In vitro antibacterial and antioxidant activity of flavonoids from the roots of Tephrosia vogelii: a combined experimental and computational study.

Tephrosia vogelii is a traditional medicinal plant used to treat hypertension, diarrhea and urinary disorders. Silica gel chromatographic separation of CH2Cl2/MeOH (1:1) roots extract of T. vogelii afforded seven compounds namely; ß-sitosterol (1a), stigmasterol (1b), 6a, 12a-dehydro-deguelin (2), tephrosin (3), maackiain (4), obovatin (5) and 6-oxo, 6a, 12a-dehydro-deguelin (6). GC-MS analysis of essential oils from the root of T. vogelii displayed a total of 17 compounds of which cis-nerolidol (41.7 %) and cadinol (19.7 %) were the major constituents. CH2Cl2/MeOH (1:1) extract, MeOH extract, maackiain (4) and obovatin (5) showed moderate inhibitory activity against Pseudomonas aeruginosa with MIC value of 0.5, 0.66, 0.83 and 0.83 mg/mL, respectively, compared to ciprofloxacin (MIC of 0.078 µg/mL). 6a, 12a-dihydro-deguelin (2), and 6-oxo, 6a, 12a-dehydro-deguelin (6) displayed significant activity against S. epidermis with MIC values of 0.66 mg/mL. Tephrosin (3) and maackiain (4) also showed moderate antibacterial activity against Staphylococcus aureus and Proteus mirabilis with MIC values of 0.83 and 0.5 mg/mL, respectively, compared to ciprofloxacin (0.312 µg/mL). The radical scavenging activity results indicated that tephrosin (3), obovatin (5) and 6-oxo, 6a, 12a-dehydro-deguelin (6) showed potent DPPH scavenging activity with IC50 values of 10.97, 10.43 and 10.73 µg/mL, respectively, compared to ascorbic acid (IC50 of 5.83 µg/mL). The docking prediction results revealed that 6a, 12a-dehydro-deguelin (2) displayed the best binding energy of -8.1 kcal/mol towards pyruvate kinase of S. aureus (PDB ID: 3T07) and -7.9 kcal/mol towards P. mirabilis urease (PDB ID: 1E9Y) and DNA gyrase B of Escherichia coli (PDB: 4F86) receptors compared to ciprofloxacin (-7.2 to -8.0 kcal/mol). Maackiain (4) and obovatin (5) displayed the minimum binding energy of -7.9 and -8.2 kcal/mol towards the LasR protein of P. aeruginosa (PDB: ID 2UV) and S. epidermidis FtsZ (PDB: ID 4M8I), respectively. The SwissADME drug-likeness and Pro Tox II toxicity prediction results indicated that compounds (2-6) obeyed Lipinski's rule of five with 0 violations and none of them were found to be hepatotoxic, mutagenic, and cytotoxic, respectively. The in vitro assessment results supported by the in silico analysis revealed that crude extracts and isolated compounds showed promising antibacterial and antioxidant activity, which proves the therapeutic potential of the roots of T. vogelii.

Ethnomedicine, antibacterial activity, antioxidant potential and phytochemical screening of selected medicinal plants in Dibatie district, Metekel zone, western Ethiopia.

BACKGROUND: Medicinal plants play a major role in the delivery of healthcare, particularly among the rural population of Ethiopia. Plant extracts and their bioactive compounds have been utilized for the treatment of several diseases. This study was aimed at evaluating the antibacterial activity, antioxidant capacity, and phytochemical content of selected medicinal plants used in Dibatie district, western Ethiopia. METHODS: Study plants were collected, shade dried, pulverized, extracted by maceration in 80% ethanol, and subjected to antibacterial, antioxidant, and phytochemical tests. Minimum inhibitory concentration (MIC) was determined using 96-well microplates and nutrient broth microdilution. Antioxidant activity was evaluated using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging assay. Phytochemical screening was conducted using standard test methods. RESULTS: The ethanolic extract of Polystachya steudneri Rchb.f. pseudobulbs was the most active against gram-negative Proteus mirabilis, Salmonella typhimurium, Klebsiella pneumoniae, Escherichia coli, and Shigella flexneri, with MIC values of 8 ± 0, 11 ± 5, 3 ± 1, 3 ± 1, and 2 ± 0 mg/mL, respectively. The ethanolic extract of P. steudneri was also the most effective against gram-positive Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus agalactiae, and Enterococcus faecalis, with MIC values of 8 ± 0, 8 ± 0, 3 ± 1, and 16 ± 0 mg/mL, respectively. Ethanolic extracts of Gnidia involucrata Steud. ex A.Rich. stems and roots were effective antioxidants, with respective 50% DPPH free radical inhibitory concentrations (IC50) of 168.68 and 181.79 µg/mL, followed by that of P. steudneri (IC50 = 203.11 µg/mL). The study plants contained alkaloids, anthocyanins, anthraquinones, cardiac glycosides, coumarins, flavonoids, phenols, saponins, steroids, tannins, and terpenoids. CONCLUSIONS: This study confirmed the antibiotic, antioxidant, and phytochemical constituents of the investigated plants and suggested further investigations that may lead to bioactive lead compounds.

Antibacterial activity and phytochemical screening of traditional medicinal plants most preferred for treating infectious diseases in Habru District, North Wollo Zone, Amhara Region, Ethiopia.

