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.

Bioactive constituents and acute toxicity of Blighia sapida capsule extracts using wistar rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Blighia sapida, commonly known as Ackee, is a plant native to West Africa, with great cultural and therapeutic value, particularly in Western Nigeria. Traditionally, Blighia sapida capsule is used in western Nigeria to treat ecthyma in sheep and goats by heating it in hot ash. This process causes the capsule to release a liquid, which is then directly applied to the entire affected area of the skin. However, there is limited information available on its phyto-constituents and medicinal effects. AIM OF THE STUDY: This work examined the bioactive constituents, acute toxicity, and sub-acute toxicity of aqueous and ethanolic extracts of Blighia sapida capsule. MATERIALS AND METHODS: Extraction of phytochemical constituents was carried out with distilled water and ethanol and was concentrated at 40 °C. The phytochemical constituents were determined using a variant 3800/4000 gas chromatography-mass spectrometry (GC-MS) machine. Lorke's method was employed to determine the acute toxicity of the aqueous and ethanolic extracts of Blighia sapida capsule. RESULTS: The GC-MS analysis revealed 15 bioactive compounds in both extracts, with kaempferol being the most abundant. Notable pharmacologically active compounds included pyrrolidin-2-ylmethanol, rutin, quinoline, apigenin, and naringenin. The study observed distinctive differences in aqueous and ethanolic extracts compound weights and peak areas. Acute toxicity study depicts that the lethal dose of aqueous and ethanolic extracts of Blighia sapida capsule is above 5000 mg/kg as no mortality was recorded in the oral administration of 10, 100, 1000, 1600, 2900, and 5000 mg/kg of aqueous and ethanolic extracts. Sub-acute toxicity results indicated no significant adverse effects on kidney and liver function, although some variations in biochemical parameters were observed. Histological analysis showed normal renal and hepatic architecture in treated animals. CONCLUSION: This study demonstrated that aqueous and ethanolic extracts of Blighia sapida capsule exhibited no acute toxicity and minimal sub-acute toxicity, suggesting they are safe for consumption at the tested doses.

In silico, in vitro, and in vivo acute and sub-acute toxicity profiling of whole plant methanol extract of Equisetum diffusum D. Don from the sub-Himalayan West Bengal, India, having ethnobotanical uses.

BACKGROUND: Equisetum diffusum D. Don commonly known as 'Himalayan horsetail', has been traditionally used in the treatment of back pain, bone fracture and dislocation, and arthritis by various tribal communities of India. Our previous study confirmed the anti-inflammatory efficacy of the plant through in silico, in vitro, and in vivo model studies. Therefore, the current research is focused on safety dose evaluation for the first-time of the whole-plant methanol extract (EDME) of E. diffusum through appropriate in silico, in vitro, and in vivo approaches. METHOD: The whole plant, along with its rhizomes, was collected, and the methanol extract was prepared. The in silico ADMET study was performed to predict the pharmacokinetics profile and toxicity of all the identified phyto-compounds of EDME previously screened by GC-MS study. In vitro cytotoxicity study of EDME was performed using two cell lines: kidney (HEK293) and liver (Huh7) cell lines. The in vivo toxicity study of EDME was validated by the acute toxicity (OECD 423, 2002) and sub-acute toxicity assays (OECD 407, 2008) in the Wistar Albino rat model. RESULTS: The in silico ADMET study of all 47 bioactives predicted good pharmacokinetic and low toxicity profiles. In vitro cytotoxicity showed higher IC50 values of EDME viz., 672 ± 15.7 µg/mL and 1698 ± 6.54 µg/mL for both kidney (HEK293) and liver (Huh7) cell lines, respectively, which were considered as low-toxic. Based on acute oral toxicity, the LD50 value of the extract was considered "non-toxic" up to a feeding range of 2000 mg/kg of body weight. The regular consumption of the extract for an extended period (28 days) was also qualified as safe based on the body and organ weight, hematological, biochemical, and histoarchitecture results in the sub-acute toxicity assay. CONCLUSION: The detailed in silico, in vitro, in vivo (acute and sub-acute oral toxicity) studies gave us a new insight to the safety dose evaluation of Equisetum diffusum, which may serve as a reliable documentation for undertaking the experimental validation of the ethnobotanical uses of the plant which would help in the field of drug development for the treatment of inflammation related complications.

