Diethyl Ether Extract Research Articles

ANTILEISHMANIAL EFFICACY OF HERACLEUM PERSICUM DESF. EX. FISCH. EXTRACTS AGAINST LEISHMANIA TROPICA PROMASTIGOTES

Background: Heracleum persicum known as “suhotu” is widely distributed in Adana, Hakkari Van and used in the production of famous Van herb cheese. The antioxidant, antiinflammatory, antidiabetic, antiepileptic, hepatoprotective, and antifungal properties were previously investigated on H. persicum extracts. Objectives: The purpose of the present study was to comparatively evaluate the in vitro antiLeishmanial potential of H. persicum extracts prepared by solvents having different polarities. Methods: The extracts of aerial parts of H. persicum were obtained by maceration, and in vitro antiLeishmanial efficacy was assayed on L. tropica promastigotes using CellTiter-Glo® Cell Viability assay. Results: The diethylether extract of H. persicum was found to be the most active extract on L. tropica promastigotes with IC50 value at 151.6 µg/ml. The n-hexane extract with 160.2 µg/ml IC50 value exhibited higher antiLeishmanial activity than methanol, ethylacetate and etanol extracts which had IC50 values at 229, 343.5, and 4708 µg/ml, respectively Conclusions: The n-hexane and the diethylether extracts of H. persicum which showed remarkable in vitro antiLeishmanial activities could constitute a valuable source for leishmanicidal compounds. Peer Review History: Received 24 May 2024; Reviewed 11 July 2024; Accepted 21 August; Available online 15 September 2024 Academic Editor: Dr. Asia Selman Abdullah, Pharmacy institute, University of Basrah, Iraq, asia_abdullah65@yahoo.com Average Peer review marks at initial stage: 6.0/10 Average Peer review marks at publication stage: 7.0/10 Reviewers: Ahmad Najib, Universitas Muslim Indonesia, Makassar, Indonesia, ahmad.najib@umi.ac.id Dr. Sangeetha Arullappan, Universiti Tunku Abdul Rahman, Malaysia, sangeetha@utar.edu.my

Investigation into the impact of certain leaf extracts derived from Inula viscosa L. on various strains of pathogenic bacteria in poultry

The study was designed to investigate the efficacy of acetone, ethanol, hexane, diethyl ether, and aqueous extracts from clay plants against four pathogenic bacteria commonly found in poultry: Escherichia coli, Streptococcus aureus, Pseudomonas aeruginosa, and Clostridium perfringens. The research took place in the microbiology and drug laboratories of the Faculty of Pharmacy at the University of Tartous, as well as the microbiology laboratory at the Faculty of Agricultural Engineering at Tishreen University. Tayon plant leaves were harvested in October and November of 2021 from the Safita region, dried, and stored for future use. The susceptibility tests were carried out on the four bacteria under investigation using the five different extracts. The findings revealed that Escherichia coli, Streptococcus aureus, Pseudomonas aeruginosa, and Clostridium perfringens were all impacted by the extracts, except for Escherichia coli which displayed resistance to acetone, ethanol, hexane, and diethyl ether extracts, but not to the aqueous extract. Pseudomonas aeruginosa also exhibited resistance to the diethyl ether extract at concentrations of 20 and 40 μl, with no inhibition zone observed. At a concentration of 80 μl, the largest average diameter of the bacterial growth inhibition zone was observed for the acetone extract (4.82, 24.95, 15.41, and 22.14 mm), ethanol (9.32, 28.11, 22.98, and 29.24 mm), hexane (6.42, 18.91, 13.23, and 17.47 mm), diethyl ether (9.33, 21.22, 9.56, and 18.75 mm), and aqueous extract (23.45, 26.22, 15.71, and 24.12 mm), respectively, in comparison to concentrations of 20 and 40 microliters and the control. These results suggest that the plant leaf extracts possess antimicrobial properties against the pathogenic bacterial strains tested, indicating their potential as natural sources of antibiotics in the future.

