Subchronic Toxicity Research Articles

Evaluation of Subchronic toxicity and phytochemical analysis of aqueous extract of Ocimum suave (Lamiciae)

Evaluation of phytochemical content, acute and sub-chronic toxicity of the aqueous leaf extract of Ocimum suave in Wistar rats was carried out. Experimental rats were administered extracts daily for 31days at dose levels of 200, 400 and 800 mg/kg. Acute toxicity and phytochemical screening of aqueous leaf extract were done according to method of Lorke’s Trease and Evans. Biochemical analysis was equally done at the end of the administration of extracts using standard methods. At the end of the study, animals were sacrificed and blood samples taken for biochemical assay. The LD50 was found to be greater than 10,000 mg/kg body weight. Results showed the presence of most phytochemicals tested for. The aqueous extract caused a decrease in ALT activity but AST and CK activities were higher in the treated groups in a dose dependent manner relative to the control. The aqueous leaf extract of Ocimum suave affected mean cell volume, mean cell hemoglobin and mean cell hemoglobin concentration. The aqueous leaf extract caused mean body weight gain, but decrease in relative organ weight. The study suggests that the aqueous extract administered at normal therapeutic doses is not likely to produce severe toxic effects on some hematological and biochemical indices in rats and the organs tested. Chronictoxicity of aqueous leaf extract of Ocimum suave should be done for histological, haematological andbiochemical evaluation. Key Words: Ocimum suave, Toxicity, Wistar Rats, Biochemical assay, Phytochemical, Extract

Toxicological Evaluation of Sub-Chronic Administration of Ethanol Leaf Extract of <i>Ocimum canum</i> on Lipid Parameters, Heart Histology and Feed Consumption of Wistar Rats

Ocimum canum is a plant that is commonly used in many parts of SubSaharan Africa to treat ailments such as infection, discomfort, and diarrhea. The study was aimed to evaluate the sub-chronic toxicity of Ocimum canum on the heart, lipid profile and feed consumption in Wister rats after oral administration of the leaf extract. Animals of either sexes were chosen. Group 1 received distilled water (10 ml/kg), while groups 2, 3, and 4 received 100, 200, and 400 mg/kg of Ocimum canum, respectively. Animals were kept in conventional cages and provided oral access to the extract, water, and food for 28 days before being weighed and slaughtered. Blood was drawn via heart puncture and immediately sent for hematological and chemopathological investigation. The plant's histological cardiotoxic potential was investigated using the haematoxylin and eosin (H&E) staining technique. There was a significant (P<0.05) drop in RBC, HGB, and MCV, but no change in neutrophils, basophils, eosinophils, or platelets. The extract did not cause a significant (P<0.05) change in cholesterol, LDL, or triglyceride levels, but it did induce a substantial increase in HDL levels. Histological examination of cardiac tissue confirms other parameters. The results indicated that the plant may not be cardiotoxic.

Evaluation of sub-chronic toxicity of melamine via systematic or oral delivery in adult zebrafish based on behavioral endpoints

Melamine-tainted products have been found in the market and raised issues about food safety. Recent studies done in rodents and humans demonstrated the toxicities of melamine, especially in causing kidney damage and bladder stone formation. However, very few studies assessed its behavior toxicity in organisms, including fish. Therefore, in this study, the researchers aim to determine whether sub-chronic exposure to melamine via oral and systematic administration could induce behavioral abnormality in zebrafish. After 14 days of systematic exposure to melamine at doses of 0.1 and 10 ppm levels, zebrafish were subjected to multiple behavioral assays. Results from both exposure routes showed that melamine indeed slightly increased fish locomotion and altered their exploratory behaviors in the novel tank assay. Furthermore, tightened shoaling formation was also displayed by the treated fish in the waterborne exposure group. However, melamine exposure did not cause any obvious alterations in fish behaviors during other behavioral tests. In addition, in comparison with previously published data on the behavior toxicities of several solvents in zebrafish, our phenomic analysis suggests the relatively low behavior toxicities of melamine via either systematic exposure or oral administration to zebrafish compared to those solvents. Nevertheless, our data indicate that the potential neurotoxicity of chronic low-dose melamine should not be ignored.

Non-clinical Safety Evaluation of Camelina Oil: Acute and 12-Week Oral Toxicities.

