The research in a newly developing field, toxicology of ageing, is highly relevant nowadays and very well interdisciplinary challenged. The improvement of eating habits might serve as an anti-ageing prevention and treatment strategy for metabolic disorders. The goal of our study was to contribute to the elucidation of the impact of ageing on immunological alterations along with prenatally conditioned metabolic syndrome in Sprague-Dawley (SD) female offspring rats. We have investigated the influence of guided two-generation nutritional intervention, specifically the effect of a high-fat diet (H) on fundamental metabolic parameters and cytokines. To mimic the dietary preferences of a western-style diet rich in high fat, we first fed the parental generation of SD rats either a control (C) standard diet or a high-fat diet (H, containing 1% cholesterol and 7.5% lard) three months prior to mating. Then, the type of diet either continued within the offspring´s generation (CC, HH) or changed (CH, HC). The non-invasive blood pressure measurement was carried out by tail-cuff plethysmography. The plasmatic levels of total cholesterol (TC), triacylglycerides (TAG), low-density lipoprotein (LDL-C), high-density lipoprotein (HDL-C), adipose triglyceride lipase (ATGL), and C-reactive protein (CRP) were assessed by ELISA kits. The serum levels of interleukins IL-1β, IL-4, IL-6, IL-10, IL-12, and tumor necrosis factor-α (TNF-α) were evaluated using magnetic bead–based immunoassays on Bio-Plex 200 systems (Bio-Rad, U.S.). Our data indicated that the inflammaging, which is age-related chronic low-grade inflammation, was present in diet-related disturbances, especially within the HH groups of SD female offspring rats. Therefore, epigenetic programming through parental diet appears to play a critical role in worsening inflammaging.
Ensuring the biocompatibility of medical devices (MDs) is essential for regulatory compliance and user safety. Under the evolving requirements of the European Medical Device Regulation (EU MDR 2017/745), products that come into contact with mucosal surfaces – such as those intended for oral use – must be supported by robust biocompatibility data. This study evaluated the in vitro cytotoxicity and irritation potential of the medical device hydropastilky in comparison with a benchmark product of similar composition and intended use, following ISO 10993 guidelines. Hydropastilky are designed to moisturise, protect, and support the regeneration of the oral and throat mucosa. Cytotoxicity was assessed using the Neutral Red Uptake (NRU) assay (ISO 10993-5). Hydro pastilky maintained cell viability above 50% at an extract concentration of up to 34.5%. Cytotoxic effects observed at higher concentrations were attributed to the strong hydrating and water-binding properties of the product, which posed challenges during extract preparation and likely impacted cell osmolarity as well. The benchmark product yielded comparable results. To generate more physiologically relevant data, irritation testing was performed using 3D reconstructed human epidermal (EpiDerm™) and oral mucosal (EpiOral™) models (ISO 10993-23). Both products demonstrated tissue viabilities above 50%, with most values exceeding 75%, confirming their non-irritant classification after prolonged exposure of 18 hours. These findings support the biocompatibility as well as biosimilarity of hydropastilky and the benchmark product and highlight the role of new approach methodologies (NAMs) in meeting modern regulatory expectations.
