Organ Weight Changes Research Articles

An acute and sub-acute toxicological assessment of Reissantia indica plant extract in male Sprague-Dawley rats: Hematological, serum biochemical and histopathology

The traditional use of medicinal plants in Sub-Saharan health management lacks thorough toxicological evaluations, particularly concerning lethal dose levels. This study aims to assess the acute and sub-acute toxicity of Reissantia indica whole-plant extract (RIE) in male Sprague-Dawley rats, with a focal point on delineating its safety profile while exploring potential therapeutic applications.RIE, obtained through precise cold maceration in 70 % ethanol, underwent rigorous analysis, revealing diverse secondary metabolites, including alkaloids, flavonoids, terpenoids, and glycosides. Renowned for antioxidant, anti-inflammatory, and anticancer properties, these compounds enhance RIE's pharmacological potential.In the acute toxicity study, RIE was orally administered at 500 and 5000 mg/kg. Sub-acute toxicity involved oral administration of the extract at various doses (5, 50 and 500 mg/kg) over 28 days, with comprehensive assessments, including hematological, biochemical, and histopathological evaluations.Results from the acute toxicity showed no mortality, suggesting a median lethal dose (LD50) exceeding 5000 mg/kg and indicating a substantial margin of safety. Sub-acute toxicity investigations, spanning 28 days revealed no significant changes in body and organ weights, hematological and biochemical parameters, or histopathological signs compared to the control group. Histological examination of kidney, liver, heart, and lung sections from treated animals showed no signs of degeneration.This study, to our knowledge, pioneers a comprehensive investigation into the toxicity profile of Reissantia indica's whole-plant ethanolic extract, addressing a significant gap in existing literature on medicinal plant safety in the Sub-Saharan region.

Sub-chronic toxicity determination of powdered Tenebrio molitor larvae as a novel food source

Tenebrio molitor larvae are the first insect species to be given a favorable assessment by the European Food Safety Authority (EFSA) as a novel food source, enabling consumption of whole insect larvae or larvae that have been powdered and processed into a variety of food products. Pressure from economic hardships and increase in population growth have paved a way for the realization of an alternative food source in Zimbabwe. This study focused on determining the potential toxicity of Tenebrio molitor larvae powder as an alternative food source for humans. To determine the sub-chronic toxicity of Tenebrio molitor, the powder was administered daily by oral gavage to Sprague-Dawley rats at dose levels of 0, 300, 1000 and 3000 mg/kg for 70 days. A toxicological assessment which included mortality, appearance of clinical symptoms, food consumption, organ and body weight changes were performed. There were no treatment-related mortalities, clinical signs, changes in food consumption, body and organ weights observed during the treatment period. The study's findings suggest Tenebrio molitor larvae to be a good alternative as it did not appear to affect the rats' normal physiological and metabolic processes hence can be considered safe for human consumption. However, further studies on hematological, histological and biochemical markers may be necessary for confirmation of these current results.

Investigations on the genotoxic potential of styrene in Fischer 344 rats using multiple endpoints.

Genotoxicity of styrene monomer was evaluated in male Fischer 344 rats using the alkaline comet assay for DNA damage, micronucleus assay for cytogenetic damage and the Pig-a assay for gene mutations. In a dose range finding (DRF) study, styrene was administered by oral gavage in corn oil for 28 consecutive days at 0, 100, 500, and 1000 mg/kg/day. The bioavailability of styrene was confirmed in the DRF by measuring its plasma levels at approximately 7- or 15-min following dosing. The 1000 mg/kg/day group exceeded the maximum tolerated dose based on body weight and organ weight changes and signs of central nervous system depression. Based on these findings, doses of 0, 100, 250, and 500 mg/kg/day (for 28 or 29 days) were selected for the genotoxicity assays. Animals were sacrificed 3-4 h after treatment on Day 28 or 29 for assessing various genotoxicity endpoints. Pig-a mutant frequencies and micronucleus frequencies were determined in peripheral blood erythrocytes. The comet assay was conducted in the glandular stomach, duodenum, liver, lung, and kidney. These studies were conducted in accordance with the relevant OECD test guidelines. Oral administration of styrene did not lead to genotoxicity in any of the investigated endpoints. The adequacy of the experimental conditions was assured by including animals treated by oral gavage with the positive control chemicals ethyl nitrosourea and ethyl methane sulfonate. Results from these studies supplement to the growing body of evidence suggesting the lack of in vivo genotoxic potential for styrene.

