Test Animals Research Articles

Differences in Blood Glucose Levels of Rats (Rattus norvegicus) Given Sugar Cane Juice and Cyclamate Solution as a Sweetener

Background: Artificial sweeteners can cause glucose intolerance, which is a condition where the glucose in the blood is abnormal. Purpose: This research was conducted to determine the difference in blood glucose levels of mice as experimental animals that were given sugarcane juice as a natural sweetener and cyclamate solution as an artificial sweetener. Methods: The research method applied was laboratory experimental followed by checking blood glucose levels using a GCU easy touch glucometer and the data analysis method used was descriptive analysis. Results: the average blood sugar before treatment was P0 (control) 110 mg/dl, P1 (test animals given sugarcane juice solution) 63 mg/dl, P2 (test animals given sodium cyclamate solution) 69.5 mg/dl . Then, the average blood sugar after treatment was, P0 (control) 77.5 mg/dl, P1 (test animals given sugar cane juice) 81.5 mg/dl, P2 (test animals given cyclamate solution) 101.5 mg/dl etc. From this data, it was obtained that the level of glucose in the blood decreased by an average of 32.5 for P0. Then, there was an increase in blood glucose levels with an average of 18.5 for P1. In addition, there was an increase in blood glucose levels with an average of 32 for P2. Blood sugar levels in the treatments given cyclamate and sugar cane juice both experienced an insignificant increase in blood sugar, but within two weeks the average increase in blood sugar was highest in the treatment given cyclamate. Conclusion: There are differences in blood glucose levels of mice as experimental animals given sugarcane juice as a natural sweetener and cyclamate solution as an artificial sweetener.

Melatonin and resveratrol alleviate molecular and metabolic toxicity induced by Bisphenol A in endometrial organoids

Bisphenol A (BPA), a widespread environmental contaminant, poses concerns due to its disruptive effects on physiological functions of the uterine endometrium. In contrast, melatonin (MT) and Resveratrol (RSV) are under scrutiny for their potential protective roles against BPA-induced damage. For the efficacy and ethical concerns in the animal test, endometrial organoids, three-dimensional models mimicking endometrium, serve as crucial tools for unraveling the impact of environmental factors on reproductive health. This study aimed to comprehensively characterize the morphological, molecular and metabolic responses of porcine endometrial organoids to BPA and assess the potential protective effects of MT and RSV. Porcine uteri were prepared, digested with collagenase, mixed with Matrigel, and incubated at 38°C with 5 % CO2. Passaging involved dissociation through trypsin-EDTA treatment and subculturing. The culture medium was refreshed every 2–3 days. To investigate the environmental impact on reproductive health, endometrial organoids were treated with BPA (0.5 µM), MT (with/without BPA at 0.1 µM), and/or RSV (10 µM). Various molecular screening using gene expression, western blotting, immunofluorescence staining, and metabolites profiling were assessed the effects of BPA, MT, and RSV in terms of cell viability, morphology, reproductivity, and metabolism alteration in the endometrial organoids. As expected, BPA induced structural and molecular disruptions in organoids, affecting cytoskeletal proteins, Wnt/β-catenin signaling, and epithelial/mesenchymal markers. It triggered oxidative stress and apoptotic pathways, altered miRNA expression, and disrupted the endocannabinoid system. The level of glucose, galactose, and essential amino acids were increased or decreased by approximately 1.5–3 times in BPA-treated groups compared to the control groups (p-value < 0.05), indicating metabolic changes. Moreover, MT and RSV treated groups exhibited protective effects, mitigating BPA-induced disruptions across multiple pathways. For the first time, our study models endometrial organoids, advancing understanding of environmental impacts on reproductive health.

MRI-compatible abdomen phantom to mimic respiratory-triggered organ movement while performing needle-based interventions.

