Brine Shrimp Lethality Bioassay Research Articles

Chemical and Biological Profiling of Derris Scandens (Roxb.) Benth

Plants have been a valuable source of bioactive phytochemicals since ancient times, providing mankind with important compounds that can be utilized in the development of innovative therapeutics for various diseases. Chemical investigation of Derris scandens (Roxb.) Benth., a plant with a long history of traditional use, led to the identification of two flavonoids such as Scandenone and Lupalbigenin. The 1H NMR spectroscopic analysis along with comparison to similar compounds in the literature, helped to elucidate the structures of these compounds. The crude methanol extract was fractionated, and different fractions were evaluated for their antioxidant, cytotoxic, and thrombolytic activities. Among the fractions, the ethyl acetate and chloroform soluble fractions showed the highest DPPH free radical scavenging activity, while the chloroform soluble fraction demonstrated significant lethality in the brine shrimp lethality bioassay. The petroleum ether soluble fraction exhibited substantial thrombolytic activity. The extraction of these important phytochemicals from the plant has brought new insights into the field of drug discovery, potentially facilitating the development and exploration of novel therapeutics. However, additional investigations are advised to gain a comprehensive understanding of their precise molecular mechanisms and potential toxicological effects. Bangladesh J. Bot. 53(2): 227-233, 2024 (June)

Antimicrobial, Cytotoxic and Phytochemical Analysis of Chloroform Extracts of Andrographis paniculata Root (Burm.F) and Parquetina nigrescens (Afzel) Aerial Part

The menace of antibiotic resistance especially multi-drug resistant bacteria is increasing in the clinical setting. Therefore, the study investigated antimicrobial activity and phytochemical constituents of chloroform extracts of Andrographis paniculata root (Burm.F.) and Parquetina nigrescens (Afzel) aerial part. The extracts were screened for cytotoxic activity using brine shrimp (Artemia salina) lethality bioassay. The antimicrobial activity of the extracts was tested against Citrobacter diversus, Klebsiella pneumoniae, Proteus vulgaris and Shigella sonnei using tetrazolium microtitre-plate method. Analysis of chemical components of the extracts was done on Agilent Technologies 7890A gas chromatograph system s. GC-MS analysis of A. paniculata root extract and P. nigrescens aerial part extracts revealed a total of 32 and 22 compounds, respectively. The major components identified in these two extracts were fatty acid esters, alkaloids and terpenes. The lowest MIC observed with A. paniculata extract was 6.25 μg/ml for Citrobacter diversus and 12.5 μg/ml for Klebsiella pneumoniae. The lowest MIC recorded for Parquetina nigrescens extract was 3.125 μg/ml for Citrobacter diversus, Shigella sonnei and Proteus vulgaris while it was 12.5 μg/ml for Klebsiella pneumoniae. The susceptibility was compared with a clinical antibiotic (0.02 μg/ml) which served as the positive control. P. nigrescens with LC50 of 89.75 ± 0.18 μg/ml showed higher lethal toxicity than A. paniculata with LC50 of 167.5 ± 0.21 μg/ml. The observed toxicity might be due to presence of fatty acids and alkaloids in the extracts. The extract of the plants investigated in our study showed promising antimicrobial activity. Dhaka Univ. J. Pharm. Sci. 23(1): 43-51, 2024 (June)

Evaluation of In-vitro Anti-Inflammatory, Anti-Fungal, Thrombolytic, Membrane Stabilizing and Cytotoxic Properties of (Camellia chrysantha Hu Tuyama)