The mechanisms underlying alcohol-induced decreased splenic size: A network meta-analysis study.

Organ weight change is widely accepted as a measure of toxicologic pathology. We and other groups have shown that excessive alcohol exposure leads to decreased spleen weight in rodents. This study explores the mechanisms underlying alcohol-induced splenic injury through a network meta-analysis. QIAGEN Ingenuity Pathway Analysis (IPA) and Mammalian Phenotype (MP) Ontology were used to identify alcohol-related molecules associated with the small spleen phenotype. Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) and IPA bioinformatics tools were then used to analyze the biologic processes and enriched signaling pathways engaging these molecules. In addition, the "downstream effects analysis" algorithm was used to quantify alcohol's effects. IPA identified 623 molecules affected by alcohol and a Venn diagram revealed that 26 of these molecules overlapped with those associated with the MP Ontology of small spleen. The 26 molecules are TGFB1, CASP8, MTOR, ESR1, CXCR4, CAMK4, NFKBIA, DRD2, BCL2, FAS, PEBP1, TRAF2, ATM, IGHM, EDNRB, MDM2, GLRA1, PRF1, TLR7, IFNG, ALOX5, FOXO1, IL15, APOE, IKBKG, and RORA. Some of the 26 molecules were also associated with the MP Ontology of abnormal white pulp and red pulp morphology of the spleen, abnormal splenic cell ratio, decreased splenocyte number, abnormal spleen physiology, increased splenocyte apoptosis, and reduced splenocyte proliferation. STRING and IPA "Core Analysis" showed that these molecules were mainly involved in pathways related to cell apoptosis, proliferation, migration, and immune responses. IPA's "Molecular Activity Predictor" tool showed that concurrent effects of activation and inhibition of these molecules led to decreased spleen size by modulating apoptosis, proliferation, and migration of splenocytes. Our network meta-analysis revealed that excessive alcohol exposure can damage the spleen through a variety of molecular mechanisms, thereby affecting immune function and human health. We found that alcohol-mediated splenic atrophy is largely mediated by increased apoptosis signaling, migration of cells, and inhibition of splenocyte proliferation.

Effects of gamal leaf extract (Gliricidia sepium Jacq.) on body weight and internal organ performance of mice (Mus musculus L.)

Gamal leaves (Gliricidia sepium Jacq.) contain toxic compounds that have the potential to act as vegetable rodenticides, such as alkaloids, tannins, dicumerol, and hydrogen cyanic acid (HCN). Coumarin compounds can bind vitamin K, disrupt the circulatory system, and damage liver cells. The compounds dicumerol and warfarin are used as rat poison because of their anticoagulant properties, which prevent blood clots in the heart. Alkaloid and tannin compounds can damage liver cells through liver necrosis and bleeding in the digestive system. This study aims to determine the effect of Gamal leaf extract on body weight and internal organ performance in mice (Mus musculus L.). The experimental research method is based on a Completely Randomized Design (CRD) pattern. A total of 16 mice weighing 20-22 grams consisting of 4 treatments, namely A0 (water control), A1 (extract 10 mg/g BW), A2 (extract 15 mg/g BW) and A3 (extract 20 mg/g BW). The extract was given orally for 14 days, and body weight was measured every 3 days to obtain data on changes in body weight (PBB). At the end of the treatment, the mice were sacrificed using chloroform and then dissected to observe the performance of the liver, kidney and heart organs in color appearance and organ weight. Quantitative data were analyzed using one-way ANOVA and further DMRT test α=0.05. The research showed that mice's mean change in body weight (PBB) decreased significantly as the dose of Gamal leaf extract increased. The percentage of liver organ weight (BOH), kidney organ weight (BOG), and heart organ weight (BOJ) decreased as the dose of Gamal leaf extract increased. The research results concluded that Gamal leaf extract caused color changes in the internal organs of mice and significantly affected the percentage change in body weight (PBB) and internal organ weight (BOD) of mice along with increasing doses of Gamal leaf extract.

