Crude Ethanolic Extract Research Articles

The Relative Quantification of Hedgehog Signalling Components of Hepatocellular Carcinoma (Hepg-2) Cells Treated With Sorghum bicolor Leaf Stalk Ethanolic Extract

Hepatocellular carcinoma (HCC) is the most frequent type of primary liver cancer. Patients with advanced HCC have a dismal prognosis due to the limited therapy choices available. Hence, this present study evaluates the cytotoxic potential of the ethanolic crude extract of Sorghum bicolor leaf stalk (SBELSE) against human hepatocellular carcinoma (HEPG2) cell line, and its effect in the expression of its Hh signaling component-GLI1 and PTCH. The leaf stalk of Sorghum bicolor (SB) was pulverized and extracted with 70% ethanol. HEPG2 cells were treated with different concentration of ethanol extract of SBELSE which was subjected to 2-fold serial dilution to achieve a graded concentration (9.77-5000 µg/ml). Cell viability was performed using CCK-8 assay. The cells were treated with 8 µg/mL and 23 µg/mL concentration of SBELSE for 48 hour, after which RNA was isolated for downstream gene expression studies using RT-qPCR. The expression level of GLI1 and PTCH genes in cell line was examined relative to the untreated group. SBELSE exerted a cytotoxic effect on HEPG-2 cells with an inhibitory concentration (IC50) of 150 µg/mL. After 48 hours of treatment and evaluation, Expression of GLI1 and PTCH genes was seen in all cell group but was increased in groups treated with 8µg/mL concentration of S. bicolor extract than in the control group. The findings of this study suggest the possibility of the extract to contain agonistic component that might support cancer cell proliferation.

Phytochemical and In vitro Antimicrobial Activities of the Fruit Extracts of Xylopia aethiopica [Dun] A. Rich. (Annonaceae)

Aim: The fruit of the plant Xylopia aethiopica has been used in ethno-medicine in southern Nigeria for treating dysentery, cough and bacterial infections. This present study investigates the phytochemical and antimicrobial activities of the different extracts of X aethiopica fruit. Method: The powdered dried fruits of Xylopia aethiopica was extracted with 95% ethanol and further fractionated into n-hexane and ethyl acetate fractions. The clinically isolated strains of Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and Candida albicans were obtained from their stock cultures and characterized using biochemical tests and then standardized with 0.5 McFarland. The antimicrobial activities of the different extracts were investigated using agar diffusion method. Results: The n-hexane extract was most effective against Pseudomonas aeruginosa, Staphylococcus aureus and Candida albicans with a minimum inhibitory concentration (MIC) of 6.25, 50 and 50 mg/ml respectively while the ethyl acetate extract was active against Pseudomonas aeruginosa and Candida albicans with MIC of 6.25 and 12.5 mg/ml respectively. The crude ethanol extract showed most activity against Pseudomonas aeruginosa with MIC of 3.125 mg/ml. The phytochemical studies on the crude extract revealed the presence of triterpenoids /steroids, carbohydrates, cardenolides and saponins. Conclusion: The antimicrobial activities of the plant justify its usage in traditional healthcare practices especially in infections involving Pseudomonas aeruginosa which showed the highest sensitivity.

Multi-Dimensional Antioxidant Screening of Selected Australian Native Plants and Putative Annotation of Active Compounds.

Acacia implexa, Eucalyptus rossii and Exocarpos cupressiformis are native plants of Australia, which were used by the First Peoples for medicinal purposes. In this study, 70% aqueous ethanol crude extracts were prepared from A. implexa bark and leaves, E. rossii leaves and E. cupressiformis leaves, and partitioned via sequential extraction with n-hexane, dichloromethane (DCM), ethyl acetate and ethanol. The crude extracts and fractions were screened for antioxidant activity using a novel, high-throughput lipid-based antioxidant assay, as well as the aqueous ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) assay and the Folin-Ciocalteu test for total phenols. In the lipid-based assay, non-polar n-hexane and DCM fractions showed higher antioxidant activity against the formation of peroxides and thiobarbituric acid reactive substances (TBARS) than the other fractions, whereas the non-polar fractions were not effective in aqueous assays. This illustrates that the high potential of the lipid-soluble n-hexane and DCM fractions as antioxidants would have been missed if only aqueous-based assays were used. In addition, the potent antioxidant compounds were putatively annotated using liquid chromatography quadrupole time-of-flight mass spectrometry (LC-qTOF-MS). Gallic acid, (+)-catechin, (-)-epicatechin and tannins were found in most crude extracts.

