. As the most anti-inflammatory effective, partially hydrogenated mar-040 pyridines (ethyl 4-({[5-cyano-6-{[2-(diphenylamino)-2-oxoethyl]thio}-4-(2-furyl)-2-methyl-1,4-dihydropyridin-3-yl]carbonyl}amino) benzoate) were shown 33-fold superior to indomethacin, 26-fold — to sodium diclofenac, 25-fold — to meloxicam and 30-fold — to sodium metamizole; mar-037 pyridines (ethyl 4-[({[3-cyano-5-({[4-(ethoxycarbonyl)phenyl]amino}carbonyl)-4-(2-furyl)-6-methyl-1,4-dihydropyridin-2-yl]thio}acetyl)amino] benzoate) — 17–23-fold superior vs. reference drugs. We also show that mаr-014 (ethyl 4-({[5-cyano-6-({2-[(3,5-dichlorophenyl) amino]-2-oxoethyl}thio)-4-(2-furyl)-2-methyl-1,4-dihydropyridin-3-yl]carbonyl}amino)benzoate) and mar033 (ethyl 2-[({[3-cyano-5-({[4-(ethoxycarbonyl)phenyl]amino}carbonyl)-4-(2-furyl)-6-methyl-1,4-dihydropyridin-2-yl]thio}acetyl)amino]-4,5,6,7-tetrahydro-1-benzothiophene-3-carboxylate) compounds are 2.7-fold more effective vs. reference drugs.Conclusion. The synthesised 1,4-dihydropyridine compounds reveal high efficacy in experimental assays. Selected novel 1,4-dihydropyridine derivatives exhibit a marked anti-inflammatory activity and offer value in future preclinical trials.

Accuracy of interferon gamma release assays for the COVID-19 immunity assessment

Emerging evidence suggests that T-cells play a significant role in COVID-19 immunity both in the context of natural infection and vaccination. Easy to use IGRA assays including QFN SARS are considered attractive alternatives to more “traditional” but laborious methods for detection of SARS-CoV-2-specific T-cell responses.In our Letter we are proposing explanations to an apparently lower than expected T-cell responses (44 % reactive individuals) reported by Krüttgen et al in a small cohort of healthy double vaccinated individuals. These results could have been affected by reporting raw optical density values instead of calculated Interferon-ɣ concentrations which is supported by unexpectedly low mitogen responses in healthy individuals.This study highlights an importance of adhering to good laboratory practice principles as well as overall importance of accurate T-cell immunity assessment using IGRA assays.

Analiza laboratorijskih parametara hronične bubrežne insuficijencije kod starijih bolesnika

Aim: The chief aim of this study was monitoring of laboratory parameters of chronic kidney failure in elderly patients. Methods: All samples were taken and processed by standard methods and according to the principles of good laboratory practice. Data were collected in an organized and systematic manner in the form of a questionnaire with respect to ethical principles and as such were analyzed by statistical tests and analyses (Student's t-test, Analysis of variance-ANOVA, Pearson's and Spearman's correlation coefficients). The limit of statistical significance was set at p < 0.05. Results: Mean values of creatinine clearance and proteinuria for the total study population were: 41.30 ± 21.43 mL/min, 1.5 ± 2.3 g/L/24 h, respectively. Hematological parameters did not significantly differ from normal values. The highest frequency of comorbidities was observed in subjects aged ≥ 80 years with an average of 2.03 comorbidities per subject. Serum creatinine and urea values as well as creatinine clearance are good indicators of disease progression. Conclusion: The results of the presented research suggest that old age is a predisposing risk factor for the development of chronic kidney disease, and that in combination with comorbidities (hypertension and/or diabetes), it contributes to poor prognosis or disease progression.

Analysis of Phthalates and Alternative Plasticizers in Gloves by Gas Chromatography-Mass Spectrometry and Liquid Chromatography-UV Detection: A Comparative Study.

Gloves represent an essential feature for hand protection because it is a requirement in the professional framework to comply with both hand hygiene standards and the principles of good laboratory practice. Despite their wide use, there is a knowledge gap regarding their composition, including phthalates. The purpose of the present study was to develop two orthogonal methods, GC–MS and HPLC–DAD, for the screening of plasticizers in gloves. Performances of these two methods were compared in terms of ease of use, number of analyzed plasticizers, and sample preparation. The two methods were validated and applied for the identification and quantification of plasticizers in ten gloves made with different materials (vinyl, nitrile, latex, and neoprene). Results revealed the presence of three main ones: DEHP, DEHT, and DINP. Additionally, the contents of plasticizers were extremely variable, depending on the glove material. As expected, the results point out a predominant use of plasticizers in vinyl gloves with an amount that should be of concern. While DEHP is classified as a toxic substance for reproduction 1B, it was, however, quantified in the ten different glove samples studied. This study provides new data regarding the plasticizers’ content in protective gloves, which could be useful for risk assessment.