Biochemical and molecular docking-based assessment of Spirulina platensis's bioactive constituents for their potential application as natural anticancer drug

Spirulina platensis and its extracts possess positive benefits for human health indications from malnutrition, antioxidant properties, and therapeutic properties. This study investigates the antioxidant activity of Spirulina platensis extracts and their cytotoxicity against the breast cancer cell line (MCF-7). The antioxidant and cytotoxicity properties of C- phycocyanin (C-PC) were directly compared with the diethyl ether extract of S. platensis (DEE). Results indicated that C-PC exhibited superior antioxidant activity compared to DEE extract. The observed values for C-PC are 1.453 μg/mL (IC50-value), 299.6 μM AA eq/mL, 86.68 μM TE eq/mL, and 179 μg/g for ABTS assay, DPPH, FRAP and total antioxidant activity, respectively. Moreover, C-PC also demonstrated enhanced cytotoxic activity against MCF-7 tumor cells with IC50 values of 100 μg/mL and 630 μg/mL for C-PC and DEE; respectively. C-PC treatment downregulated the expression of PI3K, Akt, mTOR, and Wnt. C-PC treatment to MCF-7 cells significantly induced caspase 3, caspase 8, and caspase 9. Similar effects, with lower tendency, were also observed for DEE extracts of S. platensis. Molecular docking studies suggest that S. platensis' bioactive constituents can induce apoptosis through intrinsic pathways. The current findings provide evidence for the medical importance of Spirulina platensis active constituents as promising natural anticancer drugs.

Metabolome mapping and DNA fingerprinting of Tribulus terrestris from three different Egyptian populations with the assessment of their morphological characters and antioxidant activity

Tribulus terrestris L. is a cosmopolitan annual herb that grows under various ecological conditions, reflecting the variation in its morphological, chemical, and biological features. T. terrestris was collected from three different populations (I, II, III) in Egypt and subjected to comparative investigation to evaluate the morphological, metabolomic, and genetic variations among these populations. Additionally, the antioxidant activity of the collected specimens was assessed. Significant variations in vegetative morphology were observed among the three populations. Detailed microscopic examination revealed unique traits in the foliar epidermal and pollen grains characteristics of each population. LC-ESI-MS/MS analysis of aqueous methanol extracts led to the tentative identification of 101 compounds across 17 different chemical classes from which flavonoids are predominant and represented by 28 compounds; Polymethoxylated flavones were reported for the first time in the three investigated extracts. GC/FID analysis of diethyl ether extracts identified 12 saturated and 8 unsaturated fatty acids, with linolenic acid being the major component, constituting 52.21 %, 65.71 %, and 49.52 % in populations I, II and III, respectively. The DNA fingerprinting showed variations with 54.54 % polymorphism in ISSR markers, confirming the existence of two distinct genotypes among the studied T. terrestris plants. Furthermore, the extracts from the three populations exhibited weak antioxidant activity against DPPH. The obtained data revealed considerable morphological, metabolomic, and genetic diversity among the three populations of T. terrestris. These findings suggest treating T. terrestris (I) as a distinct variety of T. terrestris and enhancing the understanding of the plant's diversity, which is essential for its effective application in traditional and modern medicinal practices.

Phenolic profile, fatty acid and mineral composition with antioxidant, antibacterial, and enzyme inhibitor activities of different extracts from Erodium Cicutarium (L.) L’Hér. consumed as a vegetable in Kilis, Turkey

This study aimed to explore the antioxidant activities, phenolic profile, fatty acid and mineral composition of the different extracts of Erodium cicutarium (L.) L’Hér. consumed as a vegetable in Kilis, Turkey. While catechine hydrate was the most abundant phenolic compounds in methanol, ethanol and diethyl ether extracts, it was chlorogenic acid in the water extract. Looking at the fatty acid profile, the amount of palmitic acid, one of the saturated fatty acids, was found to be high (34.30%), followed by stearic acid (5.10%). Total monounsaturated fatty acids are the second highest fatty acids. Total polyunsaturated fatty acids were determined as the third rank fatty acids. While the amount of linoleic acid, one of the total polyunsaturated fatty acids, was determined as 17.62%, the ratio of linolenic acid was determined as 9.60%. While the most calcium, magnesium and potassium were found among the 9 different mineral substances determined, respectively; the lowest element was found to be nitrogen. Looking at the results, it was determined that the plant is a high source of calcium (1078.503 mg/kg). Inhibitory effects against Pseudomonas aeruginosa ATCC 9027 were observed, with a zone diameter of 5.5 mm and 11 mm in methanol extract and diethyl ether extract, respectively. Ethanol and water extracts of E. cicutarium (L.) L.‘Hér. may be preferred as an alternative natural agent due to their amylase and tyrosinase activities. The results suggested that the E. cicutarium (L.) L.‘Hér. extracts may be useful for food and medicinal applications.