Toxicological evaluation of ginsenoside-rich cell culture biomass of Panax japonicus produced in a large-scale bioreactor system

An environmentally friendly, resource-efficient substitute for the production of vegetative biomass and the bioactive components of medicinal plants is provided by novel biotechnology-based suspension cells cultured in bioreactors. Despite the cultured biomass is produced in the controlled conditions, it requires detailed chemical analysis and toxicological evaluation before being approved for human consumption. This study reports the results of biochemical, toxicological, and elemental composition analysis of the suspension cell culture of a medicinal species Panax japonicus (T. Nees) C.A. Mey. cultivated in a large-scale bioreactor system. For the scaling up the process of cultivation, the suspension culture was grown sequentially in a series of laboratory (20-L), pilot (75-L), and industrial (630-L) bioreactors. The biomass productivity of cell culture was comparable or higher than of wild plants or in vitro cultured hairy roots reported in the literature. The resulting cell biomass was analyzed for ginsenoside contents using ultra-high-performance liquid chromatography-electrospray ionization-mass spectrometry (UHPLC–ESI–MS). For the elemental composition of cells, inductively coupled argon plasma mass spectrometry (ICP–MS) was used. In addition, the biomass was evaluated for acute toxicity and skin sensibilization effects on in vivo animal models. The analyses confirmed that suspension cells cultured in bioreactors accumulate a broad spectrum of ginsenosides and their malonylated derivatives at a total concentration of 7.54 % (w/w) based on dry weight and contain essential macro- (K, Ca, Mg, Na) and micro- (Zn, Mn, Fe, B, Al, Cu) elements in dietary-safe concentrations. Acute toxicity tests using the administered doses of 2000 and 5000 mg dry biomass per kg animal weight resulted in no changes in animal organ weights and no or minor changes in hematological and biochemical parameters of blood. No skin irritation or sensitizing effects were observed. Therefore, bioreactor cultivation of P. japonicus cell suspension is highly productive and provides a ginsenoside-rich and non-toxic cell biomass which is a potentially valuable component of functional foods, food additives, and natural health products.

Acute and sub-chronic toxicological profile of Musa cavendishii (Musaceae) Lamb peel extracts in New Zealand rabbit

Musa cavendishii (banana) peel is a waste product of human daily consumable fruit used traditionally in the management of ulceration and scar. This study evaluates the toxic effects of its peel in New-Zealand rabbits after topical administration. Acute and sub-chronic toxicity study of the peel extracts were carried out in 116 randomly selected rabbits using methods described by Organization of Economic Corporation and Development (OECD) 402 (2017) and 410 (2018), respectively. Changes in body and relative organ weight, hematology, serum liver and renal enzyme were investigated. Vital body organs were also processed for histopathology to observe any alteration from normal architecture. No mortality was observed during the acute toxicity with no changes in body and relative organ weight of the organs tested when compared with the control. After 28 days of topical administration of the extracts, even at the highest dose (1500 mg/kg) of the aqueous extract, the histoarchitexture of the skin’s epidermis and dermis, and other biochemical markers of the rabbits were normal. However, mild adipocyte infiltration, glandular hyperplasia and distortions in the renal parameters were seen in the hexane, ethylacetate and methanol extracts portions at the highest dose (1500 mg/kg) after 28 days topical administration. The study indicates that M. cavendishii peel is not capable of causing toxic effects in rabbits. However, it’s prolong use at higher doses may result in some harmful effects.

Anti-Atopic Dermatitis Effect of TPS240, a Novel Therapeutic Peptide, via Suppression of NF-κB and STAT3 Activation

Atopic dermatitis (AD) is a relapsing skin disease with persistent inflammation as a causal factor for symptoms and disease progression. Current therapies provide only temporary relief and require long-term usage accompanied by side effects due to persistent relapses. A short peptide, TPS240, has been tested for its potential to subside AD. In this study, we confirmed the anti-atopic effect of TPS240 in vivo and in vitro using a DNCB-induced AD mouse model and TNF-α/IFN-γ-stimulated HaCaT cells. In the AD mouse model, topical treatment with TPS240 diminished AD-like skin lesions and symptoms such as epidermal thickening and mast cell infiltration induced by DNCB, similar to the existing treatment, dexamethasone (Dex). Furthermore, skin atrophy, weight loss, and abnormal organ weight changes observed in the Dex-treated group were not detected in the TPS240-treated group. In TNF-α/IFN-γ-stimulated HaCaT cells, TPS240 reduced the expression of the inflammatory chemokines CCL17 and CCL22 and the pruritic cytokines TSLP and IL-31 by inhibiting NF-κB and STAT3 activation. These results suggest that TPS240 has an anti-atopic effect through immunomodulation of AD-specific cytokines and chemokines and can be used as a candidate drug for the prevention and treatment of AD that can solve the safety problems of existing treatments.