Ethiopia's healthcare system relies on traditional medicinal practices that use medicinal plants to treat human and livestock ailments. However, the lack of empirical validation regarding the efficacy of these treatments against specific infectious diseases necessitates rigorous scientific investigations. The objective of this study was to investigate the antibacterial activity and phytochemical screening on five selected medicinal plant species, namely Solanum somalense Franchet., Verbascum sinaiticum Benth., Rumex nervosus Vahl, Withania somnifera (L.) Dunal and Calpurnia aurea (Ait.) Benth. The plants were first identified jointly with local informants and later considering mainly their high informant consensus and fidelity level values for their efficacy in treating infectious diseases in the area. Ethanol and aqueous extracts were prepared from the plant materials, and their antibacterial activities were evaluated against standard bacterial strains, representing both gram-positive and gram-negative types. To assess the antibacterial activity of the extracts, the minimum inhibitory concentration (MIC) was determined using the broth dilution method. Additionally, phytochemical screening was performed using standard qualitative tests to identify various secondary metabolites. The results indicated antibacterial efficacy in the ethanol extracts of S. somalense, W. somnifera, and C. aurea against particular bacterial strains (S. somalense against S. agalactiae with MIC of 1.5 mg/mL; W. somnifera against S. aureus and E. coli, with MIC of 2 mg/mL; C. aurea against E. coli and K. pneumoniae, with MICs of 3 mg/mL and 3.5 mg/mL, respectively). The results of the phytochemical screening indicated the presence of steroids, alkaloids, flavonoids, saponins, and terpenoids. The selected medicinal plants demonstrated promising antibacterial activity against certain bacterial strains. The current findings support the long-standing claim of the traditional medical system of the study area for their continued use of these plants in their treatment of infectious diseases. Further investigation is required to isolate the responsible active compounds and characterize the constituents and description of their antibacterial effect for possible use in areas where these infectious diseases are major health problems.

Ethnoveterinary medicinal plants and their utilization by the people of Soro District, Hadiya Zone, southern Ethiopia.

BACKGROUND: Ethnoveterinary studies are important to maintain the sustainability of livestock health and support people's livelihoods through the provision of food, maintaining livestock health, and other biological resources. This study was carried out in Soro District, southern Ethiopia, to identify, document and analyse plant species with ethnoveterinary uses along with the associated indigenous and local knowledge. MATERIALS AND METHODS: Informants were selected using purposive (key informants) and systematic random sampling (general informants) methods. Data on ethnoveterinary plants and their uses were collected through semi-structured interviews, guided field walks, 13 focus group discussions with five to seven members in each and participant observation. Informant consensus factor and fidelity level were computed to identify the most common livestock ailment categories and the best plant species with ethnoveterinary use, respectively. Preference ranking methods were used to identify the potentially effective ethnoveterinary medicinal plants for the most frequently reported livestock ailments. The use diversity of multipurpose plants with ethnoveterinary importance was analysed using the analytical methods of ethnobotany including priority ranking, comparisons and important indices. The T-test statistic was used to compare knowledge differences among different social groups. RESULTS: A total of 132 plant species in 120 genera and 61 families were reported by informants as having ethnoveterinary uses. The plants are said to be used by the local communities in various ways to treat 50 livestock health problems. Higher number of informants (23.77%) cited Momordica foetida for the treatment of 16 livestock ailments. The highest informant consensus value for this species is associated with its use for treating blackleg in cattle; Nicotiana tabacum was cited for the treatment of 15 livestock ailments mainly recommended for the Lumpy Skin Disease/Ailment of bovines; Croton macrostachyus for treatment of 13 livestock ailments including wooden tongue, FMD in bovines; and Gymnanthemum amygdalinum for nine ailments mainly diarrhoea of all livestock types. Achyranthes aspera is claimed to provide the most effective treatment for Aspiration pneumonia (severe coughing in bovines, sheep and goats) alone, while Croton macrostachyus, Ximenia americana, Allium sativum and Juniperus procera were indicated as potential plant species to treat Lumpy Skin Disease in bovines in the order given. The fidelity level analysis showed that Datura stramonium, Dodonaea viscosa subsp. angustifolia and Asparagus africanus were potential medicinal plant species to treat the respective ailments of rabies, Peste des petits ruminants (PPR) and evil eye/spirit. Multipurpose plant species including Prunus africanus, Combretum molle and Afrocarpus falcatus have been highly threatened as indicated by direct matrix ranking mainly due to collection of fuel wood, construction materials and making household utensils, and farm implements rather than for other uses. CONCLUSION: Soro District has rich and diversified livestock herbal medicinal resources, and indigenous knowledge of remedy preparations and applications is transmitted through generation lines. This resource faces anthropogenic threats with deforestation being the leading factor. Consequently, ethnoveterinary medicinal plants continue to decline before adequate and proper scientific documentation and testing are made. There is a dire need for planning and implementation of appropriate in situ and ex situ conservation strategies and to strive towards ensuring the survival and sustainable utilization of such important plant resources of Soro District. This must be supported by further documentation of the associated indigenous knowledge and pharmacological testing of the key promising species including Balanites aegyptiaca (novel species/NS to treat specific ailment), Brugmansia suaveolens (novel species/NS reported first to treat Livestock ailments/LsAs), Euclea divinorum (NS to treat specific ailments), Grevillea robusta (NS), Hagenia abyssinica (NS for the reported specific ailment), Pentanema confertiflorum (NS), Juniperus procera (NS), Maesa lanceolata (NS), Millettia ferruginea (NS for reported specific ailments), Schrebera alata/NS, Securidaca longepedunculata, Spiniluma oxyacantha/NS, Vepris nobilis (novel species reported first to treat LsAs), Zanthoxylum asiaticum /NS and Ximenia americana (NS for specific ailments). This ethnoveterinary study attempted to fill part of the gaps concerning the prevalent livestock health problems and the associated indigenous and local knowledge in the area.