This study assessed the acute and sub-chronic toxicity of Camelina oil, a well-known oil rich in polyunsaturated fatty acids that enhance cellular immunity and human health, in Wistar rats. Wistar rats, 5 per sex per group, were randomly assigned to three groups for acute (14 days) toxicity studies and five groups for sub-chronic (90 days) toxicity studies. In the acute study, Camelina sativa oil was administered orally at a single dose of 5000 mg/kg of body weight (BW). The positive control group received a single dose of 5 000 mg/kg BW Canola oil by gavage. In the sub-chronic study, Groups III-V received 250, 500, and 1 000 mg/kg BW of Camelina oil, while Groups I and II received ultra-pure water and Canola oil at a dose of 500 mg/kg BW, respectively. Throughout the experiment, clinical signs, mortality, and body weight were monitored. At the end of the sub-chronic study, hematological, biochemical, and histopathological investigations were conducted. Administration of Camelina oil and Canola had no significant effect on daily weight gain (P > 0.05) of the test rats. Serum calcium levels decreased while phosphorous levels increased in male rats treated with Camelina oil. Other hematological and biochemical parameters showed no significant differences or dose-response effects between control and seed oil groups in both sexes (P < 0.05). Moreover, in animal necropsy, there were no apparent lesions in the liver, heart, and kidney organs in any of the doses administered. In conclusion, the results suggest that oral administration of Camelina oil is unlikely to be toxic. Therefore, the possibility for the development of future human nutrition should be considered.

The effect of the drug L-carnitine when administered intramuscularly on the blood parameters of rats

L-carnitine is a vitamin-like substance that plays an important role in the transport of fatty acids and their further use as an energy source. Most studies have identified beneficial effects of using L-carnitine for preventive and therapeutic purposes. The series of publications reviewed is devoted to the study of the results of the use of L-carnitine in metabolic disorders, diseases of various organs, organ systems, the central nervous system, as well as to elucidate the possible reasons for the decrease in the level of L-carnitine in the body. To date, L-carnitine is used in the Russian Federation only in the form of a feed additive; the purpose of our study was to study L-carnitine in the form of an injection solution. Toxicity studies were conducted on outbred rats in October 2021 in the vivarium of the St. Petersburg State University of Veterinary Medicine. The study involved females weighing 190-210 grams. When studying subchronic toxicity when administered intra-muscularly, L-carnitine was administered at 2 dose levels. 1/5 and 1/10 of the maximum tolerated dose in acute toxicity studies. In this regard, three groups of animals were formed. The first experimental group (n=10) was administered the drug intramuscularly at a dose of 0.08 mg/kg, the second group (n=10) at a dose of 0.04 mg/kg. The control group (n=10) was intramuscularly injected with a 0.09% sodium chloride solution at a dose of 0.08 mg/kg. The study was carried out over 42 days. Biological material samples were taken from 5 animals from each group the next day after the end of the drug administration (day 43), and 10 days after the last drug administration (day 53). The results of the study showed that dosages of 1/5 and 1/10 of the maximum tolerated do not cause external signs of toxicosis and death in rats. No significant changes were found in the biochemical parameters of the blood of animals in the experimental and control groups.

Androgenic Properties and Subchronic Toxicity of the Aqueous Extract of Pycnanthus angolensis (Welw.) Warb. Wood (Myristicaceae)

Aims: Pycnanthus angolensis (Welw.) Warb. also known as “Etengué” in the Baka language, is a medicinal plant used by the Baka Pygmies of Cameroon to treat erectile dysfunction. This study aimed to evaluate the androgenic activity and subchronic toxicity of the aqueous extract of Pycnanthus angolensis wood.&#x0D; Study Design: Experimental design.&#x0D; Place and Duration of Study: Laboratory of Biochemistry, Faculty of Medicine and Biomedical Sciences, University of Yaoundé I, between November 2022 and October 2023.&#x0D; Methodology: Twenty-five male Wistar strain rats were divided into 5 groups of 5 rats each including a positive control group that received testosterone enanthate (5 mg/kg) intramuscularly once a week; a normal control group that received distilled water (10 ml/kg) and three test groups which received 134 and 267 and 533 mg/kg of the aqueous extract of Pycnanthus angolensis daily per os. On the 29th day, the animals were killed, and several biochemical parameters were assessed for androgenic properties.&#x0D; Results: At a dose of 134 mg/kg, the extract significantly increased (p&lt;0.05) the levels of testicular cholesterol, testosterone and serum prostatic acid phosphatase activity compared to those of the normal control. A toxicity study showed that at doses of 134 and 267 mg/kg, the extract did not induce any significant variation (p&gt;0.05) in the creatinine level, but a significant reduction (p&lt;0.05) in of aspartate and alanine aminotransferase activities was recorded. Histopathology revealed destruction of testicular spermatozoa at a dose of 533 mg/kg compared to that of the controls.&#x0D; Conclusion: These results showed that at a dose of 134 mg/kg, the aqueous extract of Pycnanthus angolensis (Welw.) Warb. wood is capable of stimulating the production of androgens and is tolerated by the rat's body. Our results justified the traditional use of this plant for the treatment of male infertility and erectile dysfunction.