Modulatory effects of extract of Heinsia crinita against fructose/streptozotocin-induced oxidative stress in diabetic rat models

Oxidative stress plays a crucial role in the development of type 2 diabetes and the associated microvascular and cardiovascular complications. In the study, we have investigated the effects of Heinsia crinita (H. crinita) extracts on lipid peroxidation and oxidative stress responses using diabetic rats. Type 2 diabetes was induced with 10 % fructose/40 mg/kg body weight streptozotocin (STZ). H. crinita extract was administered at 200 and 400 mg/kg body weight twice daily for 21 days, in addition to metformin (MET: 500 mg/kg body weight) control. Molecular docking analysis was performed to determine the binding affinity of H. crinita extracts to the DNA binding domains of peroxisome proliferator-activated receptor (Ppar) and retinoid x receptor (Rxr) protein crystal structures, showing different binding affinities for putative active compounds from the plant. Fasting blood glucose (FBG), body and organ weight changes were determined showing that H. crinita extract induced an anti-hyperglycemic effect in the treated animals, with changes (either decrease or increase) in liver and kidney weights. A decrease in mRNA expression of peroxisome proliferator-activated receptors (ppar), sterol regulatory element-binding protein 1 (srebp-1c), liver x-receptor (lxr), retinoid x receptors (rxr), cytochrome p45041 (cyp4a1) and acyl-CoA oxidase (acox1) in diabetic animals were observed, compared to the control. A dose-specific decrease or increase in antioxidant enzymes (superoxide dismutase: SOD, catalase: CAT, reduced glutathione: GSH, glutathione peroxidase: GPx) transcripts and activity levels were also observed. We also observed exposure-specific decrease or increase of malondialdehyde (MDA) levels. Our data suggested that H. crinita extract possesses protective effects against diabetes-induced oxidative stress. These effects might be attributed to their binding and activation of nuclear receptors, indicating their cellular mode of action that is comparable to MET.

Critical assessment of the endocrine potential of Linalool and Linalyl acetate: proactive testing strategy assessing estrogenic and androgenic activity of Lavender oil main components.

The acyclic linear monoterpenes Linalool (Lin) and Linalyl acetate (LinAc) occur in nature as major constituents of various essential oils such as lavender oils. A potential endocrine activity of these compounds was discussed in literature including premature thelarche and prepubertal gynecomastia due to lavender product use. This study aims to follow-up on these critical findings reported by testing Lin and LinAc in several studies in line with current guidance and regulatory framework. No relevant anti-/ER and AR-mediated activity was observed in recombinant yeast cell-based screening tests and guideline reporter gene in vitro assays in mammalian cells. Findings in the screening test suggested an anti-androgenic activity, which could not be confirmed in the respective mammalian cell guideline assay. Mechanistic guideline in vivo studies (Uterotrophic and Hershberger assays) with Lin did not show significant dose related changes in estrogen or androgen sensitive organ weights and a guideline reproductive toxicity screening study did not reveal evident effects on sex steroid hormone sensitive organ weights, associated histopathological findings and altered sperm parameters. Estrous cycling and mating/fertility indices were not affected and no evident Lin-related steroid hormone dependent effects were found in the offspring. Overall, the initial concerns from literature were not confirmed. Findings in the yeast screening test were aberrant from follow-up guideline in vitro and in vivo studies, which underlines the need to apply careful interpretation of single in vitro test results to support a respective line of evidence and to establish a biologically plausible link to an adverse outcome.