The current study was set out to look into the anti-inflammatory, anti-fungal, cytotoxic, thrombolytic and membrane stabilizing activities of the methanolic extract from the leaf of Camellia chrysantha (Hu) Tuyama (MECCL). Primary evaluation of MECCL was performed via phytochemical screening. Phytochemical analysis of the leaf extract revealed the presence of reducing sugar, flavonoids, alkaloids, and steroids. Anti-inflammatory activity test was done using egg albumin denaturation assay. In the result of MECCL’s anti-inflammatory test showed 88.88% compared to the standard acetyl salicylic acid 98.56%. The anti-fungal activity test was performed by Disc Diffusion method using zone of inhibition against four fungi. MECCL showed moderate antifungal activity compared to standard Griseofulvin against Aspergillus niger, Saccharomyces cerevisiae (yeast), Penicillium notatum. Zones of inhibition of the fungi Aspergillus niger, Saccharomyces cerevisiae (yeast), Penicillium notatum is 11mm, 14mm and 14mm at the concentration of 700 µg/disc, which is close to the standard Griseofulvin 19mm, 20mm, and 21mm respectively at 50 µg/disc concentration. Mucor hiemalis showed mild zone of inhibition in 300, 500, and 700 µg/disc (7mm). Brine Shrimp lethality bioassay method was used to test the lethality of the methanolic leaf extract. The extract showed significant cytotoxic activity against brine shrimp nauplii with LC50 value of 0.843 µg/mL when compared with the standard Vincristine sulphate (LC50 value: 0.608 µg/mL). Comparison with positive control Vincristine sulphate signifies that cytotoxicity exhibited by MECCL and further bioactivity guided investigation can be done to find potent antitumor compounds. In thrombolytic activity test, MECCL demonstrated 95.69% thrombolysis which was significant compared to standard Streptokinase (SK) which demonstrated 91.304 % thrombolysis. While doing membrane stabilizing activity test, Heat induced hemolysis assay was performed and the percent of protection was 85.35 compared to the standard DS 73.63 which was very significant. The results highlight the potential of Camellia chrysantha (Hu) Tuyama as a valuable source of phytochemicals that possess notable anti-inflammatory, thrombolytic, and membrane-stabilizing properties. The methanolic leaf extract also shows moderate antifungal and cytotoxic effects, suggesting potential for future therapeutic advancements.

Pharmacological Profile Analysis of a Vegetable Crop Pumpkin (Cucurbita maxima Linn.) Seeds

The seeds of pumpkin (Cucurbita maxima Linn.) can be utilized as both a conventional and multifunctional dietary constituent. In this study the antioxidant, antibacterial, and cytotoxic effects of various fractions such as petroleum ether soluble fraction (PESF), chloroform soluble fraction (CFSF), dichloromethane soluble fraction (DCMSF), and aqueous soluble fraction (AQF) were obtained from crude extract (CE) of the C. maxima seeds were assessed by using 1,1-Diphenyl-2- Picrylhydrazyl (DPPH) method, disc diffusion technique, and brine shrimp lethality bioassay respectively. The results showed that all the fractions have remarkable antioxidant, antibacterial, and cytotoxic activities. The study revealed that both the AQF and PESF and the CF confirmed significant antioxidant properties. The antibacterial activity of different fractions against Gram-positive and Gram-negative bacteria was examined to determine their significant effect. PESF and CFSF fractions exhibited substantial inhibitory activity against Gram-positive bacteria, particularly Bacillus cereus. Bacillus subtilis exhibited significant susceptibility to both CFSF and AQF. Salmonella typhi and Shigella dysenteriae exhibited significant susceptibility to PESF and CFSF, while Escherichia coli was the Gram-negative bacterium most effectively inhibited by CFSF. The IC50 values revealed that the DCMSF exhibited the highest inhibitory activity, followed by other fractions. The new study emphasizes the significance of pumpkin seeds in terms of their pharmacological and biological properties, which may encourage more research on the potential benefits of this plant for human health. Jagannath University Journal of Life and Earth Sciences, 9(1): 7-18, 2023 (June)

Bioactivity and chemical screening of endophytic fungi associated with the seaweed Ulva sp. of the Bay of Bengal, Bangladesh

Abstract Several studies have shown that endophytic fungal metabolites possess vital biological activities; nevertheless, there is a lack of knowledge regarding the medicinally important marine endophytic fungi associated with the seaweeds mainly found in the Bay of Bengal, Bangladesh. In this study, six endophytic fungi, belonging to five genera and four classes, were isolated from the well-known chlorophyte, Ulva sp. and were most closely related to Chaetomium globosum, Nigrospora magnoliae, Curvularia sp., Curvularia moringae, Aspergillus terreus and Collariella sp. This is the first report of these fungi as endophytes associated with Ulva sp. from the Bay of Bengal, Bangladesh. A preliminary biological evaluation of the ethyl acetate extract of each endophytic fungal crude extract was the prime objective of this research, e.g., antimicrobial assay, 2,2-diphenyl-1-picrylhydrazyl free radical scavenging activity and brine shrimp lethality bioassay. Evaluation of test results revealed that each fungal crude extract possessed one or more relevant biological activities. Preliminary chemical screening using TLC and NMR spectroscopic analysis revealed the presence of several secondary metabolites in the crude fungal extracts. These findings suggest that the marine endophytic fungus may be a valuable source for investigating potentially bioactive chemicals or leads for novel drug candidates.