In vivo studies are often required to prove the functionality and safety of medical devices. Clinical trials are costly and complex, adding to ethical scrutiny of animal testing. Anthropomorphic phantoms with versatile functionalities can overcome these issues with regard to medical education or an effective development of assistance systems during image-guided interventions (e.g., robotics, navigation/registration algorithms). In this work, an MRI-compatible and customizable motion phantom is presented to mimic respiratory-triggered organ movement as well as human anatomy. For this purpose, polyvinyl alcohol cryogel (PVA-C) was the foundation for muscles, liver, kidneys, tumors, and remaining abdominal tissue in different sizes of the abdominal phantom body (APB) with the ability to mimic human tissue in various properties. In addition, a semi-flexible rib cage was 3D-printed. The motion unit (MU) with an electromagnetically shielded stepper motor and mechanical extensions simulated a respiration pattern to move the APB. Each compartment of the APB complied the relaxation times, dielectricity, and elasticity of human tissue. It showed resistance against mold and provided a resealable behavior after needle punctures. During long-term storage, the APB had a weight loss of 2.3%, followed by changes to relaxation times of 9.3% and elasticity up to 79%. The MU was able to physiologically appropriately mimic the organ displacement without reducing the MRI quality. This work presents a novel modularizable and low-cost PVA-C based APB to mimic fundamental organ motion. Beside a further organ motion analysis, an optimization of APB's chemical composition is needed to ensure a realistic motion simulation and reproducible long-term use. This phantom enhances diverse and varied training environments for prospective physicians as well as effective R&D of medical devices with the possibility to reduce in vivo experiments.

Reproductive and fetal toxicity studies of histamine H3 receptor antagonist DL76 used in mice to prevent maximal electroshock-induced seizure.

Introduction: Brain histamine is considered an endogenous anticonvulsant and histamine H1 receptor. H1R antagonists have, in earlier studies, been found to induce convulsions. Moreover, research during the last twodecades has provided more information concerning the anticonvulsant activities of histamine H3R (H3R) antagonists investigated in a variety of animal epilepsy models. Methods: Therefore, the in vivo anticonvulsant effect of the H3R antagonist DL76, with proven high in vitro affinity, in vitro selectivity profile, and high in vivo antagonist potency in mice against maximal electroshock (MES)-induced seizures in mice, was assessed. Valproic acid (VPA) was used as a reference antiepileptic drug (AED). In addition, DL76 was tested for its reproductive and fetal toxicity in the same animal species. Results and discussion: Our observations showed that acute systemic administration (intraperitoneal; i.p.) of DL76 (7.5mg/kg, 15mg/kg, 30mg/kg, and 60mg/kg, i.p.) provided significant and dose-dependent protection against MES-induced seizures in female and male mice. Moreover, the DL76-provided protective effects were comparable to those offered by the VPA and were reversed when animals were co-administered the CNS-penetrant selective H3R agonist R-(α)-methylhistamine (RAM, 10mg/kg, i.p.). Furthermore, the administration of single (7.5mg/kg, 15mg/kg, 30mg/kg, or 60mg/kg, i.p.) or multiple doses (3 × 15mg/kg, i.p.) of H3R antagonist DL76 on gestation days (GD) 8 or 13 failed to affect the maternal body weight of mice when compared with the control mice group. No significant alterations were detected in the average number of implantations and resorptions between the control and DL76-treated groups at the early stages of gestation and the organogenesis period. In addition, no significant differences in the occurrence of skeletal abnormalities, urogenital abnormalities, exencephaly, exomphalos, facial clefts, and caudal malformations were observed. The only significant abnormalities witnessed in the treated groups of mice were in the length of long bones and body length. In conclusion, the novel H3R antagonist DL76 protected test animals against MES-induced seizures and had a low incidence of reproductive and fetal malformation with decreased long bone lengths in vivo, signifying the potential therapeutic value of H3R antagonist DL76 for future preclinical as well as clinical development for use in the management of epilepsy.

Nanoliposomes Permeability in a Microfluidic Drug Delivery Platform across a 3D Hydrogel.