Evaluation of Subacute Toxicity of Flavonoid from Phaleria Macrocarpa Fruit Extract in Mice

Background: Traditional medicine and supplements including Phaleria macrocarpa or Mahkota Dewa are extensively utilised. Rape fruit of phaleria macrocarpa contains flavonoid as a secondary metabolite that has a beneficial effect on endometriosis treatment and other illnesses. The purpose of this study was to look into the subacute toxicity of flavonoid from phaleria macrocarpa fruit extract on the brain and heart of BALB/c female mice. Method: The BALB/c female mice aged 6-8 weeks with a body weight of 20-30gr were divided into four groups and treated for 28 days. The control group received simply water, whereas the treatment group received flavonoid at doses of 500mg/kg/day, 1000mg/kg/day, and 2000mg/kg/day. At the end of the experiment, the mice were sacrificed and important organs were extracted and assessed. The organ weight was assessed using macroscopy, and the brain and heart were assessed using microscopy (histopathological abnormalities). Result: The BALB/c female mice treated with oral administration flavonoid from phaleria macrocarpa fruit extract showed no clinical abnormalities and no change in organ weight compared to the control groups (p>0,5) in subacute toxicity experiments. There were no abnormalities in the organs of the treated mice at a dose of 500mg/kg/day, according to histopatological testing. Conclusion: As a result, flavonoid had no harmful impact in brain and heart mice after subacute therapy up to a level of 500 mg/kg/ body weight. High doses of 1000 and 2000 mg/kg/day are not recommended for long-term use.

Acute and 28-Day Repeated-Dose Oral Toxicity of the Herbal Formula Guixiong Yimu San in Mice and Sprague-Dawley Rats.

To evaluate the acute and chronic 28-day repeated-dose oral toxicity of Guixiong Yimu San (GYS) in mice and rats, high-performance liquid chromatography (HPLC) was used to determine the stachydrine hydrochloride in GYS as the quality control. In the acute toxicity trial, the mice were administered orally at a dose rate of 30.0 g GYS/kg body weight (BW) three times a day. The general behavior, side effects, and death rate were noticed for 14 days following treatment. In the subacute toxicity trial, the rats were administered orally at a dose rates of30.0, 15.0, and 7.5 g GYS/kg BW once a day for 28 days. The rats were monitored every day for clinical signs and deaths; changes in body weight and relative organ weights (ROW) were recorded every week, hematological, biochemical, and pathological parameters were also examined at the end of treatment. The results showed that the level of stachydrine hydrochloride in GYS was 2.272 mg/g. In the acute toxicity trial, the maximum-tolerated dose of GYS was more than 90.0 g/kg BW, and no adverse effects or mortalities were noticed during the 14 days in the mice. At the given dose, there were no death or toxicity signs all through the 28-day subacute toxicity trial.The oral administration of GYS at a dose rate of 30.0 g/kg/day BW had no substantial effects on BW, ROW, blood hematology, gross pathology, histopathology, and biochemistry (except glucose), so 30.0 g/kg BW/day was determined as the no-observed-adverse-effect dosage.

Safety, immunogenicity and efficacy of Relcovax®, a dual receptor binding domain (RBD) and nucleocapsid (N) subunit protein vaccine candidate against SARS-CoV-2 virus

SARS-CoV-2, severe acute respiratory syndrome coronavirus-2, causes coronavirus disease- 2019 (COVID-19). Mostly, COVID-19 causes respiratory symptoms that can resemble those of a cold, the flu, or pneumonia. COVID-19 may harm more than just lungs and respiratory systems. It may also have an impact on other parts of the body and debilitating effects on humans, necessitating the development of vaccines at an unprecedented rate in order to protect humans from infections. In response to SARS-CoV-2 infection, mRNA, viral vector-based carrier and inactivated virus-based vaccines, as well as subunit vaccines, have recently been developed. We developed Relcovax®, a dual antigen (Receptor binding domain (RBD) and Nucleocapsid (N) proteins) subunit protein vaccine candidate. Preliminary mouse preclinical studies revealed that Relcovax® stimulates cell-mediated immunity and provides broader protection against two SARS-CoV-2 variants, including the delta strain. Before conducting human studies, detailed preclinical safety assessments are required, so Relcovax® was tested for safety, and immunogenicity in 28-day repeated dose toxicity studies in rats and rabbits. In the toxicity studies, there were no mortality or morbidity, abnormal clinical signs, abnormalities in a battery of neurobehavioral observations, abnormalities in detailed clinical and ophthalmological examinations, or changes in body weights or feed consumption. In any of the studies, no abnormal changes in organ weights, haematology, clinical chemistry, urinalysis parameters, or pathological findings were observed. Immunogenicity tests on rats and rabbits revealed 100 % seroconversion. Relcovax® was therefore found to be safe in animals, with a No Observed Adverse Effect Level (NOAEL) of 20 µg/protein in rats and rabbits. In efficacy studies, Relcovax® immunised hamsters demonstrated dose-dependent protection against SARS-CoV-2 infection, with a high dose (20 µg/protein) being the most protective, while in cynomolgus macaque monkey study, lowest dose 5 µg/protein had the highest efficacy. In conclusion, Relcovax® was found to be safe, immunogenic, and efficacious in in vivo studies.

