Crude Extract Research Articles

Proteomic Analysis and Biochemical Characterization of the Nematocyst Extract of the Hydrozoan Velella velella.

The venom contained within cnidarian nematocysts has a complex composition and holds significant potential for biotechnological applications. In this context, one of the most effective methods for studying nematocyst contents is the proteomic approach, which can detect even trace amounts of compounds while minimizing the need for large-scale animal collection, thus helping to preserve ecosystem integrity. This study aimed to provide a comprehensive proteomic and biochemical characterization of the crude nematocyst extract from the common hydrozoan Velella velella. Despite not being harmful to humans, the analysis of the crude venom extract from V. velella brought to the identification of 783 different proteins, categorized into structural components, enzymes, and potential toxins, revealing a qualitative composition of the venom similar to that of other more toxic cnidarians. Biochemical assays confirmed the presence of various active hydrolytic enzymes within the extract, including proteases, phospholipases, hyaluronidases, DNases, and chitinases. These findings pave the road for future studies involving the pharmacological applications of Velella velella venom components through recombinant production and functional testing.

Olax dissitiflora Oliv. (small-fruited olax): Current status and future prospects as medicinal plant

Olax dissitiflora Oliv. (O. dissitiflora) has several medicinal uses, some of them known since prehistoric times. The present review compiles existing knowledge of the chemical and pharmacological properties, current medicinal uses, and further use potential and applications of O. dissitiflora. Multiple searches on existing literature on the medicinal, phytochemistry and pharmacological properties of O. dissitiflora were conducted in online databases such as Scopus, JSTOR, PubMed, Google Scholar and Science Direct as well as using pre-electronic literature sources obtained from the university library. This study showed that the bark, leaves, roots and twigs of O. dissitiflora are used as aphrodisiac, emetic, mosquito repellent and purgative, and as traditional medicines against dental problems, gonorrhoea, hernia, venereal diseases and wounds. The phytochemical evaluation of the plant revealed that it contains reducing sugars, alkaloids, flavonoids, phenolic compounds, quinones, saponins, steroids, tannins, terpenoids, 21-epimachaerinic acid, oleanolic acid, hederagenin, oleanolic acid 3-O-glucuronide, amphotericin, santalbic acid, exocarpic acid and octadic-9,11-diynoic acid. The pharmacological assessments showed that the crude extracts and phytochemical compounds isolated from the species have antibacterial, antifungal, antiplasmodial, larvicidal and adulticidal activities. The collected ethnomedicinal and ethnopharmacological properties of O. dissitiflora documented in this review will assist to understand the therapeutic relevance and potential of the species as well as providing baseline data required for ethnopharmacological assessments required in the discovery of novel plant-based health products. There is also need for more studies focusing on phytochemistry, pharmacological properties and toxicological evaluations, in vivo and clinical research on O. dissitiflora to compliment and validate the ethnomedicinal properties of the species.

Rapid Analysis for α-Tocopherol and Its Oxidative Products in the Pisum sativum L. Leaf Using Supercritical Fluid Chromatography-Medium Vacuum Chemical Ionization Tandem Mass Spectrometry.

A method for the rapid determination of α-tocopherol (α-T) and its oxidative products in plant tissue has been developed using supercritical fluid extraction (SFE) coupled with supercritical fluid chromatography (SFC) and medium vacuum chemical ionization (MVCI) with tandem mass spectrometry. The method is designed to study changes in levels for α-T and its oxidative products in plant cells during photosynthesis, aiming to observe the light response curves. α-T oxidation is a non-enzymatic self-defense mechanism in plant cells. Unlike enzyme-involved reactions, it cannot be stopped, so the oxidation continues in crude extracts even after extraction. Therefore, a real-time in-situ method is essential for tracking the light response curves. To optimize the selective reaction monitoring method, the reaction mixture of α-T and singlet oxygen (1O2), generated by rose Bengal under light illumination, was used as the source of oxidative products. The relative abundance changes in α-tocopherylquinone and 8a-hydroperoxy tocopherone in Pisum sativum L. (Pea) leaves under excessive light illumination have been preliminarily analyzed as part of the light response curve study. The method archives a throughput of 10-15 minutes for analyzing duplicate leaf samples. This process includes cutting off the leaf, sectioning it, placing the sample in a frozen SFE vessel, and conducting SFE/SFC analysis. Consequently, the average throughput is approximately 5-7 minutes per sample.