Toxicological profiling of methanolic seed extract of Abutilon indicum (L.) Sweet: in-vitro and in-vivo analysis.

ETHNOPHARMACOLOGICAL RELEVANCE: Abutilon indicum, a shrub of the Malvaceae family, is found abundantly in tropical countries like India. A. indicum is widely used for its high medicinal properties. Traditionally, A. indicum seed powder is consumed to treat piles, constipation, chronic cystitis, gonorrhea, gleet, and pregnancy-related problems. Despite having numerous medicinal properties and widespread traditional use of A. indicum seeds, scientific validation, and toxicity studies have yet to be documented. AIMS OF THE STUDY: The primary objective of this study is to conduct a comprehensive study on phytochemical profiling, in-vitro cytotoxicity, mutagenicity, and in-vivo acute and sub-acute toxicity, and genotoxicity on animal models of methanolic extract of A. indicum seed (MAS). MATERIALS AND METHODS: The qualitative analysis of MAS was explored through FTIR and HR LC-MS. For in-vitro cytotoxicity, the HEK-293 cell line was used, and the TA100 (Staphylococcus typhimurium) bacterial strain was used for the Ames mutagenicity test. A single oral dose of 250, 500, 1000, or 2000 mg/kg body weight of MAS was given to each male and female rat for acute toxicity study and observed for 14 days for any toxicity signs. In the sub-acute toxicity study, 250, 500, or 1000 mg/kg body weight of MAS was administered orally to each rat for 28 days. The experimental animals were weighed weekly, and general behavior was monitored regularly. After 28 days of the experiment, the rats were sacrificed, and different serum biochemical, hematological, and histological analyses were performed. The blood samples of different doses of MAS were used for genotoxicity study through comet assay. RESULTS: FTIR analysis found different functional groups, which indicated the presence of phenolics, flavonoids, and alkaloids. HR LC-MS analysis depicts several components with different biological functions. The cell cytotoxicity and Ames mutagenicity results showed minimal toxicity and mutagenicity up to a certain dose. The acute toxicity study conducted in Wistar albino rats demonstrated zero mortality among the animals, and the LD50 value for seed extract was determined to be 2000 mg/kg body weight. Sub-acute toxicity assessments indicated that the administration of seed extract resulted in no adverse effects at dosages of 250 and 500 mg/kg body weight. However, at higher doses, specifically 1000 mg/kg body weight, the liver of the experimental rats exhibited some toxic effects. In the genotoxicity study, minimal DNA damage was found in 250 and 500 mg/kg doses, respectively, but slightly greater DNA damage was found in 1000 mg/kg doses in both male and female rats. CONCLUSIONS: The consumption of A. indicum seed powder is deemed safe; however, doses exceeding 500 mg/kg body weight may raise concerns regarding use. These findings pave the path for the creation of innovative medicines with improved efficacy and safety profiles.

Antioxidant Activity and Preclinical Safety of Semen persicae Extract.

Semen persicae is the dried mature seeds of Prunus persica (L.) Batsch and P. davidiana (Carr.) Franch and is commonly used in traditional Chinese medicine (TCM) formulations because of its variety of biological effects. The present study aimed to evaluate the antioxidant activity and toxicity profiles of semen persicae extract (SPE) after determining the amygdalin content (4.95%) using HPLC. Regarding the in vitro antioxidant activity, SPE with 2 mg/mL concentration scavenged 1,1-diphenyl-2-picrylhydrazyl (DPPH), hydroxyl, and ABTS free radicals with rates of 51.78%, 55.47%, and 57.16%, respectively. The same concentration of SPE chelated 30.76% Fe2+. The in vitro cytotoxicity study revealed that SPE induced 92.45% cell viabilities of HEPG2 even at 2000 µg/mL. In the acute toxicity study, oral administration of SPE did not provoke mortality or any toxic signs at doses up to 2000 mg/kg bw. Repeated oral administration for 28 days at doses of 100, 300, and 600 mg/kg per day in rats did not show any toxicity signs or gross pathological abnormalities. The results of the present research provide basic reference data for SPE with a moderate effect on antioxidant activity and low toxicity for future screening of biological and pharmacological properties.

