Early developmental anomalies in zebrafish (Danio rerio) embryos induced by the Clematis florida Thunb.

ETHNOPHARMACOLOGICAL RELEVANCE: The C.florida. is one of the common medicines used by She population in China, with therapeutic effects of promoting blood circulation and anti-inflammatory. According to the acute toxicity grading standard of chemical substances, this herb is a low-toxicity herb. At present, the safety of C.florida., especially its impact on early embryonic development, is still unclear. AIM OF THE STUDY: This study investigated the toxic effects of C. florida. on early embryonic development using a zebrafish embryo model. MATERIALS AND METHODS: In this study, we used zebrafish embryos exposed to C.florida. at early stage to assess the early developmental toxicity by analyzing the developmental toxicity phenotype, oxidative stress, cell apoptosis, total enzyme activity, behavioral trajectory, and gene expression levels. RESULTS: Embryos of the zebrafish exposed to different concentrations of C.florida. exhibited multiple organs and systems developmental disorders, including the heart, vessels, brain, bone, liver, and so on. Especially, with the increase of drug concentration, it is observed that the developmental malformations of the cardiovascular structure and function in larvae are becoming increasingly severe. In addition, results show that the abnormalities in embryonic development may be attributed to oxidative stress induced by apoptosis and activation of immune system resulting from an imbalance in the hematopoietic system. CONCLUSIONS: This study provides a comprehensive and detailed summary of the toxic effects of C.florida. on embryonic development, which contributes to a deeper understanding of the potential adverse developmental consequences, and also prompt people to pay considerable attention to its treatment in medicinal practice.

Biomarker and adverse outcome pathway responses of Tubifex tubifex (sludge worm) exposed to environmentally-relevant levels of acenaphthene: insights from behavioral, physiological, and chemical structure-activity analyses.

Polycyclic aromatic hydrocarbons (PAHs), including acenaphthene, pose a significant threat to aquatic ecosystems by harming vital organisms such as benthic invertebrates. This study evaluated the impact of environmentally relevant concentrations of acenaphthene on Tubifex tubifex, focusing on sublethal acute toxicity and subchronic biomarker responses. Key biomarkers assessed included histopathological changes and the modulation of antioxidant enzymes: catalase (CAT), superoxide dismutase (SOD), glutathione S-transferase (GST), and malondialdehyde (MDA). Additionally, the study examined structure-activity relationships and species sensitivity distribution (SSD). Concentrations exceeding the solubility threshold of acenaphthene (3.9 mg/L) triggered distinct, concentration-dependent behavioral responses in Tubifex tubifex, such as clumping, mucus secretion, and body wrinkling. Prolonged exposure exacerbated these behavioral dysfunctions, while subchronic exposure resulted in significant histopathological alterations, including epithelial hyperplasia, inflammation, edema, fibrosis, and degenerative changes. The edematic appearance of the body wall suggested a potential immune response to exposure. Furthermore, increased activities of CAT, SOD, and GST indicated oxidative stress in the worms. The study found a 1.5-fold increase in CAT and GST activity, a fivefold increase in SOD, and a striking 100-fold increase in MDA levels compared to controls, signifying an overwhelmed antioxidant defense system and potential cellular disruption. The SSD curve revealed hazard concentrations (HC50 and HC90), indicating that Tubifex tubifex exhibited lower sensitivity to acenaphthene compared to other taxa. In silico analysis and read-across models confirmed the potential of acenaphthene to induce significant oxidative stress upon exposure. The correlation between biomarker responses and structure-activity relationship analysis highlighted the aromatic nature of acenaphthene as a key factor in generating reactive metabolites, inhibiting antioxidant enzymes, and promoting redox cycling, ultimately contributing to adverse outcomes. These findings, coupled with behavioral responses and SSD curve inferences, underscore the importance of the solubility threshold of acenaphthene as a critical benchmark for evaluating its ecological impact in aquatic environments.

Acute and multigenerational effects of short-chain chlorinated paraffins on the harpacticoid copepod Tigriopus japonicus.

