Brine Shrimp Lethality Assay Research Articles

Green Synthesis of Benzimidazoles: Sustainable Graphene Oxide Catalyst for Bioactive Compound Design and Drug Discovery

We report a novel and eco-friendly method for synthesizing a series of benzimidazoles using 1,3,5-Tris(2- hydroxyethyl) isocyanurate-Cu(II) functionalized magnetic graphene oxide (MGO-THEIC-CuII) as a catalyst under microwave irradiation in ethanol. The newly synthesized compounds were thoroughly characterized by IR and NMR spectroscopy. Furthermore, their biological activities were assessed through antimicrobial activity and Brine-Shrimp lethality assays. The results demonstrate the efficacy and versatility of the developed green synthesis approach, which could have significant implications in the design of bioactive compounds. These findings open new avenues for the development of sustainable and efficient methodologies in drug discovery and related fields. Keywords: Benzimidazole, Graphene Oxide, Biological Activity

One-Pot Synthesis of Silver Nanoparticles from Garcinia gummi-gutta: Characterisation, Antimicrobial, Antioxidant, Anti-Cancerous and Photocatalytic Applications.

Methods like the bio-synthesis of silver nanoparticles (Ag NPs) using plant extracts have become promising due to their eco-friendly approach. The study aimed to examine the utilization of Garcinia gummi-gutta fruit phytochemicals as agents in the biosynthesis of Ag NPs, evaluation of the antimicrobial, antioxidant, and anti-cancerous properties, as well as the photocatalytic ability of bio-synthesized Ag NPs against Crystal Violet (CV), a triphenylmethane dye. The characterization of the physical properties of the Ag NPs synthesized via the green route was done using UV-Vis spectrophotometry (UV-Vis), X-ray Diffraction (XRD), Fourier Transform Infrared Spectrophotometry (FTIR), Scanning Electron Microscopy (SEM), Zeta potential analysis, and Transmission Electron Microscopy (TEM). The dye degradation efficiency of CV was determined using synthesized Ag NPs under UV light by analyzing the absorption maximum at 579 nm. The antimicrobial efficacy of Ag NPs against E. coli, S. aureus, Candida tropicalis, and Candida albicans was examined using the broth dilution method. The antioxidant and anti-cancer properties of the synthesized Ag NPs were assessed using the DPPH and MTT assays. The UV analysis revealed that the peak of synthesized Ag NPs was 442 nm. Data from FTIR, XRD, Zeta potential, SEM, and TEM analysis confirmed the formation of nanoparticles. The SEM and TEM analysis identified the presence of spherical nanoparticles with an average size of 29.12 nm and 24.18 nm, respectively. Maximum dye degradation efficiency of CV was observed at 90.08% after 320 min without any silver leaching, confirming the photocatalytic activity of Ag NPs. The bio-efficiency of the treatment was assessed using the Allium cepa root growth inhibition test, toxicity analysis on Vigna radiata, and Brine shrimp lethality assay. The findings revealed the environmentally friendly nature of green Ag NPs over physical/chemically synthesized Ag NPs. The synthesized Ag NPs can effectively be used in biomedical and photocatalytic applications.

Oliveria decumbens Vent. (Apiaceae): Biological screening and chemical compositions