Acute and subacute oral toxicity evaluation of Ayurvedic formulation Tapyadi loha in rats.

Ayurveda is one of the oldest systems of traditional medicine that provides treatments for a wide range of acute and chronic health problems. It is a common myth amongst people that Ayurvedic drugs have no side effects, whereas the fact is that these drugs can cause adverse effects. Despite their wide use, the safety data of many Ayurvedic formulations are still unavailable. Tapyadi loha is an Ayurvedic formulation traditionally claimed for iron deficiency anemia in pregnant and non-pregnant patients. However, no scientific study has been conducted to evaluate its oral toxicity. Hence, the present study evaluated the acute and subacute oral toxicity of the Tapyadi loha according to the OECD test guidelines 425 and 407, respectively. Tapyadi loha did not cause mortality nor any signs of toxicity when given once orally at a dose of 2000 mg/kg. Subacute toxicity study showed no mortality as well as no behavioral, hematological, biochemical and histopathological abnormalities in rats treated with Tapyadi loha formulation at 250, 500 and 1000 mg/kg for 28 days. It is concluded that the Tapyadi loha is safe at a single dose of 2000 mg/kg and 28 days repeated dose of 1000 mg/kg by oral route in rats.

Preparation of a hydrolyzed yeast ß-glucan chromium(III) complex and evaluation of its hypoglycemic activity and sub-acute toxicity.

Yeast ß-glucan (BYG) possesses extremely low solubility that has limited its applications. In this study, we hydrolyzed BYG using snail enzyme to obtain hydrolyzed yeast ß-glucan (HBYG) with desirable water solubility and hypoglycemic activity. On the basis of HBYG, HBYG­chromium(III) complex (HBYG-Cr) was synthesized. The molecular weight of the complex was 4.41 × 104 Da, and the content of trivalent chromium was 8.95 %. The hydroxyl groups of HBYG participated in the coordination and formed the chromium complex. The space conformations of HBYG exhibited remarkable changes after complex formation. HBYG-Cr existed mainly in an amorphous state and presented good dispersibility, and the surface was uneven. The hypoglycemic activity of HBYG-Cr was studied in db/db and C57 mice. The results showed that HBYG-Cr had good hypoglycemic activity. Histopathological studies demonstrated that the liver, kidney, pancreas, and skeletal muscle in the treatment group were significantly improved compared with those in the diabetic model group. The sub-acute toxicity of HBYG-Cr was studied in KM mice and the results indicated that the complex did not cause adverse reactions or toxic side effects. This study broadened the application of yeast ß-glucan and provided an important reference for the development of hypoglycemic functional foods and drugs.

Acute Toxicity, Hematological Profile, and Histopathological Effects of MgO Nanoparticles on Gills, Muscle, Liver of Mrigal, Cirrhinus mrigala.

Nanotechnology is an advancing and emerging field of all environmental, medical, and industrial applications. Magnesium oxide nanoparticles have been widely used in medicine, consumer products, industrial products, textiles, ceramics, alleviation of heartburn, stomach ulcers, and bone regeneration. In the present study, acute toxicity (LC50) of MgO nanoparticles and hematological and histopathological changes in Cirrhinus mrigala was analyzed. The lethal concentration for 50% of MgO nanoparticles was 4.2321 mg/L. Hematological parameters such as white blood cells, red blood cells, hematocrit, hemoglobin, platelets, mean corpuscular volume, mean corpuscular hemoglobin, and mean corpuscular hemoglobin concentration, as well as histopathological abnormalities in gills, muscle, and liver were observed on the 7th and 14th days of exposure. The WBC, RBC, HCT, Hb, and platelets count increased on the 14th day of exposure compared to the control and 7th day of exposure. The MCV, MCH, and MCHC levels decreased on the 7th day of exposure compared to the control and increased on the 14th day. Histopathological changes of MgO nanoparticles in gills, muscle, and liver highly damaged in the quantity of 3.6 mg/L compared to 12 mg/L on 7th and 14th days of exposure. This study finds the level of exposure in hematology and histopathological changes in tissues in relation to the exposure of MgO NPs.

Acute and Sub-Acute Toxicity Evaluation of the Crude Methanolic Extract of Justicia schimperiana Leaf in Wistar Albino Rats.