In vitro antioxidant and antibacterial activities of Ajuga integrifolia leaf extracts obtained with different solvents

BackgroundMany diseases are increasingly recognized as public health concerns worldwide because of the increasing incidence of multidrug-resistant bacteria. Recently, interest in the use of indigenous medicinal plants to treat infectious illnesses has increased, highlighting the need to find new bioactive phytochemicals. Ajuga integrifolia is a plant commonly utilized in traditional drugs to treat a wide range of diseases, although its effectiveness has not been scientifically validated. The present study aimed to evaluate the total phenolic and flavonoid contents and assess the biological activities of A. integrifolia leaf extracts produced via different solvent systems.MethodsSoxhlet extraction was employed to obtain crude extracts from different solvents (methanol, ethanol and water). The 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ferric-reducing power assays were used to measure the antioxidant activity, and the antibacterial activity of the extract was evaluated on the basis of its minimum inhibitory concentration (MIC) against two gram-negative bacteria (Escherichia coli (ATCC-25922) and Pseudomonas aeruginosa (ATCC-43495)) and two gram-positive bacteria (Staphylococcus aureus (ATCC-25923) and Streptococcus pyogenes (ATCC-19615)) via the agar disk-diffusion technique.ResultsA significant amount of total phenolic content (TPC) and flavonoid content (TFC) were present in all the extracts. The extracts presented powerful antioxidant activity in all the assays. The disc diffusion and MIC results revealed the ability of the methanol and ethanol extracts of A. integrifolia leaves to inhibit S. aureus growth at a concentration of 3.125 mg/mL. However, the water extracts were ineffective against E. coli and P. aeruginosa.ConclusionsThese findings indicate that A. integrifolia leaf extracts have reasonable biological activities. These findings underscore the importance of A. integrifolia leaves as a source of health benefits.

Preparation of novel nanoparticle demulsifier and its demulsification for surfactant-stabilized crude oil emulison

The application of nanoparticle (NP) demulsifiers in processing the emulsified crude oil production fluid is of great significance for improving production efficiency and reducing energy consumption in crude oil extraction. In this work, a novel NP demulsifier (PPA@KAO) was prepared by chemically modifying the mineral kaolinite (KAO) nanoparticles using γ-Methacryloyl propyltrimethoxysilane (MPS) and polyether-polyquaternium (PPA). The morphology and structure of the PPA@KAO demulsifiers were characterized by FT-IR, SEM, TG, XPS and Zeta potential. The contact angle measurements, interfacial tension (IFT) characterization, and the distribution of nanoparticles in the oil-water phase showed that PPA@KAO have good amphiphilicity and interfacial activity. The demulsification performance of PPA@KAO demulsifiers for a sodium dodecyl benzene sulfonate (SDBS) stabilized crude oil emulsion was studied with bottle method. It was found that the oil-water separation could be completed within 20 min, with a demulsification efficiency of 96.75 % at a dosage of 3 g L−1，pH of 6 and temperature of 30°C. Such a result indicates that the PPA@KAO NP demulsifier can achieve rapid and efficient demulsification for the surfactant-stabilized crude oil emulsion at room temperature. The demulsification mechanism of PPA@KAO for the SDBS-stabilized crude oil emulsion was well revealed, which provides an important synthetic strategy of how obtaining an effective NP demulsifiers by regulating the amphiphilicity and surface functional groups of the nanoparticles.