Nanoliposomes are nano-sized vesicles that can be used as drug delivery carriers with the ability to encapsulate both hydrophobic and hydrophilic compounds. Moreover, their lipid compositions facilitate their internalization by cells. However, the interaction between nanoliposomes and the membrane barrier of the human body is not well-known. If cellular tests and animal testing offer a solution, their lack of physiological relevance and ethical concerns make them unsuitable to properly mimic human body complexity. Microfluidics, which allows the environment of the human body to be imitated in a controlled way, can fulfil this role. However, existing models are missing the presence of something that would mimic a basal membrane, often consisting of a simple cell layer on a polymer membrane. In this study, we investigated the diffusion of nanoliposomes in a microfluidic system and found the optimal parameters to maximize their diffusion. Then, we incorporated a custom made GelMA with a controlled degree of substitution and studied the passage of fluorescently labeled nanoliposomes through this barrier. Our results show that highly substituted GelMA was more porous than lower substitution GelMA. Overall, our work lays the foundation for the incorporation of a hydrogel mimicking a basal membrane on a drug delivery microfluidic platform.

Lambda-Cyhalothrin induced behavioural, neurotoxic and oxidative stress on vertebrate model Danio rerio (Hamilton-Buchanan 1822).

λ-cyhalothrin, a synthetic type II pyrethroid, has become increasingly popular for control of aphids, butterfly larvae, and beetles, replacing other agricultural chemicals. As a result of which, residues of this synthetic pesticide are being reported across the globe in natural water, which poses a serious threat to aquatic life. Therefore, the present study was designed to understand the toxicity effects of λ-cyhalothrin on behaviour, oxidative stress and neurotoxicity in a vertebrate aquatic model, zebrafish (Danio rerio). The fish were exposed to 0.129, 0.194 and 0.388 µg/L corresponding to 5%, 10% and 20% of 96hLC50 (1.94 µg/L) for 28 days. Upon exposure to the highest concentration (0.388 µg/L), the test animal exhibited significant alterations in behavioural patterns like number of entries to the top zone (n), decrease in average speed (m/s) and decrease in time spent in top zone (s). Moreover, the shoaling test demonstrated a significant decrease (p < 0.05) in the relative time spent by the tested fish (%) near the stimulus fish. The change in behavioural alterations might be linked to a significant decrease (p < 0.05) in the brain acetylcholine esterase activity. Furthermore, the present study also illustrates oxidative stress exerted by λ-cyhalothrin through an increase in the production of reactive oxygen species, which is again clearly depicted by a significant increase (p < 0.05) in Superoxide dismutase, Catalase and Glutathione peroxidase activities. Overall, the present study systematically demonstrates the chronic effects of λ-cyhalothrin on adult fish behaviour and physiology, which will contribute to assessing the risks of λ-cyhalothrin to organismal health.