Disinfection of hands and surfaces is a key element in preventing the spread of infections. In this study, the cytotoxic effect of chemical substances used in conventional disinfectants was assessed: n-propyl alcohol (1P), isopropyl alcohol (2P), ethyl alcohol (EtOH), phenoxyethanol (FenEtOH), glutaraldehyde (GA), didecyldimethylammonium chloride (DDAC), sodium diisocyanurate (NaDCC), potassium peroxymonosulfate (KPMS) and the ready-to-use commercially available mixture for surface disinfection Mikrozid AK liquid (MAF). The assessment was performed on two cell lines derived from the human respiratory system: BEAS-2B and A549. To assess the cytotoxic effect of the tested compounds/mixture, the MTT and NRU assays were used. Based on the obtained results, DDAC and GA were found to be the most cytotoxic, while NaDCC and KPMS exhibited slightly lower levels of cytotoxicity. The alcohols: 1P, 2P and EtOH were characterized by lower toxicity on cells of both lines. The commercially available MAF mixture was the least toxic to A549 cells. Moreover, 2P, GA and EtOH exhibited a stronger cytotoxic effect on the metabolic activity of cells assessed by the MTT in both tested respiratory system-derived cell lines, compared to their effect on cell membrane integrity, which was evaluated using the NRU test. The obtained results also indicate that BEAS-2B cells are significantly more sensitive to the cytotoxic effects of the tested compounds than A549 cells, a difference that was particularly evident upon exposure to 1P, GA, DDAC, NaDCC and MAF in both tests.
Parabens, often used as preservatives in consumer products, have raised concerns due to their endocrine-disrupting properties. The aim of this study was to quantify the levels of methyl and propyl paraben in adult urine samples and to assess potential health risks. Using high-performance liquid chromatography (HPLC), methyl and propyl parabens were detected in 20 participants at different concentrations. Methylparaben was more prevalent than propylparaben. Risk assessment was performed by calculating the estimated daily intake (EDI) and the hazard quotient (HQ), with HQ values indicating no significant health risk for the participants. Although current exposure levels appear to be safe, the long-term effects of chronic exposure remain uncertain, highlighting the need for further research. This preliminary study provides insight into paraben exposure in adults and contributes to the growing literature on the safety and prevalence of parabens.
This paper outlines the methodical approaches essential for the accurate and reproducible evaluation of foetal skeletal and visceral abnormalities in teratology studies. Teratological investigations are critical for assessing the potential of drugs, chemicals, or environmental agents to induce developmental toxicity. The established methodology involves a systematic two-part process. Firstly, visceral examinations use techniques like microdissection and serial sectioning to identify gross and microscopic changes in internal organs. Secondly, skeletal evaluations utilize staining procedures, such as Alizarin red S and Alcian blue, to visualize and analyze the ossification and structural integrity of the foetal skeleton. These standardized procedures are vital for the reliable detection and characterization of malformations and variations. A rigorous, methodical approach ensures the scientific validity of the data, which is fundamental for effective risk assessment and regulatory decision-making concerning human and environmental health.
Teratogens are substances that can significantly affect prenatal development of an embryo or foetus. Alcohol acts as a toxicant in the developing brain. It passes through the placenta in both directions based on a concentration gradient. The consequences of maternal alcohol consumption during pregnancy depend on the stage of pregnancy. Foetal alcohol spectrum disorder expresses a set of characteristic dysmorphic features, congenital behavioural disorders and changes in the development of the newborn. The authors analysed the case of a premature newborn from unmonitored pregnancy of a mother with abuse of nicotine and alcohol, of unknown paternal history. After delivery 3-chamber hydrocephalus with dysgenesis of the corpus callosum was confirmed. The child was at 5 weeks of age admitted at Neonatal Department of Intensive Medicine for neurosurgery intervention. Given the child´s age at admission, it was not possible to indicate a specific examination for the presence of alcohol metabolites. The developing brain is very vulnerable throughout prenatal development, and it is known, that alcohol causes irreversible brain damage and later leads to the development of complications. We can only assume, that abuse of nicotine and alcohol supported the origin of congenital structural anomaly of the newborn’s brain.