Hypoglycemic and antioxidant activities of Jasminum officinale L. with identification and characterization of phytocompounds

The utilization of plant-derived chemicals with anti-diabetic properties is widely promoted for its advantageous tactics in managing diabetes, as they are cost-effective and have minimal or no adverse effects. Therefore, this work investigates the medicinal plant Jasminum officinale L. leaves by extraction and bio-guided fractionation. The ethyl acetate fraction showed a higher yield of 36.4 %. A phytochemical test on Jasminum officinale confirmed flavonoids, saponins, phenols, and tannins. The highest total phenol and flavonoid contents in the ethyl acetate fraction of J. officinale are 103.01 ± 1.1 mg GAE/g and 80.29 ± 1.03 mg QUE/ value found in methanol crude extract. Furthermore, HPTLC analysis of the ethyl acetate fraction detected the existence of flavonoids (kaempferol) and phenols (gallic acid, quercetin, and rutin). The compounds detected at the greatest concentrations in the LC-M/MS analysis of the ethyl acetate fraction were cirsiliol, kaempferol, and 2-tridecanone. Additionally, J. officinale (IC50 33.845 ± 1.09 μg/mL) demonstrated the highest DPPH scavenging activity in EAF like that of ascorbic acid (IC50 22.27 ± 0.96 μg/mL). Also, in the FRAP assay, the IC50 of this fraction is 15.14 ± 0.25 μM Fe equivalents. In the range of alpha-amylase deactivating action, from 13.25 % to 74.51 %, and IC50 value (47.40 ± 0.29 μg/mL) was significantly higher in the ethyl acetate fraction of J. officinale leaf extract. Moreover, J. officinale leaf extract had a substantially higher retention of glucose level (23.92 ± 0.85 % to 87.21 ± 0.6 %), significantly higher anti-inflammatory activity with the lowest IC50 value (66.00 ± 1.84), and lipid peroxidation (IC50 value 34.67 ± 1.69) by utilizing egg yolk as a substrate for lipids. Overall, the study revealed that J. officinale has considerable anti-diabetic characteristics. However, further comprehensive research is necessary to ascertain the medicinal purposes of J. officinale and its chemical components, pharmacological effects, and clinical uses.

Synergistic action of bacteriophage and metabolites of Pseudomonas fluorescens JB3B and Streptomyces thermocarboxydus 18PM against Enterotoxigenic Escherichia coli and Bacillus cereus and their biofilm

BackgroundFoodborne disease and food spoilage are the prime public health issue and food security round the globe. Significant disease outbreaks mostly linked to the existence of pathogenic bacteria that extremely challenging due to the persistence of biofilm-forming. Proteins and bacterial metabolites have been shown to have good antibacterial activity and effectively removal bacterial biofilm. Recently, bacteriophage and their encoded lytic proteins such as lysin have attracted attention as potential alternative agent to control undesirable pathogens in human body infection, increasing food safety as advance preservations and medical treatment such as phage therapy. For these reasons, the efficacy of bacteriophage and their potential in combination with bacterial metabolites from Phyllosphere and Actinomycetes bacteria (Pseudomonas fluorescens JB3B and Streptomyces thermocarboxydus 18PM crude extracts) was the aim of this present study.ResultsIn this study, bacteriophage BC-VP (1.28 ± 0.29 × 1011 PFU/ml) and ETEC-phage-TG (8.9 ± 2.19 × 108 PFU/ml) isolated from artificial lake water from previous study showed potential activity to control Bacillus cereus (BC) and Enterotoxigenic Escherichia coli (ETEC) population. The combination of BC-VP with metabolite (P. fluorescens JB3B and S. thermocarboxydus 18PM) which were known from previous study had antibiofilm activities were able to inhibit (86.1%; 83.3%) and destruct (41%; 45.5%) biofilm formation of B. cereus respectively. Likewise, the synergy of bacteriophage ETEC-phage-TG with the same crude extract also showed promising activity against biofilm of ETEC with percentage of inhibition (81.9%; 76.4%) and percentage of destruction (54.1%; 44.4%). Application in various food, combination of BC-VP and bacterial metabolite extract (P. fluorescens JB3B; S. thermocarboxydus 18PM) were able to reduce Bacillus cereus population in mashed potato (99.6%; 99.4%) at cold temperature (4 °C) and (68.9%; 56.6%) at room temperature (28 °C), boiled pasta (99.5%; 99.4%) and (84.7%; 75.7%), also soymilk (96.9%; 96.7%) and (42.4%; 39.4%) respectively. Likewise, combination of ETEC-phage-TG and bacterial metabolite (P. fluorescens JB3B; S. thermocarboxydus 18PM) potentially reduced ETEC population after two different temperatures (4 °C and 28 °C) incubation in bean sprouts (TFTC; TFTC) and (47.5%; 49.1%), chicken meat (TFTC; TFTC) and (58.1%; 54%), also minced beef (99.5%; 99.4%) and (41.1%; 28%). GC-MS determination performed, oxalic acid, phenol, phenylethyl alcohol, N-hexadecanoic acid, and pyrolol[1,2-a]pyrazine-1,4-dione, hexadro-3-92-methylpropyl was the most active compound in P. fluorescens JB3B. 2,4-Di-tert-butylphenol, phenyl acetic acid, N-Hexadecanoic acid, pyrolol[1,2-a]pyrazine-1,4-dione, hexadro-3-92-methylpropyl, and Bis(2-ethylhexyl) phthalate was most active compound in the S. thermocarboxydus 18PM isolates.ConclusionsThe combination of isolated bacteriophages and bacterial metabolite showed promising results to be used as biocontrol candidate to overcome biofilm formed by foodborne and food spoilage bacteria using their ability to produce antibiofilm compounds and lytic activity. In addition, this combination also potentially reduces the use or replace the drawbacks of common application such as antibiotic treatment.