Ethnopharmacological relevanceOliveria decumbens Vent. (Apiaceae), a single aromatic species in Iran, is traditionally used for healing inflammation, gastrointestinal disorders, and infections. Aim of the studyRegarding the importance of O. decumbens in traditional medicine, we aimed to set out the plant's biological screening and analyze the chemical components of the active fractions. Materials and methodsAir-dried O. decumbens aerial parts were macerated by ethanol:water (70:30). Using a liquid-liquid extraction (LLE) technique, n-hexane, dichloromethane (DCM), ethyl acetate (EtOAc), n-butanol (n-BuOH), and water were successively used to fractionate the crude extract into different portions. Various biological activities were performed on the crude extract, fractions, and some experiments on pure compounds. The bioassays were as follows: antibacterial activity against Staphylococcus aureus, Bacillus subtilis, Escherichia coli, Pseudomonas aeruginosa, and Salmonella typhi (using microplate alamar blue assay; MABA), antifungal activity against Aspergillus niger, A. fumigatus, Candida albicans, C. glabarate, Fusarium lini, Microspurum canis, and Trichophyton rubrum (using agar tube dilution method), antileishmanial activity against Leishmania major and L. tropica using a 96-well serial dilution protocol, anti-inflammatory activity using the respiratory burst assay, cytotoxicity against HeLa (cervical cancer) and BJ (normal fibroblast) cells using MTT assay, insecticidal activity against Tribolium castaneum, Sitophilus oryzae, and Rhyzopertha dominica (using the contact toxicity method), larvicidal activity against Aedes aegypti, anti-DPPH• activity, and cytotoxicity against brine shrimp (Artemia salina) in a lethality assay. Eventually, the phytochemicals from the active fractions were studied by gas chromatography coupled with mass spectrometry (GC-MS). ResultsInterestingly, the DCM fraction was the most active, followed by the n-hexane fraction in the biological assays, including antibacterial (>80% inhibition), leishmanicidal (IC50 (L.major) = 29.4 μg/mL, and IC50 (L.tropica) = 30.0 μg/mL), anti-inflammatory (IC50 = 15.8 μg/mL), insecticidal (>80% inhibition), and larvicidal (100% inhibition of A. aegypti) assays. Further GC-MS analysis of the DCM and n-hexane fractions resulted in the characterization of 12 and 14 phytoconstituents, respectively, compared with the NIST library. Thymol and carvacrol were abundant in both fractions. To lesser quantities, the presence of monoterpenoids (p-cymen-8-ol, thymoquinone, 3-hydroxy-β-damascone, and 3-hydroxy-7,8-dihydro-β-ionol), phenylpropanoids (methoxyeugenol, elemicin, and 4-[(1E)-3-hydroxy-1-propenyl]-2-methoxyphanol, simple phenolics (salicylic acid and 4-methoxy-2,3,6-trimethyl-phenol), and a coumarin (6,7-dimethoxy-coumarin) were detected in the DCM fraction. On the other hand, besides a coumarin and monoterpenoids, the fatty acids (tetradecanoid acid, n-hexadecanoic acid, and linolenic acid) and a sesquiterpene (spathulenol) were observed in the n-hexane fraction. The EtOAc fraction scavenged the DPPH• radicals better than other fractions (IC50 = 41.4 μg/mL), while in brine shrimp lethality assay, the crude extract was more active than n-hexane and DCM fractions with LD50 = 385.20, 660.28, and 699.74 μg/mL, respectively. Surprisingly, the crude extract and fractions were ineffective against assayed fungal strains and tested cancer and non-cancer cell lines. ConclusionsOur findings showed that O. decumbens deserves to be a multi-bioactive medicinal plant, besides its ability for cereal protection against pests. To understand the principal mechanism of action, in silico, in vitro, and in vivo experiments may clarify the ambiguities and even figure out the synergistic behavior of the minor secondary metabolites.

Comparative Study of In vitro Cytotoxic Effect of Leaves and Stems Extracts of Clitoria ternatea by Brine Shrimp Lethality Assay

Aim: The present study aims at the comparison of cytotoxic effect of methanolic extracts of leaves and stems of Clitoria ternatea (Fabaceae) by brine shrimp lethality assay.
 Methodology: Dried leaves and stems were macerated with methanol separately and preliminary phytochemical screenings were carried out. Hatched brine shrimp nauplii were chosen for the assay. The effect was assessed by calculating % mortality of nauplii with different concentrations of the test extracts (1, 10, 25, 50, 100, 500 and 1000 μg/ml) and standard vincristine sulphate. All sets were performed in triplicate.
 Results: The extracts of leaves and stems revealed to possess chiefly alkaloids, phenols and flavonoids. Both the extracts exhibited promising outcomes in dose dependant manner while significant responses in most of the doses were also calculated. Leaves were found to be more potent (LC50 value of 276.29 µg/ml.) compared to the stems (LC50 value of 322.95 µg/ml.). All the doses of the standard Vincristine sulphate were found to display significant activity compared to the control and was calculated to have LC50 value of 11.75 µg/ml.
 Conclusion: The methanolic extracts of leaves and stems have shown potential cytotoxicity against brine shrimp which were highly comparable with standard. Further work on isolation, characterization and tests on cell lines may lead to identification of active principles.

Cytotoxicity Analysis and In Silico Studies of Three Plant Extracts with Potential Application in Treatment of Endothelial Dysfunction.