Toxicological evaluation of chronic oral administration of Justicia schimperiana (Hochst. ex Nees) T. Anderson leaf 80% methanolic extract in Wistar albino rats.

Background: Justicia schimperiana has been widely used for the treatment of various human ailments without scientific proof for chronic toxicity. Thus, this study is aimed to evaluate the chronic toxicity of 80% methanolic extracts of the leaves of Justicia schimperiana in rats. Methods: An 80% crude methanolic extract of the plant leaves was orally administered to Wistar albino rats for 6 months. The experiment was conducted in accordance with the Organization for Economic Co-operation and Development's guideline number 452. Twenty rats per group and sex were randomly assigned to three treatment groups and a control group. Daily doses of 250 mg/kg, 500 mg/kg, and 1000 mg/kg of the extract diluted with distilled water were administered orally to the rats. Rats in the control group received distilled water orally. Weekly body weight and daily food intake were measured. At the end, rats were sacrificed for histopathological, biochemical and hematological tests. The statistical analysis was done using the Kruskal-Wallis test and one-way analysis of variance. Results: Six months daily oral administration of the plant extract did not significantly affect the rats' food consumption, organ weight, and histopathology. Rats treated with 1000 mg/kg extract, however, significantly increased liver enzymes (aspartate aminotransferase, alanine aminotransferase, and alkaline phosphatase), and kidney function tests (creatinine and urea). Additionally, the high dose extract administered rats showed significantly lower red blood cell count, hemoglobin, and hematocrit compared to the control group. Conclusion: Six months oral administration in Wistar albino rats in this experiment indicated that Justicia schimperiana is relatively safe at lower and medium doses. However, increased liver enzymes, increased kidney function tests and decreased red blood cell indices was observed in rats treated at higher doses. To obtain a thorough understanding of the plant's toxicity profile, it is advised that future studies be conducted on teratogenicity and reproductive toxicity.

In vitro antimicrobial and antioxidant activities, essential oil composition, and in silico molecular modeling analysis of secondary metabolites from roots of Verbascum sinaiticum.

Verbascum sinaiticum is locally used to treat wound, stomachache, viral infection, cancer, sunstroke fever, abdominal colic, diarrhea, hemorrhage, anthrax, and hepatitis. The objective of this study was to identify the compounds and to evaluate the antimicrobial and antioxidant activity of the extracts and isolated compounds from V. sinaiticum. The 1H-NMR, 13C-NMR, and DEPT-135 were used to elucidate the structures of isolated compounds. Essential oils were extracted by hydrodistillation method and their chemical analyses were performed by GC-MS. The broth microdilution method was used to evaluate the antimicrobial activity. The radical scavenging activity of the extracts and isolated compounds were evaluated using DPPH method. Silica gel column chromatographic separation of root extracts afforded seven known compounds: 3'-(4''-methoxy phenyl)-3'-oxo-propionyl hexadecanoate (1), harpagoside (2), pulverulentoside I (3), scrophuloside B4 (4), scropolioside A (5), scropolioside-D2 (6), and harpagide 6-O-ß-glucoside (7), which are all reported from this species for the first time. The EO extracts from leaves and roots were the most susceptible to Streptococcus agalactiae, with a 2 mg/mL MIC. The EO from roots was effective against Candida albicans and Trichophyton mentagrophytes, with a MIC of 8 mg/mL. The MeOH and CH2Cl2/CH3OH (1:1) root extracts showed the maximum activity against S. epidermidis with MIC values of 0.25 mg/mL. The strongest antibacterial effects were demonstrated against Staphylococcus epidermidis, which exhibited a 0.0625 mg/mL MIC for compound 1. The strongest radical scavenging activity was exhibited by the methanol extract (IC50 = 3.4 µg/mL), and compounds 4, 6, 5, 3, 7, and 2 with IC50 values of 3.2, 3.38, 3.6, 3.8, 4.2, and 4.7 µg/mL, respectively, in comparison with ascorbic acid (IC50 = 1.3 µg/mL). The results of the molecular docking analysis of compounds revealed minimal binding energies range from -38.5 to -43.1 kJ/mol, -33.1 to -42.7 kJ/mol, -34.7 to -39.3.7 kJ/mol, -25.5 to -37.6 kJ/mol against human myeloperoxidase (PDB ID: 1DNU), murA enzyme (PDB ID: 1UAE), human topoisomerase IIß (PDB ID: 4fm9), S. epidermidis FtsZ (PDB number: 4M8I) proteins, respectively. The docking results and the in vitro antibacterial activity are in good agreement. These findings show that the isolated compounds 2-7 can act as potential antioxidants and strong antibacterials against Staphylococcus aureus and S. epidermidis. As a result, V. sinaiticum root extracts have the potential to be effective in treating diseases caused by bacteria and free radicals, as long as further investigation has been suggested for the ultimate decision of this plant's potential candidate.