Akt inhibitors prevent CyHV-2 infection in vitro

Cyprinid herpesvirus 2 (CyHV-2) is a double-stranded DNA virus that infects goldfish (Carassius auratus) and crucian carp (C. carassius), resulting in substantial mortality rates and significant epidemiological implications. To gain deeper insights into CyHV-2-host interactions and identify potential therapeutic targets, quantitative proteomics analysis was conducted on CyHV-2-infected Ryukin goldfish fin (RyuF-2) cells. Our findings revealed significant alterations in the expression of proteins associated with the PI3K/Akt signaling pathway, which were up-regulated upon viral infection. Building on these observations, we employed LY294002, a specific inhibitor of PI3K, to investigate its impact on viral replication by inhibiting the PI3K/Akt pathway in GiCF cell line derived from the caudal fin of Carassius auratus gibelio (Bloch). Our results demonstrated the inhibition of both CyHV-2 replication and Akt phosphorylation within this pathway. Quercetin, a plant-derived analogue of LY294002, was further investigated for its anti-CyHV-2 effects in vitro as well as its underlying mechanism. The results suggested that quercetin exhibits antiviral properties against CyHV-2 and may exert its effects through mechanisms similar to those observed with LY294002. Given that aquaculture water serves as a vector for aquaculture viral diseases and the release of chemical compounds can lead to pollution of the aquatic environment, our study shifted focus to crude extracts obtained from plants. We confirmed crude quercetin extract derived from Cuminum anisum has antiviral activity against CyHV-2 in vitro. Therefore, based on our identification of the involvement of PI3K/Akt signaling pathway in CyHV-2 replication, along with verification of its antiviral mechanism using LY294002, we propose natural dietary quercetin as a promising candidate for development into a novel anti-CyHV-2 drug.

Optimizing germination dynamics in seven key industrial and medicinal hemp varieties through seed priming techniques: An initial study for hemp cultivation in Pakistan

Seed germination is crucial for studying factors influencing production conditions since it represents the initial physiological stage of the plant's life cycle. Hemp seeds are grown to obtain cannabinoids; however, these varieties, hybrids, and feminized seeds are costly in the market with low germination rates. Therefore, optimizing the germination of hemp seeds can play an important role in maintaining plant populations and crop establishment and can reduce seed costs. This study was aimed at evaluating the germination potential of seven hemp varieties/cultivars (Cannabis sativa L.) in response to priming with Indole Butyric Acid (IBA), moringa leaf crude extract, hydro-priming, and a control. Results showed that IBA at 1000 ppm significantly improved germination rates; while the hydro-priming exhibited the fastest germination across all varieties/cultivars. Two cultivars viz., Santhica-70 and Fedora-17 exhibited the highest germination percentages of 83% and 77%, respectively with IBA treatment; while CBD Pink Kush exhibited a germination rate of 63%. This study represents the first comprehensive investigation into seed germination and seedling growth of hemp varieties, offering insights into optimized priming agent selection for industrial and medicinal hemp crop cultivation in Pakistan, highlighting the importance of variety-specific traits and priming techniques for maximizing crop yield.