Endothelial dysfunction is the basis of the physiopathological mechanisms of vascular diseases. In addition to the therapeutic activity of plant extracts, cytotoxicity is significant. This research evaluates the cytotoxicity of three vegetal extracts (Calendulae flos extract-CE, Ginkgo bilobae folium extract-GE, and Sophorae flos extract-SE). In vitro evaluation was performed using an endothelial cell line model (Human Pulmonary Artery Endothelial Cells-HPAEC) when a dose-dependent cytotoxic activity was observed after 72 h. The IC50 values were calculated for all extracts: Calendulae flos extract (IC50 = 91.36 μg/mL), Sophorae flos extract (IC50 = 68.61 μg/mL), and Ginkgo bilobae folium extract (IC50 = 13.08 μg/mL). Therefore, at the level of HPAEC cells, the cytotoxicity of the extracts follows the order GE > SE > CE. The apoptotic mechanism implied in cell death was predicted for several phytocompounds using the PASS algorithm and molecular docking simulations, highlighting potential interactions with caspases-3 and -8. In vivo analysis was performed through brine shrimp lethality assay (BSLA) when lethal, behavioral, and cytological effects were evaluated on Artemia salina larvae. The viability examined after 24 h (assessment of lethal effects) follows the same sequence: CE > SE > GE. In addition, the predicted cell permeability was observed mainly for GE constituents through in silico studies. However, the extracts can be considered nontoxic according to Clarckson's criteria because no BSL% was registered at 1200 µg/mL. The obtained data reveal that all three extracts are safe for human use and suitable for incorporation in further pharmaceutical formulations.

Toxicity Studies and in vivo Antiplasmodial Evaluation of Leaf and Stem Bark Extracts of Terminalia avicennioides

Malaria is one of the most serious infectious diseases found in the tropics and subtropics. P. falcifarum resistance to almost all anti-malarial drugs has necessitated the search for anti-malarial compounds. The aim of this study is to investigate the antiplasmodial effects of crude leaf and stem bark extracts of T. avicennioides, a plant utilized by traditional healers in Northern Nigeria to treat malaria and other ailments. Brine shrimp lethality (cytotoxicity) assay and acute toxicity were also evaluated, which are considered a useful tool for preliminary assessment of toxicity in plant extracts. The median lethal dosage (LD50) was calculated by administering different doses of the extract (100–4000 mg/kg) intraperitoneally to three groups of three mice each for 24 hours and observing the animals for physical sign of toxicity. Three models were employed to assess the extracts antiplasmodial effectiveness in vivo against the rodent malaria parasite Plasmodium berghei: suppressive, curative, and repository. The leaf and stem bark extracts were found to show significant toxicity to brine shrimp larvae of Artemia salina in the brine shrimp lethality test with LC50 values of >1000 and 29.6 respectively. Excitation, paw licking, and death were symptoms of acute toxicity of the extracts based on physical and behavioral observations. Death was recorded at 1000 mg/Kg of the stem bark extract and the LD50 is ≤100 mg/Kg body weight while for the leaf extract it was estimated to be ≤600 mg/Kg with no death record. In vivo evaluation revealed that the extracts significantly (P <0.05) reduced parasite count in all models (suppressive, curative and prophylactic), with high average percentage inhibition of parasitaemia (54.68, 58.67 and 65.61%) seen in both T. avicennioides leaf and stem bark extracts, which was comparable to that produced by chloroquine and artesunate, respectively (64.31&93.1; 70.08&80.67; 63.40&75.20%). Both extracts had moderate antiplasmodial activity and were non-toxic to mice and brine shrimps. This finding validates the plant's traditional use in malaria treatment.

In vitro Antibacterial and Cytotoxic Activities of Various Fractions of Lippia alba (Mill.)

Lippia alba (Mill.) (Family: Verbenaceae) is widely known due to both ethnobotanical uses and chemical diversity. The present work was designated to determine the antimicrobial and cytotoxic activities of the aqueous ethanolic extract and various fractions of this plant. The whole plant was made into coarse powder and was extracted with 5% aqueous ethanol. The concentrated ethanolic extract (abbreviated as LAE) was successively partitioned with n-hexane (LAH), chloroform (LAC), ethyl acetate (LAA), and the remaining part was designated as water (LAQ). The antimicrobial and cytotoxic activities of extract and all these fractions were evaluated in vitro using disc diffusion technique and brine shrimp (Artemia salina) lethality assay, respectively. In antimicrobial assay, five Gram-positive and five Gram-negative bacteria and standard antibiotic ciprofloxacin (10 μg/disc) were used. All the samples were tested at four concentrations (200, 400, 600 and 800 μg/disc). Our study showed that the extractive and different fractions of L. alba had antibacterial activity against all the tested bacteria except Pseudomonas aeruginosa. Among the extractives, LAE and LAH had significant antibacterial activity when compared to standard ciprofloxacin. The zone of inhibition of LAE and LAH were found to be 14.4, 24.4, 13.4, 15.2, 18.5, 24.1, 16.1, 23.5, 23.2 and 14.5, 20.1, 12.5, 17.1, 17.5, 20.1, 12.5, 19.4, and 18.16 mm against Bacillus cereus, Staphylococcus aureus, Streptococcus-β- haemolyticus, Bacillus subtilis, Sarcina lutea, Escherichia coli, Klebsiella pneumonia, Salmonella typhi and Shigella dysenteriae, respectively, at 800 μg/disc. The median lethal concentration (LC50) in brine shrimp lethality bioassay was determined by extrapolation from graph and the values were found to be 41.9, 23.9, 21.3, 88.7, 6.7 and 7.5 μg/ml for LAQ, LAH, LAC, LAA, LAE and standard vincristine sulfate, respectively. In the study, different mortality rate was observed at different concentration and all the samples showed positive response, indicating the presence of cytotoxic components in L. alba. Phytochemical screening revealed that L. alba contains saponins, tannins, glycosides, steroids, alkaloids, terpenoids and flavonoids. The results of the study indicate that L. alba might be a good source of potent antibiotic and anticancer drugs. Bangladesh Pharmaceutical Journal 26(2): 150-156, 2023 (July)