Acute and Subchronic Toxicological Study of the Cocktail Extract from Curcuma xanthorrhiza Roxb, Phyllanthus niruri L. and Morinda citrifolia L.

Curcuma xanthorrhiza Roxb, Phyllanthus niruri L., and Morinda citrifolia L. are Indonesian medicinal herbs used empirically as traditional therapeutics for maintaining health. The cocktail extract of these three plants (CECPM) had been developed and demonstrated immunostimulant activity in rats. This study aimed to evaluate the acute and subchronic toxicity of CECPM in vivo. The acute toxicity assay was conducted by orally administering a range dose of CECPM (313, 625, 1250, 2500, or 5000 mg/kg body weight (bw) on female mice once and then evaluating the toxic symptom every day for 14 days later. The chronic toxicity test was carried out by giving various doses of CECPM (600, 800, and 1000 mg/kg·bw) to female and male rats orally continuously for 90 consecutive days. The signs of toxicities were evaluated at the 90- and 28 days postadministration. The acute oral toxicity assays showed that there was no toxic syndrome and mortality found during the period of the experiment. The lethal dose level (LD50) of CECPM was more than 5000 g/kg, which was categorized as practically non-toxic. Meanwhile, in the sub-chronic toxicity study, some parameters tested at 90 days postadministration and after 28 days of withdrawal, such as the body weight, relative organ weight, food intake, hematological and biochemical blood parameters, and also histopathological examination of five primary tissues (heart, liver, kidney, spleen, and lung) revealed no abnormalities. There was no-observed adverse effect level (NOAEL) for the present study of CECPM 1000 mg/kg·bw of the rat. Therefore, it is concluded that the orally administered CECPM was relatively nontoxic during acute and subchronic toxicology studies.

Evaluation of acute, 28-day, 13-week repeated dose oral toxicity and genotoxicity of a herbal extract (HemoHIM G) from Angelica sinensis, Ligusticum chuanxiong, and Peaonia lactiflora.

HemoHIM G is a functional food ingredient composed of a triple herbal combination of Angelica sinensis, Ligusticum chuanxiong, and Paeonia lactiflora, to improve impaired immune function. Considering the pharmacological benefits of its constituent herbal components, HemoHIM G is anticipated to have various health benefits; however, its toxicity has not been thoroughly evaluated. Here, we conducted a comprehensive study to assess the safety of HemoHIM G in terms of acute oral toxicity, 13-week repeat-dose toxicity, and genotoxicity. In the oral acute toxicity study, Sprague-Dawley rats were orally administered a single dose of HemoHIM G at 5000mg/kg/day, the limit dose for the acute study. No abnormal findings or adverse effects were observed in this study, as confirmed by gross pathology. A 13-week repeated-dose toxicity study was conducted with HemoHIM G at doses of 1250, 2500, and 5000mg/kg/day to examine the subchronic toxicity in both male and female rats after 28days of dose-range finding study. No test substance-related clinical signs or mortality was observed at any of the tested doses. Gross pathology, hematology, blood chemistry, and histopathology were within normal ranges, further supporting the safety of HemoHIM G. Therefore, the NOAEL of HemoHIM G was considered to be at 5000mg/kg/day for both sexes of rats. Bacterial reverse mutation tests, a chromosome aberration test in human peripheral blood lymphocytes, and a mouse micronuclei test were conducted to identify the potential genotoxicity of HemoHIM G. HemoHIM G is non-mutagenic and non-clastogenic. Collectively, these findings provide valuable evidence for the safe use of HemoHIM G as a functional food ingredient.