Anti-nutritional and gas chromatography-mass spectrometry (GC-MS) analysis of ethanol root extract and fractions of Sphenocentrum jollyanum

Abstract Objectives The anti-nutritional composition and gas chromatography-mass spectrometry (GC-MS) analysis of ethanol root extract and fractions of Sphenocentrum jollyanum (SJ) were carried out. Methods The anti-nutritional factors and GC-MS were analysed using standard methods. Key findings The anti-nutritional constituents of the samples were in the order of phenols > terpenoids > flavonoids > tannins > glycosides > alkaloids > hydrogen cyanide > saponins > steroids. The phytochemicals were significantly (P < 0.05) higher in extract than fractions, the terpenoids value obtained as 1904.72 mg/100 g from ethanol extract is significantly higher (P < 0.05) than the values of 935.80 mg/100 g and 968.92 mg/100 g gotten from it ethyl acetate and methanol fractions, respectively. There is no significant difference (P > 0.05) for the ethyl acetate 0.74 mg/100 g and methanol 0.79 mg/100 g fractions of steroids. However, the methanol fraction of tannins, phenols, glycosides, saponins and hydrogen cyanide were significantly higher than its ethyl acetate fractions. Conclusion Chromatogram of GC-MS analysis of the samples of SJ showed 49, 43 and 24 peaks for crude ethanol extract, ethyl acetate and methanol fractions, respectively. GC-MS analysis of the crude ethanol root extract and fractions as shown in Tables 2–4 contain hexadecanoic acid, oleic acid and nonanoic acid methyl ester. The ethyl acetate and methanol fractions of SJ contain 16.9 and 10.1% of 1, 2-benzenedicarboxylic acid, bis (8-methylnonyl) ester, respectively.

Unrivalled insight into potential biopharmaceutical application of Allardia tridactylites (Kar. & Kir.) Sch. Bip.: Chemodiversity, in vitro bioactivities and computational analysis

This research work offers unrivalled insight into the chemodiversity, antioxidant and enzyme inhibition activities of Allardia tridactylites substantiated by computational analysis. Flowers and leaves of A. tridactylites were extracted separately with hexane, chloroform, ethyl acetate and ethanol to prepare four extracts of each part. Highest antioxidant potential for the ATFA and ATFLA fractions was observed in the CUPRAC (cupric ion reducing antioxidant capacity) assay as 115.57 ± 1.16 and 112.79 ± 1.8 mg of trolox equivalents per gram of the extract (mg TE/g extract) respectively, followed by chloroform and ethyl acetate extracts of the leaves and the flowers. In tyrosinase inhibition assay, the ATFE was found most active (28.92 ± 1.42 mg of Kojic acid equivalents per gram of the extract (mg KAE/g extract), while against α-amylase and α-glucosidase, the ATLC (0.63 ± 0.02 mmol of acarbose equivalents per gram of the extract (mmol ACAE/g extract) and ATFH (1.10 ± 0.01 mmol ACAE/g) were the most active respectively. Overall, ATLA and ATFA were rich in phenolics and flavonoids and also exhibited higher antioxidant activities, while in enzyme inhibition assay, the low polar extracts were more active. UHPLC-MS analysis of crude ethanolic extract result in identification of 125 secondary metabolites of various classes. The findings from various multidirectional investigations indicated that this plant held significant importance for the biopharmaceutical industries, as it served as a valuable source for the development of multiple formulations that were effective in targeting major biological enzymes.