Synthesis, In-Silico, In Vitro and DFT Assessments of Substituted Imidazopyridine Derivatives as Potential Antimalarials Targeting Hemoglobin Degradation Pathway

Malaria is a serious illness transmitted through the bite of an infected mosquito, which is caused by a type of parasite called plasmodium and can be fatal if left untreated. Thus, newer antimalarials with unique mode of actions are encouraged. Fused pyridines have been vastly reported for numerous pharmacological activities including but not limited to analgesics, antitubercular, antifungal, antibacterial and antiapoptotic agents. In a current study, a series of substituted Imidazo[1,2-a]pyridine-3-carboxamides (IMPCs) (SM-IMP-01-13) along with some hydrazides (DA-01-DA-02) were synthesized and characterized by Fourier-transform infrared spectroscopy (FTIR), 1H-/[Formula: see text]C-NMR (proton/carbon nuclear magnetic resonance), elemental analyses and mass spectra. These synthesized analogies were subjected for in vitro biological activities such as Brine Shrimp lethality (BSL), and assay of [Formula: see text]-hematin formation inhibitions. The BSL assay results suggested that compounds, SM-IMP-09, SM-IMP-05 were found to be less toxic and they also had comparable toxicity as of 5-Flurouracil (control) ((e.g., at 10 [Formula: see text]g/ml: 20% deaths of nauplii). Derivatives SM-IMP-02, and DA-05 inhibited [Formula: see text]-hematin formation: IC[Formula: see text]: 1.849 and 0.042 [Formula: see text]M, respectively). Our molecular docking analysis on plasmodial cysteine protease falcipain-2 indicated that compound DA-05 (–9.993 kcal/mol) had highest docking score and it was comparable to standard Chloroquine (–7.673 kcal/mol). The most active molecule, DA-05 was also retained with lower HOMO–LUMO energy gap as 3.36 eV. Further, we have also analyzed MEP, and other global reactivity indexes for all IMPCs using DFT. Finally, our in-silico pharmacokinetic analysis suggested that all compounds were having good% human oral absorption values ([Formula: see text]100%), good Caco-2 cell permeabilities (>1600 nm/s), and non- carcinogenic profiles.

Analysis of phytochemical, antioxidant, and α-amylase inhibition capacity of methanol extract of six plants from Kaski, Gulmi and Rupandehi districts of Nepal

This study was designed to explore phytochemicals and biological activities of methanolic extract of six traditional medicinal plants (Clerodendrum trichotomum, Mallotus philipinensis, Dioscorea-bublifera, Rubia cordifolia, Viscum articulatum) of Nepalese origin. Plant extracts were prepared in methanol through cold percolation. Analysis of phytochemical constituent, antioxidant capacities and brine shrimp lethality assay of the test plant samples were carried out using standard methods. The 1,1-diphenyl-2-picryl hydrazyl (DPPH) method was used to study antioxidant activity of different plant extracts. Furthermore, starch-iodine method was used to study the inhibition effect of extract on α-amylase. Phytochemical analysis showed the presence of phytochemicals like alkaloids, flavonoids, phenolic content, glycosides, reducing sugars, etc. in six medicinal plants. Brine shrimp lethality assay suggested the presence of pharmacologically active compounds. Total phenolic content and total flavonoid content of C. trichotomum’s leaves extract were found to be higher with 212.742 mg GAE/g and 112 mg Q/g respectively with strong antioxidant activity. Similarly, the α-amylase inhibition of R. cardifolia’s root extract and C. trichotomum‘s leaves extract was found to be 252.44±0.55 µg/mL and 293.33± 0.81 µg/mL comparative with IC50 value 119.063±0.73 µg/mL of standard acarbose that showed remarkable antidiabetic property among six samples. The results, obtained here suggested that six medicinal plants i.e. C. trichotomum, M. philipinensis, D. deltoidea, D. bublifera, R. cordifolia, and V. articulatum, Nepal origin showed biological activity by targeting multiple drug targets which justifies their traditional uses. This is the first finding that C. trichotomum‘s leaves can be a promising source for the development of natural antioxidant and antidiabetic agents.