Efficacy of Trigonella foenum-graecum Linné in an animal model of particulate matter-induced asthma exacerbation

Ethnopharmacological relevanceThe seeds of Trigonella foenum-graecum Linné (TFG) has traditionally been used in Central Asia to relieve inflammation. Aim of the studyThis study investigated the efficacy of TFG in a bronchial cell model and an animal model of asthma exacerbation caused by PM. MethodsBEAS-2B bronchial epithelial cells were simultaneously treated with tumor necrosis factor-α/interleukin (IL)-4 and PM, and the expression of inflammatory cytokines, DNA damage, and autophagy mechanisms were analyzed. In an animal model of asthma exacerbation, we analyzed changes in organ weight, distribution of inflammatory cytokines and inflammatory cells in the bronchoalveolar lavage fluid, and intra-tissue mucus production. ResultsIn the cell model, TFG suppressed the expression of the inflammatory cytokines IL-6, granulocyte-macrophage colony stimulating factor, monocyte chemoattractant protein-1, and IL-8; reactive oxygen species levels and DNA damage; and the phosphorylation of ERK, JNK, P38, AKT, and mTOR. In the animal model, TFG significantly reduced weight gain of the liver, lung, and spleen; IgE, IL-6, and IFN-γ levels; and bronchial mucus secretion and smooth muscle thickness. ConclusionTFG alleviated the PM-exacerbated inflammatory response by inhibiting the MAPK and autophagy signaling pathways; it is expected to be an effective treatment for asthma.

Toxicological Evaluation of <i>Vaisvanara churna</i> an Ayurvedic Formulation

Vaisvanara churna an ayurvedic formulation was screened for its toxicity profile. The study was carried out based on CCRAS Protocols. For the acute oral toxicity study, Group I served as vehicle control, Vaisvanara churna at a single oral dose of 6500 mg/kg b.w., was administered to 5 male and 5 female Swiss albino mice (Group II) and observed for 14 consecutive days for clinical signs, body weight and mortality. For 90 days repeated dose toxicity study eighty animals (40 male and 40 female) were distributed into four groups. Group I served as vehicle control. Groups II, III, and IV were administered with 450, 2250 and 4500 mg/kg b.w., respectively for 90 days orally. During this period, feed intake, body weight, signs of toxicity, haematological and biochemical parameters were recorded at periodical intervals. After 90 days animals were sacrificed for necropsy, vital organs were weighed and preserved for histopathological analysis. In an acute toxicity study, a single dose of Vaisvanara churna at 6500 mg/kg b.w., caused no mortality in animals, suggesting that the median lethal dose is greater than 6500 mg/kg b.w. In 90-day, repeated dose toxicity study, administration of Vaisvanara churna for 90 days, up to 4500 mg/kg b.w., caused no significant changes in body weights or organ weights of rats in the treated groups when compared with the control group. In addition, haematological and biochemical parameters were within the physiological limits. Histopathological analysis revealed no signs of degeneration. These results showed that Vaisvanara churna is apparently safe as indicated therapeutically for long-term administration for chronic ailments.