GREEN MARKETING PRACTICES IN THE CONTEXT OF ENVIRONMENTAL SUSTAINABILITY: A CASE STUDY OF AN ECO-FRIENDLY CLEANING FIRM

Purpose- Having started of climate change’s impact shaped businesses’ production process directly in 21st century. After consumers’ attitudes and behaviours had affected marketplace, businesses started to show themselves in aspect of corporate social responsibility. Since 1970s, sustainability has been highlighted as a topic reflected to corporate’s politics. Environmental sustainability is a variety of sustainability takes environment on center and keep providing products or services for this perspective. By using of green marketing mix, aim of a lot of brands to get profit take environment on center. From producing of packets to types of electric energy that use in factories, many effects started turning to metaphorses with corporates’ environmental sustainability politics. In context of ethical production and environmental sustainability, while animal testing is being removed vegan product production is tried to develop. Aim of this research is to understand on how environmental sustainability affects firms’/brands’ production process. Methodology- In this research, case study is applied as a types of qualitative research designs. By Turkey’s representative of an eco-friendly cleaning firm that from Denmark, semi-structured in depth interview were conducted. Then, the semi-structure in depth interview transcripted and highlighted to topic which turned into codes. The codes were got together and tied to themes. Findings- In result of the analysis reveals that 4 main themes which are 1) Scandinavian Morality and Ethical Production; 2) Green Sustainability; 3) Vegan Product; 4) Organic Product. To the analysis, Scandinavian morality has a big impact on ethical production. The firm’s practices are related to green sustainability; almost all production processes become “green”. In context of climate change, vegan perspective is rising day by day in the marketplace; producing animal-derived resources evaluate a part of climate change. Organic production is assessing an aspect of ecological life which is related to environmental sustainability. Conclusion- In green marketing perspective, environmental sustainability is one of the most indispensable issue, especially, while climate change gaining visibility. Basic marketing mix turning to “green marketing mix”. When brands and firms go green by politically, their production process goes directly green, as well. Climate change could be assessed like ethical issue in marketplace in context of environmental degradation. In addition, firms and brands caring about vegan and organic products, due to green conditions, are being positioned in environmental sustainability can be evaluated as a method of differentiation.

Acute toxicity testing of a mixture of ethanol extracts of ginger rhizome (Zingiber officinale Rosc.), lemongrass (Cymbopogon citratus (DC.) Stapf.), turmeric (Curcuma longa L.), and habbatussauda seeds (Nigella sativa L.) on the anatomy of mice

Abstract Immunostimulators are a collection of certain substances that are used to improve the immune system, especially when the body is sick. The use of herbs as immunostimulators is an option because it has been proven to be relatively safer if consumed correctly and appropriately. Four types of plants that are thought to have benefits as immunostimulators and have been used by some Indonesian people for generations are ginger, lemongrass, turmeric and habbatussauda. This study aims to determine the toxicity effects of a mixture of ethanol extracts based on observations of toxic symptoms, anatomical changes in mice (Mus musculus L.) and determine the level of toxicity through calculating the LD50. This research was experimental using 30 male mice divided into 6 treatments and 5 replications. Treatment consisted of control, mixed extracts of ginger, lemongrass, turmeric and black seed at doses of 1000 mg/kgBB, 2000 mg/kgBB, 3000 mg/kgBB, 4000 mg/kgBB and 5000 mg/kgBB given orally. The results showed that the ethanol extract of a mixture of ginger, lemongrass, turmeric and black seed had no significant effect on the average weight of the lungs, liver, heart and spleen. But it has a significant effect on the average weight of the kidney. The results of this study also showed changes in the mice’s body weight after 14 days of administration of the extract which tended to increase. The results of the research showed that no deaths occurred in the test animals so it was declared with a pseudo LD50 and was included in the practically non-toxic category so that the mixture of ginger, lemongrass, turmeric and black cumin was safe to use.

1H NMR spectroscopic characterisation of HepG2 cells as a model metabolic system for toxicology studies

The immortalised human hepatocellular HepG2 cell line is commonly used for toxicology studies as an alternative to animal testing due to its characteristic liver-distinctive functions. However, little is known about the baseline metabolic changes within these cells upon toxin exposure. We have applied 1H Nuclear Magnetic Resonance (NMR) spectroscopy to characterise the biochemical composition of HepG2 cells at baseline and post-exposure to hydrogen peroxide (H2O2). Metabolic profiles of live cells, cell extracts, and their spent media supernatants were obtained using 1H high-resolution magic angle spinning (HR-MAS) NMR and 1H NMR spectroscopic techniques. Orthogonal partial least squares discriminant analysis (O-PLS-DA) was used to characterise the metabolites that differed between the baseline and H2O2 treated groups. The results showed that H2O2 caused alterations to 10 metabolites, including acetate, glutamate, lipids, phosphocholine, and creatine in the live cells; 25 metabolites, including acetate, alanine, adenosine diphosphate (ADP), aspartate, citrate, creatine, glucose, glutamine, glutathione, and lactate in the cell extracts, and 22 metabolites, including acetate, alanine, formate, glucose, pyruvate, phenylalanine, threonine, tryptophan, tyrosine, and valine in the cell supernatants. At least 10 biochemical pathways associated with these metabolites were disrupted upon toxin exposure, including those involved in energy, lipid, and amino acid metabolism. Our findings illustrate the ability of NMR-based metabolic profiling of immortalised human cells to detect metabolic effects on central metabolism due to toxin exposure. The established data sets will enable more subtle biochemical changes in the HepG2 model cell system to be identified in future toxicity testing.