Purpose: This study evaluates the acute and sub-acute toxicity of 80% methanolic extracts of the leaves of Justicia schimperiana in Wistar albino rat models. Methods: Dried powdered leaves of Justicia schimperiana were macerated in 80% methanol. The experiment was conducted in accordance with the Organization for Economic Co-operation and Development guideline 423 for acute and 407 for sub-acute toxicity testing. A single dose of 5000 mg/kg extract was orally administered to three female rats for the acute toxicity study. The plant extract was administered orally for 28 days in daily dosages of 250, 500, and 1000 mg/kg for the sub-acute study. Animals in a control group were given distilled water. A total of 40 rats (5 rats/group/sex) were used for the sub-acute toxicity testing. Daily food intake and weekly body weight measurements were done. The rats were sacrificed at the end of the 28-day treatment period for hematological, biochemical, and histopathological tests. One-way analysis of variance and Kruskal-Wallis tests were employed for the analysis. Results: The single-dose oral administration of the plant resulted in no deaths or serious morbidity. The median lethal dose was >5000 mg/kg. The 28-day oral treatment of the plant extract had no significant effect on general behavior, food intake, organ weight, biochemical parameters, or the majority of the hematological markers, with the exception of the decrease in hemoglobin and hematocrit in the 1000 mg/kg extract-treated groups compared to the controls. Both sexes experienced significant weight increases at all dosage levels. With the exception of minor alterations in a few of the organs, no significant histological change was identified. Conclusion: It is concluded that the single-dose and repeated-dose 28-day oral administration of the methanolic leaf extract of Justicia schimperiana is relatively safe.

In Vivo Toxicity of Industrial Biocide Containing 2,2-Dibromo-3-nitrilopropionamide in Adult and Zebrafish Larvae.

2,2-Dibromo-3-nitrilopropionamide (DBNPA) is a widely used biocide with potential environmental implications due to its toxicity. This study aimed to investigate the in vivo toxicity of DBNPA in zebrafish (Danio rerio), a model organism in environmental toxicology. Both adult and larval zebrafish were exposed to varying concentrations of DBNPA, and significant morphological changes and mortality rates were observed. The study found that even relatively low concentrations of DBNPA can have detrimental effects on zebrafish embryonic development, and high concentrations resulted in rapid mortality in adult zebrafish and larvae. The LC50 values calculated from this study were 9.3 ppm for adults and 9.1 ppm for larvae, indicating the high toxicity of DBNPA to these organisms. These findings underscore the potential environmental impact of DBNPA and highlight the need for further research into its effects on aquatic ecosystems.

Estudios de Seguridad Preclínica del extracto hidroalcohólico de semillas de Opuntia soehrensii, una especie alto-andina con actividad anti-Herpes Virus, en ratas Sprague Dawley / Preclinical safety of the hydroalcoholic extract of Opuntia soehrensii seeds with anti-herpes virus activity, in SD rats

Introducción. Alrededor de 3700 millones menores de 50 años con infección por VHS-1 y 491 millones de personas de 15 a 49 años cursan con infección por VHS-2 en el mundo; sus síntomas, vesículas o ulceras dolorosas reaparecen periódicamente. El tratamiento convencional disminuyó su efectividad en cepas resistentes e inmunodeprimidos. Alternativas terapéuticas con extractos de plantas medicinales y potencial antiviral, como Opuntia soehrensii Brito conocida como "ayrampù" en Bolivia, utiliza infusión de sus semillas como analgésico, antidiabético, hipotensor y febrífugo. En vapores por inhalación para afecciones respiratorias; como tintura tópica en lesiones dérmicas de viruela, sarampión y herpes labial. Objetivo. Evaluar la seguridad preclínica de un gel que contiene el extracto hidro-alcohólico de semillas de Opuntia soehrensii en diferentes dosis, aplicado en la mucosa vaginal de ratas Sprague Dawley. Material y métodos. Se ejecutaron protocolos de toxicidad aguda y subaguda para evaluar la respuesta sistémica, a través de marcadores bioquímicos y de comportamiento, y la respuesta local en mucosa vaginal, mediante estudios histopatológicos, en grupos de animales a los que se aplicó el gel con diferentes concentraciones del extracto de Opuntia soehrensii, comparados con un grupo control y otro que recibió solo el vehículo. Resultados. Se encontró que los indicadores sistémicos de comportamiento y ganancia de peso no mostraron diferencias entre grupos. Los indicadores hematológicos y bioquímicos mostraron resultados fisiológicamente esperados y sin cambios en los grupos de estudio. La citología expuso conservación del fenotipo celular para las fases del ciclo estral en todos los grupos. Los indicadores histológicos de reacción local e integridad celular se distribuyeron de igual manera en los todos los grupos. Conclusión. La aplicación de un gel de Opuntia soehrensii no muestra niveles apreciables de toxicidad local y sistémica, lo que permite recomendar la iniciación de estudios de aplicación clínica.