Abstract Cardiotoxicity is one of the most serious adverse effects of exposure to arsenic. It has been shown that cytidine 5′-diphosphocholine or citicoline exerts a variety of biological functions, including anti-inflammatory and antioxidant activities. The aim of current study was to investigate the effect of citicoline on arsenic trioxide (ATO)-induced cardiotoxicity in H9C2 cells. H9C2 cardiac cells exposed to ATO for 24 h to induce cytotoxicity. The cells were pre-incubated with citicoline for 72 h prior to the exposure to ATO. The effect of citicoline on cell viability was investigated by MTT method. Malondialdehyde (MDA), total thiol groups, and total antioxidant power (TAP) levels were also evaluated. The results indicated that citicoline could effectively enhanced cell viability of H9C2 cells exposed to ATO. Treatment with ATO increased MDA levels, while pretreatment with citicoline reduced lipid peroxidation. Moreover, it could enhance total thiol contents and TAP levels. Taken together, our data suggest that citicoline has protective effects against ATO-induced oxidative stress in H9C2 cells, that could be attributed to inhibition of oxidative stress and its antioxidant effects.
Abstract Iodine is a trace element indispensable for thyroid hormone biosynthesis but its deficiency is a global problem. Programs of iodine prophylaxis are frequently based on salt iodization with the use of potassium iodate (KIO3), which, however, reveals potential prooxi-dative properties. Indole-3-propionic acid (IPA), deamination product of tryptophan, is an effective antioxidant without prooxidative properties. We have shown in our previous study that melatonin is able to reduce lipid peroxidation caused by KIO3. Taking into account the similarity between melatonin and IPA we decided to evaluate potential protective effects of this compound against KIO3-induced lipid peroxidation in porcine thyroid homogenates. In the current study thyroid homogenates were incubated in the presence of different concentrations of KIO3 without/with IPA. Six experiments were performed with different concentrations of IPA. Malondialdehyde+4-hydroxyalkenals (MDA+4-HDA) concentration (LPO index) was measured spectrophotometrically. IPA reduced in concentration-dependent manner (with statistical significance for concentrations of 10 mM, 7.5 mM, 5.0 mM) KIO3-induced lipid peroxidation, but only when KIO3 was used in concentrations of 10 mM and 7.5 mM, which are close to physiological iodine concentration in the thyroid (i.e. ~10 mM). In conclusion, IPA might be considered as a pharmacological agent to be used for protection against prooxidative effects of iodates.
Abstract Diabetes mellitus (DM) is one of the most common metabolic disorders in pregnancy. In infants of diabetic mothers, the morbidity and incidence of complications are increased in the prenatal, perinatal and neonatal periods, but also in the later period of life. The aim of this study was to analyse prenatal and postnatal neonatal complications. The study consisted of 96 newborns who were hospitalized from January 1st 2017 to June 30th 2020. 81.3% of newborns were born to mothers with gestational diabetes. 61.5% were born by Caesarean section. Macrosomia was diagnosed in 15%, birth trauma occurred in 24% of cases. Respiratory distress syndrome developed in 34.4% of neonates, syndrome of persistent pulmonary hypertension in 10.4%, congenital heart disease was present in 35.4%. The most common was persistent ductus arteriosus in 18.75%. Hypertrophic cardiomyopathy was diagnosed in 7 newborns (7.3%), of which 6 times without obstruction and 1 time with obstruction of the outflow tract of the left ventricle. 15.6% had congenital malformation of the digestive system and 12.5% had anomaly of the urinary and genital system. The largest proportion of the congenital malformations of CNS had meningomyelocele (6.25%). Hypoglycaemia was confirmed in 22% and hyperbilirubinemia in 30% of newborns. The obtained results were a source for creating the manual that could help neonatologists or paediatricians in practice, to get a comprehensive picture of a newborn whose mother had DM during pregnancy.