Although the measurement of short-chain chlorinated paraffins (SCCPs) in aquatic ecosystems has increased, limited information is available on their toxic effects on aquatic animals. To evaluate the harmful effects of SCCPs, we assessed their acute impact on 24-h survival and biochemical parameters, as well as their chronic effects on growth and reproduction over three generations in the harpacticoid copepod Tigriopus japonicus. Dose-dependent increases in mortality were observed, with an LC50 value of 74.6 µg L-1 for 24 h. Acute exposure to the LC10 value for 24 h significantly reduced feeding behavior, accompanied by a notable decrease in acetylcholinesterase enzymatic activity. Simultaneously, the intracellular levels of reactive oxygen species increased, along with elevated malondialdehyde contents. Glutathione level was increased by the LC10 value of SCCPs with the induction of enzymatic activities of antioxidant defense components, including glutathione S-transferase, catalase, superoxide dismutase, glutathione peroxidase, and glutathione reductase. When T. japonicus was continuously exposed to 1/10 of the NOEC and NOEC values for 12 days across three generations (F0-F2), growth retardation was observed in the F2 generation, with delay in the developmental periods from nauplius to adult. Although the total number of nauplii per brood was not significantly altered across generations, a significant delay in the onset of reproduction was observed in the F2 generation. Our findings suggest that even sublethal concentrations of SCCPs can negatively affect the health of copepod populations with consistent exposure.

Toxicity and safety profile evaluation of Shenfu injection in a murine sepsis model.

AIM: This study aimed to evaluate the preclinical safety of Shenfu injection for the treatment of sepsis. Tests were designed and conducted to determine the acute and long-term toxicity of Shenfu injection in rats, based on the recommended indications and dosage for human use. MATERIALS AND METHODS: Rats were administered 22.5 g of raw drug/kg/day via tail vein injection. Toxicity symptoms were monitored for 14 days following the intravenous injection of Shenfu injection, and target organs affected by toxicity were analyzed. To assess long-term toxicity, rats were given 12, 9, or 6 g of raw drug/kg/day by intraperitoneal injection, equivalent to 12, 9, and 6 times the daily clinical dose for adult sepsis patients (3.3 mL of stock solution per 1 g of raw drug/kg/day), for 30 consecutive days. This was followed by a 28-day recovery period after withdrawal of the drug. During the administration and recovery periods, signs of toxicity were observed and compared with those in the control (stromal fluid) group. The aim was to predict potential clinical adverse reactions, including the nature and severity of these reactions, dose-response and time-response relationships, and the reversibility of the effects. Additionally, the study sought to identify the target organs or tissues potentially affected by repeated administration and suggest clinical indicators that should be monitored during the product's use. Furthermore, the safety of co-administration with commonly used chemical medications for the treatment of sepsis was investigated. RESULTS: In the acute toxicity test, administration of the maximum dose of Shenfu injection (75 mL of stock solution/22.5 g of raw drug/kg/day) via tail vein injection resulted in transient symptoms, including piloerection (vertical hair response), weight loss, and reduced food intake. In the long-term toxicity experiments, rats received intraperitoneal injections of 0.3 g/mL (stock solution), 0.225 g/mL, and 0.15 g/mL Shenfu injection per day, which corresponded to 12, 9, and 6 times the daily clinical dose for adults with sepsis. The injections were administered twice daily for 30 days, followed by a 28-day drug withdrawal period for recovery. After 28 days, no significant toxicological changes were observed, apart from a hemodilution effect caused by the excessive volume of the drug and a slight increase in alkaline phosphatase and total bilirubin levels. The effects were reversible upon drug discontinuation. CONCLUSIONS: A single intravenous injection of 22.5 g of raw drug/kg/day and long-term intraperitoneal administration of up to 12 g of raw drug/kg/day are considered safe doses for rats.

Unveiling phytochemical diversity and safety profile of hot water extract from Tetrapleura tetraptera fruit.

BACKGROUND: Tetrapleura tetraptera, a widely used medicinal plant in West Africa, has been traditionally employed for various ailments. Despite its folkloric significance, scientific validation of its safety and potential neuroactive properties remains limited. OBJECTIVES: This study aimed to investigate the acute and subchronic toxicity of Tetrapleura tetraptera hot water extract (HWETTF) in rats and to elucidate its phytochemical composition. METHODS: Acute oral toxicity was assessed in mice using the OECD guideline 423, while a 14-day repeat-dose toxicity study was conducted in rats. The phytochemical analysis included HPLC, FT-IR, and GC-MS. RESULTS: HWETTF exhibited no significant toxicity in acute or subchronic studies, even at high doses. Phytochemical analysis revealed a diverse array of compounds, including those with potential GABAergic and CNS depressant activities. CONCLUSION: Tetrapleura tetraptera demonstrated a favourable safety profile in rodents and possesses a rich phytochemical composition. Further research is warranted to explore its potential neuroactive properties and develop therapeutic applications.