Study of Aspir-35 toxicity

Preclinical research on veterinary pharmaceuticals is crucial and required when creating new dosage forms. Preclinical research aims to ascertain the test substance's therapeutic efficacy, toxicity, future dose form, impact on the body's fundamental systems, and identification of potential side effects. The objective of this study was to assess the potential toxicity of Aspir-35 following short- and longer-term exposure in rats. The acute toxicity of the drug was studied on 30 white mice weighing 18–20.5 g and 15 white rats weighing 180–215 g. The animals were kept in a vivarium in accordance with sanitary rules and on a standard diet taken in the vivarium using compound feed. The drug “Aspir-35” was administered to rats and mice once in the morning on an empty stomach orally through a probe with a cannula in doses of 1250, 2500 and 5000 mg/kg of body weight. Animal feeding was started two hours after drug administration. The study of the toxicity of the drug “Aspir-35” during long-term subcutaneous administration was studied on rats with an initial body weight of 180–230 g divided according to the principle of analogues into two groups of 6 heads each. The animals were kept in similar conditions as in the study of the acute toxicity of the drug. The drug “Aspir-35” was administered to the rats of the experimental group subcutaneously daily for 18 days at a dose of 0.5 ml/kg of body weight. The drug “Aspir-35” did not cause the death of experimental rats and mice when administered once orally in doses of 1250, 2500 and 5000 mg/kg of body weight. On this basis, the drug “Aspir-35” can be attributed to the 4th class of danger according to the International Standard GOST 12.1.007-76, or to category 5 according to the International Global Classification of the Global Harmonized System, (GHS), since the LD50 of the drug “Aspir-35” when taken orally will exceed 5000 mg/kg of body weight. The drug “Aspir-35” in a dose of 0.5 ml/kg of body weight when administered subcutaneously for 18 days did not cause negative and harmful effects on the body of experimental rats, did not affect their growth and development, did not cause changes in the relative weight of internal organs and did not led to changes in hematological indicators in experimental animals. The analysis of the results of the conducted research indicates the relative harmlessness of the potential drug for veterinary medicine and allows us to predict that the drug “Aspir-35” can be classified as a low-risk substance, which justifies the feasibility of its further study and implementation in practice.

Toxicological Evaluation and Pain Assessment of Four Minor Cannabinoids Following 14-Day Oral Administration in Rats.

Introduction: Despite growing consumer interest and market availability, the safety of minor cannabinoids, generally present in low concentrations in Cannabis sativa L., is not well understood. Materials and Methods: Cannabichromene (CBC; 3.2, 10, 17, 22, 32, or 100 mg/kg-bw/day), cannabinol (CBN; 1, 3.2, 10, 17, 32, or 100 mg/kg-bw/day), delta-8-tetrahydrocannabinol (D8-THC; 0.32, 1, 3.2, or 10 mg/kg-bw/day), tetrahydrocannabivarin (THCV; 3.2, 10, 17, 22, 32, or 100 mg/kg-bw/day), and vehicle (medium-chain triglyceride oil) preparations were administered via oral gavage once daily for 14 days to Sprague Dawley rats. Changes in behavior, body weight, food consumption, clinical pathology, organ weights, body temperature, and thermal pain sensitivity (tail flick assay) were assessed. Select organ tissues were collected at terminal necropsy and fixed for histopathological examination. Results: No treatment-related deaths were observed throughout the study, and cannabinoids were generally well tolerated. While some significant trends in body weight differences from controls (increases and decreases) were observed, these occurred independently of food consumption. Overall, differences in serum chemistry and hematology parameters between cannabinoid groups and their respective control groups were considered to occur due to biological variation among rats. No treatment-related gross abnormalities were observed in examined organs. Significant changes in absolute and relative organ weights occurred primarily in males and were generally of negligible magnitude. There were no biologically significant histopathological observations. While pain tolerance was significantly improved in animals treated with D8-THC (3.2 and 10 mg/kg-bw/day, day 14), results across minor cannabinoids were inconsistent and warrant further study. Conclusion: Minor cannabinoids were well tolerated across 14 days of daily oral administration at the doses assessed. Modest, dose-dependent trends in relative organ weights and serum chemistry parameters warrant exploration at higher oral doses. These data will assist in dose selection for future studies investigating the long-term safety and effects of CBC, CBN, D8-THC, and THCV.

Anti-Diabetic and Anti-Adipogenic Effect of Harmine in High-Fat-Diet-Induced Diabetes in Mice.