Antimicrobial activity, cytotoxicity, and qualitative phytochemical composition of aqueous and methanolic leaf extracts of Physalis peruviana L. (Solanaceae)

Background: The high morbidity and mortality associated with microbial infections and the ineffectiveness of conventional antibiotics due to inappropriate use and resistance warrant alternative stratagems. Aim: We investigated the antimicrobial, cytotoxicity, and qualitative phytochemical composition of the aqueous and methanolic leaf extracts of Physalis peruviana L. (Solanaceae) based on its ethnomedicinal information. Methods: The antimicrobial activity of the extracts was investigated using the disk diffusion technique. The brine shrimp lethality assay method was used to determine the cytotoxicity of the study extracts on brine shrimp nauplii. Qualitative phytochemistry of the study extracts was performed using standard procedures. Results: The two extracts did not possess antimicrobial activity against P. aeruginosa and were either inactive or slightly inactive against the other microbes at concentrations of ≤ 50 µg/ml. Notably, the aqueous extract exhibited high to very high activities against E. coli and S. aureus and very high to remarkable activities against B. cereus. The methanolic extract showed remarkable activity at concentrations of ≥200 µg/ml against E. coli, moderate to very high against S. aureus, high to very high against B. cereus, and moderate to high against C. albicans. The aqueous extract's minimum inhibitory concentrations (MICs) were 50 μg/ml (E. coli and B. cereus), 100 μg/ml (S. aureus), and 200 μg/ml (C. albicans). The minimum bactericidal concentration concentrations (MBCs) of the aqueous extract were 100 μg/ml (E. coli and B. cereus), 200 μg/ml (S. aureus), while the minimum fungicidal concentration (MFC) for C. albicans was 400 μg/ml. The MICs of the methanolic extract were 25 μg/ml for E. coli, S. aureus, and B. cereus and 100 μg/ml for C. albicans, and the MBCs were 100 μg/ml for E. coli, S. aureus, and B. cereus, and the MFC for C. albicans was 200 μg/ml. The aqueous was non-toxic, while the methanolic extract was slightly toxic to brine shrimp nauplii, indicating their relative safety. Various phytochemicals were detected in the study extracts, depicting broad pharmacologic activity, including antimicrobial activity. Conclusion: The aqueous and methanolic leaf extracts of P. peruviana may serve as a source of efficacious and safe novel antimicrobial agents upon further evaluation.

Study of the Bioactive Properties, Radical Scavenging Activity and Phenolic Profiles of Extracts Crocus cancellatus subsp. mazziaricus (Herb.) B. Mathew, 1982

Human beings have used plants as medicine for various health cause of years. Plants are widely used in the traditional medicine of different countries and are a source of strong drugs. To evaluate research some bioactive properties of the extract of the Crocus cancellatus subsp. mazziaricus (Herb.) B. Mathew, 1982 the antioxidant activities of these extracts were analyzed by means of cleaning methods (DPPH, ABTS scavenging activity), β-carotene/linoleic acid test system and FRAP activity. Phenolics and flavonoid contents as the equivalents of gallic acid, quercetin respectively. The phenolic content of the extracts was analyzed using HPLC. Brine shrimp lethality test was applied to analyze cytotoxic activity. The obtained results indicated that the highest phenolic compound ferulic acid with 2376.2±0.00 µg/g was in extracts. The highest total phenolic and flavonoid were found in the ethanolic extracts. The aerial part-methanol extract exhibited the highest antioxidant capacity and the corm acetone extract the highest amount of ferric reducing power activity. Among the four different extracts, the acetone extract showed the highest amount of radical scavenging both corm and aerial parts. The brine shrimp lethality assay of bulb ethanol extract has showed good cytotoxic with LC50 of 320.535 μg/mL. This study will be a source for future studies.

Antibacterial activity of herbal formulation against common oral pathogens.

The development of antibiotic resistance in microorganisms is a global challenge for the clinicians, pharmacist and research scientists leading to the development of new medicinal formulations that are effective and easily consumable. The plant yielding essential oil with chief constituent as eugenol has been identified as an important compound with strong inhibition of bacteria, and storage fungi. Ocimum gratissimum and Ocimum sanctum is an aromatic shrub occurring in warm tropical regions has been used in traditional medicine in India to cure various ailments in general and as an antimicrobial agent in particular. The aim of this present study is to assess the antimicrobial and cytoxic activity of the formulation against oral pathogens. The formulation of O. gratissimum and O. sanctum plant extract was prepared and filtered. Antimicrobial activity was done by agar well diffusion method, minimum inhibitory concentration assessment was determined by broth dilution method and cytotoxicity was assessed by brine shrimp lethality assay. Agar well diffusion method against S. mutans, Enterococcus faecalis, C. albicans, Lactobacillus sp, and S. aureus revealed no zone of inhibition but at 100µL concentration at every time interval, the study formulation showed more bacteriostatic activity than positive control and the standard used. The formulation showed very minimal cytotoxicity. The formulation of O. gratissimum and O. sanctum synergistically showed more antibacterial, antifungal and cytotoxic activity and more research has to be done in invivo environment.