Antibacterial and phytochemical analysis of traditional medicinal plants: An alternative therapeutic Approach to conventional antibiotics.

The purpose of this study was to carry out antibacterial and phytochemical analyses on six selected medicinal plants that have been traditionally used by the local people to treat and control different diseases. The antibacterial activities of methanolic extracts of these plants were assessed using the Agar well diffusion and Microtiter broth dilution methods. The root extract of Andrachne aspera showed significantly (p < 0.05) highest mean zone of inhibition at concentrations of 100 mg/ml (33 ± 0.17) and 200 mg/ml (33.5 ± 0.84) against S. epidermidis. The second highest mean zone of inhibition (24.8 ± 0.41) was recorded by Dichrostachys cinerea leaf extract against S. epidermidis at 200 mg/ml concentration. The minimum inhibitory concentrations 1.0 ± 0.0 was recorded by Andrachne aspera against E. faecalis and 2.0 ± 0.0 against S. aureus by Dichrostachys cinerea. The preliminary phytochemical analysis showed that Andrachne aspera and Dichrostachys cinerea contained strong concentration of Polyphenols and Flavonoids. Therefore, these two medicinal plant species have promising potential for further detailed investigations, including safety tests, characterization and isolation of bioactive secondary metabolites for the development of alternative drugs.

Evaluation of Anti-Inflammatory Activity of the Methanol Extracts of Premna schimperi Engl (Lamiaceae) Leaves in Rats.

Background: Even though it is a protective reaction, inflammation continues to be one of the most challenging medical disorders. The current conventional anti-inflammatory drugs have many undesirable health effects and are in need of newer drugs. The purpose of this study was to evaluate the anti-inflammatory effects of an aqueous methanol crude extract of Premna schimperi leaves. Methods: Premna schimperi leaf was extracted with 80% methanol and concentrated; the concentrated extract was used to evaluate the acute toxicity and anti-inflammatory effects. For the acute toxicity study, a single dose of Premna schimperi extract at a dose of 2000 mg/kg was administered and observed for 14 days. Acute, sub-acute, and chronic anti-inflammatory models were employed to evaluate the anti-inflammatory effect of the extract compared to the standard drug. Data were analyzed with SPSS V. 27, and the significance was established with a one-way ANOVA followed by a post hoc Tukey's test. Results: Acute oral toxicity testing at a dose of 2000 mg/kg did not show any sign of toxicity. According to the phytochemical study, the plants contained flavonoids, terpenoids, tannins, cardiac glycosides, steroids, phenolics, and anthraquinones. The extract doses of 200 mg/kg, 400 mg/kg, and 800 mg/kg of extracts effectively (p<0.001) reduced paw edema in the acute and sub-acute models of inflammation. When compared to the negative control group, all tested doses in the chronic model significantly (p<0.05) decreased the production of exudates and the amount of granuloma tissue. Conclusion: Premna schimperi displayed significant anti-inflammatory activity. The tested doses inhibit the formation of edema, granulomas, and exudates.

Antidiarrheal Activities of the Methanol Leaf Extracts of Olinia rochetiana (Oliniaceae) Against Castor Oil-Induced Diarrhea in Mice.

Background: Olinia rochetiana has been used traditionally to cure diarrheal disease. Therefore, this study aimed to investigate the acute toxicity and antidiarrheal effect of O. rochetiana leaf extracts. Methods: Cold maceration was used to extract plant leaf powder with 80% methanol. The extract's antidiarrheal action was tested against a castor oil-induced diarrheal model, a charcoal meal test, and enteropooling tests at doses of 100, 200, and 400 mg/kg. Negative controls received the vehicle at 10 mL/kg, while positive controls received loperamide at 3 mg/kg. Results: From the study, no apparent toxicity was observed when a single dose of 2000 mg/kg was administered. In the castor oil-induced model, the extract delayed the onset of diarrhea, reduced stool frequency, and decreased wet feces weight and number in a dose-dependent manner at 200 mg/kg (p < 0.05) and 400 mg/kg (p < 0.01). The percent reduction in moist feces at 100, 200, and 400 mg/kg was 54.2, 23.97, and 18.26%, respectively, indicating a significant dose-dependent decrease. In a charcoal meal test, the extracts at 200 and 400 mg/kg revealed a peristaltic index of 65 and 46%, respectively, with considerable inhibition of charcoal transport at 23 and 39%. The weight and volume of intestinal contents dropped significantly at a dose of 400 mg/kg (p < 0.01), which is 0.43 mg/kg, in the enteropooling test when compared with the tested dose. The computed in vivo antidiarrheal index revealed diarrheal inhibition values of 46.06 and 71.06% at 200 and 400 mg/kg, respectively. Conclusion: In the current investigation, O. rochetiana showed significant antidiarrheal activity with no symptoms of toxicity in mice.

Antibacterial Activity of a Phenylpropanoid from the Root Extract of Carduus leptacanthus Fresen.