Possible Causes of Extreme Variation of Benzo[a]pyrene Acute Toxicity Test on Daphnia magna.

There are enormous differences in benzo[a]pyrene (BaP) acute toxicity tests on Daphnia magna, according to previous publications. The explanations of the reasons for this extreme variation are necessary. In this context, the acute toxicity tests of different experiment conditions (light/dark, culture medium, and solvent) were conducted on Daphnia magna with BaP as the toxicant of concern. Based on the experiments above, molecular dynamics (MD) simulations were employed to investigate the mechanisms of action. According to our results, the significant influence of light exposure on the acute toxicity test of BaP (p < 0.05) on D. magna was recorded. On the basis of the MD simulations, it was possible that BaP may not affect the normal operation of Superoxide Dismutase and Catalase directly, and it could be quickly transferred from the body through Glutathione S-transferase and Cytochromes P450. Therefore, when exposed to light, the oxidative stress process intensifies, causing damage to Daphnia magna. Apparently, the ecotoxicity tests based on inhibition for D. magna cannot adequately reflect the toxic effects of BaP.

Apodanthera glaziovii (Cucurbitaceae) Shows Strong Anti-Inflammatory Activity in Murine Models of Acute Inflammation.

Background/objectives:Apodanthera glaziovii is an endemic species from the semi-arid Brazilian, which has limited toxicological and pharmacological studies. This species belongs to a well-studied family known for its bioactive compounds used in treating inflammatory. This study aimed to identify secondary metabolites in the stems from A. glaziovii, evaluate toxicity, and investigate the anti-inflammatory potential of the stem hydroalcoholic extract (SHE-Ag). Methods: qualitative and quantitative assays were employed to identify secondary metabolites, along with chromatographic analyses and 1H and 13C NMR. Toxicity was assessed through in vitro hemolytic toxicity, in vivo genotoxicity, and oral acute toxicity tests before the pharmacological assays were conducted. Results: phytochemical screening, HPLC and NMR analyses suggested the presence of saponins of the norcucurbitacin class. The SHE-Ag exhibited no hemolytic activity and no mutagenic potential. However, in vivo toxicity at a dose of 2000 mg/kg revealed hematological and biochemical alterations, while the 500 mg/kg dose was safe. In the anti-inflammatory assays, SHE-Ag at 100 mg/kg reduced paw edema by 55.8%, and leukocyte and neutrophil migration by 62% and 68% in the peritonitis model, respectively; inflammatory cell migration by 70% in the air pouch model, outperforming indomethacin, which showed a 54% reduction. Conclusions: these findings indicate that SHE-Ag is rich in saponins, confirmed through HPLC and 1H and 13C NMR analyses. The SHE-Ag also demonstrated low toxicity. The inflammation models used showed a reduction in inflammation, pro-inflammatory cells, and edema, highlighting the significant anti-inflammatory activity of hydroethanolic extract A. glaziovii stems.

Insights into Clematis cirrhosa L. Ethanol Extract: Cytotoxic Effects, LC-ESI-QTOF-MS/MS Chemical Profiling, Molecular Docking, and Acute Toxicity Study.