Introduction. Around 3.7 billion people under 50 years of age are infected with HSV-1 and 491 million people between the ages of 15 and 49 are infected with HSV-2 in the world; his symptoms, vesicles or painful ulcers recur periodically. Conventional treatment decreased its effectiveness in resistant and immunosuppressed strains. Therapeutic alternatives with extracts of medicinal plants and antiviral potential, such as Opuntia soehrensii Brito known as "ayrampù" in Bolivia, uses infusion of its seeds as an analgesic, antidiabetic, hypotensive and febrifuge. In vapors by inhalation for respiratory conditions; as a topical tincture in skin lesions of smallpox, measles and cold sores. Objectives . To evaluate the preclinical safety of a gel containing the hydroalcoholic extract of Opuntia soehrensii seeds in different doses, applied to the vaginal mucosa of Sprague Dawley rats. Material and Methods. Acute and sub-acute toxicity protocols were carried out to evaluate local response in the vaginal mucosa, through histo pathological studies, and systemic responses, through biochemical and behavioral markers, in groups of animals to which the gel with different concentrations of the extract of Opuntia soehrensii was applied, compared with a control group and another that received only the vehicle. Results. It was found that the histological indicators of local reaction and cell integrity were equally distributed in all groups. Cytology showed conservation of the cell phenotype for the phases of the estrous cycle in all groups. The systemic indicators of behavior and weight gain did not show differences between groups. Hematological and biochemical indicators showed results ranged in physiologic parameters, without changes in the study groups. Conclusion. The application of a gel from Opuntia soehrensii does not show appreciable levels of local and systemic toxicity, which makes it possible to recommend the initiation of clinical application studies.

Evaluation of Acute and Sub-Acute Toxicity, Oxidative Stress and Molecular Docking of Two Nitrofuranyl Amides as Promising Anti-Tuberculosis Agents.

Tuberculosis (TB) remains a widespread infectious disease and one of the top 10 causes of death worldwide. Nevertheless, despite significant advances in the development of new drugs against tuberculosis, many therapies and preventive measures do not lead to the expected favorable health results for various reasons. The aim of this study was to evaluate the acute and sub-acute toxicity and oxidative stress of two selected nitrofuranyl amides with high in vitro antimycobacterial activity. In addition, molecular docking studies were performed on both compounds to elucidate the possibilities for further development of new anti-tuberculosis candidates with improved efficacy, selectivity, and pharmacological parameters. Acute toxicity tests showed that no changes were observed in the skin, coat, eyes, mucous membranes, secretions, and vegetative activity in mice. The histological findings include features consistent with normal histological architecture without being associated with concomitant pathological conditions. The observed oxidative stress markers indicated that the studied compounds disturbed the oxidative balance in the mouse liver. Based on the molecular docking, compound DO-190 showed preferable binding energies compared to DO-209 in three out of four targets, while both compounds showed promising protein-ligand interactions. Thus, both studied compounds displayed promising activity with low toxicity and can be considered for further evaluation and/or lead optimization.

Acute and sub-acute toxicity study reveals no dentrimental effect of formononetin in mice upon repeated i.p. dosing.