α-GLUCOSIDASE AND TYROSINASE INHIBITORY GUIDED ISOLATION, IDENTIFICATION OF MAJOR COMPOUND AND IN SILICO STUDY FROM EDIBLE MUSHROOM COPRINUS COMATUS

Coprinus comatus is an edible and medicinal mushroom. The crude hexane, EtOAc and methanol extracts were extracted successively from C. Comatus. All the crude extracts were evaluated for anti-a-glucosidase and tyrosinase. The results showed that %inhibition valued for the crude hexane, EtOAc and methanol extracts of anti-a-glucosidase were 98.58 ± 0.09, 98.90 ± 0.21, and 6.68 ± 0.79, respectively. The anti-tyrosinase for the crude hexane, EtOAc and methanol extracts were 97.31 ± 2.77, 97.31 ± 3.34 and 64.94 ± 3.34, respectively. Notably, the bioassay-guided showed that the crude hexane and EtOAc were the highest for anti-a-glucosidase and tyrosinase activities, which led to the interest in purifying the major compound. The isolation and purification were successively done using chromatographic techniques. The pure compounds were determined by spectroscopic analysis (UV, FTIR, and NMR) and comparison with previously reported. The isolation led to three compounds (1-3), ergosterol (1), indole-3-carbaldehyde (2), and 4-hydroxybenzoic acid (3). The molecular docking of compounds 1-3, reported as α-glucosidase and tyrosinase inhibitors, was done by the AutoDockTools 1.5.6 (ADT) software. The results revealed that ergosterol (1) provides the highest binding free energy (-9.91 kcal/mol) to the active site of a-glucosidase, indicating that it is quite specific to this enzyme. Moreover, ergosterol (1) has strong binding energy (-9.64 kcal/mol) to tyrosinase but not significant tyrosinase specificity. This evidence supports that C. Comatus would be applicable for a functional food for controlling glucose blood levels.

Tyrosinase Inhibitory Activity of Crude Procyanidin Extract from Green Soybean Seed and the Stability of Bioactive Compounds in an Anti-Aging Skin Care Formulation

Green soybean (Glycine max L.) seed contains a high procyanidin content and high antioxidant activity. Moreover, ultrasound-assisted extraction (UAE) has proved to be advantageous in providing high extraction efficiency. Hence, this study aimed to extract procyanidins from green soybean seeds (GSSs) using UAE. This study also evaluated the inhibitory activities of tyrosinase and the cytotoxic effects of crude procyanidin extract. The extract exhibited maximum levels of bioactive components and antioxidant capacity when subjected to a temperature of 15 °C and an extraction time of 20 min. The crude procyanidin extract at a concentration of 10 mg/mL inhibited the tyrosinase enzyme by more than 60%, and the half-maximal inhibitory concentration (IC50) value obtained for the extract was 6.85 ± 0.81 mg/mL. This result was much greater than the IC50 value obtained for kojic acid (0.089 ± 0.08 mg/mL), which was used as a positive control. For the cytotoxicity assessment, the results indicated that the crude procyanidin extract showed no cytotoxicity and actually stimulated the growth of human skin fibroblast cells. More than 80% of the bioactive compounds (total phenolic content (TPC), total flavonoid content (TFC), procyanidin content (PC)) and antioxidant activities (DPPH and FRAP) of the crude extract powder were retained at 38.68 ± 0.01 mg GAE/g, 16.07 ± 0.01 mg CAE/g, 9.24 ± 0.01 mg PC/g, 359.8 ± 0.72 μM Trolox eq/g, and 1640 ± 2.86 μM Trolox eq/g, respectively, after 12 weeks of storage at 25 °C. The crude procyanidin extract powder was then included in a facial serum formulation and tested for pH value and physical evaluation. The stability of the crude procyanidin extract facial serum was shown to be greater for bioactive compounds and antioxidant activity when stored at a temperature of 4 °C than when stored at a temperature of 25 °C. These results suggest that the GSS extracts obtained via ultrasonication show promise for use in cosmeceutical formulations for whitening skincare products.