Background: The emergence and rapid spread of antimicrobial drug-resistance microorganisms exacerbate the treatment of infectious diseases, underscoring the importance of finding new, safe, and effective drugs. In Ethiopia, the roots of Carduus leptacanthus have traditionally been employed to treat microbial infectious diseases The aim of this study was to evaluate the antibacterial activity of the root extract and its primary components against six bacterial strains (Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus agalactiae, Escherichia coli, Pseudomonas aeruginosa, and Klebsiella pneumonia). Methods: The extraction involved maceration of air-dried and powdered roots of C. leptacanthus with 80% methanol. The compound was isolated from the root extract using silica gel column chromatography and recrystallization in CHCl3/MeOH (9 : 1) and was characterized using ESI-MS and 1D-NMR spectroscopy. Antibacterial activity of the extract was assessed using agar well diffusion and broth microdilution methods. Results: Syringin, a phenylpropanoid, was isolated and characterized from the extract of C. leptacanthus. The extract showed the most substantial efficacy against S. epidermidis (MIC = 5.33 mg/ml and inhibition zone diameter of 24 mm at 200 mg/m). Syringin also elicited antibacterial activity against S. aureus (MIC = 13.33 mg/ml), S. epidermidis (MIC = 16 mg/ml), and S. agalactiae (MIC = 16 mg/ml). Despite being tested up to a maximum concentration of 16 mg/ml, syringin did not exhibit antibacterial activity against the Gram-negative bacteria (P. aeruginosa, E. coli, and K. pneumonia). Conclusions: In conclusion, the findings suggest that syringin exhibits partial involvement in the root extract's antibacterial activity, thereby potentially supporting the traditional medicinal use of the plant.

Toxic effect of Syzygium guineense ethanolic extract on female reproduction in rats: An evidence from a 10 week repeated-dose toxicity study.

Introduction: It has been reported that some herbal products affect reproduction. To date, reproductive toxicity of Syzygium guineense has not been investigated although the plant is widely used in treating fertility related problems. Thus, the objective of the current study was to investigate the toxic effects of 70% ethanol extract of S. guineense leaves on the reproductive function and histopathology of reproductive organs in female rats. Methods: Eighty female Wistar albino rats were randomly divided into four groups where each group consisted of 20 rats. Rats in the first three groups were treated with S. guineense extract at doses of 250, 500 and 1000 mg/kg body weight, respectively. The fourth group served as a control group. The rats were treated for ten consecutive weeks. The length of estrous cycle, reproductive indices, pregnancy outcomes, and number of postnatal deaths were recorded. At necropsy, organ weight was measured, gross and histopathological examinations of ovaries, uterus, and vagina were conducted. Results: Treatment of rats, with high dose (1000 mg/kg) of S. guineense, significantly prolonged the duration of estrous cycle and reduced weight of uterus and ovaries as well as the number of total and live birth pups. However, there were no significant changes observed in reproductive indices and gross morphology as well as histopathology of ovaries, uterus, and vagina. Conclusion: Administration of high doses of S. guineense could be toxic to some aspects of the reproductive system of female rats and might also affect reproduction. Therefore, consuming high dose of S. guineense leaves is not recommended.

Acute and developmental toxicity of embelin isolated from Embelia schimperi Vatke fruit: In vivo and in silico studies.

Background: Embelin is a hydroxybenzoquinone constituent of the Embelia species that has anti-disease properties. However, its toxicity, particularly the in silico, acute, and developmental toxicity profiles, has yet to be thoroughly investigated. Hence, this study aims to assess these toxicity profiles. Materials and Methods: In silico and in vivo experimental studies were conducted on embelin isolated from the fruits of Embelia schimperi Vatke. In silico toxicity predictions were computed using the ProTox model. The in vivo experiment was done by administering 5000 mg/kg of embelin to a single female albino Wistar rat, followed by three female rats in the absence of death, to determine the mean lethal dose (LD50). Afterwards, three groups of pregnant rats were treated with embelin at doses of 250 mg/kg, 500 mg/kg, and 1000 mg/kg for the developmental toxicity test. Vehicle and ad libitum control groups were used to compare the acute and developmental toxicity variables. Results: In silico toxicity predicted that embelin is free from hepatotoxic, carcinogenic, mutagenic, and cytotoxic effects. No inhibitory effect on hERG channels was observed. It has an immunotoxic property and an inhibitory effect on the CYP2D6 enzyme. Since mortality and signs of toxicities were not observed after treatment with 5000 mg/kg, the mean lethal dose (LD50) is determined to be > 5000 mg/kg. There was no significant difference in the morphological scores or number of somites among experimental animals. None of the embryonic systems possessed developmental delays. Nevertheless, the crown-rump length of the high-dose group became significantly shorter. Maternal food intake and weight gain exhibited significant dose-dependent differences between embelin-treated animals and controls. The number of implantations was significantly low in the treatment group, accompanied by a higher frequency of prior resorption. Conclusion: Embelin is predicted to have a high probability of immunotoxicity potential and affect drug metabolism by inhibiting CYP2D6. In addition, it affects food intake, weight gain, and the number of implantations in pregnant rats. Therefore, it is highly recommended not to take embelin and embelin-rich plants during pregnancy. Further in vitro and in vivo studies need to be conducted to understand the mechanism behind the toxicity of embelin.

Toxic effects of 70% ethanol extract of Moringa stenopetala leaf (Baker f.) Cufod. (Moringaceae) on fetus and placenta of pregnant Wistar rats.