One of the most important health issues facing the world today is obesity. It is an important independent risk factor for developing type 2 diabetes. Harmine offers various pharmacological effects, such as anti-inflammatory and anti-tumor effects. The current study aims to investigate Harmine's anti-diabetic and anti-adipogenic properties in albino mice after inducing low-grade inflammation with a high-fat diet (HFD). About forty-eight male albino mice were divided into four groups. Group 1: control mice were injected with daily saline and fed a normal chow diet of 21% protein for 5 months. Group 2: mice were treated daily with IP-injected Harmine (30 mg/kg body weight) and were fed a normal chow diet for 5 months. Group 3: mice were fed HFD to induce type 2 Diabetes Mellitus (T2DM) for 5 months. Group 4: mice were fed HFD for 14 weeks and treated with Harmine for the last 6 weeks. A figh-fat diet caused a significant increase in body and organ weight, lipid profiles, and destructive changes within the pancreas, kidney, and liver tissue. The administration of Harmine led to a remarkable improvement in the histological and ultrastructural changes induced by HFD. The findings indicate that mice cured using Harmine had lower oxidative stress, a higher total antioxidant capacity, and a reduced lipid profile compared to HFD mice. Harmine led to the hepatocytes partly restoring their ordinary configuration. Furthermore, it was noticed that the pathological incidence of damage in the structure of both the kidney and pancreas sections reduced in comparison with the diabetic group. Additional research will be required to fully understand Harmine and its preventive effects on the two forms of diabetes.

CD19 chimeric antigen receptor T cell therapy in leukemia xenograft mouse: Anti-leukemic efficacy, kinetics, and 4-week single-dose toxicity

CD19 Chimeric antigen receptor T (CAR-T) cell therapy has shown a promising response rate for relapsed/refractory B-cell malignancies. However, serious side effects such as cytokine release syndrome and immune effector cell-associated neurotoxicity syndrome arose in early case reports. Though several preclinical and clinical studies of CAR-T cell therapy have been reported, there is a lack of toxicological assessments. This study was carried out as a preclinical assessment of CD19 CAR-T cell therapy, including the anti-leukemic efficacy, kinetics in peripheral blood, and 4-week single-dose toxicity evaluation in leukemia xenograft mice. Leukemia xenograft mice model was established by injecting 1.0 × 105 cells/mouse of luciferase-labeled human B cell acute lymphoblastic leukemia (B-ALL) cell line via the tail vein, and after 3 days, 2.0 or 4.0 × 106 cells/mouse of CD19 CAR-T cells were injected intravenously. CD19 CAR-T cells showed significant anti-leukemic efficacy, showing inhibition of tumor progression in the bioluminescence-based in-vivo imaging system. In the kinetics study using qPCR, CAR-T cells peaked in peripheral blood on day 60 in males and day 30 in females. In a 4-week single-dose toxicity study, CD19 CAR-T cell injected groups showed no mortality and toxicological signs, or changes in body weight, food/water consumption, hematology, clinical chemistry, organ weights, and histopathology compared to control groups. These results suggested that 4.0 × 106 cells/mouse of CD19 CAR-T cells were effective in B-ALL xenograft mice without serious side effects, so the no-observed adverse effect level (NOAEL) was estimated to be higher than 4.0 × 106 cells/mouse, under the condition examined in the current study.

Preclinical evaluation of general toxicity and safety pharmacology of a receptor-binding domain-based COVID-19 subunit vaccine in various animal models.

The coronavirus disease 2019 pandemic has resulted in the introduction of several naïve methods of vaccine development, which have been used to prepare novel viral vectors and mRNA-based vaccines. However, reluctance to receive vaccines owing to the uncertainty regarding their safety is prevalent. Therefore, rigorous safety evaluation of vaccines through preclinical toxicity studies is critical to determine the safety profiles of vaccine candidates. This study aimed to evaluate the toxicity profile of HuVac-19, a subunit vaccine of SARS-CoV-2 utilizing the receptor-binding domain as an antigen, in rats, rabbits, and dogs using single- and repeat-dose study designs. Repeat-dose toxicity studies in rats and rabbits showed transient changes in hematological and serum biochemical parameters in the adjuvant and/or vaccine groups; however, these changes were reversed or potentially reversible after the recovery period. Moreover, temporary reversible changes in absolute and relative organ weights were observed in the prostate of rats and the thymus of rabbits. Gross examination of the injection sites in rats and rabbits treated with the adjuvant- and HuVac-19 showed discoloration and foci, whereas histopathological examination showed granulomatous inflammation, inflammatory cell infiltration, and myofiber degeneration/necrosis. This inflammatory response was local, unassociated with other toxicological changes, and resolved. In a pharmacological safety study, no toxicological or physiological changes associated with HuVac-19 administration were observed. In conclusion, HuVac-19 was not associated with any major systemic adverse effects in the general toxicity and safety pharmacology evaluation, demonstrating that HuVac-19 is a vaccine candidate with sufficient capacity to be used in human clinical trials.