Effect of crude ethanolic seed extract from Mucuna pruriens on proliferation, apoptosis, and cell cycle arrest in gastric adenocarcinoma (AGS) cells

Abstract Background Gastric cancer is a prevalent form of malignancy among many common carcinoma cases globally. This study was designed to assess the anticancer potential of the crude ethanolic seed extract from Mucuna pruriens against the gastric cancer cell line (AGS). Various assays were employed to assess the anticancer properties, including examinations of cell viability, nuclear morphology, apoptosis using AO/EB staining, changes in mitochondrial membrane potential, lactate dehydrogenase activity, DNA fragmentation, and cell cycle arrest. Results The crude extract exhibited significant anticancer activity against the human gastric cancer cell line (AGS), as determined by the MTT assay, with an inhibition concentration (IC50) of 600 µg/mL at 24 h. Distinct cellular and nuclear morphological changes were observed with different concentrations of crude ethanolic seed extract. The LDH release assay reveals cell death in AGS cells, as evidenced by a significant increase in the release of LDH enzyme. DNA fragmentation analysis and flow cytometry results indicate that the extract induces chromatin condensation, apoptotic cell death, and cell cycle arrest at the G0/G1 phase in the AGS cancer cell line. These results highlight the potential therapeutic advantages of Mucuna pruriens seed extract against gastric cancer cells. Conclusion This study could pave the way for identifying diverse natural bioactive compounds sourced from Mucuna pruriens seed, leading to the development of novel drug with potential anticancer properties.

The Insecticidal Activity of Secondary Metabolites Produced by Streptomyces sp. SA61 against Trialeurodes vaporariorum (Hemiptera: Aleyrodidae).

Trialeurodes vaporariorum Westwood poses a significant threat to vegetable and ornamental crops in temperate zones, resulting in notable reductions in yield and substantial economic burdens. In order to find compounds with high insecticidal activity against T. vaporariorum, five compounds were isolated and identified from the crude extract of Streptomyces sp. SA61. These include three new polyketides, named strekingmycins F-H (1-3); one new diterpenoid, named phenalinolactone CD8 (4); and one known compound, strekingmycin A (5). Their structures were analyzed using high-resolution electrospray ionization mass spectrometry and one-dimensional and two-dimensional nuclear magnetic resonance spectroscopy data and by comparing them with previously reported data. The insecticidal activities of compounds 1-5 against T. vaporariorum were evaluated. Among them, compound 5 exhibited the highest insecticidal activity, with an LC50 of 6.949 mg/L against T. vaporariorum at 72 h using the leaf-dip method. Lower insecticidal activities were found in compounds 1-4, with LC50 values of 22.817, 19.150, 16.981 and 41.501 mg/L, respectively. These data indicate that strekingmycin could be a potential candidate for a novel insecticide to control T. vaporariorum.

Antimicrobial activity of propolis agents on some pathogenic microbes

The study aims to investigate the antimicrobial activity of propolis obtained from different regions of Iraq compared with that of propolis obtained from Iran. Samples were investigated for their antimicrobial activity against Staphylococcus aureus, Pseudomonas aeruginosa, Eschericha coli, Klebsiella pneumoniae, Bacillus cereus , Staphylococcus epidermidis and Candida albicans using standard antimicrobial assays. Marked variations in the antimicrobial activity of the different propolis samples were observed, the method of extraction selected gives the highest antimicrobial activity and the best alcohol concentration using in the extraction of propolis , then the crude extract of propolis showed synergistic effect with some antibiotics in inhibition of pathogenic microorganisms.

Determination of Potential Anti Quorum Sensing Activity of Eleusine indica Ethanolic Crude Extract against Escherichia coli and Staphylococcus aureus

Determination of Potential Anti Quorum Sensing Activity of Eleusine indica Ethanolic Crude Extract against Escherichia coli and Staphylococcus aureus

Antibacterial and anticancer potential of bioactive compounds and secondary metabolites of endophytic fungi isolated from Anethum graveolens.