Exploring antioxidative, cytotoxic and neuropharmacological insights into Bixa orellana leaves: Experimental and in silico approaches

Background StudyThe aim of this research was to examine possible antioxidant, cytotoxic and neurological activity of methanol and n-hexane extracts of Bixa orellana leaves. Additionally, we aimed to identify potential lead compounds through in-silico analysis. MethodsIn-vitro antioxidative properties were investigated through different assays, including: total phenolic content assay (TPC), total flavonoid content assay (TFC), DPPH free radical scavenging assay and reducing power assay. Also, the cytotoxic effect of the samples was assessed using the brine shrimp lethality test. In addition, anxiolytic, locomotor, and CNS depressant activities were assessed utilizing various established methods. Moreover, reported compounds were used in the in silico study to explore the best-fit phytoconstituents against gamma-aminobutyric acid (GABAA) receptor. ResultsMBOL displayed substantial antioxidative activities in various established assays compared to NBOL. In brine shrimp lethality bioassay, both MBOL and NBOL revealed cytotoxic activity in a concentration-dependent approach. Again, in Elevated Plus Maze test, 200 and 400 mg/kg of NBOL and MBOL demonstrated significant anxiolytic activities evident from time spent in open arms. In addition, maximum number of head dipping was demonstrated by MBOL at 400 mg/kg (53.90 ± 1.16) in Hole Board test. NBOL and MBOL at both doses significantly diminished the magnitude of movements from the 2nd to 5th observation periods in Open Field test. Furthermore, in Hole Cross test, MBOL remarkably dwindled the locomotor activity at 120 min and 180 min (3.60 ± 0.40 and 2.40 ± 0.51) at 400 mg/kg. Finally, in silico analysis revealed 13 compounds as promising leads with strong binding affinity to GABAA receptor along with good pharmacokinetics and toxicity profiles. ConclusionTherefore, the present study's findings advocate the traditional usage of this plant and recommend both MBOL and NBOL as as a potential source of therapeutic candidate for the management of neurological disorders.

Variability in Chemical Composition and Biochemical Activities of Mentha x piperita L. Essential Oil, in Response to Mycorrhizal Symbiosis and Heavy Metal Stress.

The present paper highlights the effect of Pb/Cd-stress and/or mycorrhizal colonization by Glomus Intraradices on yield, chemical composition, cytotoxicity and antimicrobial activity of Mentha x piperita L. essential oil. Our findings showed that mycorrhizal colonization could be used to improve the essential oil yield of M. x piperita, either in non-stressed or Pb/Cd-stressed plants. GC-MS analysis revealed three chemotypes: linalool/pulegone (32.6/30.8 %) chemotype in essential oils of non-mycorrhizal Pb-stressed plants, menthone/menthyl acetate (30.3/25.1 %) chemotype in essential oils of non-mycorrhizal Cd-stressed plants and menthol (44.6 %) chemotype in essential oils of non-mycorrhizal non-stressed plants, mycorrhizal non-stressed plants and mycorrhizal Pb/Cd-stressed plants. The cytotoxicity of M. x piperita essential oil, evaluated by brine shrimp lethality bioassay, was increased in presence of Pb/Cd-stress (from 379.58 to 72.84 μm/mL) and decreased in mycorrhizal plants (from 379.58 to 482.32 μm/mL). The antimicrobial activity of M. x piperita essential oil, evaluated by disc diffusion method and determination of Minimum Inhibitory Concentration against ten microorganisms, was enhanced by the mycorrhizal colonization and deceased by the Pb/Cd-stress. In conclusion, the inoculation of medicinal plants with mycorrhizal fungi is a real avenue for alleviating abiotic stress and/or increasing the quantity and quality of secondary metabolites in terms of biological activities.