Vaginal irritation testing-prospects of human organotypic vaginal tissue culture models.

Personal lubricants intended for local or systemic delivery via the vaginal route can induce vaginal irritation, damage the vaginal epithelial barrier which can enhance microbial entry, induce inflammation, and alter the microbiome of the vaginal ecosystem. Therefore, manufacturers of personal lubricants and medical devices are required to show biocompatibility and safety assessment data to support regulatory decision-making within a specified context of use. Furthermore, due to ethical concerns and the introduction of the 7th amendment of the European Council Directive which bans animal testing for cosmetic ingredients and products coupled with the Food and Drug Administration modernization Act 2.0 guidelines, there is a wave of drive to develop alternative test methods to predict human responses to chemical or formulation exposure. In this framework, there is a potential to use three-dimensional organotypic human vaginal-ectocervical tissue models as a screening tool to predict the vaginal irritation potential of personal lubricants and medicaments. To be physiologically relevant, the in vitro tissue models need to be reconstructed using primary epithelial cells of the specific organ or tissue and produce organ-like structure and functionality that recapitulate the in vivo-like responses. Through the years, progress has been made and vaginal tissue models are manufactured under controlled conditions with a specified performance criterion, which leads to a high level of reproducibility and reliability. The utility of vaginal tissue models has been accelerated in the last 20years with an expanded portfolio of applications ranging from toxicity, inflammation, infection to drug safety, and efficacy studies. This article provides an overview of the state of the art of diversified applications of reconstructed vaginal tissue models and highlights their utility as a tool to predict vaginal irritation potential of feminine care products.

Development and characterisation of a novel complex triple cell culture model of the human alveolar epithelial barrier

Owing to increased pressure from ethical groups and the public to avoid unnecessary animal testing, the need for new, responsive and biologically relevant in vitro models has surged. Models of the human alveolar epithelium are of particular interest since thorough investigations into air pollution and the effects of inhaled nanoparticles and e-cigarettes are needed. The lung is a crucial organ of interest due to potential exposures to endogenous material during occupational and ambient settings. Here, an in vitro model of the alveolar barrier has been created in preparation for use in the quasi-air liquid interface (qALI) and (aerosol) air–liquid interface (ALI) exposures. The model consists of an alveolar type 1-like cell line (TT1), an alveolar type 2-like cell line (NCI-H441) and a model of (alveolar) macrophages (dTHP-1). The model formulates a complex, multi-cellular system, cultured at the air–liquid interface, that mimics the apical layer of the alveolar epithelial region in the human lung. Characterisation data has shown that both TT1 and NCI-H441 epithelial cells are able to be cultured together in addition to dTHP-1 cells through imaging (morphology), pro-inflammatory response and viability measurements. This dataset also demonstrates evidence of a reasonable barrier created by the cell culture in comparison to negative controls. Furthermore, it shows that while maintaining a low baseline of (pro)-inflammatory mediator expression during normal conditions, the model is highly responsive to inflammatory stimuli. This model is proposed to be suitable for use in toxicology testing of inhaled exogenous agents.