In‐vitro Investigation of Anti‐Parasite, Anti‐Oxidant Activities, Cytotoxicity and GC/MS analysis of Ehanolic Extracts for Rhynchosia memnonia var. memnonia and Sonchus oleraceus used in Sudanese Ethnomedicine

AbstractThis study aims to investigate the anti‐parasite, anti‐oxidant activities, cytotoxicity and chemical contents of Rhynchosia memnonia var. memnonia and Sonchus oleraceus aerial parts. The study showed the efficacy of the ethanolic extracts of R. memnonia var. memnonia and S. oleraceus against Giardia lamblia. R. memnonia var. memnonia was a high effect of (76 %) at a concentration of 500 ppm after 72 h, while the highest result of the crude extract of S. oleraceus was (86 %) at the same concentration after 72 h. The antioxidant activity of S. oleraceus ethanolic crude extract showed high activity (72 %). At the same time, the ethanolic extract of R. memnoniavar. memnonia showed weak activity (36 %). This was done using the 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH) assay. The results of cytotoxicity testing revealed that the ethanolic extract of R. memnonia var. memnonia is highly toxic, while the toxicity was slight for the ethanolic extract of S. oleraceus used to brine shrimp. In addition, the cytotoxicity (MTT‐assay) of these plants′ ethanolic extracts against the normal Vero cell line verified the safety of the examined extracts with an IC50 less than 100 μg/ml. In order to detect the constituents of crude extracts, GC‐MS was used. The analysis revealed the presence of 22 compounds in the ethanolic extract of R. memnonia var. memnonia and eleven compounds in the S. oleraceus crude extract. From R. memnonia var. memnonia and S. oleraceus, the main compounds detected were n‐Hexadecadnoic acid and Lup‐20(29)‐en‐3b‐ol, respectively. Also, many other compounds were identified at low area percent from these extracts.

Wound Healing Potential of Couroupita guianensis Aubl. Fruit Pulp Investigated on Excision Wound Model.

Wound care management aims at stimulating and improving healing process without scar formation. Although various plants have been reported to possess wound healing properties in tribal and folklore medicines, there is a lack of scientific data to validate the claim. In this aspect, it becomes inevitable to prove the efficacy of naturally derived products at pharmacological levels. Couroupita guianensis as a whole plant has been reported to exhibit wound healing activity. The leaves and fruit of this plant have been utilized in folkloric medicine to cure skin diseases and infections for many years. However, to the best of our knowledge, no scientific studies have been conducted to verify the wound healing properties of C. guianensis fruit pulp. Therefore, the present study seeks to investigate the wound healing potential of C. guianensis fruit pulp using an excision wound model in Wistar albino male rats. This study indicated that the ointment prepared from crude ethanolic extract of C. guianensis fruit pulp facilitated wound contraction that were evidenced by a greater reduction in the wound area and epithelialization period and increased hydroxyproline content. The experimental groups treated with low and mid dose of C. guianensis ethanol extract (CGEE) ointments had shown a wound closure of 80.27% and 89.11% respectively within 15days, which is comparable to the standard betadine ointment which showed 91.44% healing in the treated groups. Further, the extract influenced the expression of genes VEGF and TGF-β on post wounding days that clearly explained the strong correlation between these genes and wound healing in the experimental rats. The animals treated with 10% CGEE ointment showed a significant upregulation of both VEGF and TGF-β as compared with other test and standard groups. These findings provide credence to the conventional application of this plant in the healing of wounds and other dermatological conditions, and may represent a therapeutic strategy for the treatment of wounds.

In vitro Antibacterial Activities of Selected Medicinal Plants Used by Traditional Healers for Treating Urinary Tract Infection in Haramaya District, Eastern Ethiopia.