Evaluation of in vitro anthelmintic, cytotoxic, antimicrobial and thrombolytic activities of <i>Ipomoea hederifolia</i> stem

<i>Ipomoea hederifolia</i>, a member of the convolvulaceae family, is an annual climbing ornamental vine with wide range of pharmacological activities. The methanolic extract of powdered <i>Ipomoea hederifolia</i> stem was subjected to evaluate the<i> in-vitro</i> anthelmintic, cytotoxic, antibacterial and thrombolytic potentials. The disc-diffusion technique explored the antibacterial activity of <i>Ipomoea hederifolia</i> stem extract (IHSE) against six detrimental microorganisms. Anthelmintic and thrombolytic potentials were assessed using earthworms and human erythrocytes as test samples. Moreover, a brine shrimp lethality bioassay procedure was applied to determine cytotoxic activity. The methanolic IHSE demonstrated statistically significant (<i>p < 0.05</i>) anthelminthic potential in a dose-dependent fashion. In the brine shrimp lethality assay, IHSE depicted a sharp soar in the death rate of brine shrimp nauplii, and 50% (LC<sub>50</sub>) of nauplii died at 4.544 µg/mL of IHSE. Moreover, the IHSE revealed moderate clot lysis activity in a dose-dependent manner; the highest clot lysis was 37.167± 2.40% at 20 mg/mL (<i>p < 0.05</i>). However, no significant antimicrobial activity was observed for IHSE. Based on the findings, our investigation suggests that the methanol fraction of IHSE possesses substantial anthelmintic, cytotoxic and thrombolytic potentials, and the plant could be deemed as a probable hub for future drug discovery.

ZIKV Inhibitors Based on Pyrazolo[3,4-d]pyridazine-7-one Core: Rational Design, In Vitro Evaluation, and Theoretical Studies.

The Zika virus (ZIKV) is believed to cause birth defects, and no anti-ZIKV drugs have been approved by medical organizations to date. Starting from antimicrobial lead compounds with a pyrazolo[3,4-d]pyridazine-7-one scaffold, we synthesized 16 derivatives and screened their ability to interfere with ZIKV infection utilizing a cell-based phenotypic assay. Of these, five compounds showed significant inhibition of ZIKV with a selective index value greater than 4.6. In particular, compound 9b showed the best anti-ZIKV activity with a selectivity index of 22.4 (half-maximal effective concentration = 25.6 μM and 50% cytotoxic concentration = 572.4 μM). Through the brine shrimp lethality bioassay, 9b, 10b, 12, 17a, and 19a showed median lethal dose values in a range of 87.2-100.3 μg/mL. Compound 9b was also targeted to the NS2B-NS3 protease of ZIKV using molecular docking protocols, in which it acted as a noncompetitive inhibitor and strongly bound to five key amino acids (His51, Asp75, Ser135, Ala132, Tyr161). Utilizing the pharmacophore model of 9b, the top 20 hits were identified as prospective inhibitors of NS2B-NS3 protease, and six of them were confirmed for their stability with the protease via redocking and molecular dynamics simulations.

Assessment of antibacterial activity and cytotoxic effects of in vitro and in vivo plant parts of a medicinal plant Gynura procumbens (Lour.) Merr.

The goal of this study was to evaluate the antibacterial and cytotoxic effects of both the in vitro and in vivo plant part extracts of the medicinal plant Gynura procumbens. An effective protocol for regeneration and callus formation was developed using nodal segments and regenerated leaf explants, respectively. The highest fresh and dry weight calli were produced after four weeks of culture on Murashige and Skoog (MS) medium containing 2.0 mg/L BAP and 2.0 mg/L NAA, while the most shoots were produced on MS medium containing 1.0 mg/L BAP and 0.5 mg/L IAA. The in vitro shoots developed roots on MS media with 0.1 mg/L IBA. The antibacterial activity of extracts against various bacteria was examined to determine their significance (p < 0.05). The least significant difference (LSD) test results showed that the regenerated leaf extract had the highest antibacterial activity while the callus extract had the lowest. The minimum bactericidal concentration (MBC) and the minimum inhibitory concentration (MIC) were also established. Regenerated leaf extract had the highest toxicity and the lowest lethal concentration (LC50) value (1.21 ± 0.03 μg/mL) in a brine shrimp lethality bioassay. In contrast, callus extract had the lowest toxicity and the highest LC50 (11.09 ± 0.4 μg/mL). In addition, the in vitro cytotoxicity test results revealed that the callus and field leaf extracts had anti-cell-proliferative properties. The regenerated leaf and stem extract, however, could induce cell growth.