Test Of The Effectiveness Of Oral Administration Of Red Dragon Fruit (Hylocereus Polyrhizus) In Preventing Dyslipidemia In Male Wistar Rats (Rattus Norvegicus) Given A High High-Cholesterol Diet

Hyperlipidemia can result from obesity, which causes excess fat to build in the tissue under the skin and spread to other organs and tissues. Indonesia has an alarming dyslipidemia rate. Diet and exercise are vital. Hypolipidemia is in red dragon fruit (Hylocereus spp.) The study examined whether oral red dragon fruit extract prevented dyslipidemia in male Wistar rats (Rattus norvegicus) fed a high-cholesterol diet. This research used 24 mice as test animals. The research procedures included acclimation of the test animals, dragon fruit extract, phytochemical tests, preparation of the test animals, treatment with four groups, dyslipidemia parameter checks, histopathological preparations, and observations. Normal tests for total cholesterol, LDL, LSD, and HDL yielded p-values&gt; 0.05. Homogeneity tests with a 5% significance level yielded p values &lt; 0.05. This study's one-way Anova test yields a significance value of 0.000, or &lt;0.05, whereas the post-hoc LSD test yields &lt;0.05. The best red dragon fruit extract dose for lowering total and LDL cholesterol and raising HDL is 120 mg/mL. The phytochemical experiments on red dragon fruit extract (Hylocereus polyrhizus) show promise for improving the lipid profile.

Number of Erythrocytes in Male Mice (Mus musculus) Exposed to Cigarette Smoke After Treatment with Ethanol Extract of Red Pomegranate Peel (Punica ganatu)

Plant extracts have the potential to serve as natural therapeutic agents that can counteract the negative effects of various toxins, including those found in cigarette smoke. One such plant with this potential is the red pomegranate (Punica granatum). This study aims to determine the effect of administering Red Pomegranate (Punica ganatum) peel extract on the number of erythrocytes and hemoglobin levels in white mice exposed to cigarette smoke. This type of research is experimental laboratory post test only group design. Mice were divided into 4 treatment groups, namely: P0, control group, test animals treated with distilled water; treatment groups P1, P2 and P3, which were treated with neem leaf ethanol extract at a dose of 10 each; 20; 40 mg/head/day and mice were exposed to cigarette smoke. The research results showed that the higher the extract given, the higher the HB and erythrocyte levels. From this research it can be concluded that administration of Red Pomegranate rind extract can increase the number of erythrocytes and hemoglobin levels but it is not significant (p&gt;0.05). The contribution of this research provides important data on the interaction between toxins (in this case, cigarette smoke) and protective agents (red pomegranate peel ethanol extract). This information is valuable in the fields of toxicology and pharmacology for understanding cellular protection mechanisms and potential new treatments.

Sitagliptin ameliorates L-arginine-induced acute pancreatitis via modulating inflammatory cytokines expression and combating oxidative stress.

Acute pancreatitis (AP) is an inflammatory condition that resolves spontaneously, but occasionally, develops into systemic inflammation, organ failure and mortality. Oxidative stress and activation of inflammatory pathways represent major players in AP pathogenesis. Current management of AP relies on attenuating injuries to the pancreas and putting the inflammatory process under control. In this study, we investigated the role of sitagliptin in modulating L-arginine-induced AP in rats. Swiss rats were subdivided into a healthy control group, AP group (a single dose of L-arginine 250mg/100g, intraperitoneal), and sitagliptin + L-arginine-treated group (10mg sitagliptin/kg body weight/day, orally). Sitagliptin treatment started 1hour after L-arginine injection and continued for 3days. Biochemical and histopathological investigations were performed on serum and tissue samples collected from test animals. L-arginine increased pancreatic meyloperoxidase and serum amylase- and lipase activities and serum levels of TNF-α, LT-α, IFN-γ, IL-1α/β, IL-6, IL-10, IL-12, and IL-15. AP animals showed elevated MDA and NO and decreased GSH and serum calcium levels. Histopathological changes were observed by H&E staining. Sitagliptin treatment significantly ameliorated these biochemical and histological changes diminishing the signs of AP. Sitagliptin treatment was effective in ameliorating L-arginine-induced AP which can be regarded to its anti-inflammatory and antioxidant effect.

Predicting Chemical Immunotoxicity through Data-Driven QSAR Modeling of Aryl Hydrocarbon Receptor Agonism and Related Toxicity Mechanisms.