Fungal endophytes are known to produce bioactive chemicals and secondary metabolites that are often identical to those produced by their host plants. The main objective of the current study was to isolate and identify endophytic fungi associated with the medicinal plant Anethum graveolens, and to investigate their potential antibacterial and anticancer properties. The ethyl acetate extracts from the isolated endophytic fungi, as well as the host plant A. graveolens, were subjected to bioactivity assays to evaluate their antibacterial and anticancer potential against multi-drug resistant bacterial strains and the human hepatocellular carcinoma cell line HepG2. The endophytic fungi isolated and identified from the A. graveolens samples included Diaporthe, Auxarthron, Arthrinium, Aspergillus, Microsporum, Dothiorella, Trichophyton, Lophiostoma, Penicillium, and Trichoderma species. The minimum inhibitory concentration (MIC) assay revealed that the A. graveolens extract exhibited the strongest antibacterial activity, with an MIC value of 4 μg/ml, followed by the Trichoderma sp. (5 μg/ml) and Penicillium sp. (6 μg/ml) extracts. Additionally, the crude extracts of Trichoderma sp., Penicillium sp., and Fusarium sp. demonstrated high anticancer activity against HepG2 cells, with inhibition rates ranging from 89 to 92% at a concentration of 50 μg/ml. Interestingly, the A. graveolens extract showed the most potent anticancer activity, with a 95% inhibition rate against HepG2 cells at the same concentration. These findings highlight the significant potential of endophytic fungi associated with A. graveolens, as a source of bioactive compounds with promising antibacterial and anticancer properties. The results reinforce the hypothesis that medicinal plants and their endophytic fungi can serve as an attractive alternative for the development of novel therapeutic agents, potentially offering a more sustainable and less harmful approach to disease management compared to traditional chemical-based methods.

UPLC-HRMS/MS Guided Isolation and NMR Investigation of Major Flavonoids from Enarthrocarpus strangulatus Boissier (Brassicaceae) with In Vitro Enzymes Inhibitory Potential.

Various members of the family Brassicaceae are economically importantand traditionally used to treat many disorders. Among the family members, Enarthrocarpus strangulatusBoissier is a common Egyptian species that was rarely studied. Consequently, the current study aims to characterize its phytochemical composition and assess its potential bioactivity comprehensively. A metabolomics approach integrating UPLC-HRMS/MS-based molecular networking enabled the dereplication of 91 metabolites, including primary (i.e. organic acid, amino acids, fatty acids, and phospholipids) and secondary metabolites (i.e. glucosinolates, phenolic acids, and flavonoids). Among the 91 annotated features, 13 major metabolites were fully characterized following their isolation and purification. Exclusive of only three flavonoids, all the detected metabolites are described for the first time within this species. Furthermore, the crude extract and four major isolated flavonoids were subjected to in vitro biological screening, including antioxidant, radical scavenging, anti-diabetic, anti-Alzheimer's, and anti-inflammatory activities. It was noticed that nobiletin (61) exhibited the highest antioxidant and anti-Alzheimer's activities. At the same time, isorhamnetin 3-O-glucose (51) showed the highest anti-diabetic activity compared to the other isolated flavonoids and the total extract itself. Regarding the anti-inflammatory activity, no obvious differences were detected numerically among all studied flavonoids.

In-vitro scientific validation of anti-inflammatory activity of Punica granatum L. on Leukemia monocytic cell line

Background: The induction of the inflammatory cascade results in the production of a number of inflammatory mediators, including prostaglandin E2 (PGE2), nitric oxide (NO), and proinflammatory cytokines like TNF-, IL-, and IL-6. This study examined the cytotoxicity and anti-inflammatory properties of a methanolic crude extract of Punica granatum L. peel (PPM) on monocytic leukaemia cell line (THP-1). Materials and methods: The PPM along with Quercetin as reference was used to assess the cytotoxic effect on THP-1 cells and describe its effect on pro-inflammatory cytokines such as COX-2, TNF-α, IL-6 against cancer cell line by flow cytometry. Results: The percentage of viable cells significantly decreased which correlates to non-toxicity whereas quercetin was found to be highly toxic, the IC50 could not be calculated because of drug precipitation. There was a significant decrease in the expressions of inflammatory cytokines upon pre-treatment of the cells with PPM prior to LPS stimulation. Conclusion: Our findings indicate that no cytotoxicity was observed after the treatment of THP-1 cells with PPM (25-400 µg/ml), but at higher concentration (400µg/ml), the cell viability decreased to 84% and attenuated the expression level of inflammatory cytokines. The inhibitory effect of the extract on pro-inflammatory factors production may provide a theoretical source on upcoming treatment of inflammation. Keywords: Punica granatum L; Monocytic leukaemia cell line; cytotoxicity; anti-inflammation.