BACKGROUND: Moringa stenopetala leaves (Baker f.) Cufod. (Moringaceae) are used as a staple food and traditional medicine for treating various diseases like malaria, hypertension, stomach pain, diabetes, elevated cholesterol, and removing the retained placenta. Its prenatal toxicity study is minimal. Thus, this study aimed to assess the toxic effects of a 70% ethanol extract of Moringa stenopetala leaf on the fetuses and placentas of pregnant Wistar rats. METHOD: Fresh leaves of Moringa stenopetala were collected, dried at room temperature, ground to powder, and extracted using 70% ethanol. For this study, five groups of animals, each containing ten pregnant rats, were used. Groups I-III were experimental groups and treated with 250, 500, and 1000 mg/kg body weight of Moringa stenopetala leaf extract, respectively. Groups IV and V were pair-fed and ad libitum control groups. The extract was given during gestation days 6 to 12. The fetuses were recovered at day 20 of gestation and examined for the presence of developmental delays, gross external malformations, skeletal and visceral defects. Gross and histopathological changes in the placenta were also evaluated. RESULTS: Compared to the pair-fed control group, maternal daily food intake and weight gain were reduced in the 1000 mg/kg-treated group during the treatment and post-treatment periods. A significantly higher number of fetal resorptions was also seen in the 1000 mg/kg treatment group. The crown-rump length and fetal and placental weights were all significantly reduced in pregnant rats given 1000 mg/kg. However, there were no visible malformations in the visceral organs as well as external genitalia in all the treatment and control groups. About 40.7% of the fetuses in the 1000 mg/kg treated rats had no proximal hindlimb phalanges. In addition, light microscopic investigations of the placenta in the high-dose treated rats revealed structural changes in the decidual basalis, trophoblastic zone, and labyrinthine zones. CONCLUSION: In conclusion, consumption of M. stenopetala leaves at a higher dose may have toxic effects on the development of rat fetuses. At a higher dose, the plant extract increased the number of fetal resorptions, reduced the number of fetuses, decreased the fetal and placental weights, and alter the placental histopathology. Thus, it is recommended to limit the excess feeding of M. stenopetala leaves during gestation.

Analgesic and Anti-Inflammatory Activities of 80% Methanol Extract and Solvent Fractions of Ehretia cymosa Thonn (Boraginaceae) Leaves in Rodents.

Background: Ethnobotanical studies in various districts of Ethiopia reported that Ehretia cymosa (E. cymosa) is used for the management of headache, abdominal pain, arthritis and rheumatism. However, there is no scientific investigation done so far to confirm these traditional claims. Thus, the aim of this study was to assess the analgesic and anti-inflammatory effects of the 80% methanol extract and fractions of E. cymosa leaves. Methods: The dried and pulverized leaves of E. cymosa were soaked with 80% methanol to obtain a crude extract. Fractionation was done using chloroform, ethyl acetate and water by a soxhlet apparatus. The analgesic effects of the crude extract and solvent fractions were assessed using acetic acid-induced writhing and hot plate tests whereas anti-inflammatory activities were investigated using carrageenan-induced paw edema and cotton-pellet-induced granuloma models. Results: In all the tested doses, the 80% methanol extract and solvent fractions revealed substantial (p < 0.001) analgesic activities in acetic acid induced writhing test. In the hot plate method, all the tested doses of E. cymosa crude extract and the solvent fractions produced significant analgesic activities (p < 0.05). In the carrageenan-induced acute inflammation model, all tested doses of the crude extract and solvent fractions resulted in a significant decline in paw edema. The 80% methanol extract and solvent fractions of E. cymosa at all the tested doses significantly reduced inflammatory exudates and granuloma mass formations (p < 0.001). Conclusion: From the results of this investigation, it can be stated that 80% methanol extract, aqueous, ethyl acetate and chloroform fractions of E. cymosa exhibited considerable analgesic and anti-inflammatory activities, supporting the plant's traditional use as a remedy for a variety of painful and inflammatory conditions.

Nutrient Density and Microbial Safety of Open-Air-Dried Beef Meat and Its Biochemical and Organ Histopathology Effects in Albino Rats: A Promising Ingredient for Complementary Food Formulation.

Introduction: Dried beef meat is a major source of essential fatty acids, minerals, and vitamins that are digestible and absorbable, thus could be a potential source of nutrients in complementary food formulations. Composition, microbial safety, and organ function tests were analyzed, and histopathological effect of air-dried beef meat powder was determined in rat model. Methods: Three groups of diets were given for the three groups of animals: (1) standard rat diet, (2) meat powder+standard rat diet (1 : 1 formulation), and (3) dried meat powder. A total of 36 Wistar albino rats (18 males and 18 females) of 4-8 weeks old were used and randomly assigned to the experiments. After acclimatization for one week, the experimental rats were followed for 30 days. Microbial analysis, nutrient composition, organ histopathology (liver and kidney), and organ function tests were conducted from serum samples taken from the animals. Results: Protein, fat, fiber, ash, utilizable carbohydrate, and energy contents of meat powder on a dry weight basis were 76.12 ± 3.68, 8.19 ± 2.01, 0.56 ± 0.38, 6.45 ± 1.21, 2.79 ± 0.38 g/100 g, and 389.30 ± 3.25 kcal/100 g, respectively. Meat powder could be also a potential source of minerals such as potassium (766.16 ± 77.26 mg/100 g), phosphorus (150.35 ± 16.26 mg/100 g), calcium (18.15 ± 7.80 mg/100 g), zinc (3.82 ± 0.10 mg/100 g), and sodium (123.76 ± 32.71 mg/100 g). Food intakes were lower in MP group compared to the others. According to organ histopathology results, animals fed with the diet have shown normal values, except rise in alkaline phosphatase (ALP) and creatine kinase (CK) in groups fed with meat powder. The results of organ function tests were all within the acceptable ranges and comparable with their counterpart control groups. However, some of the microbial contents of the meat powder were not within the recommended level. Conclusion: Dried meat powder has a higher amount of nutrients, which would be a potential recipe in complementary food preparation that can support to reduce child malnutrition. However, further studies need to be conducted on the sensory acceptability of formulated complementary foods containing dried meat powder; also, clinical trials are aimed at observing the effect of dried meat powder on child linear growth.