Despite the presence of antibacterial agents for urinary tract infection treatment, most of the uropathogenic bacteria reveal multi-drug resistance. Health and economic loss due to these represent a rising burden worldwide which necessitates serious action at regional, national and global levels. Thus, alternative approaches to overcome this problem by using bioactive compounds from traditional medicinal plants are required. This study was designed to evaluate the in-vitro antibacterial activity of Punica granatum fruit peels, Nigella sativa seeds, and Echinops kebericho used in the traditional treatment of urinary tract infections. An experimental study was employed to evaluate the in vitro antibacterial activity of methanol and ethanol crude extract of Punica granatum fruit peels, Nigella sativa seeds, and Echinops kebericho roots of six dilutions (25, 50, 100, 125, 250, and 500) mg/mL. Disc diffusion and macro broth dilution methods were used to determine antimicrobial activity test and minimum inhibitory concentration respectively against E. coli, P. aeruginosa, K. pneumoniae, P. mirabilis, and S. aureus bacterias. Antibacterial activities of ethanol and methanol crude extract of Punica granatum fruit peels against E. coli ATCC25922, P. aeruginosa ATCC27853, S. aureus ATCC25923, K. pneumoniae UK5099 and P. mirabilis UK5999 had highest inhibition zones among tested plants. All tested bacteria were highly sensitive to Punica granatum extract. The second most active plant extract in inhibiting the growth of tested bacteria was Nigella sativa while Echinops kebericho showed the smallest efficacy against tested bacteria. The inhibition zone diameter produced by the methanol extract of each screened plant had higher inhibition zones than ethanol extract. The crude extracts of Punica granatum fruit peels, Nigella sativa seeds, and Echinops kebericho roots have promising antibacterial activity against tested uropathogenic bacteria.

Exploration of anthelmintic potency of Pisonia alba

Helminthiasis or worm infestation is one of the most prevalent disease and one of the most serious problems in the world. Pisonia alba (Ceodeus Grandis) plant was selected for exploration of anthelmintic activity because Nyctaginaceae family plants such as Boerhavia diffusa, Bougainvillea glabra, and Mirabilis jalapa L., already tested for the anthelmintic activity and they have potent anthelmintic activity. The Phytochemical test performed in this ethanolic extract of Pisonia alba. This study indicated that the crude ethanol extract produces anthelmintic activity against the Indian earthworm Pheretima Posthuma. Ethanolic extract of Pisonia alba [20, 40, 60 mg/ml] compared with standard drug (Albendazole). The extract showed significant potency when compared with standard drug Albendazole. The plant explored significant anthelmintic activity at 60 mg/ml concentration in time taken for paralyse /death of the earthworms The current investigation indicates the leaves of Pisonia alba have potent anthelmintic activity the further study is needed to explore its full potential.

The Extract of Perilla frutescens Seed Residue Attenuated the Progression of Aberrant Crypt Foci in Rat Colon by Reducing Inflammatory Processes and Altered Gut Microbiota.

Perilla frutescens (PF) seed residue is a waste from perilla oil production that still contains nutrients and phytochemicals. This study aimed to investigate the chemoprotective action of PF seed residue crude ethanolic extract (PCE) on the inflammatory-induced promotion stage of rat colon carcinogenesis and cell culture models. PCE 0.1 and 1 g/kg body weight were administered by oral gavage to rats after receiving dimethylhydrazine (DMH) with one week of dextran sulfate sodium (DSS) supplementation. PCE at high dose exhibited a reduction in aberrant crypt foci (ACF) number (66.46%) and decreased pro-inflammatory cytokines compared to the DMH + DSS group (p < 0.01). Additionally, PCE could either modulate the inflammation induced in murine macrophage cells by bacterial toxins or suppress the proliferation of cancer cell lines, which was induced by the inflammatory process. These results demonstrate that the active components in PF seed residue showed a preventive effect on the aberrant colonic epithelial cell progression by modulating inflammatory microenvironments from the infiltrated macrophage or inflammatory response of aberrant cells. Moreover, consumption of PCE could alter rat microbiota, which might be related to health benefits. However, the mechanisms of PCE on the microbiota, which are related to inflammation and inflammatory-induced colon cancer progression, need to be further investigated.

Renoprotective Impacts of Inonotus obliquus Ethanol-Ethyl Acetate Extract on Combined Streptozotocin and Unilateral Nephrectomy-Induced Diabetic Nephropathy in Mice.