Development of Loop-Mediated Isothermal Amplification (LAMP) Assay for In-Field Detection of American Plum Line Pattern Virus.

American plum line pattern virus (APLPV) is the most infrequently reported Ilarvirus infecting stone fruit trees and is of sufficient severity to be classified as an EPPO quarantine A1 pathogen. In late spring, yellow line pattern symptoms were observed on leaves in a few flowering cherries (Prunus serrulata Lindl.) grown in a public garden in Northwest Italy. RNA extracts from twenty flowering cherries were submitted to Ilarvirus multiplex and APLPV-specific RT-PCR assays already reported or developed in this study. One flowering cherry (T22) with mixed prunus necrotic ringspot virus (PNRSV) and prune dwarf virus (PDV) infection also showed infection with APLPV. Blastn analysis of PCR products of the full coat protein (CP) and movement protein (MP) genes obtained from flowering cherry T22 showed 98.23% and 98.34% nucleotide identity with reference APLPV isolate NC_003453.1 from the USA. Then, a LAMP-specific assay was designed to facilitate the fast and low-cost identification of this virus either in the laboratory or directly in the field. The developed assay allowed not only the confirmation of APLPV (PSer22IT isolate) infection in the T22 flowering cherry but also the identification of APLPV in an asymptomatic flowering cherry tree (TL1). The LAMP assay successfully worked with crude flowering cherry extracts, obtained after manually shaking a single plant extract in the ELISA extraction buffer for 3-5 min. The developed rapid, specific and economic LAMP assay was able to detect APLPV using crude plant extracts rather that RNA preparation in less than 20 min, making it suitable for in-field detection. Moreover, the LAMP assay proved to be more sensitive in APLPV detection in flowering cherry compared to the specific one-step RT-PCR assay. The new LAMP assay will permit the estimation of APLPV geographic spread in the territory, paying particular attention to surrounding gardens and propagated flowering cherries in ornamental nurseries.

Screening phytochimique et étude de l’activité antioxydante de trois plantes médicinales de la pharmacopée traditionnelle du Niger : Vernonia amygdalina Del., Tephrosia purpurea (L.) et Momordica ba

Considering the potential role in food industry and human health, antioxidants are gaining popularity all across the globe. Antioxidants are defined as a substance that even in small amounts, is capable of preventing or delaying the oxidation of easily oxidizable materials. In this work, different parts of three plant species of the Niger Pharmacopoeia (Vernonia amygdalina Del., Tephrosia purpurea (L..) and Momordica balsamina (L..) were studied. Tube characterization reactions were used for phytochemical screening. The Folin-Ciocalteu and aluminium trichloride methods were used to determine the total polyphenol and flavonoid content. The antioxidant activity of the various crude extracts was determined through DPPH tests. Two types of extraction were used for these tests : maceration and decoction, and raw extracts were used for the work. The results of phytochemical screening showed on the different plants parts indicate the presence of various chemical groups, including polyphenols, tannins, saponins, flavonoids, alkaloids, saponosides, and others.. The compounds were quantified by spectrophotometry. Quantification of the compounds by spectrophotometry showed that the aqueous extracts of the bark and leaves of Vernonia amygdalina were the richest in polyphenols (57.05±0.33 mg EAG/g and 47.062±0.88 mg EAG/g, respectively), while the hydroethanolic and ethanolic extracts of the leafy branches of Tephrosia purpurea were the richest in total flavonoids (13.79±0.28 mg EQ/g and 13.73±0.12 mg EQ/g respectively). In terms of antioxidant power, the results obtained showed that the extract of Vernonia amygdalina bark exhibited the highest antioxidant activity with an inhibitory concentration (CI50=0.0023mg/ml). Our findings showed that the aqueous extracts of these plants have antioxidant properties; which substantiated their utilization in the conventional management of specific ailments. Key words : Vernonia amygdalina, Momordica balsamina, Tephrosia purpurea, Antioxidants, Niger.