BACKGROUND: In Jordanian traditional medicine, Clematis cirrhosa is commonly employed for the management of different diseases. Numerous investigations have documented the cytotoxic properties of different Clematis species against numerous types of cancer. Previously, we demonstrated the potential cytotoxicity of Clematis cirrhosa against HT-29 colorectal cancer cells. Extending our work, the current research aimed to explore the possible mechanisms underlying its antiproliferative activity with a plant safety evaluation. METHODS: This study evaluates the extract's impact on the cell cycle, apoptosis, and cell migration through in vitro assays, LC-ESI-QTOF-MS/MS analysis, docking studies, and an acute toxicity evaluation. RESULTS: The Clematis cirrhosa ethanol extract (CEE) induced G2/M phase cell cycle arrest (19.63%), triggered significant apoptosis (41.99%), and inhibited cell migration/wound healing by 28.15%. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) analysis revealed increased expression of the proapoptotic markers BAX (6.03-fold) and caspase-3 (6.59-fold), along with the reduced expression of the antiapoptotic BCL-2, in CEE-treated cells. Moreover, CEE significantly restrained angiogenesis by reducing VEGF mRNA expression by 63.9%. High-resolution LC-ESI-QTOF-MS/MS studies identified 26 metabolites, including phenolic compounds, fatty acids, and triterpenoids. Docking studies suggested that manghaslin had the highest binding affinity for VEGFR-2, followed by calceolarioside B, quercetin 7-O-rhamnopyranoside, luteolin, and quercetin-3,7-O-diglucoside. On the other hand, salvadoraside exhibited the highest binding affinity for the inhibition of caspase-3, followed by quercetin-3,7-O-diglucoside, kaempferol-3,7-O-α-L-dirhamnoside, manghaslin, and tectoridin, supporting the observed apoptotic effects. Interestingly, the outcomes further indicate that a single oral administration of up to 5000 mg/kg CEE is safe for consumption. CONCLUSIONS: These outcomes point to the potential of Clematis cirrhosa as a promising candidate for further exploration in cancer therapy.

Toxicological assessment of the aqueous extract of Juniperus oxycedrus L. on acute and subacute toxicities in rats.

Juniperus oxycedrus L. (J. oxycedrus) has a rich historical background in herbal remedies to treating digestive system abnormalities. However, no comprehensive evaluation of its potential toxic effects has been conducted. The current investigation aimed to evaluate the acute and subacute toxicity of an aqueous extract of J. oxycedrus (AEJO). AEJO was prepared by the conventional Moroccan methods by decoction the arial part of the plant. The acute and subacute toxicity tests were conducted in mice and rats, respectively. Acute toxicity tests showed that the extract was not toxic even at high doses of 5000 mg/kg. In the subacute study, no detectable indications of toxicity or mortality were observed and there were no notable deviations in food intake or water consumption among all rats. However, changes in body weight of animals treated with 1000 and 2000 mg/kg underwent a significant decrease. AEJO administration decreased platelet number, elevated levels of alanine aminotransferase and alkaline phosphatase, and reduced albumin levels. Histological examination revealed normal renal parenchyma despite increased creatinine. It also showed binucleation, and hepatocyte vacuolation. The results indicate that AEJO has considerable tolerance for consumption, but repeated use can affect hepatocytes and kidneys. Therefore, additional analyses, such as subchronic, chronic, and neurotoxic studies, are required before using this plant in clinical research.

Assessment of pharmacological effects and abuse potential of 5F-EDMB-PICA, CUMYL-PEGACLONE, and NM-2201 in mice.

RATIONALE: The newly emerging synthetic cannabinoids (SCs) 5F-EDMB-PICA, CUMYL-PEGACLONE, and NM-2201 have been observed to produce effects by activating cannabinoid type 1 (CB1) receptors. Nevertheless, the pharmacological effects and potential for abuse of these three substances remain to be studied. These substances have yet to be regulated in many countries. OBJECTIVES: We investigated the safety, pharmacological effects, rewarding effects, and cannabinoid withdrawal of 5F-EDMB-PICA, CUMYL-PEGACLONE, and NM-2201. METHODS: This study evaluated the drug safety and the cannabinoid-specific pharmacological effects of the three substances through acute toxicity experiments (in which the LD50 of each substance was obtained) and tetrad experiments (comprising assessments of hypothermia, analgesia, locomotion inhibition, and catalepsy). Furthermore, the conditioned place preference (CPP) experiments and withdrawal experiments were conducted to evaluate the rewarding effect and cannabinoid withdrawal potential of the substances in question. RESULTS: The results demonstrated that all three drugs exhibited certain acute toxic effects and could potentially induce tetrad effects. The data were analyzed using non-linear regression, and the corresponding ED50 values and 95% confidence intervals (CI) were obtained. The rank order of potency was determined to be CUMYL-PEGACLONE > 5F-EDMB-PICA > NM-2201. In the CPP experiments, it was demonstrated that 5F-EDMB-PICA significantly induced an increase in CPP score at a dose of 0.3 mg/kg, while NM-2201 caused an increase in CPP score and a significant aversion effect at a dose of 2 and 3 mg/kg, respectively. It is noteworthy that all three types of SCs were observed to produce a significant biphasic effect, indicating that CPP scores were biphasic for all compounds. Following the administration of the CB1 receptor antagonist rimonabant, a notable increase in head twitches and paw tremors was observed, indicating that these three SCs induce cannabinoid withdrawal through the mediation of CB1 receptors. CONCLUSIONS: The results of this study indicated that these SCs possess cannabinoid-specific pharmacological effects and abuse potential, which provides substantial experimental data to support the future regulation of these substances.