Antioxidant, antibacterial, cytotoxic and thrombolytic activities of flowers of Mirabilis jalapa L: Possible role of phenolics and flavonoids

Mirabilis jalapa L. belongs to Nyctaginaceae family is an ornamental plant and domestically known as Sandhamaloti in Bangladesh. In this report we evaluated phenol and flavonoid content of cold methanol extractives of flower of M. jalapa and their role in antioxidant, antibacterial, cytotoxic and thrombolytic potentials. This study utilized HPLC to identify qualitative phenolic and flavonoid compounds. Gallic acid and Kaempferol were employed as standards for phenolic and flavonoid identification, respectively. Consequently, evaluating the overall phenolic and flavonoid content in the plant extract was considered a logical step. Flower of M. jalapa is an abundant source of phenols and flavonoids. The antioxidant activities in the phosphomolybdenum assay are differed in distinct extractives and have been found as 334.78 ± 0.62, 142.53 ± 0.51, 398.04 ± 0.81, 470.29 ± 0.55 and 480.90 ± 0.59 mg of ascorbic acid equivalent per gm of crude methanol extract (MSF), petroleum ether fraction (PSF), carbon tetrachloride fraction (CTF), chloroform fraction (CSF) and ethyl acetate fraction (ESF), respectively. The IC50 values for DPPH radical scavenging have been evaluated to be 13.70 ± 0.32, 49.05 ± 0.47, 6.76 ± 0.22, 9.30 ± 0.41 and 6.94 ± 0.31 μg/mL for MSF, PSF, CTF, CSF and ESF, respectively. The cytotoxicity was evaluated against brine shrimp lethality bioassay and LC50 values of MSF, PSF, CTF, CSF and ESF have been found 11.83 ± 0.32, 35.50 ± 0.54, 5.00 ± 0.18, 11.30 ± 0.30 and 4.61 ± 0.15 μg/mL, respectively in comparison to a standard vincristine sulphate 2.50 ± 0.11 μg/mL. Plant polyphenols and flavonoids having antioxidant properties exhibit potential antibacterial activity and our results revealed that the entire tested fractions of vegetation of M. jalapa exhibit potential antibacterial activity. On this observation, the thrombolytic activity, is the first strive in M. jalapa to investigate for more modern thrombolytic and its ethyl acetate fraction was shown to substantial (53.81 ± 0.52%) thrombolysis property. A high correlation was observed between antioxidant, cytotoxicity, thrombolytic and antibacterial activity with polyphenol and flavonoid contents (r2 = 0.903 to 0.996, p < 0.05). These data suggest the role of polyphenols and flavonoids in the bioactivities. Further study of this plant may lead to isolation of bioactive polyphenols and flavonoids which will be effective in the management of many chronic diseases.