Aqueous Extract of Clitoria ternatea Attenuates the Growth of Streptococcus mutans

In the human oral cavity, Streptococcus mutans is often observed and is a major contributor to tooth decay. Increased S. mutans levels may be linked to progressively more severe forms of periodontal disease because root exposure in people with periodontitis increases caries rates. Hence, a new potential antibacterial compound needs to be searched to combat this pathogenic bacterium. The butterfly pea, or Clitoria ternatea is an ornamental plant that has been reported to exhibit antibacterial properties against several bacteria. Thus, the goal of this investigation was to determine how well C. ternatea aqueous (CTA) extract inhibited S. mutans. The disk diffusion assay was performed to access the antibacterial properties of the CTA extract. The efficiency of the extract against the test bacterium was then determined through MIC/MBC determinations and a time-kill study. Meanwhile, the toxicity of the extract was tested using a brine shrimp lethality assay (BSLA). The CTA extract demonstrated substantial antibacterial activity against the test bacterium at a concentration of 200 mg/ml, with a diameter of the inhibition zone of 13.4±0.4 mm, according to the disc diffusion assay. The aqueous extract’s MIC and MBC values were found to be 100 and 400 g/mL, respectively. Time-kill analysis revealed the CTA extract exerted a strong bactericidal effect on S. mutans and this activity was dose-dependent. A scanning electron microscope (SEM) exhibited the bacterial cells experienced severe damage after being exposed to CTA extract including formation cavities, irregular shape, and crumpled cells. Thus, the present study suggested the potential of CTA extract as an antibacterial agent against oral cavity bacteria and can be used in the formulation of natural mouthwash due to no toxicity effect.

Antioxidant and Ameliorative Effects of Basella alba L. On Letrozole-Induced Polycystic Ovarian Syndrome in Rats

Basella alba L. (BA) is utilized in Nigeria to manage Polycystic Ovarian Syndrome (PCOS), according to an ethnobotanical study. However, this claim has not been supported by any scientific data. This study investigated the effect of the methanol leaf extract of the plant in alleviating polycystic ovary conditions in rats. The methanol extract of BA was subjected to a brine shrimp lethality assay. Total Phenolic Content (TPC), Total Flavonoid Content (TFC), and the DPPH assay were used to measure antioxidant activity. To induce PCOS in thirty female Wistar rats (180-200 g), letrozole (1 mg/kg) was used. For 15 days, BA leaf methanol extract (100 mg/kg body weight) and Clomiphene citrate (1 mg/kg body weight), a standard medication, were given. Microscopically, the ovaries underwent histopathological investigation. ELISA was used to assess luteinizing hormone (LH), follicle-stimulating hormone (FSH), and estradiol levels. One-way ANOVA was used to analyze the data, and P < 0.05 was set as the threshold for Dunnett's Multiple Comparison Test. LC50 value for the extract was 17.29±0.15 μg/mL. The DPPH radical scavenging activity of the ethyl acetate fraction was the highest. When PCOS animals were given BA, the ovarian stroma was normal and the granular cells were leutinized. The level of estradiol increased but the level of LH was not significantly different between the BA-treated group (0.23±0.03 mIU/mL) and the untreated PCOS group (0.23±0.01 mIU/mL). Basella alba leaf displayed ameliorative effects on polycystic ovary conditions in rats. This support the use of this plant in the management of PCOS.

Effect of altitude and solvent on Psidium guajava Linn. leaves extracts: phytochemical analysis, antioxidant, cytotoxicity and antimicrobial activity against food spoilage microbes