A New Rare Halogenated Depside from Lichen and Study of its Anti-Proliferative Activity.

Lichens are a symbiotic association of algae and fungus, belonging to the family Parmeliaceae. Some lichen species are edible and used as an active ingredient for preparation of exotic spices as well as folklore medicine to cure different kinds of ailments. A specimen of lichen was collected from Munner in the Kerala State of South India for chemical profiling. Chemical analyses of the diethyl ether extract of the defatted lichen led to the isolation of six phenols 1-6 with variation of relative abundance. Amongst them, the relative abundance of compound 3 was the greatest (1 % of crude extract) and it was identified as atranorin. The structures of known compounds were confirmed by comparison of their 1H-NMR, 13C NMR, and mass data with published values available in the literature. In vitro bioassay for anti-proliferative activity of these compounds has been conducted against various human cancer cell lines in comparison with paclitaxel as control using SRB assay. Interestingly, a new compound 5 was found along with previously reported compounds from this lichen. This new compound was designated as fluoroatranorin 5 which was reported for the first time herein. The structural characterization of a new depside was determined by spectral methods such as 1H-NMR, 13C NMR, 19F NMR, IR, LC-HRESI-MS, and LC-MS/MS study. Its structure was confirmed by single crystal X-ray diffraction study. This new compound was designated as fluoroatranorin 5 which was reported first time herein. Anti-proliferative activity of all these compounds was evaluated against six different cancer cell lines. The inhibitory activity, IC50 value of compounds 1-3 and 5 exhibited at 99.64, 102.04, 109.20, 53.0 and 2.4 μM on cancer cell lines HT-29 (colon), Hela (cervical), HT-29, HPAC (pancreas) and A2780 (ovarian cancer cell line) respectively in comparison with paclitaxel as control. The new compound 5 exhibited significant activity with IC50 value 2.4 μM on A2780 ovarian cancer cell line.

Immunomodulatory Effects of Mallotus japonicus Extract on Innate Immune Responses in HeteroTilapia buttikoferi Infected with Aeromonas hydrophila

The intensive use of antibiotics in modern aquaculture has been linked to the emergence of multidrug resistance in human health. Despite continuous caution against such use by farmers, antibiotics serve as the primary means of disease control in aquaculture. In this study, we explore the potential of medicinal plant extracts as immunomodulant in enhancing the innate immune defense of a fish host, HeteroTilapia buttikoferi, infected with the prevalent fish pathogenic bacteria Aeromonas hydrophila. Our previous research reveals the bactericidal activity of the medicinal plant, Mallotus japonicus, against the pathogen, but its effectiveness as a prophylactic in enhancing host immune defense has not yet been investigated. Therefore, the objective of this study is to address this knowledge gap.By our findings, it was uncovered that the supplementation of the diethyl ether extract of the plant’s bark as an additive in the fish diet significantly improved the innate immune defense by upregulating the total levels of lysozyme, serum bactericidal activity, globulin and albumin levels, total protein, phagocytosis, respiratory burst activity and overall survival. A dose-dependent increase in immunological parameters was recorded, and this was in sharp contrast to the control group.These findings suggest that M. japonicus has the potential of use to enhance the innate immune defense of H. buttikoferi against the pervasive fish pathogenic bacteria, Aeromonas hydrophila. The improvements in immune parameters indicate the ability of M. japonicus extract to strengthen the hosts to combat bacterial infections.

Eco-friendly bio-larvicidal and antimicrobial activity of isolated bioactive compound from Kurthia gibsonii (Bacillales)

The targeted organisms include mosquito vectors, bacterial pathogens and non-targeted organisms. Preliminary mosquito larvicidal activity was conducted using cell-free supernatants (CFSs) from 11 gut bacteria. Among them, the bacterium SS11 exhibited promising results and was identified as Kurthia gibsonii based on its 16S rRNA sequence (1350 bp). The diethyl ether extract (DEE) of K. gibsonii demonstrated significant larvicidal effects, with LC50 values of 5.59 µL/mL and 8.59 µL/mL for 3rd instar larvae of Aedes aegypti and 2nd instar larvae of Anopheles stephensi, respectively. Analysis of the DEE using FT-IR, and GC-MS revealed the presence of 16 functional groups, and 7 bioactive compounds, respectively. A molecular docking study identified GC-MS compounds against odorant receptors from A. aegypti and odorant-binding proteins from A. stephensi was performed to assess the interaction and binding affinity. Overall, these findings suggest that the bioactive compounds 2, 4, 6-tribromoaniline from the DEE of K. gibsonii hold potential as an environmentally compatible alternative for biocontrol purposes, and compounds 9-tricosene and didecyl phthalate can be used for mosquito traps.