In-vitro Pharmacological Investigation of Ludwigia adscendens

Aims: This research objective was to investigate some in-vitro properties of methanol-extracted plant extracts of Ludwigia adscendens (the water primrose). The primary method of assessment for the methanolic extract of Ludwigia adscendens (MELA) was investigated for phytochemical screening. It was determined to look into the plant's potential for in vitro activities, such as antioxidant, antimicrobial, anti-inflammatory anti-arthritis, cytotoxic, thrombolytic and membrane stabilizing activity due to the pharmaceutical interest in its component parts.&#x0D; Methodology: Phytochemical screening was performed utilizing a battery of test reagents. Antioxidant, antimicrobial, anti-inflammatory, anti-arthritic, cytotoxic, thrombolytic and membrane stabilizing activities were measured at various doses using the DPPH Free Radical Scavenging Assay, the Disk Diffusion Method, Egg Albumin Protein Denaturation Assay, the Inhibition of Protein Denaturation Assay, the Brine Shrimp Lethality Bioassay, Clot lysis assay and heat induced and hypotonic induced haemolysis method.&#x0D; Results: The findings of research on MELA's phytochemical screening showed the presence of a wide range of various phytochemical components. The higher antioxidant activity of MELA is shown by its IC50 value of 85.76 µg/mL. Significant antibacterial action was shown by MELA. As compared to the standard ascorbic acid, this extract showed significantly greater anti-arthritic and anti-inflammatory effects. MELA exhibited much higher LC50 value which is 1.147 µg/mL than the typical vincristine-sulphate in a lethality test with brine shrimp. Comparatively speaking, this herb has quite powerful thrombolytic action. Significant membrane stabilizing action has also been shown by MELA.&#x0D; Conclusion: To wrap it up, it is clear that this plants phytochemical can be used for wide range of drug discovery field due to its potent pharmacological actions.

Phytochemical Evaluation of Different Solvent Mediated Extracts and Brine Shrimp Lethality Bioassay of Psychotria Octosulcata W. A Talbot

Plants’ bioactive chemicals have been utilized throughout human history as supplements and medicines to support the body’s natural physiological processes. When their function is discovered, many substances are added for medical uses. Herbs are known to contain a variety of organic substances. These crucial herbs are the main ingredients in many pharmaceutical and healthcare products. Traditional folk medical practices offer essential information for the extraction and creation of numerous medications and activities as well. The production of a few highly active pharmaceuticals is justified by the extraction and characterization of a variety of bioactive chemicals from green factories. This study uses quantitative analysis of ethyl acetate, ethanol, n-hexane, chloroform and aqueous extract along with established methodologies to assess the phytochemicals. Alkaloids, flavonoids, reducing sugars, proteins, amino acids, saponins, tannins, terpenoids and glycosides were discovered in the quantification and phytochemical screening results. Due to the presence of several phytochemical substances, the entire plant of Psychotria octosulcata W. A. Talbot demonstrated a diversity of medicinal potential. The cytotoxicity activity of the plant extracts was predicted using brine shrimp lethality assay (BSLA). The results showed that the shrimp in the ethanolic extract of Psychotria octosulcata W. A. Talbotare extremely hazardous in higher concentrations. Only 10 shrimps are discovered to be lethal at lowest concentrations even after 24 h. This can be due to the presence of bioactive substances.

Comparative Pharmacological Evaluation of Mangrove Plant Xylocarpus mekongensis Pierre and Associated Fungus

Xylocarpus mekongensis commonly known as Poshur, is an evergreen mangrove plant originating from Asia, Indonesia, the Southwestern Pacific Islands, and northern Australia. This study was aimed at compiling information on the comparative pharmacological properties of the methanolic bark extract of X. mekongensis and the fungal endophyte based on their antioxidant, antimicrobial, and cytotoxic activity. The plant extract showed more phenolic (277 mg GAE/g) and flavonoid (140 mg QE/g) content than the fungal extract (45 mg GAE/g and 76 mg QE/g, respectively). The bark extract exhibited better DPPH scavenging capacity (IC50 = 28.27 µg/ml) than endophyte XMSF-I (IC50 = 143.46 µg/ml) extract. Furthermore, we observed that the endophytes associated with this plant showed more significant antimicrobial activity against E. coli, S. aureus, B. subtilis, and A. brasiliensis than its bark extract. In the brine shrimp lethality bioassay, the bark extract and endophyte revealed diminutive lethality (214 and 286 µg/ml, respectively) in comparison with standard vincristine sulfate (0.44 µg/ml). Hence, methanolic bark extract showed more positive reviews than associated fungi.

Antimicrobial, antioxidant, and cytotoxic properties of endophytic fungi isolated from Thysanolaena maxima Roxb., Dracaena spicata Roxb. and Aglaonema hookerianum Schott.