BackgroundGuava (Psidium guajava Linn.) has been traditionally used in the treatment of a wide range of diseases due to its rich content of secondary metabolites.AimThis study was aimed to evaluate the effect of altitude and solvent systems on guava leaves crude extract’s phenolics and flavonoid content, antioxidant, antimicrobial, and toxicity nature.MethodsGuava leaves were collected from three different geographical locations in Nepal while solvents with an increasing polarity index were used for extraction. The yield percentage of extracts was calculated. Total Phenolic Content, Total Flavonoid Content, and antioxidant activity were determined by the Folin-Ciocalteu method, Aluminium chloride colorimetric method, and DPPH (2,2′-Diphenyl-1-picrylhydrazyl) assay respectively. The quantification of fisetin and quercetin was performed using the HPLC with method validation. The antimicrobial activity of the extracts was tested against bacteria and fungus isolated from spoiled fruits and vegetables and identified through 16s and 18s rRNA sequencing. Finally, Brine Shrimp Lethality Assay (BSLA) was used for testing the toxicity of the extracts.ResultsThe phenolic and total flavonoid content was found to be higher in ethanol extract (331.84 mg GAE/g dry extract) and methanol extract (95.53 mg QE/g dry extract) from Kuleshwor respectively. Water extract of guava leaves from Kuleshwor (WGK) did not show significantly different antioxidant activity when compared to methanol and ethanol extracts. Fisetin and quercetin were higher in WGK (1.176 mg/100 g) and (10.967 mg/100 g) dry extract weight respectively. Antibacterial activity against food spoilage bacteria was dose-dependent and found to be highest for all the extracts from different solvents and altitudes at higher concentrations (80 mg/ml). Similarly, methanol and ethanol guava extracts from all locations showed antifungal activity against Geotrichum candidum RIBB-SCM43 and Geotrichum candidum RIBB-SCM44. WGK was found to be non-toxic.ConclusionOur study concludes that the antioxidant and antimicrobial activity of WGK was found to be similar statistically to that of methanol and ethanol extracts of Bishnupur Katti and Mahajidiya. These results suggest the possibility of using water as a sustainable solvent to extract natural antioxidant and antimicrobial compounds which can further be used as natural preservatives to extend the shelf life of fruits and vegetables.

ANTI-BACTERIAL, CYTOTOXICITY, AND ANTIOXIDANT PROPERTIES OF THE ISOLATED FLAVONOIDS EXTRACT FROM WHITE DRAGON FRUIT (HYLOCEREUS UNDATUS) PEELS AND FLESH

The presence of flavonoids in white pitaya’s flesh and peel was confirmed using the Thin Layer Chromatography Screening method. Flavonoid was extracted using solvent fractionation and then subjected to the three assays. Using the Kirby-Bauer method, better antibacterial activity against E. coli was found using 100% peel extract, partially active at 10.28 mm. The flesh flavonoid extract was inactive against both E. coli and S. aureus at concentrations of 100%, 75%, and 50%. The flavonoid peel and flesh have the potential as anticancer agents since they are active, toxic, and potent, with LC50 of less than 1000 ppm in Brine Shrimp Lethality Assay. Using a UV VIS spectrophotometer at 517 nm, both the flesh flavonoid extract and the peel flavonoid extract have antioxidant potential. However, the flesh with a percent Radical Scavenging Activity close to the catechin control is a better antioxidant. This study proved that the peel and flesh of White Pitaya, particularly its flavonoid, have medical benefits, can be a potential source of income for farmers, and decrease waste in the environment.

Toxicity evaluation of some plants used in ethnofishery in Arunachal Pradesh, India

Plant-based natural products have shaped traditional knowledge systems due to their numerous biological and pharmacological utility. However, in addition to their extensive utilization, some of these plants are also known for their toxicological properties. Since the foundation of human civilization, these toxic plants were used for fishing, hunting and safeguarding against enemies. In this study, a total of five plant species used in ethnofishery were considered for the toxicological study. The toxicities of these plants were evaluated using Brine Shrimp Lethality Assay (BSLA) and LC50 values were estimated. All the plants have displayed the toxic effect and have been graded to be highly toxic (i.e., LC50 of 0–100 μg/ml or LC50 < 1000 μg/ml). Furthermore, with the lowest LC50 (0.14 in 12 h of incubation), the plant species D. scandens was reported to be the most toxic. The toxic plants have an important role in rural livelihood; however, if the concentration of these compounds from poisonous plants reaches a high concentration in the vulnerable biological system, it could bring a devastating health-related issue for the frequent consumers.

Structural, vibrational, optical, and anticancer properties of Hyphaene thebaica‐reduced nano‐lanthanum oxide (La2O3)