Sub-chronic oral toxicity study of the alkaloid rich fraction from Luffa cylindrica fruit in Sprague-Dawley rats

The loofah/sponge gourd Luffa cylindrica (L.), a member of the Cucurbitaceae family, is one of the neglected medicinal plants. Traditionally, Luffa cylindrica is prescribed for inducing labor. It has a long history of use in China for the treatment of fever, diabetes, dyspnea, and dysentery. This study investigated the toxicity profile of the alkaloid-rich fraction of Luffa cylindrica (ARF-LC) for the first time in Sprague Dawley rats. A total of 80 rats (40 male and 40 female rats) aged 13 weeks old and weighing 200–220 g were selected for this study. In SD rats, sub-chronic oral toxicity was investigated at doses of 100, 200, and 400 mg/kg/d for a total of 90 days, followed by a 30-day recovery period. The results showed no variation in body weight among the three dose groups compared to the control group. Treatment-related adverse events, such as alterations in hematology and serum biochemistry parameters and the histology of the liver were sporadic in the high-dose rats but within the reference range. However, these changes disappeared after the doses were withdrawn during the recovery period. In conclusion, the “no observed adverse effect level” (NOAEL) of oral administration of ARF-LC in SD rats was considered 400 mg/kg/d and can be studied for its potential in further in vivo chronic investigations.

A study on the subchronic toxicity of triclocarban to the early-life development of oryzias melastigma and focused on the analysis of osmoregulatory regulation mechanisms

Triclocarban (TCC), a novel antimicrobial agent found in personal care products, has been extensively detected in marine environments. However, research on the toxic effects of TCC on marine organisms remains inadequate. This study delved into the subchronic toxic effects of TCC on the early life stages of marine medaka (Oryzias melastigma, O. melastigma), revealing that TCC could reduce embryo heart rate and hatching rate while diminishing the survival rate of larvae. Biomarker assays indicated that TCC could inflict damage on the embryos' antioxidant and nervous systems. Transcriptomic analysis suggested that TCC could impact cell growth, reproduction, and various life processes, activating cancer signaling pathways, increasing the likelihood of cancer, and exerting toxic effects on the immune and osmoregulatory systems. To validate and enhance our understanding of TCC's unique toxic impact on the osmoregulatory system of O. melastigma, we conducted homology modeling and molecular docking analyses on the protein involved in osmoregulation. The study intuitively revealed the potential binding affinity of TCC to sodium/potassium-transporting ATPase subunit alph (ATP1A1), indicating its ability to disrupt osmotic balance in marine fish by affecting this target protein. In summary, the results of this study will further enhance our comprehension of the potential toxic effects and mechanisms of TCC on the early stages of marine fish, with a specific focus on its unique toxic effects in osmoregulation.

Oral Safety Evaluation of the Aqueous Fruit Extract of Annona Muricata in Wistar Rats

Annona muricata fruit (i.e. Soursoup or custard apple a tropical plant species known for its edible fruit)has been attributed with numerous medicinal benefits (including antimicrobial, anti-inflammatory, anti-protozoan, antioxidant, anxiolytic, anti-stress, anti-ulcerogenic, wound healing, hepato-protective, anti-icteric, hepatoprotective, anticancer and antihyperglycemic activities) as well as some toxicological effects (neurotoxicity and neurodegeneration). Despite its ancestral use and wide applications in human health, scientific information on its oral safety remains scanty. In this study, the oral safety of 100 mg/kd/day, 200 mg/kg/day and 400 mg/kg/day of the aqueous fruit extract of Annona Muricata (AFAM) was evaluated in young adult, male and female white albino Wistar rats using standard acute and 42-days sub-chronic oral toxicity testing guidelines on anthropometric, biochemical, hematological and histo-pathological endpoints. In addition, preliminary qualitative and quantitative analyses of AFAM were undertaken using standard procedures. Results of the study showed that the estimated LD50 value for the acute oral toxicity study of AFAM calculated to be greater than 5 g/kg body weight/oral route although the testing was associated with transient but reversible behavioral toxicities. For its sub-chronic oral toxicity testing, AFAM treatment resulted in profound %weight gain, decreases in the serum triglycerides and very low density lipoprotein cholesterol and liver enzymes. Similarly, prolonged oral AFAM treatments caused significant decrease and increase in the differential neutrophils and platelet counts, respectively while the hist-opathological features of hepatic steatos is and renal tubule collapse in the AFAM-treated livers and kidneys, respectively suggested possible increased intrahepatic lipids biosynthesis and nephrotoxicity. The preliminary phytochemical analyses of AFAM showed the presence and relative amount of flavonoids, alkaloids, tannin, glycosides, saponin, and reducing sugars while the Gas Chromatography-Mass spectroscopy showed the relative abundance of thirteen compounds. Composite analyses conducted on the extract showed the presence of carbohydrate (64.65%), protein (2.14%), moisture content (8.07%), ash value (6.73%), lipid (14.22%) and fiber (4.19%). Overall, the study suggested that the prolonged AFAM oral treatments could predispose to the development of fatty liver disease from de novo intrahepatic biosynthesis of triglycerides, nephrotoxicity and hematotoxicity. In conclusion, the results of this study showed that prolonged consumption of AFAM should be with a great caution as it could pose serious health concerns.