Abstract Polypyrrole is one of the most promising polymers for biomedical applications, and its low solubility in most common solvents led to the development of soluble forms of this conducting polymer. In this study, the physicochemical properties and the in vivo toxicity of soluble polypyrrole were evaluated, aiming to validate its potential for biomedical applications. The pH, density, and miscibility in different solvents were determined in two soluble polypyrrole samples synthesized on different dates. Two nonclinical studies on the toxicity of soluble polypyrrole were conducted with Swiss mice distributed into groups according to the dose administered. The single-dose toxicity study (acute toxicity) consisted of a single intraperitoneal application of a soluble polypyrrole solution at the doses of 500 and 1,000 mg/kg, followed by observation during 14 days. The repeated-dose toxicity study lasted for 28 days with daily intraperitoneal applications of the soluble polypyrrole solution, at the doses of 250, 500, and 1,000 mg/kg. Parameters such as mortality, behavioral screening, water and ration intake, body mass evolution, organ indices, and the macroscopic evaluation of the organs were observed in both studies. Additionally, hematological and histopathological analyses were performed in the repeated-dose toxicity study. In the acute toxicity study, no mortality was verified during the 14 days of observation at any of the doses tested, indicating that soluble polypyrrole has an LD50>1,000 mg/kg. The main alterations observed in the repeated-dose toxicity study involved the liver changes, such as in the increase of its relative weight, the presence of necrosis, and granulomatous inflammation. Considering the high doses administered and the time of application, the results indicate low toxicity of soluble polypyrrole when administered by the intraperitoneal route.
Abstract Andrographis paniculata is a plant herb that is used to treat various disease conditions because of its medicinal properties. This study sought to assess the antioxidant potentials of A. paniculata in vitro and evaluate its toxicological effects Wistar albino rats. Aqueous extract of A. paniculate, on which the following antioxidant assays, azino-bis-3-ethylbenzo-thiazoline-6-sulfonate radical (ABTS*), 1, 1-diphenyl-2- picrylhydrazyl radical (DPPH*), hydroxyl radical (OH*), ferric reducing antioxidant property (FRAP) and Fe2+ chelation were carried out, was prepared. For the toxicological study, the extract was administered orally for 14 days at different doses of 5, 50, 500, and 2000 mg/kg body weight. Afterwards, the liver function markers [aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), albumin (ALB), total and direct bilirubin], as well as kidney function markers [creatinine, urea, and uric acid] were evaluated. Results obtained from this study showed that A. paniculata extract possess antioxidant activities in vitro and also revealed that the plant extract is safe at the various doses administered as there was no record of death or any observable sign of skin pathology and also possess antioxidant properties. Therefore, aqueous extract of A. paniculata is non-toxic and could be used for medicinal and therapeutic purposes.
Abstract Naphazoline nitrate is a medicine with a number of side effects, which should be taken into account not only in therapeutic practice, but also when developing preventive measures in pharmaceutical production. The purpose of our work was to study the effect of naphazoline nitrate on redox processes and the duration of survival of bull spermatozoa. Bull semen samples were divided into groups: control – without adding naphazoline nitrate and experimental – with adding naphazoline nitrate. The respiratory activity of spermatozoa was determined by the polarographic method, reducing capacity – potentiometrically, succinate dehydrogenase activity – photo-metrically; sperm survival – visually. The respiratory activity of spermatozoa is significantly inhibited under the effect of all studied doses of naphazoline nitrate (η = 0.762 and 0.840) and the degree of effect for the highest dose reaches 85.5 %. The reducing capacity changes significantly, starting from a dose of 1/10 LD50 and above, the dose-indicator correlation is low, the dose of LD50 causes changes in the indicator by 229.0% with a change of charge of the medium. SDH activity significantly decreases under the effect of 1/10 LD50 and above, the dose-indicator correlation is moderate, the dose of LD50 causes changes in the indicator by 49.5 %. The substance affects the duration of survival only in the maximum dose, reducing the survival time by 18 %. Conclusion: The substance has the maximum effect on the respiratory activity of germ cells and on the reducing capacity, it has a lesser effect on the SDH activity and slightly affects the survival time. The dose-dependent nature of the effect of naphazoline nitrate on certain indicators of redox processes in spermatozoa confirms the prospects of using the latter as alternative test objects in studying the harmful effects of chemicals.