Diabetes nephropathy (DN) is one of the most common causes of end stage renal disease (ESRD) globally. Medication options to stop or slow the progression of chronic renal disease (CKD) are limited, and patients with DN remain at a high risk of developing renal failure. Inonotus obliquus extracts (IOEs) of Chaga mushroom have been shown to have anti-glycemic, anti-hyperlipidemia, antioxidant, and anti-inflammatory effects against diabetes. In this study, we examined the potential renal protective role of an ethyl acetate layer after water-ethyl acetate separation from Inonotus obliquus ethanol crude extract (EtCE-EA) from Chaga mushrooms in diabetic nephropathy mice after preparation with 1/3 NT + STZ. Our data showed that treatment with EtCE-EA can effectively regulate blood glucose, albumin-creatinine ratio, serum creatinine, and blood urea nitrogen (BUN) levels, and it can improve the renal damage in 1/3 NT + STZ-induced CRF mice with an increase in concentration (100, 300, and 500 mg/kg). In the immunohistochemical staining test, EtCE-EA can effectively reduce the expression of TGF-β and α-SMA after induction according to the increase in the concentration (100 mg/kg, 300 mg/kg), thereby slowing down the degree of kidney damage. Our findings demonstrate that EtCE-EA could provide renal protection in diabetes nephropathy, possibly due to the decreased expression of transforming growth factor-β1 and α-smooth muscle actin.

Rapid and cost-efficient microplate assay for the accurate quantification of total phenolics in seaweeds

Brown seaweeds (Phaeophyceae) are a rich source of polyphenols (up to 20% dry weight) with a structure based on phloroglucinol (1,3,5-trihydroxybenzene). To-date the determination of total phenolics content (TPC) involves a redox reaction with the Folin-Ciocalteu (FC) reagent. However, side reactions with other reducing substances preclude accurate, direct measurement of TPC. This research reports a novel microplate assay involving a coupling reaction between phloroglucinol with Fast Blue BB (FBBB) diazonium salt, at basic pH, to form a stable tri-azo complex with maximum absorbance at 450 nm. Linear regression correlation values (R2) were ≥0.99 with phloroglucinol as standard. Direct quantification of TPCs (phloroglucinol equivalents, PGEs) in crude aqueous and ethanolic extracts from A. nodosum demonstrated that the new FBBB assay is not subject to side-redox interference and provides a more accurate estimate of TPC (1.2–3.9-fold lower than with the FC assay) in a relatively rapid (30 min), cost-effective (0.24€/test) microplate format.

Action of crude ethanol extract of Hymenaea martiana leaf, gallic acid, and polypyrrole (PPy) against Aeromonas hydrophila.

Pisciculture represents one of the industries with the fastest growth rates worldwide. However, it presents obstacles to its development, such as bacteriosis, which is conventionally treated with antibiotics. The indiscriminate and inappropriate use of antibiotics can lead to bacterial resistance, thus alternatives to the use of antibiotics have been researched. The study aimed to analyze the potential of crude ethanol extract (CEE) from Hymenaea martiana leaf, gallic acid (GA), and polypyrrole (PPy) against Aeromonas hydrophila. Tests were performed to determine the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the compounds individually and in synergy (checkerboard) against A. hydrophila and in silico tests between the compounds evaluated. The CEE of H. martiana leaf and PPy were effective against A. hydrophila with MBC results of 3125 μg/mL for the CEE of H. martiana and 125 μg/mL for PPy. Evaluating the GA, a MIC and MBC of 125 μg/mL was obtained. In the interaction tests (checkerboard, using PPy/CEE and PPy/GA), there was a significant reduction in individual introductions. Thus, for the PPy/CEE tests, we had a reduction of MIC/MBC to 1.95 and 781.25 μg/mL, and for the synergy tests between PPy/GA to 7.8125 and 31.125 μg/mL, respectively. The synergy tests are encouraging, and it is possible to verify a decrease of up to 98% in the introduction of PPy, 75% in CEE for H. martiana and 75.1% for GA, when compared to their individual tests. The tests with GA are encouraging due to GA's effectiveness as an antimicrobial agent and high synergy with polypyrrole, both in vitro results and molecular docking experiments showed the actions at the same activation site in A. hydrophila. In vivo tests evaluating isolated components of CEE from H. martiana in synergy with PPy should be performed, to verify the quality of the interactions and the improvement of the immune responses of the animals. It was evidenced that gallic acid, a substance isolated from the extract, tends to have more promising results. This is relevant since the industry has been developing these compounds for different uses, thus providing easier access to the product. Thus, the present study indicates an efficient alternative in the use of bioactive compounds as substitutes for conventional antimicrobials.