Evaluation of the antidiabetic activity and toxicological properties of Jaundea pinnata (P. Beauv.)

BackgroundJaundea pinnata (JP), is a new plant native to North Africa, East Africa, Southern Nigeria, and Guinea. It is traditionally used as an anti-rheumatic, anti-inflammatory, antibacterial and antidiabetic herb, thus making it a potential source for modern antidiabetic medications. ObjectiveThis investigation aimed to assess the toxicological profile and the antidiabetic potentials of the methanol extract of the Jaundea pinnata (JP) leaf in mice and rats. MethodsPreliminary phytochemical screening of JP leaf was conducted using standard procedures. The plant extract's acute and sub-chronic toxicity was evaluated using a modified OECD recommendation. In normal and streptozotocin-induced diabetic rats, its anti-diabetic action was tested over 28 days at doses of 100 mg/kg, 200 mg/kg, and 400 mg/kg, with glibenclamide (10 mg/kg) and distilled water serving as the positive and negative controls, respectively. Relevant organs' histopathological analysis was also assessed. ResultsAccording to the preliminary phytochemical analysis of the leaf extract, JP contains flavonoids, saponins, alkaloids, tannins, terpenoids, deoxy-sugars, and anthraquinones. The extract demonstrated an LD50 of above 2000 mg/kg in mice. On days 14, 21, and 28 in streptozotocin-induced diabetic rats, the extract showed a dose-dependent antidiabetic effect; nevertheless, the activity of the extract at 400 mg/kg on day 28 was barely comparable to that of glibenclamide (10 mg/kg). At 250 mg/kg, 500 mg/kg, and 1500 mg/kg, respectively, the extract was appreciably non-toxic to the liver and kidney, while inducing a significant change in haematological indicators. ConclusionThe study found that JP presented antihyperglycemic properties and was relatively safe at normal dosages (< 2000 mg/kg).

Biosafety and chemical solubility studies of multiscale crystal-reinforced lithium disilicate glass-ceramics.

Lithium disilicate (Li2 Si2 O5 ) glass-ceramics are currently a more widely used all-ceramic restorative material due to their good mechanical properties and excellent aesthetic properties. However, they have a series of problems such as high brittleness and low fracture toughness, which has become the main bottleneck restricting its development. Therefore, in order to compensate for these shortcomings, we propose to prepare a reinforced glass-ceramics with better mechanical properties and to test the biosafety and chemical solubility of the material. Li2 Si2 O5 whiskers were synthesized by a one-step hydrothermal method, and multi-scale crystal-enhanced Li2 Si2 O5 glass-ceramics were prepared by reaction sintering. The biosafety of multi-scale crystal-reinforced Li2 Si2 O5 glass-ceramics was investigated by in vitro cytotoxicity test, rabbit pyrogen test, mice bone marrow micronucleus test, skin sensitization test, sub-chronic systemic toxicity test, and chronic systemic toxicity test. Additionally, the chemical solubility of multi-scale crystal-reinforced Li2 Si2 O5 glass-ceramics was investigated. The test results showed that the material was non-cytotoxic, non-thermogenic, non-mutagenic, non-sensitizing, and non-systemic. The chemical solubility, determined to be 377 ± 245 μg/cm2 , complied with the ISO 6872 standard for the maximum solubility of ceramic materials. Multi-scale crystal-reinforced Li2 Si2 O5 glass-ceramics' biosafety and chemical solubility met current normative criteria, and they can move on to mechanical property measurements (such as flexural strength test, fatigue life test, friction and wear property study, etc.) and bonding property optimization, which shows promise for future clinical applications.