Assessment of experimental conditions affecting spontaneous mutation level of Salmonella strains used in the Ames test

Introduction. The bacterial reverse gene mutations test (the Ames test) is widely used to assess chemicals’ mutagenic activity. The spontaneous mutation level of test strains is a mandatory characteristic that has to be monitored in a laboratory performing mutagenicity studies using the Ames test. In this regard, it is important to assess the factors affecting the spontaneous mutation level in the experiment and, therefore, on the general conclusion on the test item mutagenicity. Material and methods. A plate incorporation test version was used both in the presence and absence of a metabolic activation system. Results. We summarized the historical control data obtained in the laboratory in 2016-2020, determine the fluctuation limits in the number of revertant colonies for each strain, and identify the factors affecting the negative control variability. No significant differences were found in the spontaneous background of test strains when using DMSO or water as solvents, polypropylene or polystyrene tubes, as well as Petri dishes of different types. In the case of the TA1535, TA102 and TA100 cultures, no influence of the presence of the S9 mixture on the spontaneous reversion range was revealed (p≤0.05). Statistically significant differences in the number of spontaneous revertants (at + S9 or -S9) were found for the strains that allow detecting frameshift mutations, TA97 and TA98. It has been shown that the volume of the selective medium and the brand of gelling agent in its composition are important factors leading to the variability of the historical negative control. Conclusion. To ensure the quality of experiments according to the principles of good laboratory practice and the reliability of the data obtained using the bacterial reverse mutation method, it is necessary to standardize the operations in advance of experiments.

Comparative evaluation of chitosan-based multifunctional wound dressings: a multistage randomised controlled experimental trial

Background. Wounds of various aetiology are among the most frequent traumatic injuries. A prospective route to improve treatment of this nosology is the development of novel or advancement of the already on-stage dressing materials.Objectives. A comparative experimental assay of novel chitosan-based wound dressings in the healing of soft tissue wounds of different genesis using ultrasound to control biodegradation of the developed dosage form.Methods. Soft tissue wounds were modelled in experimental animals, conventional and lineage male rats, 275 (± 25) g body weight, and male rabbits, 2,900 (± 150) g weight, using a proprietary methodology (Patent on invention RU No. 2703709 dated 23 August, 2018).Two wound dressing samples selected as most promising and applicable in large-volume soft tissue aseptic wounds were tested in the final step of experiment. Ultrasound imaging of the model wound area during the sample material biodegradation was used to control the wound process in more detail. At all steps of experiment, animal models were handled in compliance with GOST 33044-2014 “Principles of Good Laboratory Practice”.Histomorphological assays were carried out with common protocols. Variation statistics was used for the data analysis, including the mean (M) and standard mean error (± m) estimation. The level of statistical significance was p < 0.05.Results. The animal assays and comprehensive evaluation of the obtained data revealed a high healing efficacy of chitosan-based wound coatings. The analyses of wound dressing samples have confirmed their predesigned properties: scaffold stability, capillarity, biodegradability and matrix capacities for the carried drug delivery.Additionally, ultrasound was proved effective to estimate dynamics of the coated wound healing and biopolymer biodegradation processes.Conclusion. The developed chitosan-based wound dressings exhibited efficacy in an experimental model of the soft tissues wound process.The use of ultrasound to study dynamics of the wound process and coating biodegradation provided an adequate congruence between the imaging and tissue histomorphology data, as well as the structure and properties of coating material.

SARS-CoV-2 inactivated vaccine (Vero cells) shows good safety in repeated administration toxicity test of Sprague Dawley rats

The outbreak of COVID-19 has posed a serious threat to global public health. Vaccination may be the most effective way to prevent and control the spread of the virus. The safety of vaccines is the focus of preclinical research, and the repeated dose toxicity test is the key safety test to evaluate the vaccine before clinical trials. The purpose of this study was (i) to observe the toxicity and severity of an inactivated SARS-CoV-2 vaccine (Vero cells) in rodent Sprague Dawley rats after multiple intramuscular injections under the premise of Good Laboratory Practice principles and (ii) to provide a basis for the formulation of a clinical trial scheme. The results showed that all animals in the experimental group were in good condition, no regular changes related to the vaccine were found in the detection of various toxicological indexes, and no noticeable stimulating reaction related to the vaccine was found in the injected local tissues. The neutralizing antibodies in the low- and high-dose vaccine groups began to appear 14 days after the last administration. In the negative control group, no neutralizing antibodies were observed from the administration period to the recovery period. Therefore, the repeated administration toxicity test of the inactivated SARS-CoV-2 vaccine (Vero cells) in Sprague Dawley rats showed no obvious toxic reaction. It was preliminarily confirmed that the vaccine can stimulate production of neutralizing antibodies and is safe in Sprague Dawley rats.

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.

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.

Evaluation of the in vitro and in vivo genotoxicity of a Dioscorea Rhizome water extract.