Evaluating the Effects of Some Selected Medicinal Plant Extracts on Feed Degradability, Microbial Protein Yield, and Total Gas Production In Vitro

This study evaluates the effect of 22 crude ethanolic plant extracts on in vitro rumen fermentation of Themeda triandra hay using monensin sodium as a positive control. The experiment was run independently three times at 16 and 48 h of incubation periods using the in vitro gas production techniques. Fermentation parameters were determined at both hours of incubation. Plant extracts influenced gas production (GP) in a varied way relative to control at both hours of incubation, and GP is consistently highly significant (p < 0.0001) at 16 and 48 h. Microbial protein yield (MY) was not significantly affected at 16 h (p > 0.05), but it was at 48 h (p < 0.01). Higher MY was recorded for all treatments except for A. sativum and C. intybus at the early incubation stage (16 h) relative to 48 h of incubation. Compared to the control group at 48 h, all plant extracts have higher MY. After 48 h of incubation, the result shows that plant extracts have an effect on fermentation parameters determined; ruminal feed degradation, gas production, microbial protein yield, and partitioning factor in varied manners. All the plant extracts improve the MY which is the major source of amino acids to ruminants and has significant importance to animal performance. C. illinoinensis, C. japonica, M. nigra, P. americana, C. papaya, and A. nilotica (pods) were the most promising plant extracts, but further study is recommended to validate the in vitro observation in vivo.

Phytochemical Characterization of Cannabis sativa L. Roots from Northeastern Brazil.

This article describes the phytochemical study of Cannabis sativa roots from northeastern Brazil. The dried plant material was pulverized and subjected to exhaustive maceration with ethanol at room temperature, obtaining the crude ethanolic extract (Cs-EEBR). The volatile compounds were analyzed by gas chromatography coupled with mass spectrometry (GC/MS), which allowed to identify 22 compounds by comparing the linear retention index (LRI), the similarity index (SI) and the fragmentation pattern of the constituents with the literature. By this technique the major compounds identified were: friedelan-3-one and β-sitosterol. In addition, two fractions were obtained from Cs-EEBR by classical column chromatography and preparative thin layer chromatography. These fractions were analyzed by NMR and IR and together with the mass spectrometry data allowed to identify the compounds: epifriedelanol, friedelan-3-one, β-sitosterol and stigmasterol. The study contributed to the phytochemical knowledge of Cannabis sativa, specifically the roots, as there are few reports on the chemical constituents of this part of the plant.

Antioxidant Activities of Marine Green Algae 4 Species of a Genus Enteromorpha from Mandapam Coast, South India

The present study focused on determining the antioxidant properties of the Enteromorpha compressa (Linnaeus) Nees, Enteromorpha flexuosa (Wulfen) J. Agardh, Enteromorpha intestinalis (Linnaeus) Nees, and Enteromorpha prolifera (Müller) J. Agardh, ethanol crude extracts, from the green algae. The evaluation of antioxidant properties was estimated and determined by using ten standard methods namely 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2’-azino-bis 3-ethylbenzthiazoline-6-sulphonic acid (ABTS), metal chelating activity, hydrogen peroxide radical assay, superoxide radical scavenging (SOD), reducing power, hydroxyl radical assay, nitric oxide (NO) scavenging assay, ferric reducing antioxidant power (FRAP) assay and total antioxidant activity assay. The tested green algae also had antioxidant activity like algae diatom which are showing good antioxidant potential. These algae had a lesser radical scavenging ability than standard ascorbic acid. This study suggests that ethanol crude extracts contain different potential antioxidant compounds capable to scavenge different types of free radicals.