Isolation of a novel quercetin derivative from Terminalia chebula and RT-PCR-assisted probing to investigate its DNA repair in hepatoma cells.

DNA damage can lead to carcinogenesis if replication proceeds without proper repair. This study focused on the purification of a novel quercetin derivative present in Terminalia chebula fruit and studied its protective role in hepatoma cells due to H2O2-DNA damage. The pure compound obtained from the silica gel column was subjected to structural characterization using spectroscopic techniques. MTT assay was employed to select a non-toxic concentration of the isolated compounds on HepG2 and Chang liver cells. The antigenotoxic property of the compound on HepG2 and Chang liver cells was carried out by alkaline comet assay. Analyses of expression levels of mRNA for two DNA repair enzymes, OGG1 and NEIL1, in HepG2 and Chang liver cells, were carried out using the RT-PCR method. The pure compound obtained from the fraction-5 of diethyl ether extract was identified as a novel quercetin derivative and named 7-(but-2-en-1-yloxy)-2-(4(but-2-en-1-yloxy)-3-hydroxyphenyl)-3- (hexa-2,4-dien-1-yloxy)-6-hydroxy-4H-chromen-4-one. This compound recorded modest toxicity at the highest concentration tested (percentage cell viability at 100 μg/mL was 64.71 ± 0.38 for HepG2 and 45.32 ± 0.07 for Chang liver cells). The compound has demonstrated noteworthy protection against H2O2-induced DNA damage in both cell lines. Analyses of mRNA expression levels for enzymes OGGI and NEIL1 enzymes in HepG2 and Chang liver cells asserted the protective role of the isolated compound against H2O2-induced DNA damage. The protective effect of a novel quercetin derivative isolated from T. chebula in the hepatoma cells is reported here for the first time.

Phytochemical characterization and antifungal activity of Marchantia polymorpha L. against Rhizoctonia solani

IntroductionMarchantia polymorpha L. is a common liverwort in the family Marchantiaceae. It is used in traditional Chinese herbal medications to heal cuts, scalds, snake bites, fractures, burns and open wounds. Chinese people used it to treat jaundice and inflammation also. Polyphenols (bis-bibenzyls and flavonoids), long-chain polyunsaturated fatty acids and terpenoids have been identified as the primary metabolites in M. polymorpha by phytochemical studies. The present study was aimed to explore the phytochemicals and the fungicidal properties of M. polymorpha extracts against Rhizoctonia solani by Poisoned Food Technique. MethodsThe plant was extracted in four different solvents (Acetone, Methanol, Hexane and Di-ethyl ether) for assessing antifungal activity by Poisoned Food Technique and for phytochemical characterization by GC–MS and LC-MS. The extract with best antifungal activity was further examined for its mode of action using FE-SEM and Fluorescence microscopy. ResultsSignificant fungal growth inhibition was caused by M. polymorpha extracts. The highest inhibition was shown by di-ethyl ether extract (PI= 77.25±0.68). Field Emission Scanning electron microscopy and Fluorescence microscopy depicted the ultrastructural changes in the fungal species induced after treatment with the plant extract. GC–MS analysis revealed the presence of various bioactive compounds in di –ethyl ether extracts such as n- Hexadecanoic acid, Stigmasterol and Phytol with reported pharmacological properties. LC-MS revealed the presence of three antifungal compounds: Marchantin A, Marchantin B. and Plagiochin E. DiscussionThe antifungal properties of M. polymorpha were equivalent to the standard, Amphotericin, which had significant effects against R. solani, indicating good efficacy of the plant extract. The damaging effect of plant extract was further confirmed by FE-SEM and Confocal microscopy. Thus, the present study potentially introduces a solution to the existing dilemma caused by antifungal agrochemicals.

Selective peripheral tissue response to high testosterone levels in an infertile woman without virilization signs.