Oral toxicity assessment and the mitigation of lung carcinogenesis by phytol and α-bisabolol combination treatment in swiss albino mice: insights into redox enzyme modulation and caspase-dependent cell death mechanisms.

This study examined the safety and potential anti-lung cancer effects of combinations of phytol and α-bisabolol in Swiss albino mice. Both acute and subacute toxicity assessments showed that the combination of phytol and α-bisabolol is safe, with no adverse effects observed at higher concentrations. Hematological, biochemical, and histopathological tests showed no signs of toxicity in the heart, lungs, liver, spleen, and kidneys. The LD50 was greater than 2000 mg/kg, indicating a large safety margin. Histopathological analysis confirmed cancer induction in the B(a)P-induced group, which had significantly altered relative lung weights. Lung weight increased slightly pre and post-treatment, but histopathology showed normal alveolar epithelium. GSH and SOD levels increased significantly in B(a)P-exposed groups, indicating an adaptive antioxidant response. CAT levels increased significantly in the post-treatment group, demonstrating the role of combination of phytol and α-bisabolol in protecting against B(a)P-induced oxidative damage. Upregulation of Bax and downregulation of Bcl-2 caused a pro-apoptotic environment, suggesting a way to inhibit malignant cell survival. Modulation of caspase-3 and caspase-9 showed the complexity of carcinogen-induced apoptotic signaling. In conclusion, phytol and α-bisabolol were found to be safe and organ-protective, and demonstrated no acute or subacute toxicity. They modulate antioxidant defenses and apoptotic pathways, which may help prevent and treat lung cancer.

Assessing the efficacy of Stemona collinsiae roots extract against third-stage larvae of Gnathostoma spinigerum and its safety profiles.

Gnathostomiasis, caused by the advanced third-stage larvae of Gnathostoma spinigerum, demands novel treatment avenues. The ethanolic root extract of Stemona collinsiae has been postulated to have anthelminthic properties, suggesting its potential as an alternative remedy. In this study, S. collinsiae roots were collected, identified, and extracted with 95 % ethanol. The crude extracts were standardized using didehydrostemofoline as chemical marker. The efficacy of the S. collinsiae root extract against third-stage larvae of G. spinigerum and its toxicity to Wistar rats were evaluated. Both in vitro and in vivo tests were performed, where the in vitro tests assessed the anthelminthic potential of S. collinsiae extract against G. spinigerum larvae, while in vivo tests examined the extract's efficacy against G. spinigerum larvae in infected Wistar rats and the efficacy was compared with albendazole. Parallelly, Wistar rats underwent acute and sub-chronic toxicity tests to establish the safe dosage of the extract. The in vitro tests showcased significant anthelminthic activity, marked by discernible morphological alterations in the exposed larvae. Acute toxicity proved fatal at 2000 mg/kg body weight, while a dose of 300 mg/kg proved non-toxic. Using the Globally Harmonized Classification System, an LD50 of 500 mg/kg was determined. In vivo trials revealed a pronounced decline in G. spinigerum larvae among rats treated with the S. collinsiae extract. The larvae were also observed to be encysted post-treatment, while those treated with albendazole were not encysted. The S. collinsiae extract, with its noteworthy in vitro efficacy and favorable safety metrics in rodents, can be a potential anthelminthic agent. The diminished inflammatory response compared to albendazole hints at S. collinsiae being a safer gnathostomiasis treatment alternative. The promising results in these preliminary trials warrant a deeper investigation to determine the root extract's optimal dosing, suitable delivery methods, and its broader clinical implications.

Fumaric acid per se and in combination with methotrexate arrests inflammation via moderating inflammatory and oxidative stress biomarkers in arthritic rats.