Cytotoxic and genotoxic properties of silver nanoparticles synthesized by ethanolic extract of Salacia chinensis

In this study, in vitro and in vivo methods were used to evaluate the cytotoxicity and genotoxicity properties of silver nanoparticles (Ag-NPs) made from a crude ethanolic extract of Salacia chinensis. The test Ag-NPs had no cytotoxicity on the fibroblast cell line at a concentration of 100 μg/mL, according to the MTT assay results. The Chinese hamster ovary (CHO) cell line treated with varied concentrations of test Ag-NPs, with a maximum concentration of 200 μg/mL, did not exhibit any appreciable genotoxic activity, either by comparing the results with positive controls of genotoxicity caused by Methyl methane sulfonate and Benzo (a) pyrene at the concentration of 20 μg/mL, the lack of genotoxicity was established. An in vivo study in Swiss albino mice using various concentrations (250, 500, and 1000 mg/kg) of test Ag-NPs, which were compared with positive controls, further confirmed this in vitro result pattern. Contrary to the genotoxicity caused by the positive control, mouse bone marrow micronucleus testing findings revealed the absence of genotoxicity. These findings imply that at the measured doses, the Ag-NPs produced from the crude ethanolic extract of Salacia chinensis do not exhibit any cytotoxicity or genotoxicity.

Microalga Chlorella sp. extract induced apoptotic cell death of cholangiocarcinoma via AKT/mTOR signaling pathway

Cancer is the leading cause of death worldwide. Drug resistance and relapse after current standard treatments frequently occur; thus, alternative and effective treatments are required. Algae and cyanobacteria are abundant organisms that serve as bioresources of nutrients/metabolites, which are attractive sources of numerous bioactive compounds for drug discovery. In the present study, we, therefore, investigated anti-cancer activities of crude polysaccharide and ethanolic extracts from Chlorella sp., Sargassum spp., and Spirulina sp. against cell lines of five top-leading cancers including lung cancer (A549), cervical cancer (Hela), breast cancer (MCF7), hepatocellular carcinoma (Huh7), and cholangiocarcinoma (CCA; KKU213A). Only ethanolic extracts of Chlorella sp. showed consistent inhibition of growth of all cancer cell types. CCA was the most sensitive to Chlorella sp. ethanolic extract with CC50 of 277.4, 400.5, and 313.4 µg/mL for KKU055, KKU100, and KKU213A cells, respectively. Flow cytometric analysis demonstrated that CCA cell death was triggered via apoptosis pathway in accompany with lowering procaspase-3, -8, and -9 and increasing caspase enzymatic activity in addition to reducing anti-apoptosis Bcl-2 protein. Interestingly, the treatment of the extract at 400 µg/mL greatly inhibited the AKT/mTOR survival signaling as evidenced by significant reduction of phosphorylated-AKT and phosphorylated-mTOR proteins. The presence of reported bioactive compounds, gallic acid, and lutein, were confirmed in Chlorella sp. extract by high-performance liquid chromatography. Gallic acid and lutein treatment caused a significant reduction of KKU055, KKU100, and KKU213A cell viability. This study demonstrated the anti-cancer effect of Chlorella sp. ethanolic extract to promote cancer cell death via inhibition of AKT/mTOR pathway.