BackgroundEndophytic fungi have recently been recognized as an impressive source of natural biomolecules. The primary objective of the research was to isolate fungal endophytes from Thysanolaena maxima Roxb., Dracaena spicata Roxb. and Aglaonema hookerianum Schott. of Bangladesh and assess their pharmacological potentialities focusing on antimicrobial, antioxidant, and cytotoxic properties.MethodsThe fungal isolates were identified up to the genus level by analyzing their macroscopic and microscopic characteristics. Ethyl acetate extracts of all the fungal isolates were screened for different bioactivities, including antimicrobial (disc diffusion method), antioxidant (DPPH scavenging assay), and cytotoxic (brine shrimp lethality bioassay) activities.ResultsAmong the thirteen isolates, Fusarium sp. was the most recognized genus, while the others belonged to Colletotrichum sp. and Pestalotia sp. Comparing the bioactivity of all the extracts, Fusarium sp. was shown to be the most effective endophyte, followed by Colletotrichum sp. and Pestalotia sp. In the antimicrobial study, two isolates of Fusarium sp. (internal strain nos. DSLE-1 and AHPE-4) showed inhibitory activity against all the tested bacteria and the highest zone of inhibition (15.5 ± 0.4 mm) was exerted by AHPE-4 against Bacillus subtillis. All the fungal isolates produced mild to moderate free radical scavenging activity, where the highest antioxidant activity was revealed by one isolate of Fusarium sp. (internal strain no. AHPE-3) with an IC50 value of 84.94 ± 0.41 µg/mL. The majority of Fusarium sp. isolates exhibited notable cytotoxic activity, where AHPE-4 exhibited the highest cytotoxicity, having the LC50 value of 14.33 ± 4.5 µg/mL.ConclusionThe findings of the study endorsed that the fungal endophytes isolated from T. maxima, D. spicata, and A. hookerianum hold potential as valuable origins of bioactive substances. Nevertheless, more comprehensive research is warranted, which could develop novel natural compounds from these endophytes to treat various infectious and cancerous diseases.

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)

Evaluation of Antioxidant, Cytotoxic, Thrombolytic and Membrane Stabilizing Activities of Canna indica L. Leaves (Family: Cannaceae)

Canna indica L., a perennial plant known as Indian shot and African arrowroot, is a member of the family Cannaceae. Historically, this plant was used to treat menstrual cramps, diarrhea and dysentery. The purpose of this investigation was to assess the antioxidant, cytotoxic, thrombolytic and membrane stabilizing activities of the methanol extract of C. indica leaf and its organic soluble fractions. The DPPH free radical scavenging method and the total phenolic content determination were utilized to evaluate the antioxidant capacity, whereas the brine shrimp lethality bioassay was implemented to study the cytotoxic potential. The ability of the extractives to prevent the hemolysis of red blood cells (RBC) in response to the application of heat and hypotonic solutions was used to estimate of their membrane stabilizing potential. In all of these experiments, the methanolic leaf extract (ME) displayed moderate activity, while the petroleum ether soluble fraction (PSF) and the dichloromethane soluble fraction (DSF) demonstrated the most promising antioxidant and cytotoxic activities among the extractives. However, the strongest thrombolytic and membrane stabilizing activities were observed by the chloroform soluble fraction (CSF). Additional research may be necessary to determine which phytoconstituents are most likely to be responsible for the aforementioned qualities. Bangladesh Pharmaceutical Journal 26(2): 162-166, 2023 (July)

Antihyperglycemic, Neuropharmacological, Cytotoxic, Anticoagulant, and Anti-inflammatory Pharmacological Evaluations of Vernonia elaeagnifolia DC Leaves Secondary Bioactive Metabolites