Total testosterone, which is peripherally converted to its biologically active form dihydrotestosterone (DHT), is the first-line hormone investigation in hyperandrogenic states and infertility in premenopausal women. Polycystic ovary syndrome (PCOS), the most common cause of hyperandrogenism and infertility in young women, is often associated with mild elevations of total testosterone. Whereas very high levels of total testosterone (>2-3 SD of normal reference), are most often associated with hyperandrogenic signs, menstrual irregularity, rapid onset of virilization, and demand a prompt investigation. Herein, we report a case of a 32-year-old woman who was referred to the endocrinology outpatient clinic due to secondary amenorrhea and extremely high testosterone levels without any virilization signs. We initially suspected pitfalls in the testosterone laboratory test. Total serum testosterone decreased after a diethyl-ether extraction procedure was done prior to the immunoassay, but testosterone levels were still elevated. An ovarian steroid-cell tumor (SCT) was then revealed, which was thereby resected. Twenty-four hours post surgery, the total testosterone level returned to normal, and a month later menstruation resumed. This case emphasizes that any discrepancy between laboratory tests and the clinical scenario deserves a rigorous evaluation to minimize misinterpretation and errors in diagnosis and therapeutic approach. Additionally, we describe a possible mechanism of disease: a selective peripheral target-tissue response to high testosterone levels that did not cause virilization but did suppress ovulation and menstruation. Total testosterone is the most clinically relevant hormone in investigating hyperandrogenic states and infertility in premenopausal women. Very high total testosterone levels in women (>2-3 SD of normal reference) are most often associated with hyperandrogenic signs, menstrual irregularities, and a rapid onset of virilization. In women with very elevated testosterone levels and the absence of clinical manifestations, laboratory interference should be suspected, and diethyl ether extraction is a useful technique when other methods fail to detect it. Ovarian steroid cell tumors (SCT) encompass a rare subgroup of sex cord-stromal tumors and usually secrete androgen hormones. SCTs are clinically malignant in 25-43% of cases. A selective response of peripheral target tissues to testosterone levels, with clinical manifestations in some tissues and no expression in others, may reflect differences in the conformation of tumor-produced testosterone molecules.

Effects of Cultivation Stress on the Glaucophyte Cyanophora paradoxa and Bioactive Potential in Human Cancer Cell Lines

ABSTRACT The propensity to accumulate high-value metabolites under specific conditions makes microalgae promising candidates for commercial applications. Here, the freshwater microalga Cyanophora paradoxa CCAP 981/1 was cultivated under varying conditions (control, nitrogen limitation, NaCl supplementation and blue light exposure) and the biological activity of the corresponding biomass extracts was assessed against human breast (MCF-7) and lung (A549) cancer cell lines. The stress treatments hindered the growth of C. paradoxa, the control displaying the highest specific growth rate (0.16 d−1 ± 0.01). Biomass screening by GCMS and HPLC identified diverse bioactive compounds including pigments (chlorophylls, xanthophylls, carotenes) and fatty acids (e.g., EPA). The control cultivation conditions returned the highest yields of biomass (724 mg), zeaxanthin (6.8 mg g−1), EPA (1.1 mg g−1) and phycocyanin (113.9 mg g−1), while the blue light treatment enhanced the proportions of unsaturated forms of C18 (34.3%). Extracts of dried biomass obtained by cold maceration using solvents of increasing polarity were used in the treatment of MCF-7 and A549 cancer cells. Ethyl acetate extracts from nitrogen-limited cultures reduced MCF-7 viability (~70%) after 72 h. However, the diethyl ether and methanol extracts showed no impact on MCF-7 and A549 cell viability. Furthermore, acridine orange staining revealed no significant DNA damage in these cell lines upon exposure to C. paradoxa extracts. Overall, the minimal impacts on cell viability observed here supports considering C. paradoxa as a promising source of natural bioactive compounds with applications in various biotechnological sectors.

Phytochemical composition of the diethyl ether extract of Artemisia lactiflora Wall. ex DC and its antimetastatic activity in human lung cancer cells

Artemisia lactiflora Wall. ex DC. is a traditional Chinese medicinal plant used in the treatment of menstrual and hepatic disorders due to its antioxidant and anti-inflammatory properties. However, its anti-metastatic activity, which is the clinical challenge of lung cancer treatment, has not yet been reported. From the diethyl ether extract of Artemisia lactiflora, the four terpenoids, including dihydroactinidiolide, megastigmatrienone, alpha-curcumene, and dehydrovomifoliol, were the most intense peaks observed using LC-MS/MS, whereas bis (2-ethylhexyl) phthalate was a contaminant. In a transwell assay, the A. lactiflora diethyl ether extract (32 μg/ml) and dihydroactinidiolide (250 μg/ml) markedly inhibited the migration and invasion of non-small cell lung cancer (NSCLC) cells, similar to the standard anti-metastatic drug (capmatinib). Western blot analysis revealed that mesenchymal N-cadherin is downregulated in NSCLC cells under the treatment conditions. The potential anti-metastatic property of dihydroactinidiolide is promising as a new candidate anti-metastatic agent for lung cancer treatment.