Prenatal developmental toxicity study of herbal tea of Moringa stenopetala and Mentha spicata leaves formulation in Wistar rats.

Background: Moringa stenopetala and Mentha spicata have long been used to treat diabetes, hypertension, asthma, and other ailments. Herbal tea of M. stenopetala and Mentha spicata leaves formulation showed better antidiabetic and antihypertensive activities. This study investigated the prenatal developmental toxicity potential of the herbal tea of M. stenopetala and M. spicata leaves blend in rats. Methods: Wistar pregnant rats were randomly distributed into four groups (n = 8). Group I (control) dams received distilled water. Group II-IV dams were treated with 559.36, 1118.72, and 2237.44 mg/kg of herbal tea of M. stenopetala and Mentha spicata leaves formulations, respectively, during days 5-19 of gestation. Maternal mortality, clinical signs, body weight changes, and food consumption were recorded. On gestation day 20, cesarean sections were performed, and maternal parameters of systemic toxicity (e.g., body weight, serum biochemistry, organ weight, and macro-pathology) as well as reproductive toxicity (e.g., number of corpora lutea, implantations, resorptions (early/late), pre/postimplantation losses, number of fetuses (live/dead), and fetal body weights, length, and their sex ratio) were evaluated. Fetuses were further examined for external, soft tissue, and skeletal alterations. Results: No herbal tea-related maternal deaths or overt toxic symptoms were observed. The measured maternal systemic and reproductive toxicity parameters showed no herbal tea-associated significant alterations at any dosage levels. Moreover, there were no overt toxic effects of the herbal tea on the fetal external, visceral, or skeletal prenatal growth and development. Conclusion: The study findings demonstrated that the herbal tea of M. stenopetala and M. spicata leaves blend could be relatively safe/low toxic to pregnant rats and developing fetuses. The no-observed-adverse-effect level (NOAEL) of herbal tea for maternal toxicity, fetotoxicity, and teratogenicity in rats is estimated to be > 2237.44 mg/kg/day.

Teratogenic Evaluation of 80% Ethanol Extract of Embelia schimperi Vatke Fruits on Rat Embryo and Fetuses.

Introduction: Embelia schimperi Vatke (family Myrsinaceae) is a commonly consumed anthelminthic plant in Ethiopia. The plant has significant efficacy in treating intestinal worms. However, there are limited data about the safety/toxicity of the plant. Moreover, the teratogenic effect of the plant is not yet well studied despite significant number of Ethiopian mothers consuming herbal medication during their pregnancy. Purpose: This study aimed to evaluate the teratogenic effect of the hydroalcoholic extract of E. schimperi fruit on rat embryos and fetuses. Methods: Pregnant albino Wistar rats were treated with 80% hydroalcoholic fruit extract of E. schimperi at 250 mg/kg, 500 mg/kg, and 1000 mg/kg dosage, whilst the controls were pair-fed and ad libitum groups. Maternal food intake, maternal weight gain, number of implantations, number of prior resorptions, fetal viability, fetal weight, fetal and embryonic crown-ramp length, placental weight, placental gross morphology and histopathology of placental tissue, number of somites, embryonic system, gross/visceral morphological malformations, and ossification centers were evaluated as teratogenicity indices. Results: The crude extract of E. schimperi did not exhibit a significant difference in most developmental indices including the development of a circulatory system, nervous system, and musculoskeletal systems among treated animals and the controls. However, histopathological evaluation of placentas from the treatment groups showed that inflammatory reactions and calcifications compared to the pair-fed and ad libitum controls. Conclusion: Administration of the 80% hydroalcoholic extract of E. schimperi fruit during the period of organogenesis in rats did not show a significant toxic effect on embryonic and fetal developmental indices. However, it might affect the structural integrity of the placenta as it is evidenced by inflammatory reactions and calcifications of decidua basalis of rat placenta.

Synthesis and antimicrobial activity of knipholone analogs.

In the present study, we use knipholone as a prototype molecule to identify new anti-infective agents. Since knipholone is insoluble in water, which would have a detrimental effect on its bioavailability and efficacy, we synthesized knipholone Mannich base derivatives (2-4) that have better predicted solubility and investigated their in vitro antimicrobial activity against eight pathogenic bacterial and fungal strains. The chemical structures of compounds 1-4 were elucidated from their 1H and 13C NMR data, and their antimicrobial activity evaluation was carried out by a broth microdilution MTT assay. Compound 3 exhibited the strongest efficacy against Staphylococcus epidermidis, with MIC value of 9.7 µg/mL. While 4 exhibited the best activity against Staphylococcus aureus, with an MIC value of 19.5 µg/mL, and was the only one to significantly inhibit the fungus Trichophyton mentagrophytes (MIC = 78.2 µg/mL). The study provides evidence for the antibacterial activity of aminoalkyl derivatives of knipholone.