AIM: Formononetin is a phytoestrogen which possess different pharmacological activities. The intraperitoneal route permits the identification of target organs involved in toxicity without compromising the molecule's bioavailability. The current study investigated the safety profile of intraperitoneal formononetin in Swiss albino mice. MATERIAL AND METHODS: For acute toxicity study, formononetin administered intraperitoneally to mice at the doses of 5, 50, 100, 150, 200, and 300 mg/kg for 14 days. For the subacute toxicity study, mice were intraperitoneally administered with formononetin (12.5, 25, and 50 mg/kg) daily for 28 days. RESULTS: During the acute study, no deteriorating effect was observed on body weight, food and water intake, no behavioral changes were observed in animals. The lethal dose 50% (LD50) of formononetin was determined to be 103.6 mg/kg of BW, with a no observed adverse effect level (NOAEL) of 50 mg/kg of BW. Mortality was observed in the 300 mg/kg dose group and histopathological changes such as a mild degree of diffuse granular degeneration in the liver but for rest all doses did not have any adverse effect. In subacute study, no signs of adverse effects, mortality, no changes in body weight, food and water intake, and hematological and biochemical parameters were observed. Histopathology of subacute study indicates, formononetin did not have any noxious effect on organs. CONCLUSION: Formononetin shows mortality at acute dose 300 mg/kg and LD50 at 103.6 mg/kg of BW, with a NOAEL of 50 mg/kg of BW, rest all doses for acute and sub-acute are safe when given intraperitoneally.

Quercetin and Ferulic Acid Elicit Estrogenic Activities In Vivo and In Silico.

In this study, we examined the sub-acute toxicity of quercetin and ferulic acid and evaluated their effects on protein, cholesterol, and estrogen levels in vivo. Six groups of female Wistar rats were fed by gavage. The first and second groups represent the positive (Clomiphene citrate 10 mg/kg) and negative (NaCl 0.9%) control groups, while the other groups received quercetin and ferulic acid at doses of 5 and 10 mg/kg/day for 28 days. The sub-acute toxicity was monitored by examining the weights, biochemical parameters (AST, ALT, ALP, urea, and CREA), and histological changes in the kidneys and liver of the treated animals. Furthermore, the in vivo estrogenic effects were studied in terms of the serum and ovarian cholesterol levels, serum estradiol, and uterine proteins. Finally, Docking studies were conducted to evaluate the binding affinity of quercetin and ferulic acid for alpha and beta estrogen receptors. Results showed that both compounds were devoid of any signs of nephrotoxicity or hepatotoxicity. Additionally, quercetin and ferulic acid caused significant estrogenic effects evidenced by an increase of 8.7 to 22.48% in serum estradiol, though to a lesser amount than in the reference drug-treated group (64.21%). Moreover, the two compounds decreased the serum cholesterol levels (12.26-32.75%) as well as the ovarian cholesterol level (11.9% to 41.50%) compared to the negative control. The molecular docking in estrogen alpha and estrogen beta active sites showed high affinity of quercetin (-10.444 kcal/mol for estrogen alpha and -10.662 kcal/mol for estrogen beta) and ferulic acid (-6.377 kcal/mol for estrogen alpha and -6.3 kcal/mol for estrogen beta) to these receptors. This study provides promising insights into the potential use of quercetin as a therapeutic agent for the management of female fertility issues.

Acute and sub-acute toxicity evaluation of Merremia tridentata (L.) stem extract on mice.

The acute and sub-acute toxicity studies were performed in male and female Swiss albino mice as per the guidelines mentioned in OECD. The oral administration of M. tridentata stem extract (MSE) showed no treatment-related mortality and body weight change in mice up to the single dose of 30,000 mg/kg body weight in the acute toxicity study and up to a dose of 30,000 mg/kg/day body weight in the sub-acute toxicity study. Moreover, the clinical signs, body weight, gross pathology, organ weight, hematology (except for platelet count), biochemical analysis, and histopathology did not show significant variation at a medium dose of 15,000 mg/kg/day body weight compared to the control group. However, toxicological signs in behavior, very mild interstitial nephritis, as well as significant variation in platelet count and total protein parameters were observed at a dose of 30,000 mg/kg/day in the 28-day oral toxicity study. Thus, the no-observed-adverse-effect level was determined at a dose of 15,000 mg/kg/day body weight. Based on study results, it is concluded that MSE showed LD50 of greater than 5000 mg/kg/day body weight. Hence, this could be a potential candidature as a future safe pharmaceutical product.

Revealing the most effective anticonvulsant part of Malvaviscus arboreus Dill. Ex Cav. and its acute and sub-acute toxicity.