The current investigation was carried out to confirm the antihyperglycemic, cytotoxic, neuropharmacological, anticoagulant, and anti-inflammatory effects of the ethanolic extract of Vernonia elaeagnifolia leaves in different experimental models. In the oral glucose tolerance test, the plant extract demonstrated significant (p &lt; 0.05) antihyperglycemic activity by reducing the elevated blood glucose level in mice at both doses (250 mg/kg and 500 mg/kg) when compared to the control groups. The crude ethanolic leaf extract, however, didn’t afford to block the α-glucosidase enzyme up to 0.5 mg/mL. The experimental mice showed a noticeable (p &lt;0.001) decrease in locomotor and exploratory activities in the hole cross and open field tests, as well as head dipping in the hole-board test model in the neuropharmacological activity assay at both doses (250 mg/kg and 500 mg/kg). The extracts also showed cytotoxic activity in a brine shrimp lethality bioassay with an LC50 of 91.929 μg/mL and showed significant concentration-dependent anticoagulant activity (p&lt;0.001). Moreover, the extract (10–500 mg/kg) caused a significant (P&lt;0.01 – 0.001) dose-dependent reduction of inflammation induced in rats by using two different models: formalin-induced paw edema and egg-albumin-induced paw edema. From the above results, the crude extract of V. elaeagnifolia DC leaves have significant pharmacological potential in different in vitro and in vivo study model. So, it could be a source of a substance for isolating lead that could be used to treat several diseases.

Essential Oils from Different Parts of Azorean Cryptomeria japonica (Thunb. ex L.f.) D. Don (Cupressaceae): Comparison of the Yields, Chemical Compositions, and Biological Properties

Azorean Cryptomeria japonica is widely used for local wood production, generating large amounts of aerial-part biomass residues that can be a sustainable source of value-added bioproducts. This comparative study aimed (i) to determine the yield and chemical profile of the essential oils (EOs) extracted by hydrodistillation from Azorean C. japonica foliage, leaves, male cones (MCs), and female cones (FCs), and (ii) to investigate the antimicrobial and brine shrimp lethality (BSL) effects of the obtained EOs and some major components. The EOs yield revealed a wide range (ca. 1–3%, w/d.w.) and their chemical composition, analysed by GC–MS and GC–FID, showed the presence of seventy-one components. Monoterpene hydrocarbons (38–71%) dominated in all the studied EOs, mainly α-pinene (17–45% of total EOs), decreasing as follows: cone EOs &gt; leaf EOs &gt; foliage EOs. Oxygen-containing monoterpenes (mainly terpinen-4-ol) also dominated in cone EOs. Contrariwise, oxygen-containing sesquiterpenes (mainly elemol) and diterpene hydrocarbons (mainly phyllocladene) dominated in foliage/leaf EOs. The studied EOs exhibited activity against Gram-positive bacteria but no activity against Gram-negative bacteria. A similar trend was displayed by α-pinene. Only the cone EOs showed antifungal activity against Penicillium chrysogenum, but this was significantly lower than those of α-pinene and terpinen-4-ol. Moreover, FC EO should also be considered for further investigation due to its high toxicity on BSL bioassay. In conclusion, this study demonstrated the variability of yield, chemical profile, and bioactivities in the EOs from different parts of the Azorean C. japonica, expanding the knowledge of the potentialities of C. japonica aerial-part EOs, particularly the ones from the cone samples.

A New Annulenol from Aconitum heterophyllum with Bioactivity and Cytotoxicity against Vero and Lung Cell Line

In this research work, a detailed phytochemical and biological investigation of Aconitum heterophyllum was accomplished. The cold methanolic root extract of A. heterophyllum was subjected to VLC for fractionation over silica gel, which led to the isolation of two compounds 1,1,8,8-trimethyl-4-methylene-decahydro- 1H-cyclopenta-[8]-annulen-1-ol (1) and β-stigmasterol (2). The compounds were characterized by analysis of their 1D and 2D NMR spectral data. Compound 1 is first reported from this plant. The crude extract was screened for antimicrobial activity against several gram-positive and gram-negative bacteria, and fungi using disk diffusion method where significant inhibitory activity was observed. The extract was also tested for cytotoxic property using brine shrimp lethality bioassay, which revealed moderate activity in comparison with the standard anticancer drug, vincristine sulphate. Antioxidant property of the crude methanol extract of A. heterophyllum was determined by the scavenging of the DPPH (2,2-diphenyl-1-picrylhydrazyl) free radical and found moderate antioxidant activity (IC50 = 53.25 μg/mL) when compared to standard antioxidant. Total phenolic and flavonoid content and polyphenolic compound content of the crude extract were determined by HPLC. The isolated compounds 1 and 2 showed dosedependent antiproliferation activity against the vero and lung A549 cells. Dhaka Univ. J. Pharm. Sci. 22(1): 1-10, 2023 (June)