Evaluation of Wound Healing and Anti-Inflammatory Activity of Hydroalcoholic Leaf Extract of Clematis simensis Fresen (Ranunculaceae).

Introduction: Clematis simensis is one of the most widely used medicinal plant for the treatment of wound traditionally. However, its claim was not scientifically tested, the current study therefore assessed the anti-inflammatory and wound healing properties of 80% methanol leaf extract of C. simensis. Methods: The dried and powdered leaf of C. simensis was macerated with 80% methanol. The topical ointment was then made in two concentrations (5% and 10% w/w), and two models, excision and incision, were used to test the extract's capacity to treat wounds in mice. In addition, anti-inflammatory test was also conducted using carrageenan-induced hind paw edema model in three doses (100, 200, and 400 mg/kg) in rats. The DPPH test was used to determine the extract's anti-oxidant properties where ascorbic acid was used as standard agent. Results: When wounds were treated with ointments containing 5% and 10% (w/w) extract, the rate of wound contraction, the length of time it took for the epithelium to form, and the strength of the skin to break were all significantly increased (p < 0.05). There was no discernible difference in wound healing activity between the 10% (w/w) and 5% (w/w) extracts. Moreover, they had also similar impact as that of the positive control nitrofurazone in wound healing activity. Compared to the negative control, C. simensis extract considerably (p < 0.01-p < 0.001) reduced inflammation. The extract also demonstrated antioxidant activity with IC50 values of 0.7 mg/mL for the extract and 1.04 mg/mL for ascorbic acid, respectively. Conclusion: Overall, it is plausible to draw the conclusion that C. simensis 80% methanol extract possesses wound healing activity, perhaps as a result of its anti-inflammatory and antioxidant effects.

Composition of the Essential Oil Thymus schimperi and Evaluation of Its Acute and Subacute Toxicity in Wistar Albino Rats: In Silico Toxicity Studies.

BACKGROUND: In Ethiopian traditional medicine, the aerial part of Thymus schimperi is widely used to treat diseases such as gonorrhea, cough, liver disease, kidney disease, hypertension, stomach pain, and fungal skin infections. However, there is insufficient investigation on the toxic effect of the essential oil of T. schimperi. The aim of this study was, therefore, to evaluate the acute, subacute, and in silico toxicity of Thymus schimperi essential oil in the Wistar albino rats. METHOD: Essential oil of the aerial part of T. schimperi extracted by hydrodistillation was analyzed by GC-MS. The oil was subjected to toxicity studies. In the acute toxicity study, rats were randomly divided into seven groups (n = 5). The control group received only distilled water with 2% of tween 80, whereas the experimental groups received single doses of 300, 600, 900, 1200, 1500, and 2 000 mg/kg of the oil. In the subacute toxicity study, rats were randomly divided into four groups (n = 10). The control group received distilled water with 2% of tween 80, whereas the experimental groups received 65 mg/kg, 130 mg/kg, and 260 mg/kg of the oil orally for 28 days. At the end of the experiment, blood samples were collected for hematology and clinical chemistry evaluation. Gross pathology and histopathology of the liver and the kidneys were also evaluated. For the in silico toxicity study, PubChem CID numbers of GC-MS identified bioactive compounds in the essential oil of T. schimperi obtained from PubChem. Chemdraw (8.0) was used to construct two-dimensional structures of the compounds. The Swiss ADMET web tool was used to convert the two-dimensional structures into a simplified molecular-input line input system (SMILES). In addition, the toxicity parameters were predicted via vNN and ADMET servers. RESULTS: In this study, the LD50 of the essential oil of T. schimperi was found to be 1284.2 mg/kg. According to the World Health Organization, the oil is classified as moderately hazardous in its oral administration. In the subacute toxicity study, rats showed no significant changes in behavioral indices, gross pathology, body weight, biochemical, and most hematological parameters. However, hematological profiles showed a significant decrement in WBC counts and a significant increment of MCV in high dose (260 mg/kg) groups as compared to the control group. Furthermore, no significant differences were observed between the control and essential oil-treated groups, observed in the gross histopathology of the liver and the kidneys. In the in silico toxicity study, all compounds derived from the essential oil showed no cardiac toxicity (h-ERG Blocker), AMES (Ames Mutagenicity), and cytotoxicity via ADMET and vNN-ADMET toxicity predictors. However, by using these servers, about 8.6% of the compounds showed hepatotoxicity, only 3.45% caused drug-induced liver injury, and only 1.75% were potentially toxic to the mitochondrial membrane. CONCLUSION: From the results of this study, oral administration of the essential oil T. schimperi up to a dose of 130 mg/kg is not harmful. However, in the high-dose (260 mg/kg) group, the WBC count was significantly decreased and the MCV was significantly increased. In the in silico toxicity study, most of the components of the oil were found to be nontoxic, although a few of the compounds showed hepatotoxicity and mitochondrial membrane potential toxicity. It is, therefore, essential to conduct chronic toxicity of the essential oil as well as its components, which showed toxicity in the in silico study before using preparations containing the essential oil of T. schimperi.