Acute and Sub-Chronic Toxicity Profile of Aqueous Fruit Extract of Spondias Purpurea Linn (Anacardiaceae): In Wistar Rats

This study aimed to investigate the toxicological profile of aqueous fruit extract of Spondias purpurea (AFES) in Wistar rats. The AFES was subjected to preliminary phytochemical screening, acute and sub-chronic toxicity studies which were conducted using standard methods. Sub-chronic (90 days) toxicity studies were carried out on four (4) groups. The first group received distilled water (1 ml/kg). Groups 2, 3 and 4 received AFES (250, 500 and 1,000 mg/kg respectively). The effect of the extract on body weight, relative organ weights, haematological and biochemical (kidney function, liver function and lipid profile) parameters as well as histopathological studies of the organs (liver, kidney, lungs, brain, heart, stomach and testes) was investigated. Preliminary phytochemical screening revealed the presence of alkaloids, cardiac glycosides, flavonoids, saponins, sterols, triterpenes, steroids and tannins. The oral median lethal dose (LD50) was greater than 5,000 mg/kg. Sub-chronic administration of AFES did not produce significant (p &gt; 0.05) changes in body weights, relative organ weights, haematological and biochemical parameters of treated rats. However, there was a significant (p &lt; 0.05) increase in urea at 250 mg/kg, a decrease in sodium ion at 500 and 1,000 mg/kg in a dose-dependent manner and decrease in chloride ion as well as decrease in triglycerides at 1,000 mg/kg treatment groups. Histopathology investigation revealed some levels of damage to the organs; liver, kidney, testis, lung and heart of the extract treated animals. These findings suggest that aqueous fruit extract of S. purpurea is relatively safe on acute administration. However, sub-chronic administration could result in mild toxic effects.

Effects of nano-cerium dioxide on intestinal microflora in rats by oral subchronic exposure.

To investigate intestinal toxicity in rats and the effects of Nano-cerium dioxide on intestinal flora in rats after oral sub-chronic exposure. Forty healthy male SD rats were randomly divided into four groups: a control group (deionized water) and three groups treated with different doses of Nano-ceria (e.g., 20 mg/kg, 100 mg/kg, and 500 mg/kg), with 10 rats in each group. The rats were given intragastric administrations (every other day) for 90 days. After the last intragastric administration, fresh fecal samples were collected by pressing the abdomen, and the animals were sacrificed. Jejunum, ileum and cecum tissues were retained for pathological analysis by Hematoxylin-eosin staining. The stool samples of rats were sequenced by the Illumina NovaSeq sequencing platform, and the sequencing results were further analyzed by QIIME2 software. The histopathology results show that compared with the control group, in the middle- and high-dose groups, epithelial tissue was shed, lamina propria glandular structures were damaged or disappeared, and large numbers of inflammatory cells were distributed in the mucosa. The intestinal flora results show that there were no significant differences in the α-/β-diversities in each Nano-ceria-treated group compared with the control group (P>0.05). Compared with the control group, the intestinal pathogenic bacteria, Mucispirillum and Streptococcus increased significantly after Nano-cerium dioxide ingestion, while Weissella decreased. The abundances of Akkermansia in all Nano-ceria-treated groups were higher than those in the control group, but the abundances decreased with increasing dose. MetagenomesSeq analysis show that, compared with the control group, the abundances of S24-7, Lactobacillus and Clostridiales in all experimental groups significantly decreased. The sub-chronic toxicity of Nano-cerium dioxide to rats can affect the structure and abundance of intestinal microflora, long-term exposure to high doses (>100 mg/kg) causes enteritis, but there was no significant difference in the diversity of gut microbiota. Therefore, we infer that the enteritis in rats may be associated with the relative ratios of the pathogenic bacteria and intestinal probiotics, and increased of the intestinal pathogenic bacteria can disrupted intestinal homeostasis.