Abstract Iodinated radiographic contrast media (IRCM) is currently being used in radiography for disease diagnosis. The aim of the present study was to examine three IRCMs (i.e., iohexol, iopamidol, and iodixanol) in terms of their impact on morphology, osmotic fragility, and hemolysis of red blood cells. Blood was shortly treated with IRCMs (1, 5, 10, and 50 mgI/ml) at 37 °C for 5 minutes. The morphology of red blood cells was then observed under a light microscope and the number of burr cells (echinocytes) was recorded. The osmotic fragility and hemolysis of red blood cells were determined by using spectrophotometric technique. The results showed no statistically significant change in the number of burr cells, osmotic fragility, and hemolysis of red blood cells after an in vitro exposure to various concentrations of IRCMs when compared to the corresponding non-exposed control groups. These findings suggested that iohexol, iopamidol, and iodixanol did not exhibit any short-term effects on morphology, osmotic fragility, and hemolysis of red blood cells.
Abstract This study aimed to access the cytotoxicity, on the human conjunctival cell line, of different agents (neutral soap, 4% chlorhexidine, Efferdent effervescent tablets, 1% triclosan, and citronella essential oil) used for cleaning ocular prosthesis. Human conjunctival cell line was cultivated on acrylic resin specimens and the cytotoxicity of the studies cleaning agents was evaluated by MTT, Neutral Red, and RTPCR (Real-time reverse transcription polymerase chain reaction) for the gene expression of type IV collagen, transforming growth factor β, myeloid cell leukemia sequence 1, matrix metalloproteinase 9, and caspases 3 and 9. Data were submitted to the two-way analysis of variance (ANOVA) and Bonferroni (p<0.05) tests. All cleaning agents showed biocompatibility for the human conjunctival cells. This, immersion in 4% chlorhexidine, effervescent tablets, and 1% triclosan could be the best protocols indicated for ocular prosthesis cleaning due to their biocompatibility.
Abstract The aim of the research was to verify the theory of the free radical interaction of manganese compounds by assessing the activity of the enzymes: catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR) and glutathione S-transferase (GST) in an experimental rat model of exposure to Mn. Forty five male Buffalo rats were divided into: control, group I (MnCl2; 5 mg/kg b.w.) and groUp II (MnCl2; 7.5 mg/kg b.w.). The activity of the enzymes: CAT, GPx, GR and GST activity were determined in the supernatants of the kidneys and liver. The histopathological alterations were evaluated in tissue sections. In rats intoxicated with Mn (group I and II) the decreased CAT and GR activities was observed in the liver, with a significantly lower the all enzymes activities measured in the kidneys. The the lowest CAT, GPx, GR and GST activity were observed in the kidneys of group II rat. An examination of hepatic tissues revealed moderate (group I) and severe (group II) histopathological lesions, with a insignificant lesions oserved in the renal tissues. It has been shown that manganese intoxication has cytotoxic activity in the kidneys and liver, causing an decrease in antioxidant enzymes activity. The observed histopathological changes in the liver tissue suggest sensitivity to oxidative stress in this organ. In the kidneys, despite a significant weakening of the antioxidant defense, insignificant histopathological lesions were found, which suggests a different defense mechanism independent of the antioxidant enzymatic defense.