Anti-lipidemic and protein restoration potential of Monodora myristica (Gaertn.) in rats fed with cassava containing crude oil

Consumption of cassava meal affected by crude oil has significant effects on lipid and protein metabolism. The hepatoprotective action of spices is mostly attributed to the suppression of lipid oxidation and protein breakdown. This study examined the protein restoration and anti-lipidemic potential of Monodora myristica (Gaertn.) in rats fed with cassava contaminated with crude oil.
 The research involved 36 albino rats separated into six groups (n = 6). Group 1 (control) consumed cassava without crude oil. Group 2 received cassava with crude oil. Groups 3, 4, and 5 fed on cassava with crude oil and various extracts of M. myristica, i.e., aqueous, ethanol, and diethyl ether, respectively. Group 6 received non-ionic synthetic surfactant Tween 80. The experiment relied on standard methods.
 Blood serum and liver obtained from the rats of Group 2 showed a significant (p < 0.05) increase in total cholesterol, low density lipoprotein cholesterol, triacylglycerol, and malondialdehyde, as well as a decrease in total protein, albumin, and high-density lipoprotein cholesterol. The groups that received M. myristica extracts showed a significant increase (p < 0.05) in total protein, albumin, and high-density lipoprotein cholesterol. They also had lower total cholesterol, low density lipoprotein cholesterol, triacylglycerol, and malondialdehyde as compared to Group 2, which dieted on cassava contaminated with crude oil without additives.
 In this research, crude oil-contaminated cassava affected proteins and lipids in rats. Diethyl ether extract of M. myristica demonstrated the best anti-lipidemic and protein restoration.

Recent Updates Regarding the Antiproliferative Activity of Galium verum Extracts on A375 Human Malignant Melanoma Cell Line.

The biological activity of Galium verum herba was exerted on various tumor cell lines with incredible results, but their potential effect on malignant melanoma has not been established yet. Therefore, the current study was structured in two directions: (i) the investigation of the phytochemical profile of diethyl ether (GvDEE) and butanol (GvBuOH) extracts of G. verum L. and (ii) the evaluation of their biological profile on A375 human malignant melanoma cell line. The GvDEE extract showed an FT-IR profile different from the butanol one, with high antioxidant capacity (EC50 of GvDEE = 0.12 ± 0.03 mg/mL > EC50 of GvBuOH = 0.18 ± 0.05 mg/mL). The GvDEE extract also showed antimicrobial potential, especially against Gram-positive bacteria strains, compared to the butanol extract, which has no antimicrobial activity against any bacterial strain tested. The results regarding the antitumor potential showed that both extracts decreased A375 cell viability largely (69% at a dose of 55 µg/mL of the GvDEE extract). Moreover, both extracts induce nuclear fragmentation by forming apoptotic bodies and slight chromatin condensation, which is more intense for GvDEE. Considering the results, one can state that the Galium verum herba possesses antitumor effects on the A375 human malignant melanoma cell line, a promising phytocompound for the antitumor approach to skin cancer.

Effect of solvents on aniseed aerial plant extraction using soxhlet and ultrasound methods, regarding antimicrobial activity and total phenolic content

Pimpinella anisum, commonly known as aniseed or anise, is a flowering plant whose fruits, characterized by distinct aroma and flavor, are widely used in Mediterranean gastronomy. This study aimed to evaluate the effect of the extraction method and solvent on the antimicrobial properties of aniseed extracts. For this purpose, hexane, diethyl ether, ethanol and aqueous extracts of aniseed aerial parts were obtained through Soxhlet and Ultrasound-Assisted Extraction (UAE) methods. The extraction yield was calculated and the antimicrobial activity against food spoilage non-pathogenic microorganisms was measured through Disc Diffusion Assay (Kirby Bauer Test). The study indicated that the Soxhlet extraction method results in higher yield and lower total phenolic content than UAE. All the extracts exhibited antimicrobial activity against certain strains, however, the activity of UAE samples were more significant. In contrast, the ethanolic extract delivered by UAE was the only one that inhibited the growth of all the tested microorganisms (bacteria, yeast and fungi). The analysis of phytochemicals revealed that ethanolic extracts contained petroselinic acid. Finally, regarding the antioxidant activity, water extract with UAE presented the strongest, that was similar with that of ascorbic acid. The observed effects provided a good starting point for further studies on Pimpinella anisum extracts for future applications in the food industry.