Phytoextracts modulated synthesis of nano‐scaled spherical lanthanum oxide nanoparticles (La2O3 NPs) are reported using the aqueous extracts of Hyphaene thebaica as a bioreducing and chelating agent. X‐ray diffraction (XRD) pattern revealed sharp peaks and single‐phase purity. The Debye–Scherrer approximation indicated an average crystallite size of 37 nm, whereas W‐H plot revealed a crystallite size of 59 nm, complemented by the high‐resolution transmission electron microscopy (HR‐TEM). The infrared spectroscopy revealed characteristic La‐O band at 426 and 648 cm−1. K‐M function revealed the bandgap of 5.6 eV, whereas broad intense emission peaks between 345 and 750 nm. The Raman spectra revealed peaks at 182, 325, 384, and 491 cm−1. Elemental composition was confirmed using energy‐dispersive X‐ray spectroscopy (EDS) spectrum, and the spotty rings from selected area electron diffraction (SAED) indicated the crystalline nature of the HT‐La2O3 NPs. In addition, the HT‐La2O3 NPs were investigated for in vitro antioxidant, anticancer, antiangiogenic, and cytotoxic potentials. DPPH free radical scavenging activity was highest, that is, 49.54 ± 0.88% at 1000 μg/mL. MTT assay on MCF‐7 and 3T3 cell lines revealed a median lethal concentration of 114 and 43 μg/mL, respectively. The antiangiogenic effect examined by chick chorioallantoic membrane (CAM) assay indicated inhibition of 57.14 ± 0.96% at 1000‐μg/mL concentration with an IC50 of 551 μg/mL. IC50 for the brine shrimp lethality assay was 314.2 μg/mL. Our study indicates cytotoxicity of HT‐La2O3 NPs, and it further concluded that HT‐La2O3 NPs were more potent against 3T3 cells as compared with MCF‐7 cells. Further research on the biological synthesis and anticancer studies are recommended for the large‐scale applications of the La2O3 NPs.

Safety evaluation of Amphora sp., a marine microalgal diatom isolated from the Cape coastal waters, South Africa

Recent studies have highlighted the huge potential of a marine diatom, Amphora sp. as an affordable food supplement with significant therapeutic activities, however, the safety of the microalgae for human consumption has not been evaluated. Thus, in this study the in vitro and in vivo toxicity profiles of Amphora sp. biomass as well as its extracts were investigated to facilitate its use in human nutrition. The in vitro toxicity profile was assessed through the brine shrimp lethality, cytotoxicity and mutagenicity assays while the in vivo study was carried out using a rat model. The microalgal extracts displayed no cytotoxicity on the PBMC and MCF-7 cell lines neither did it display any evidence of mutagenicity on the Salmonella typhimurium strains. Similarly, results from the acute toxicity assessment demonstrated no remarkable changes in body and organ weights, the biochemical parameters as well as in the histopathology of the murine liver, heart and kidneys. Specifically, it was recorded that the oral administration of Amphora sp. to a dosage level of 4000 mg/kg/BW was relatively non-toxic, thus providing a scientific basis for selecting a safe dose in future studies aimed at highlighting the potentials of the diatom in human nutrition.

An environmentally safe approach for the facile synthesis of anti-mutagenic fluorescent quantum dots: Property investigation and the development of novel antimicrobial applications

Quantum dots as nanocrystals of semiconductor materials are synthesized with different methods. The green synthesis of fluorescent quantum dots using natural stabilizing agents has attracted much attention. This research aims to study the ability of Teucrium polium L. extract to green synthesize fluorescent silver quantum dots (AgQDs). The green synthesis of AgQDs was proved using UV–Visible spectrophotometer, Fourier Transform Infrared spectroscopy (FTIR), X-Ray Diffraction (XRD), Transmission Electron Microscopy (TEM), Scanning Electron Microscopy (SEM), Energy-Dispersive X-ray spectroscopy (EDX), Photoluminescence (PL) analysis, and Dynamic Light Scattering (DLS).According to the findings, the shape of fluorescent AgQDs was spherical, and the maximum abundance of particle size distribution was between 3 and 5 nm.Thereupon, antimicrobial, anti-mutagenicity, anticancer, and antioxidant activities were evaluated to distinguish the biological features of AgQDs. Gram-positive bacteria (S. epidermidis, MIC: 31.25 μg/ml) and fungi (C. albicans, MIC: 31.25 μg/ml) were the most susceptible to AgQDs. Moreover, the AgQDs solution had higher antimicrobial activity than AgNO3. The Ames test demonstrated that AgQDs have considerable anti-mutagenic activities (% mutagenicity inhibition greater than 40) and are not mutagenic. The anticancer activity of AgQDs was distinguished using MTT and brine shrimp lethality (BSL) assays. The analysis indicated that AgQDs had a strong potential for cytotoxicity (BSL: LC50 = 2.4 μg/ml, MTT: IC50 = 0.8 μg/ml). Further, AgQDs were used for bioimaging human ovarian OVCAR3 cells, and their biocompatibility was remarkable. In the DPPH assay, AgQDs displayed high antioxidant activity (89.9% inhibition). Using AgQDs to coat materials to develop antibacterial agents revealed significant activity against C. albicans and S. epidermidis strains. Our results indicate that T. polium is a promising source for synthesizing AgQDs. Due to the potent bioactivities of AgQDs, they can be used in medicine and industry.