ACUTE AND SUB-CHRONIC TOXICITY ASSESSMENT OF PYRIDOSTIGMINE BROMIDE 90 MG EXTENDED- ELEASE TABLETS IN EXPERIMENTAL ANIMALS

Purpose: This study aimed to assess the acute and subchronic toxicity of Pyridostigmine bromide 90 mg extended-release tablets in experimental animals. Subjects and methods: Pyridostigmine bromide 90 mg extended-release tablets were prepared complying in-house specifications. Acute oral toxicity (LD50) was determined in white mice using the Litchfield-Wilcoxon method. Subchronic toxicity was assessed in rabbits following OECD guidelines. Results: The acute toxicity study revealed an LD50 of 17.9 mg/kg when Pyridostigmine bromide 90 mg extended-release tablets were orally administered to mice. In the subchronic toxicity assessment, rabbits were treated with Pyridostigmine bromide tablets for 28 days. Doses of 7.2 mg/kg/day and 21.6 mg/kg/day showed no significant alterations in hematological parameters (red blood cells, hemoglobin, hematocrit, mean corpuscular volume, white blood cells, platelets), blood biochemical parameters (AST, ALT, total protein, creatinine, urea), and did not cause damage to the histopathology of the liver, spleen, and kidneys in rabbits. However, the dose of 21.6 mg/kg/day resulted in a statistically significant difference in rabbit weight gain compared to those administered a dose of 7.2 mg/kg/day and those in the control group.

Toxicological study of “Mendim me zõn”: A traditional tea made with Solanum aethiopicum Shum berries

Mendim me zõn is a traditional tea made with Solanum aethiopicum Shum berries; a plant belonging to the Solanaceae family. It is a Bantu beverage consumed for its revitalizing properties. Powdered berries of this plant are now distributed and consumed in many countries around the world. Bioactive substances such as glycoalkaloids, galactosamine and fucose, among others which are potentially toxic, can be found in Mendim me zõn. However, very few studies have looked in the safety of this increasingly popular beverage. The present work aimed therefore to carry out a toxicological study of this drink, focusing on acute and sub-chronic toxicity. To do this, 50 g of powdered berries obtained in the locality of Ngaoundere, Adamawa Region of Cameroon. They were infused in 10 L of boiling water for 10 min. The extract was used to study acute and subchronic toxicity using rats as animal model, during 14 and 42 days respectively. Clinical, biochemical and histopathological analyses were carried out on male and female rats given progressive doses of the extract. The results showed that there was no death at any dose. Signs of toxicity including vomiting, diarrhoea, abdominal pain, self-mutilation, apathy, agitation, continuous grooming, drowsiness, stupor, tremors, hair erection, difficulty in breathing and anorexia were not observed in rats. Repeated intake of Mendim me zõn increased appetite and proved to be laxative. Microscopic sections of the liver and kidney showed normal architecture. Blood parameters remained within normal ranges. However, at high doses (1000 mg/kg/day), extract increased erythrocytes while white blood cells decreased in males. In female platelets and mean corpuscular volume increased. With the exception of bilirubin, consumption of extract statistically reduced the parameters linked to renal and hepatic function. These results might suggest that Mendim me zõn is safe beverage.

EVALUATION OF ACUTE AND SEMI-CHRONIC TOXICITY OF THE REMEDY “BO PHE DINH SUYEN QY” ON EXPERIMENTAL ANIMALS

Purpose: To evaluate the acute toxicity and semi-chronic toxicity of the remedy “Bo phe dinh suyen QY” on experimental animals. Subjects and methods: The remedy “Bo phe dinh suyen QY” meeting basic standards was used to evaluate for acute toxicity in Swiss white mice, and for semi-chronic toxicity in Wistar white rats. Acute toxicity was assessed according to the Litchfield-Wilcoxon method. Sub-chronic toxicity was assessed according to the guidance of the Ministry of Health of Vietnam. Results: The remedy “Bo phe dinh suyen QY” at a dose of 375 g/kg mouse body weight/24 hours did not cause acute toxicity or death of experimental mice. For 28 consecutive days, rats administered “Bo phe dinh suyen QY” at doses of 18.2 g/kg/day and 54.6 g/kg/day without affecting their general condition and hematological indices. Conclusions: The remedy “Bo phe dinh suyen QY” was safe in the assessment of acute toxicity (on mice) and semi-chronic toxicity (on rats).