Abstract Erythrocyte death by eryptosis or erythronecrosis may induce erythrocyte shrinking or swelling with increase in osmotic resistance or fragility as indication of cytotoxicity. We investigated heterogeneous cytotoxic outcomes during in vitro exposure of goat erythrocytes to aluminium chloride, lead acetate or mercuric chloride using erythrocyte osmotic fragility (EOF) testing. The metallic salt solution (MSS) was added to 4.0 μL of high (100 mosmol/L) and low (250 mosmol/L) hyposmolar sucrose media at 0.3 or 0.4 mosmol/L concentration during testing of the osmotic fragility of 5.0 μL of blood from 10 goats. Hemolysis induced in the media (with and without MSS) was estimated in the supernatant with spectrophotometer at 540 nm. Osmotic stabilization or destabilization was calculated with probability for each test. Inducible osmotic resistance (IOR) was the ratio of mean stabilization to destabilization in both high and low hyposmolar media. Each MSS induced both osmotic resistance (stabilization) and fragility (destabilization) in varied media concentrations, with greater likelihood (P) of stabilization (0.93) or destabilization (0.77) in high or low media hyposmolarity, respectively. The EOF outcomes of the goats diverged within the group. High IOR induced by mercuric chloride (2.90) and low IOR by lead acetate (0.07) and aluminium chloride (0.04) reflected high stabilizing and destabilizing outcomes, respectively. In conclusion, MSS induced dual EOF outcomes (stabilization or destabilization) on the fragility domain, suggesting occurrence of both eryptosis (as stabilization) and erythronecrosis (as destabilization) at low exposure level, whereby biphasic, nonmonotonic or hormetic response to MSS toxic action might exist.
Abstract Estrogen deficiency and excessive alcohol intake are associated with numerous adverse effects on immune functions and cognitive abilities. There is lots of evidence for the role of cytokines in the functions of the central nervous system like emotions, learning and memory. The aim of the present study was to investigate the effects of the opioid antagonist naltrexone on IFN-γ serum levels and spatial memory of ovariectomized (OVX) rats with repeated alcohol intake. Female Wistar rats (200–200 g) were ovariectomized and randomly divided in four groups: 1 – “control” (physiological solution 1 ml/100 g p.o. for 4 weeks after OVX); 2 – “alcohol” (ethanol 30%, 1 ml/100 g, p.o. for 4 weeks after OVX); 3 – “alcohol+naltrexone” (ethanol 30%, 1 ml/100 g, p.o. for 4 weeks after OVX and naltrexone 2.5 mg/kg, i.p. for 3 weeks after OVX); 4 – “naltrexone” (naltrexone 2.5 mg/kg, i.p. for 3 weeks after OVX). Our results suggested a correlation between IFN-γ levels and learning and memory abilities of the experimental rats. Moreover, we demonstrated that naltrexone reduced the adverse effects of OVX and excessive ethanol on IFN-γ levels and cognitive performance. Our data suggest that naltrexone may have beneficial therapeutic effects on immunodeficiency and emotional and intellectual deficits in women with estrogen insufficiency and excessive alcohol intake.
Abstract Vipoxin is the main toxic component isolated from the venom of long-nosed viper Vipera ammodytes meridionalis, which population is widely distributed on the Bulgarian area. The neurotoxin consists of two subunits – unstable and toxic phospholipase A2 (PLA2) and non-toxic acidic component without enzymatic activity (Vipoxin’s acidic components, VAC). In the current study, the action of neurotoxic complex was examined by neurophysiological in vivo experimental model on anaesthetized rats. Vipoxin produces neuromuscular blockade in a dose-dependent manner with non-depolarizing post-junctional site of action. It was found that isolated PLA2 displays significantly lower blocking activity in comparison with Vipoxin when both have been applied in equimolar concentration. That implies the stabilizing role of VAC and its significance for the toxicity of the whole complex. The Vipoxin-induced neuromuscular blockade is completely reversed when antivenom is administrated.
Abstract This present report reveals the important aspects of sodium and potassium nitrite-induced genotoxicity during the development of a model organism, Drosophila melanogaster. Dietary sodium nitrite at ≥20 mM concentration caused significant nuclear deformation of two important larval cells, namely hemocytes and neuroblasts, that play important roles in immunity as well as growth and metamorphosis of the fly. In contrast, potassium nitrite induced significant nuclear deformities only in hemocytes at 30 mM concentration. Due to the unavailability of any report regarding the developmental genotoxic effects of nitrite food preservatives in immune and neural stem cells, results of our study deserve special mention as these provide first evidences of genotoxicity in these cells of the developing Drosophila larvae by these food additives.