Computational modeling has emerged as a time-saving and cost-effective alternative to traditional animal testing for assessing chemicals for their potential hazards. However, few computational modeling studies for immunotoxicity were reported, with few models available for predicting toxicants due to the lack of training data and the complex mechanisms of immunotoxicity. In this study, we employed a data-driven quantitative structure-activity relationship (QSAR) modeling workflow to extensively enlarge the limited training data by revealing multiple targets involved in immunotoxicity. To this end, a probe data set of 6,341 chemicals was obtained from a high-throughput screening (HTS) assay testing for the activation of the aryl hydrocarbon receptor (AhR) signaling pathway, a key event leading to immunotoxicity. Searching this probe data set against PubChem yielded 3,183 assays with testing results for varying proportions of these 6,341 compounds. 100 assays were selected to develop QSAR models based on their correlations to AhR agonism. Twelve individual QSAR models were built for each assay using combinations of four machine-learning algorithms and three molecular fingerprints. 5-fold cross-validation of the resulting models showed good predictivity (average CCR = 0.73). A total of 20 assays were further selected based on QSAR model performance, and their resulting QSAR models showed good predictivity of potential immunotoxicants from external chemicals. This study provides a computational modeling strategy that can utilize large public toxicity data sets for modeling immunotoxicity and other toxicity endpoints, which have limited training data and complicated toxicity mechanisms.

Development of a Zebrafish Embryo-Based Test System for Thyroid Hormone System Disruption: 3Rs in Ecotoxicological Research.

There is increasing concern regarding pollutants disrupting the vertebrate thyroid hormone (TH) system, which is crucial for development. Thus, identification of TH system-disrupting chemicals (THSDCs) is an important requirement in the Organisation for Economic Co-operation and Development (OECD) testing framework. The current OECD approach uses different model organisms for different endocrine modalities, leading to a high number of animal tests. Alternative models compatible with the 3Rs (replacement, reduction, refinement) principle are required. Zebrafish embryos, not protected by current European Union animal welfare legislation, represent a promising model. Studies show that zebrafish swim bladder inflation and eye development are affected by THSDCs, and the respective adverse outcome pathways (AOPs) have been established. The present study compared effects of four THSDCs with distinct molecular modes of action: Propylthiouracil (PTU), potassium perchlorate, iopanoic acid, and the TH triiodothyronine (T3) were tested with a protocol based on the OECD fish embryo toxicity test (FET). Effects were analyzed according to the AOP concept from molecular over morphological to behavioral levels: Analysis of thyroid- and eye-related gene expression revealed significant effects after PTU and T3 exposure. All substances caused changes in thyroid follicle morphology of a transgenic zebrafish line expressing fluorescence in thyrocytes. Impaired eye development and swimming activity were observed in all treatments, supporting the hypothesis that THSDCs cause adverse population-relevant changes. Findings thus confirm that the FET can be amended by TH system-related endpoints into an integrated protocol comprising molecular, morphological, and behavioral endpoints for environmental risk assessment of potential endocrine disruptors, which is compatible with the 3Rs principle. Environ Toxicol Chem 2024;00:1-18. © 2024 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Multi-Endpoint Acute Toxicity Assessment of Organic Compounds Using Large-Scale Machine Learning Modeling.

In recent years, alternative animal testing methods such as computational and machine learning approaches have become increasingly crucial for toxicity testing. However, the complexity and scarcity of available biomedical data challenge the development of predictive models. Combining nonlinear machine learning together with multicondition descriptors offers a solution for using data from various assays to create a robust model. This work applies multicondition descriptors (MCDs) to develop a QSTR (Quantitative Structure-Toxicity Relationship) model based on a large toxicity data set comprising more than 80,000 compounds and 59 different end points (122,572 data points). The prediction capabilities of developed single-task multi-end point machine learning models as well as a novel data analysis approach with the use of Convolutional Neural Networks (CNN) are discussed. The results show that using MCDs significantly improves the model and using them with CNN-1D yields the best result (R2train = 0.93, R2ext = 0.70). Several structural features showed a high level of contribution to the toxicity, including van der Waals surface area (VSA), number of nitrogen-containing fragments (nN+), presence of S-P fragments, ionization potential, and presence of C-N fragments. The developed models can be very useful tools to predict the toxicity of various compounds under different conditions, enabling quick toxicity assessment of new compounds.