Evaluation of anticancer, immunomodulatory and anti-inflammatory potential of antioxidant rich Anzia ornatoides, a lichen species from eastern Himalayan region

The lichen Anzia ornatoides was identified by its morpho-anatomical and chemical features. The hexane, diethyl ether, ethyl acetate, methanol, and water extracts of the lichen were assessed for their antioxidant capacities. The presence of various phyto-compounds of the methanol extract were run through a GC–MS and identified three major compounds viz. methyloxyolivetol, imidazole 2-t‑butyl‑1,4-dimethyl-5-phenyl and benzoic acid 2,4-dihydroxy-3,6-dimethyl-methyl ester. The crude extraction of a lichen sample was carried out by Soxhlet apparatus. Total phenolic content was determined by Folin-Ciocalteau assay while flavonoid content was estimated using aluminium chloride assay. The antioxidant activity of the extracts was analysed by phosphomolybdenum assay, FRAP, DPPH, ABTS, and lipid peroxidation inhibition. To see the immunomodulatory and anti-inflammatory potential of the extracts, cell viability assay of macrophage was studied. In in-vitro anticancer activity, the extracts were explored using cancer cell line of different organ and tissue origin it includes PC-3, OVCAR-3, hep-G2, h-1299, HeLa, and MCF-7. Apoptotic potential is studied using the OVCAR-3 cell line. In-vitro cytotoxicity assay was conducted with the non-cancer cell lines HEK-293, L-929, hMSCs, and MCF-10A. Using mouse model, in-vivo animal toxicity was also carried out. The lichen extracts showed a good amount of phenolic and flavonoid content and high antioxidant potential. Based on the findings of the experiment, methanolic extracts of A. ornatoides showed higher immunomodulatory, anti-inflammatory and anticancer capabilities. In an in-vitro cytotoxicity test against the cancer cell line IC50 value ranged from 38 to 78 µg/ml. Based on in-vivo toxicological evaluation and histopathological analysis, no significant toxicity was detected in-vitro cultured non-cancerous cells or in the liver or kidney tissue of mice.

Phytochemical characterization, in vivo and in vitro biological activities of Atriplex lindleyi Moq. subsp. inflata

Ethnopharmacological relevanceAtriplex lindleyi Moq. subsp. inflata from Algeria is known for its traditional use for therapeutic properties. Aim of the studyWe analysed quantitatively and qualitatively the infusion and diethyl ether, n-butanol and ethereal extracts of A. lindleyi, as well as assessed their acute toxicity, anti-inflammatory, antinociceptive and antioxidant activities. Materials and methodsThe total phenolic and flavonoid contents were quantified. The anti-inflammatory effect was assessed using a carrageenan-induced paw oedema assay and the antinociceptive effect was evaluated using an acetic acid-induced writhing method. In addition, antioxidant activity was examined by three tests: DPPH, reducing power and phenanthroline assays. ResultsThe quantity of total phenols in the roots of A. lindleyi was larger than in the aerial parts, while the amounts of total flavonoids were larger in the aerial parts than in the roots. The HPLC profiles allowed us to identify 32 compounds belonging to the phenolic acid and flavonoid classes. Intraperitoneal administration of the infusions and phenolic extracts in mice did not cause any symptoms of toxicity or mortality. Results revealed that the aerial parts and roots of A. lindleyi had potential anti-inflammatory and antinociceptive activities and were higher than diclofenac and paracetamol, respectively. The diethyl ether extract (DEE) of aerial parts and roots showed the greatest antioxidant activity in comparison to the other tested extracts. ConclusionThe phenolic composition attempted to be identified by HPLC confers this plant's pharmacological potential. The antioxidant potential may be due to active ingredients, including ascorbic acid, in the two studied parts of A. lindleyi, which is a bioactive molecule with strong antioxidant properties. Also, detecting salicylic acid, the active antinociceptive ingredient of aspirin, gallic and ferulic acids may justify this subspecies' antinociceptive and anti-inflammatory potentials.