Dioscorea Rhizome is commonly used in traditional herbal medicines for the treatment of diabetes, hyperthyroidism, liver damage, neuropathy, and asthma. Here, we investigated the genotoxicity potential of D. Rhizome water extract (DRWE) using three standard battery systems in accordance with the test guidelines of the Organisation for Economic Cooperation and Development and Ministry of Food and Drug Safety as well as the principles of Good Laboratory Practice. A bacterial reverse mutation test (Ames test) was performed using the direct plate incorporation method in the presence or absence of a metabolic activation system (S9 mixture). The tester strains used included four histidine auxotrophic strains of Salmonella typhimurium, TA100, TA1535, TA98, and TA1537, along with a tryptophan auxotrophic strain of Escherichia coli, WP2 uvrA. An in vitro chromosome aberration test was performed using CHL/IU cells originally derived from the lung of a female Chinese hamster in the presence or absence of the S9 mixture. An in vivo mouse bone marrow micronucleus test was performed using male ICR mice. The micronucleus was confirmed after observation of the micro-nucleated polychromatic. The Ames test showed that DRWE did not induce gene mutations at any dose level in any of the tested strains. Additionally, DRWE did not result in any chromosomal aberrations specified in the in vitro chromosomal aberration and in vivo micronucleus tests. These results showed that DRWE exhibited neither mutagenic nor clastogenic potential in either the in vitro or in vivo test systems.

Evaluation of Subchronic Oral Dose Toxicity of Freeze-Dried Skimmed Powder of Zophobas atratus Larvae (frpfdZAL) in Rats.

Zophobas atratus (Coleoptera: Tenebrionidae), the giant mealworm beetle, is known as an edible insect containing a high protein content which may serve as new sources of human food and animal feed. However, potential toxicity and food safety analyses of Z. atratus have not been previously investigated. Therefore, in this study, we aimed to evaluate toxicity of freeze-dried skimmed powder of Z. atratus larvae (frpfdZAL), known as the super mealworm. Toxicological assessments were performed at the doses of 1250, 2500, and 5000 mg/kg/day in a 2- and a 13-week oral repeated-dose toxicity study of frpfdZAL in male and female Sprague-Dawley (SD) rats in accordance with the Organisation for Economic Co-operation and Development (OECD) guidelines and the principles of Good Laboratory Practice (GLP). No toxicological changes in clinical signs, body weights, water and food consumption, urinalysis, hematology, clinical biochemistry, gross findings, and histopathological examinations were observed. In conclusion, the no-observed-adverse-effect level (NOAEL) of frpfdZAL was 5000 mg/kg/day and target organ was not identified in both sexes of rats. In addition, frpfdZAL did not induce increases of serum ImmunoglobulinE (IgE), an identifier of allergic reactions in rats. Collectively, these results suggest that frpfdZAL is safe with no adverse effects, and able to be applied as an edible ingredient or other biological uses.

Building a Quality Management System in a Core Facility: A Genomics Core Case Study

Core facilities are key resources supporting the academic research enterprise, providing access to innovative and essential technologies and expertise. Given the constraints placed on core facilities as recharge centers and the ever-changing research environment, an important competitive differentiator that can support rigorous and reproducible approaches in core labs is the implementation of a quality management system (QMS). This paper describes a systematic approach to building a QMS in a genomics core facility at the University of North Carolina School of Medicine. This model is based on principles of the International Organization for Standardization 9001 system with initiatives focused on process mapping, training (communication, customer service, performance management, development of standard operating procedures, and quality audits), root cause analysis, visual control boards, mock quality audits, and continuous improvement through metrics tracking and "voice of the customer" exercises. The goal of this paper is to share practical step-by-step recommendations and outcomes of this core facility QMS that are generally applicable to academic core facilities, regardless of technical focus. Application of these good laboratory practice principles will foster "competitiveness through compliance" and promote outstanding interdisciplinary research between academic cores and their nonacademic pharmaceutical and federal research partners. Additionally, implementation of the QMS qualified this core to apply for federally funded contracts, thereby diversifying its types of projects and sources of revenue.

Toxicological evaluation of exosomes derived from human adipose tissue-derived mesenchymal stem/stromal cells

Several studies report that the therapeutic mechanism of action of mesenchymal stem/stromal cells (MSCs) is mainly mediated by paracrine factors that are released from MSCs such as exosomes. Exosomes are nano-sized extracellular vesicles that are transferred to target cells for cell-to-cell communication. Although MSC-derived exosomes (MSC-exosomes) are suggested as novel cell-free therapeutics for various human diseases, evaluation studies for the safety and toxicity of MSC-exosomes are limited. The purpose of our study was to evaluate the toxicological profile, including skin sensitization, photosensitization, eye and skin irritation, and acute oral toxicity using exosomes derived from human adipose tissue-derived MSCs (ASC-exosomes) in accordance with the OECD guidelines and the principles of Good Laboratory Practice. The ASC-exosomes were classified as a potential non-sensitizer in the skin sensitization test, UN GHS no category in the eye irritation test, and as a skin non-irritant in the skin irritation test, and did not induce any toxicity in the phototoxicity test or in acute oral toxicity testing. Our findings are the first to suggest that ASC-exosomes are safe for use as a topical treatment, with no adverse effects in toxicological testing, and have potential application as a therapeutic agent, cosmetic ingredient, or for other biological uses.