Effect Of Anting-Anting Leaf Extract (Acalypha Indica L.) On Lowering Blood Glucose Rats (Rattus Norvegicus) Induced By Sukrosa

Anting-anting leaves (Acalypha indica L.) are weeds that have various properties, one of which is as a lowering blood glucose levels. The chemical content contained in the form of flavonoids that function as inhibitors of α-glucosidase, maltase, α-amylase enzymes, tannin substances are also known to function as astrigents that can inhibit glucose intake is not excessive. The purpose of this study is to determine whether there is an effect of anting-anting leaf extract (Acalypha indica L.) on reducing blood glucose levels in rats (Rattus norvegicus) induced by sucrose. This research used 20 rats (Rattus norvegicus) as test animals which were divided into 5 groups. The group consisted of negative control group, positive control and 3 dose groups. The doses used were 500 mg/kgBB, 1000 mg/kgBB and 1200 mg/kgBB. Mice were made hyperglycemia with sucrose induction which was then given anting-anting leaf extract (Acalypha indica L.) according to lower glucose levels in mice. The results obtained were continued with the two-way ANOVA test and obtained significant results indicated by p &lt; 0.05 (0.005) which showed there was a significant effect of anting-anting leaf extract (Acalypha indica L.) on reducing blood glucose levels in sucrose-induced rats (Rattus norvegicus). Significant results were continued with the post hoc test and obtained significant results.

Guidelines for Manufacturing and Application of Organoids: Skin.

To address the limitations of animal testing, scientific research is increasingly focused on developing alternative testing methods. These alternative tests utilize cells or tissues derived from animals or humans for in vitro testing, as well as artificial tissues and organoids. In western countries, animal testing for cosmetics has been banned, leading to the adoption of artificial skin for toxicity evaluation, such as skin corrosion and irritation assessments. Standard guidelines for skin organoid technology becomes necessary to ensure consistent data and evaluation in replacing animal testing with in vitro methods. These guidelines encompass aspects such as cell sourcing, culture techniques, quality requirements and assessment, storage and preservation, and organoid-based assays.

A miniaturised semi-dynamic in-vitro model of human digestion

Reliable in-vitro digestion models that are able to successfully replicate the conditions found in the human gastrointestinal tract are key to assess the fate and efficiency of new formulations aimed for oral consumption. However, current in-vitro models either lack the capability to replicate crucial dynamics of digestion or require large volumes of sample/reagents, which can be scarce when working with nanomaterials under development. Here, we propose a miniaturised digestion system, a digestion-chip, based on incubation chambers integrated on a polymethylmethacrylate device. The digestion-chip incorporates key dynamic features of human digestion, such as gradual acidification and gradual addition of enzymes and simulated fluids in the gastric phase, and controlled gastric emptying, while maintaining low complexity and using small volumes of sample and reagents. In addition, the new approach integrates real-time automated closed-loop control of two key parameters, pH and temperature, during the two main phases of digestion (gastric and intestinal) with an accuracy down to ± 0.1 °C and ± 0.2 pH points. The experimental results demonstrate that the digestion-chip successfully replicates the gold standard static digestion INFOGEST protocol and that the semi-dynamic digestion kinetics can be reliably fitted to a first kinetic order model. These devices can be easily adapted to dynamic features in an automated, sensorised, and inexpensive platform and will enable reliable, low-cost and efficient assessment of the bioaccessibility of new and expensive drugs, bioactive ingredients or nanoengineered materials aimed for oral consumption, thereby avoiding unnecessary animal testing.