Objective: Fumaric acid is a dicarboxylic acid that belongs to the phenolic class enriched in fruits and vegetables that are traditionally used for the treatment of various ailments. The research was planned to find out the anti-inflammatory and anti-arthritic activities of fumaric acid using in-vitro and in-vivo assays. Moreover, safety study was also done.Materials and methods: The 0.1 ml complete Freund's adjuvant was injected in left hind paw in all Wistar rats except normal rats at day 1 to induced arthritis. The treatment with fumaric acid at 10, 20, 40, and fumaric acid 40 mg/kg together with methotrexate (MTX) was administered to immunized rats at 8th day via oral gavage and continued till 28th day though, MTX was administered as standard control.Results: The fumaric acid notably (p < 0.0001) lessened the paw edema and arthritic scoring, reinstated body and immune organ weight, and oxidation status in treated rats. Fumaric acid notably restored altered C-reactive protein, rheumatoid factor, liver function tests, ESR, WBCs, RBCs and Hb levels in treated rats. The fumaric acid in combination noticeably (p < 0.01-0.0001) suppressed the expression of TNF- α, IL-6, IL-1ß, NF-kß, and COX-2, and over expressed IL-4, and IL-10 in contrast to other treated groups. Fumaric acid had presented a dose-dependent antioxidant, anti-inflammatory and anti-arthritic activities while notable activity exhibited by fumaric acid in combination with MTX. The fumaric acid exhibited non-significant clinical signs of toxicity and mortality in acute toxicity study. The LD50 was more than 2000 mg/kg.Conclusion: Fumaric acid in combination can be used as disease-modifying anti-rheumatic drug but it will need extensive pre-clinical and clinical studies.

Insight into didecyl dimethyl ammonium bromide toxicity following acute exposure in pullets (Gallus gallusdomesticus).

Didecyl dimethyl ammonium bromide (DDAB) is a quaternary ammonium compound used for the sanitation of drinking water of poultry and water pipelines in farms. There is scarcity of information on the toxicology of DDAB in poultry. This study set out to profile the acute toxicity of DDAB in poultry. Issa brown pullets (n = 34) as experimental birds were orally administered varying doses of DDAB, using a syringe, after 12 h fasting, and observed for toxicity over 14 days. Control birds (n = 10) were similarly given normal saline orally. Toxic signs in the experimental birds were depression, anorexia, adipsia, vocalization with foamy salivation, later emaciation and death. The LD50 was calculated as 458.00 mg/kg. Birds given 2151 mg/kg DDAB died within 24 h, while those treated with 516 mg/kg succumbed on Day 14. At necropsy, grossly, there were necrosis and sloughing of the oesophagus and intestines, pale and friable liver, congested and necrotic lungs, friable popped out kidneys and emaciated carcasses. Microscopically, desquamation and necrosis of the oesophagus, crop, proventriculus and intestines and disruption of the koilin membrane of the gizzard were observed. The lungs, liver and kidneys were congested with mononuclear cellular infiltration plus loss of architecture in the lungs and liver. In conclusion, at high doses, DDAB caused significant toxicity in chickens and these findings provide new information which could serve as a guide in the diagnosis of quaternary ammonium toxicity in chicken. The results could be extrapolated to other quaternary ammonium toxicities in related avian species.

Mild histological distortions in rat organs after a 14-day oral exposure to the slime extract of African giant land snails.

Objective: Snail slime possesses various pharmacological activities that are becoming attractive for zootherapy, thereby necessitating the profiling of its safety and toxicity. Therefore, using OECD 425 guidelines, this study assessed the acute toxicity of Archachatina marginata slime extract and performed a histological analysis of the vital organs. Methods: Eighteen (18) Wistar rats were assigned randomly into three groups: control, 2000 mg/kg, and 5000 mg/kg bw slime extract. The dosing of the animals with 2000 mg/kg bw and 5000 mg/kg bw was done according to the limit test procedure, after which the animals were observed for 14 days. During the observation period, clinical and behavioral changes were recorded. The rats were euthanized after 14 days of monitoring, and their essential organs were excised for gross histological examination. Results: There was no mortality during the observation period, and the LD50 of A. marginata slime extract was determined to be greater than 5000 mg/kg bw. Although there were no behavioral alterations in the rats after oral exposure to the slime extract, the histological examination revealed mild cellular distortions in the rat organs. Furthermore, a preliminary chemical analysis of the slime extract revealed the presence of flavonoids and phenolics. Conclusion: A. marginata slime extracts may be grouped as low toxic substance based on the results obtained (LD50 > 2000 - 5000 mg/kg). However, the histological distortions in rat organs following acute oral exposure to the snail slime extract not only warrant further, in-depth toxicological investigations but also caution in its use for traditional medicinal purposes.