ETHNOPHARMACOLOGY RELEVANCE: Different parts of Malvaviscus arboreus Dill. Ex Cav. (M. arboreus) are traditionally used in the West Region of Cameroon to treat many diseases, including epilepsy. AIM OF THE STUDY: To determine which part of M. arboreus offers the best anticonvulsant effect, and to assess the acute and sub-acute toxicity of the part of interest. MATERIALS AND METHODS: the anticonvulsant effect of the aqueous lyophilisate of the decoction of flowers, leaves, stems and roots of M. arboreus at various doses was evaluated and compared on the model of acute epileptic seizures induced by pentylenetetrazole (PTZ) (70 mg/kg), injected 1 h after oral administration of the various extracts. Out of these plant parts, the leaves were then selected to prepare the hydroethanolic extract and its anticonvulsant effect against PTZ at the doses of 122.5, 245 and 490 mg/kg, as well as its acute toxicity were compared with those of the aqueous lyophilisate of the leaves. The anticonvulsant effect of the aqueous lyophilisate of M. arboreus leaves was further evaluated on models of acute epileptic seizures induced by picrotoxin (PIC) (7.5 mg/kg), strychnine (STR) (2.5 mg/kg) and pilocarpine (350 mg/kg). The 28 days sub-acute toxicity, as well as the quantitative phytochemistry and the in vitro antioxidant potential (FRAP, DPPH, ABTS+) of the aqueous lyophilisate of the leaves of M. arboreus were also evaluated. RESULTS: M. arboreus leaves showed the best anticonvulsant effect and the aqueous lyophilisate was the best extract. The latter significantly protected the animals against convulsions induced by PTZ (71.43%) (p < 0.01), PIC (57.14%) (p < 0.05) and STR (42%) and had no effect on pilocarpine-induced seizures. Furthermore, it showed no acute or sub-acute toxicity, and revealed a high content of flavonoids, saponins, tannins and alkaloids, and antioxidant activity in vitro. CONCLUSION: The aqueous lyophilisate of the leaves of M. arboreus offers the best anticonvulsant effect on the extraction solvent used, and it would act mainly via a potentiation of the inhibitory systems of the brain (GABA, Glycine). In addition, its richness in bioactive compounds gives it an antioxidant potential, and it is not toxic in acute and sub-acute toxicity. All this justifies at least in part its empirical uses, and makes M. arboreus a candidate for the alternative treatment of epilepsy.

Sub-acute toxicity of graphene oxide (GO) nanoparticles in male mice after intraperitoneal injection: Behavioral study and histopathological evaluation.

Graphene oxide (GO) is a graphene derivative used for numerous applications in which biomedical uses are significant. However, for this application, the security of GO is doubtful. In this work, we synthesized this nanoparticle to assess its toxicity in male mice. In addition, we studied the effects of this nanomaterial on behavior by administering GO intraperitoneally to mice at different doses (2 mg/kg and 5 mg/kg) for five days. Subsequently, we performed biochemical analyses of blood serum and measured peroxidase and malondialdehyde (MDA) activity. Then, we performed histological sections to evaluate the brain's and liver's pathological and morphological changes. The data showed that the open field tests did not alter the locomotor activity. Furthermore, the elevated cross-maze tests showed no anxiety effect in the GO doses in the animals. The biochemical analyses indicated that GO influenced the level of biochemical parameters. Although, the oxidative stress assay showed an increase in peroxidase and MDA activity after GO intoxication. However, histopathological analysis of liver sections showed that GO caused liver inflammation, whereas, at the brain level, GO did not affect neuronal cells. The results indicate that GO caused toxic effects and that its toxicity could be mediated by oxidative stress.

Phytochemical analysis, in silico study and toxicity profile of Cycas pectinata Buch.-Ham seed in mice.