Analysis of Requirements for Inhalation Toxicity Studies of Chemicals

SCIENTIFIC RELEVANCE. Studies of the inhalation administration of chemicals are associated with challenges in designing experiments. The parameters to be selected include the experimental animal species, the inhalation chamber, and the mode of inhalation (dynamic or static).AIM. This study aimed to analyse the practical application of regulatory requirements to non-clinical studies of the inhalation toxicity of chemicals.DISCUSSION. This review compares international and Russian standards for studying the inhalation toxicity of chemicals, including GOST 32542-2013, GOST 326432020, GOST 32636-2020, GOST 32383-2013, and GOST 2646-2014. The improvement of the legal and regulatory framework correlates with adopting the Good Laboratory Practice and the risk-based approach to categorising test substances into hazard classes. Hazard classes are determined in rodents without dose extrapolation to humans. The authors present the differences between the main guidelines on inhalation exposure in rodent studies of acute, subacute, subchronic, and chronic toxicity. The article describes current approaches to assessing the inhalation toxicity of chemicals, which allow researchers to replace animal studies with in vitro tests.CONCLUSIONS. According to the current regulatory standards, inhalation toxicity is studied in rats/mice, which have anatomical differences from humans. As an alternative to animal studies, researchers are developing and validating in vitro methods, which yet require regulatory review and approval.

Investigation of acute and sub-chronic oral toxicity and effects of Allium ascalonicum L. extract on Triton WR1339-induced hyperlipidemia on Swiss albino mice

IntroductionMetabolic disorders are increasingly rapidly becoming a burden on global healthcare systems. Among these, hyperlipidemia frequently leads to cardiovascular diseases such as myocardial infarction and cerebral stroke, posing significant life-threatening risks. The pharmacological drugs, which are often costly and may cause a range of adverse side effects detrimental to health. Allium ascalonicum L., a member of the Alliaceae family, a widely used species of the Allium genus, is commonly employed as a spice and food ingredient in Asia. Allium ascalonicum (AS) is rich in triterpenoids, phenolic acids, flavonoids, thiosulfates, and anthocyanins, whose therapeutic effects have been documented. In Chinese, Allium ascalonicum L. is known as 葱 (Cōng), which is used for various types of onions and similar plants in Chinese, including shallots. Ancient Chinese medicine documented that AS has a sweet and mildly spicy taste with a slight astringency and is warm in nature. In traditional Chinese medicine, AS is prepared as infusions, juices, and poultices to harness their health benefits, and commonly used in cooking for their nutritional and medicinal purposes. Modern medicine has discovered that this type of bulb possesses antibacterial and anti-inflammatory properties, acts as a diuretic, treats diarrhea, reduces blood sugar levels, lowers blood pressure, reduces cholesterol, and among its other roles. Currently, there is a lack of in vivo study on the cholesterol-regulating potential of AS. MethodsFor our experiments, 24 and 32 Swiss albino mice were employed to investigate the acute and sub-chronic oral toxicity according to the Guidelines for preclinical and clinical trials of Traditional Medicine and Herbal Medicines by the Vietnam Ministry of Health and OECD. To investigate the effects on hyperlipidemia, 40 mice of both sexes were divided into five groups (n = 6–8 each). Tyloxapol (Triton WR1339, 400 mg/kg, i.p.) was used to induce hyperlipidemia. Atorvastatin (60 mg/kg, p.o.) was used as the standard, and AS extract was conducted at the doses of 500 mg/kg and 1000 mg/kg. Hyperlipidemic activity was assessed using a variety of parameters. ResultsAllium ascalonicum (AS) extract showed no acute toxicity and sub-chronic toxicity. The AS showed a significant decrease in total cholesterol, triglycerides and LDL-c but showed no changes in HDL-c. ConclusionOur results indicated that AS is safe to use for a longtime and has a potential as an antihyperlipidemic agent, possibly due to its ability to inhibit the elevation of total cholesterol, triglycerides and LDL-c. The AS is rich in phytochemicals like triterpenoids, flavonoids and saponins, which may contribute to its hypolipidemic effect.