Development of toxicity tests for Polyurethane foams.

Polyurethanes (PUs) are a special class of polymeric materials that differ significantly from most other types of plastic in many aspects. They can be utilized in a wide range of products, including paints, coatings, elastomers, insulators, elastic fibers, and foams. PU foams are especially important as part of various convenience products. PU products often end up in landfills when they are no longer useful and can release toxic compounds when damaged by humans or microbes. Therefore, the ecotoxicological assessment of PU foams is essential. In this paper, five PU foam samples were prepared with different NCO indices (NCO-0.8, 0.9, 1.0, 1.1, and 1.2) and together with the Control sample (a previously tested non-toxic foam sample) were applied to develop toxicity tests procedure, while intentionally prepared Toxic foam has been used to verify the accuracy of the developed testing procedure. Two test organisms were successfully applied, Sinapis alba (white mustard) seeds and Escherichia coli (non-pathogenic) bacterial model organisms, and toxicity tests were adapted for the examination of PU-derived substances. Regarding Sinapis alba test, the highest NCO index (NCO-1.2) significantly reduced root length by 9.8 % compared to the Control sample. In the bacterial test, it was observed that the samples containing NCO-1.1 and NCO-1.2 had lower colony numbers (5.0 × 108 and 4.9 × 108 CFU/mL respectively) in comparison to the Control plate (9.6 × 108 CFU/mL). All in all, two toxicity tests were successfully adapted for PU foams, and both are applicable in their ecotoxicological assessment.

Evaluation of the chronic oral toxicity of the classical ancient prescription Kai-Xin-San.

ETHNOPHARMACOLOGICAL RELEVANCE: The Kai-Xin-San (KXS), as an ancient classic prescription, has been used for the treatment of amnesia for thousands of years. Modern clinical and non-clinical pharmacological studies have found that it has significant therapeutic effects on dementia and depression, but there are relatively few studies on its safety. AIM OF THE STUDY: Subacute and chronic toxicity studies were conducted to investigate the symptoms, severity, target organs, development and recovery of toxic reactions, as well as the toxic dose. These studies provide technical data for ensuring the safety of KXS. MATERIALS AND METHODS: In the sub-acute toxicity study, rats were orally administered KXS at doses of 0.80, 1.61, 3.22, and 6.43 g/kg body weight for a duration of 4 weeks. In the chronic toxicity study, rats were orally administered KXS at doses of 0.27, 0.81, and 2.43 g/kg body weight for a duration of 26 weeks, and a withdrawal study was conducted for a period of 4 weeks after the treatment.The rats were observed daily for clinical signs and mortality. Changes in body weight, food consumption, and water consumption were periodically monitored. Additionally, urinalysis results, hematological and biochemical parameters, relative organ weights, and pathology were monitored at specific observation time points. RESULTS: In the sub-acute toxicity study, necropsy of dead and moribund rats revealed evident distension and swelling of the gastrointestinal tract, as well as thinning of the intestinal wall. The main adverse reactions observed included flatulence, piloerection, abnormal breathing sounds, and emaciation. Doses of 1.61 g/kg and below did not cause animal death. The gastrointestinal system is the main target organ of toxicity. In the chronic toxicity study, the no-observed-adverse-effect-level (NOAEL) of KXS was 0.27 g/kg, and its toxic effects were primarily concentrated in the gastrointestinal system. This led to secondary pathological changes in the immune system, hematopoietic system, and heart, suggesting that relevant indicators should be monitored when large doses are used clinically for an extended period of time. CONCLUSIONS: During the rodent toxicity evaluation, severe gastrointestinal damage was observed when KXS, powdered with crude drugs, was administered. The NOAEL for rats was found to be 0.27 g/kg/day.

Analysis of Differential Gene Expression under Acute Lead or Mercury Exposure in Larval Zebrafish Using RNA-Seq.