Antioxidant, Anti-inflammatory, Antimicrobial and Thrombolytic Activities of Eclipta alba L. Growing in Bangladesh

Eclipta alba L. (Family: Asteraceae) is an evergreen herb of ethnomedicinal significance in the Indian subcontinent. The present study was carried out to determine the in vitro antioxidant, anti-inflammatory, antimicrobial and thrombolytic activities of n-hexane, chloroform and aqueous soluble fractions of ethanol crude extract of E. alba whole plant. Preliminary phytochemical screening was performed by qualitative tests which revealed the presence of alkaloids, steroids, glycosides, tannins, flavonoids, saponins and amides in E. alba. During the DPPH (2,2-diphenyl-1-picrylhydrazyl) scavenging assay, the aqueous soluble fraction of E. alba at a dose of 100 μg/ml showed maximum 87.5% inhibition in scavenging the DPPH radical with IC50 value of 7.86 μg/ml. Anti-inflammatory activity was evaluated by egg albumin denaturation technique and RBC membrane stabilizing method. In both experiments, all the test samples of E. alba showed a concentration-dependent anti-inflammatory potential while the aqueous soluble fraction at the dose of 500 μg/ml demonstrated the maximum 90.12% inhibition of egg albumin denaturation and 78.42% inhibition of RBC membrane hemolysis against hypotonic solution. Antimicrobial sensitivity was evaluated by disc diffusion assay against several bacterial and fungal species. All the test samples at a concentration of 500 μg/disc exhibited noticeable antibacterial (zone of inhibition = 11-23 mm) and antifungal (zone of inhibition = 10-21 mm) activities when compared to the respective standard drug. The aqueous soluble fraction showed maximum activity against Lactobacillus casai (zone of inhibition = 23 mm). During antifungal test, the maximum activity was exhibited by the n-hexane fraction against Pityrosporum ovale (zone of inhibition = 21 mm). During in vitro thrombolytic assay, the aqueous soluble fraction revealed 56.84% lysis of blood clot, as compared to the standard streptokinase (84.60%). The results of our study suggest that E. alba can be considered for further investigation in order to discover the pharmacologically active natural products. Bangladesh Pharmaceutical Journal 26(1): 20-27, 2023 (January)

Chemometric-Based Analysis of Metabolomics Studiesof Bioactive Fractions of Pleurotus osteratus and Their Correlation with In Vitro Anti-Cancer Activity.

Richness in nutrients with an ample of the myco-bioactive molecules makes Pleurotus osteratus preferential mushroom. In this paper, we conducted a preliminary study on bio-assay-guided fractionation of dichloromethane:ethanol crude extract(1:1, v/v) of P. osteratus (CD) against human breast cancer cell line (MDA-MB-231). Later, CD and its potent hexane (H) and ethyl acetate (EA) fraction were screened against a panel of a human cancer cell lines. H fraction possesses higher cytotoxicity followed by EA and CD. Literature review revealed that polyphenol and ergosterol are the biomarkers found in P. osteratus and could responsible for its cytotoxic potential. Accordingly, hyphenated liquid chromatography with tandem mass spectrometry (LC-MS/MS)-based polyphenol and ergosterol-targeted myco-metabolite profiling of CD, H, and EA fractions were carried out. Despite being significantly rich in polyphenol and ergosterol content, EA fraction showed moderate cytotoxicity. Considering this, liquid chromatography-hybrid quadrupole time-of-flight mass spectrometry (LC-QTOF/MS)-based untargeted myco-metabolite profiling of CD, H and EA fractions was further conducted to identify a new biomarker. Tentatively, 20 myco-metabolites were identified, belonging to the class of steroids, alkaloid, terpenoid, fatty alcohol, and polyketide. The myco-metabolite variabilities among potent samples in correlation to their in vitro anti-cancer activity was explored using the different chemometric tools: principal component analysis (PCA), hierarchical clustering analysis (HCA), and partial least square (PLS). A probable synergistic action among identified myco-metabolites (betulin, solanocapsine, ophiobolin F, linoleoyl ethanolamide, (13R,14R)-7-labdene-13,14,15-triol, asterosterol, cholest-5-ene, (3b,6b,8a,12a)-8,12-epoxy-7(11)-eremophilene-6,8,12-trimethoxy-3-ol, beta-obscurine, myxalamid B, momordol, andavocadyne 4-acetate) may be responsible for the observedcytotoxicity potential of H fraction of P. osteratus.