Neuroprotective and Antioxidant Activities of Algerian Punica granatum L. Peel Extracts

Background: Oxidative stress is implicated in many neurodegenerative disorders, such as Alzheimer’s disease. As an alternative, Punica granatum peels are widely used in the treatment of several ailments. To date, this is the first study on the anti-acetyl/butyrylcholinesterase effects of diethyl ether extract from Algerian pomegranate peels. Objective: This research aimed to evaluate the content of total phenolics, flavonoids, flavonols, and tannins in Punica granatum peel crude extract and its fraction diethyl ether, as well as to assess its antioxidant and neuroprotective activities. Methods: The total phenolic, flavonoid, flavonol, and tannin contents in the extracts were evaluated using spectrophotometric methods. The inhibitory effects on acetyl/butyrylcholinesterase were assessed using the in vitro models. In addition, several methods were used to investigate the antioxidant capacity of extracts, including 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline- 6-sulfonate) (ABTS) free radicals scavenging, phenanthroline, reducing power and cupric ion reducing antioxidant capacity (CUPRAC) assays. Results: Phytochemical analysis revealed that the diethyl ether extract recorded the highest content of total phenolics, flavonoids, flavonols, and tannins with values of 643.3 ± 9.96 μgGAE/mg, 78.97 ± 9.97 μgQE/mg, 31.32 ± 1.79 μgQE/mg and 96.6 ± 13.65 μgTAE/mg, respectively. Moreover, the diethyl ether extract exhibited the strongest activity in the DPPH scavenging assay (IC50=3.36 ± 0.11 μg/mL), ABTS scavenging assay (IC50=1.46 ± 0.10 μg/mL), CUPRAC assay (CA0.5=3.62 ± 0.36 μg/mL), phenanthroline assay (CA0.5=1.04 ± 0.10 μg/mL), reducing power assay (CA0.5=4.14 ± 0.32 μg/mL), antiacetylcholinesterase assay (IC50=114.86 ± 13.66μg/mL), and anti-butyrylcholinesterase assay (IC50=147.00 ± 10.41 μg/mL). Conclusion: The present study has shown the extracts from Punica granatum peel to be a rich source of bioactive compounds, potent antioxidants, and cholinesterase inhibitors. They could be considered as natural agents that can be used in anti-neurodegenerative pharmaceutical formulations.

Characterization of ACE inhibitory activity in Dioscorea alata cv and its implication as a natural antihypertensive extract

Ethnopharmacological relevanceYam (Dioscorea sp.) extracts have been shown to possess a vast array of medicinal properties such as antihypocholesterolemic, antiatherogenic and antihypertensive bioactivity. However, the compounds conferring its antihypertensive bioactivity have not been fully explored. Aim of the studyThe objective of this study was to identify extractable bioactive fractions and associated compounds in Jamaican Renta Yam (Dioscorea alata) that contribute to its antihypertensive properties, using an activity driven chemoinformatic profiling method. Materials and methodsA diethyl ether extract of Dioscorea alata was obtained by sequential Solid-Liquid extraction coupled to SPE-HPLC fractionation and its chemical composition was analyzed by GC-MS analysis. Its influence on hypertension was evaluated through a combination of in vitro ACE-Inhibitory activity assays and by molecular docking of the identified compounds to the ACE enzyme. ResultsSLE revealed the presence of potent antihypertensive activity (ACE IC50 41.99 μg/mL) in the diethyl ether extract (DR2). GC-MS analysis of DR2 indicated the presence of small organic compounds (95.1 g/mol to 200 g/mol) with 2-Phenyl-1,3-oxazol-2-ine (2PO) being the most predominant small organic compound present in the bioactive extract. The binding affinity of 2PO was assessed using molecular docking of 2PO to the ACE enzyme and showed strong binding affinities forming two (2) hydrogen bonds with Tyr135 and Trp220 in the active site of the enzyme. The in vitro effect of DR2 using human umbilical vein endothelial cell lines (HUVECs) revealed; a significant dose-dependent ACE-Inhibitory activity, a stimulating of nitric oxide (NO) release and no toxicity towards these cells. ConclusionOverall, this study identified Jamaican Renta Yam (Dioscorea alata) as an alternative source of antihypertensive compounds which may address the toxicity seen with known synthetic antihypertensive agents.