This study was first conducted to investigate the effects of acute lead exposure on developing zebrafish embryos or larvae from 24 to 120 h post-fertilization (hpf). Our data showed that treatment with 50-200 µM lead significantly affected larval survivability and morphology compared to the respective control. Second, we chose 120 hpf larvae treated with 12.5 µM lead for RNA sequencing due to its exposure level being sufficient to produce toxic effects with minimum death and lead bioaccumulation in developing zebrafish. A total of 137.45 million raw reads were obtained, and more than 86% of clean data were mapped to the zebrafish reference genome. Differential expression profiles generated 116 up- and 34 down-regulated genes upon lead exposure. The most enriched GO terms for representative DEGs were ion transport and lipid metabolism. Third, a comparison with the dataset of mercury-regulated gene expression identified 94 genes (64 up-regulated and 30 down-regulated) for exposure specific to lead, as well as 422 genes (338 up-regulated and 84 down-regulated) for exposure specific to mercury. In addition, 56 genes were co-regulated by micromolar mercury and lead treatment, and the expression of thirteen genes, including mt2, ctssb.1, prdx1, txn, sqrdl, tmprss13a, socs3a, trpv6, abcb6a, gsr, hbz, fads2, and zgc:92590 were validated by qRT-PCR. These genes were mainly associated with metal ion binding, proteolysis, antioxidant activity, signal transduction, calcium ion or oxygen transport, the fatty acid biosynthetic process, and protein metabolism. Taken together, these findings help better understand the genome-wide responses of developing zebrafish to lead or mercury and provide potential biomarkers for acute exposure to toxic metals.

The physiological effects of acute and sub-lethal exposure to phenol on antioxidant enzyme activity in the freshwater sludge worm Tubifex tubifex.

The current study investigates the severe effects of commonly employed chemicals, such as phenol, on the freshwater bottom-dwelling annelids of Tubifex tubifex. In an acute toxicity test, phenol's 96-hour LC50 value against Tubifex tubifex was identified to be 221.552 mg/L. Using the GUTS simulation, which places the GUTS-SD model on top of the GUTS-IT model, it was possible to confirm that the test organism would survive an acute exposure to phenol overall. After 14 days of treatment with 10 % and 20 % of the phenol's 96-hour LC50 values, long-term bioassays revealed changes in protein levels and in oxidative stress enzyme levels. Total protein concentration dropped during the bioassay, but levels of antioxidant enzymes (CAT, GST, SOD, and MDA) increased. The Pearson correlation matrix and the Integrated Biomarker Response (IBR) index were used for examining the relationship between biomarkers, toxicants, and phenol-induced stress. The results show that exposure to phenol is detrimental to the survival and general health of Tubifex tubifex.

Evaluation of arsenic induced genotoxicity and its impact on life processes of Daphnia magna.

Heavy metals like arsenic is ubiquitously present in the environment. Geologic and anthropogenic activities are the root cause behind high concentration of arsenic in natural water bodies demanding strict monitoring of water quality prior to human consumption and utilization. In the present study, we have employed Daphnia magna for studying the biological effects of environmentally relevant high concentration of arsenic in water. In acute toxicity study, the LC50 value for 24hr exposure was found to be 2.504 mg/L, which gradually decreased with increase in time period (24hr- 96hr) to 2.011 mg/ L at 96hr. Sub-chronic toxicity was evaluated over 12 days using sub-lethal concentrations (5 %, 10 %, 15 %, and 20 % of the 24-hr LC50). Survivability in Daphnia showed a decreasing trend from 96 % to 91 % with increase in arsenic concentrations from 5 % of LC50 24 hr value to 20 % of LC 50 24hr value respectively. Alongside decreased survivability, there was a significant reduction in body size, with organisms exposed to the highest concentration of arsenic measuring 0.87±0.01 mm compared to 1.51±0.10 mm in the control group. Reproductive potential declined concentration dependently with exposure, with the highest reduction observed at 20 % of LC50 24hr value, where offspring numbers decreased to 7±1 from 23±5 in the control. Heart rate decreased in concentration and time-dependent manners, with the lowest rates observed at the highest arsenic concentration (279±16 bpm after 24hr and 277±27 bpm after 48hr). Comet assay and micronucleus assay conducted after 48 hrs of exposure revealed concentration-dependent genotoxic effects in Daphnia magna. Our results indicate negative impact on physiology and reproduction of Daphnia magna at environmentally existent concentration of arsenic. Also Daphnia magna could serve as a sensitive test system for investigating arsenic contamination in water bodies.