Fruit of Cycas pectinata Buch.-Ham has been used as medicine by the local community in some parts of the north eastern state of India. Despite its uses for different purposes, the safety assessment study has not been conducted. Therefore, we have evaluated the acute and the sub-acute toxicity of methanolic extract of C. pectinata fruit (CPFE) in a mice model via oral route of administration. Phytochemicals analysis was carried out by liquid chromatography-mass spectroscopy (LC-MS), nuclear magnetic resonance (NMR), and Fourier-transform infrared spectroscopy (FTIR). The acute toxicity study was performed at a single dose of 1000, 3000 and 5000 mg/kg and the sub-acute toxicity study at a dose of 100, 300 and 500 mg/kg was administered daily for 28 days. The calculated Lethal dose 50 (LD50) of CPFE was found to be 4000 mg/kg. Both acute and sub-acute studies showed that 5000 mg/kg and 500 mg/kg dose was toxic to the mice. The results of acute toxicity showed CPFE could have a mild toxic effect on the kidney at a dose of 3000 and 5000 mg/kg, as some deteriorated changes in the kidney along with increase creatinine levels were observed. Acute toxicity also showed an increase in white blood cells (WBC) at a dose of 3000 mg/kg.However, sub-acute toxicity studies do not show any detrimental effects on liver, kidney and hematological parameters. Thus, it can be suggested that CPFE at a dose of 100 and 300 mg/kg would be safe for consumption. The phytochemicals analysis by LC-MS, NMR and FTIR showed the presence of 32 major chemical compounds with certain biological activity like anti-neoplastic, antioxidant, and possible modulator of steroid metabolism (cholesterol antagonist and agonist of testosterone 17ß-dehydrogenase) as predicted by PASS analysis.

Acute and sub-acute toxicity study of ethanol extract from Nectandra leucantha Nees & Mart. (Lauraceae) barks.

Nectandra leucantha has been used in traditional medicine. Several metabolites isolated from N. leucantha extracts displayed immunomodulatory, antileishmanial properties, but the determination of the toxicological profile in mammals has not previously been performed. In this study, the ethanol extract from N. leucantha barks (EENl) was characterized by HPLC/HRESIMS. To study acute toxicity, female mice received EENl in a single dose of 100, 300, 1000, or 2000 mg/kg bw. Later, sub-acute toxicity was introduced in female and male mice by oral gavage at 100, 500 or 1000 mg/kg bw for 28 consecutive days. Hematological and biochemical profiles from the blood as well as histological analysis from the liver and kidney were performed. The HPLC/HRESIMS analysis of the EENl revealed the presence of six neolignans chemically related to dehydrodieugenol B. In the oral acute and sub-chronic studies, EENl did not produce in all doses evaluated any alteration in behavior, biochemical, hematological, body weight gain and food intake or sudden death in Swiss mice. In addition, histopathological data did not reveal any disturbance in liver and kidney morphology after 28 days of EENl treatment. Our results indicate that EENl at dosage levels up to 2000 mg/kg bw is non-toxic and can be considered safe for mammals.

The sub-acute toxicity of kavalactone in rats: a study of the effect of oral doses and the mechanism of toxicity in combination with ethanol.

Kava is a herbal supplement and beverage made from the Piper methysticum plant, which is known for its recreational use as a mood enhancer, relaxation, as well as pain relief for centuries. Kava is widely used among alcoholics, but it is dangerous and potentially fatal. The objectives of this study were to examine the sub-acute toxicity effects of different doses of 70% kavalactone (KL) in rats by oral application, as well as to elucidate the mechanisms of toxicity alone and in combination with ethanol (EtOH). The most common side effects observed were abnormal breathing, ataxia, lethargy, loss of appetite, indigestion, and loss of coordination, especially in the 800 mg/kg bw, po bodyweight dosage of kava treatment group alone, and in combination with EtOH. In the sub-acute study, there were dose-related decreases in body weight, feed intake, and water consumption rates. Gross and histopathological findings revealed that the liver was abnormal in color, size, consistency, and the weight significantly increased at a dose of 800 mg/kg bw, po, with KL alone and a greater increase in combination with EtOH. Hepatocellular hypertrophy (HP) and necrosis with Kupffer cells hyperplasia were observed in the periacinar zone of all rats dosed with KL (800 mg/kg bw, po) alone, and extensive changes were observed in combination with EtOH. The periportal (Z1) and mid-zonal (Z2) areas of hepatocytes were less affected as compared to the periacinar zone. These results demonstrate that EtOH exacerbated the sedative and hypnotic activity of KL, and markedly increased toxicity. The histopathological results supported the clinical and biochemical findings and the severity of hepatic damage in a dose-dependent manner.