GAS CHROMATOGRAPHY-MASS SPECTROMETRY ANALYSIS AND ANTIOXIDANT ACTIVITY OF PUNICA GRANATUM L. PEELS AND ITS ROLE AS IMMUNOSTIMULANT AGAINST SCHISTOSOMA MANSONI INFECTION IN BIOMPHALARIA ALEXANDRINA

Objective: To evaluate the antioxidant activity and chemical composition of Punica granatum L. and test it as immunostimulants against Schistosoma mansoni infection to Biomphalaria alexandrina snails.Methods: Antioxidant activity was evaluated by measuring the free radical scavenging activity of the 90% defatted methanol extract (90% DM) of P. granatum peels and its sub-derived fractions was evaluated via 2,2'-diphenyl-1-picrylhydrazyl and its chemical constituents were identified via gas chromatography-mass spectrometry (GC-MS) analysis. B. alexandrina snails were exposed to pomegranate extracts (PEs) for 1 month before their challenging with S. mansoni miracidia. Infection rates, immunological and histological parameters were, then, evaluated in PE-exposed snails and compared to controls.Results: The antioxidants activities of PE, expressed as scavenging concentration at 50%, were in the following order; 90% DM (12.45) ˃n-butanol(15.59) ˃ethyl acetate (21.36) ˃water (49.16) µg/ml, compared to 7.50 µg/ml for ascorbic acid. The infection rates of PE-exposed snails were 20%,50%, 60%, 70%, and 80%, respectively, for 90% DM, n-butanol, ethyl acetate, water, and dichloromethane extracts compared to 95% in control snails. The number of amoebocytes showed a significant increase, clear differentiation, and size increment in exposed snails compared to controls. Moreover, hermaphrodite glands histology shows a full maturity in the formation of reproductive cells in PE-exposed snails. The GC-MS analysis of the 90% DM extract revealed the presence of 36 compounds representing 93.1% of the total composition. Piperidin-4-ol, 1,3-dimethyl-2,4,6-triphenyl (19.87%), and 6,11-dihydroxy-5,12 naphthacenequinone-1-carboxylic acid (7.80%) were the major components.Conclusion: The identified compounds in 90% DM extract of P. granatum may be responsible for the high antioxidant activity of the fruit and it may account for its immunostimulatory effect against S. mansoni infection in B. alexandrina.Keywords: Punica granatum L., Antioxidant activity, Biomphalaria alexandrina, Schistosoma mansoni, Infection rate, Gas chromatography-mass spectrometry

Bio-fabricated zinc oxide nanoparticles mediated by endophytic fungus Aspergillus sp. SA17 with antimicrobial and anticancer activities: in vitro supported by in silico studies

IntroductionIn recent years, the world’s attention has been drawn to antimicrobial resistance (AMR) because to the frightening prospect of growing death rates. Nanomaterials are being investigated due to their potential in a wide range of technical and biological applications.MethodsThe purpose of this study was to biosynthesis zinc oxide nanoparticles (ZnONPs) using Aspergillus sp. SA17 fungal extract, followed by characterization of the produced nanoparticles (NP) using electron microscopy (TEM and SEM), UV-analysis, X-ray diffraction (XRD), and Fourier-transform infrared spectroscopy (FT-IR).Results and DiscussionThe HR-TEM revealed spherical nanoparticles with an average size of 7.2 nm, and XRD validated the crystalline nature and crystal structure features of the generated ZnONPs, while the zeta potential was 18.16 mV, indicating that the particles’ surfaces are positively charged. The FT-IR was also used to identify the biomolecules involved in the synthesis of ZnONPs. The antibacterial and anticancer properties of both the crude fungal extract and its nano-form against several microbial strains and cancer cell lines were also investigated. Inhibition zone diameters against pathogenic bacteria ranged from 3 to 13 mm, while IC50 values against cancer cell lines ranged from 17.65 to 84.55 M. Additionally, 33 compounds, including flavonoids, phenolic acids, coumarins, organic acids, anthraquinones, and lignans, were discovered through chemical profiling of the extract using UPLC-QTOF-MS/MS. Some molecules, such pomiferin and glabrol, may be useful for antibacterial purposes, according to in silico study, while daidzein 4’-sulfate showed promise as an anti-cancer metabolite

Insights about clinically approved and Preclinically investigated marine natural products

Since the early research efforts focusing on bioactive marine substances such as spongouridine, from the marine sponge Cryptotethya crypta, marine natural products (MNPs) have arisen as a robust and sustainable supplier for bioactive drug leads. Marine natural products present definite, unprecedented structural diversifications and varieties of interesting biomedical potentialities with novel mechanisms of action. Until today, eight clinically approved marine natural products-based drugs by two worldwide medical organizations (including U.S. FDA, European Medicines Agency (EMEA)), have been developed for the treatment of different forms of carcinoma, pain, Alzheimer's disease and other current medical challenges. Recent clinical trial analysis disclosed that the current clinical pipeline contains more than twenty listed drug candidates in different clinical trials in phase III, II, or I. Herein, we present recent insights centered to clinically approved and preclinically investigated marine bioactive compounds, as well as sampling techniques, extraction & identification tools, classification of MNPs, some reported biological activities, and challenges faced during MNPs development

CYTOTOXIC SCREENING OF THREE EGYPTIAN PLANTS USING BRINE SHRIMP LETHALITY TEST

Objective: Brine shrimp (Artemia salina L.) bioassay is considered as a preliminary screening for the presence of antitumor compounds and used to determine the toxicity of plant extracts. Different extracts of three Egyptian plants, philodendron selloum (Araceae), Bougainvillea alba (Nyctaginaceae) and Ulmus parvifolia (Ulmaceae) were screened for their preliminary cytotoxicity. Methods: The dry powdered leaves of the three plants under investigation (500 g from each plant) were extracted with 85% methanol; then undergo successive fractionation via petroleum ether, chloroform and ethyl acetate. The brine shrimp lethality test (BSLT) was carried out via taking 50 mg of Artemia salina Leach eggs in a hatching chamber and serial concentrations from each extract were used. Results: The chloroform extract of Philodendron selloum showed the most potent cytotoxic activity at sub-lethal concentration (LC50=16 µg/ml) followed by its methanol extract (LC50=26 µg/ml). The methanol, chloroform and petroleum ether extracts of Bougainvillea alba plant showed the same cytotoxic effect at LC50= 40 µg/ml. Ulmus parvifolia showed the least cytotoxic effect at LC50 =380 & 300 µg/ml for methanol and ethyl acetate extracts respectively. Conclusion: It may be concluded that the tested plants possess strong cytotoxic activies; also this work suggests that plants extracts could possibly be used locally as naturally occurring cytotoxic agents

Phytochemical analysis of Eucalyptus camaldulensis leaves extracts and testing its antimicrobial and schistosomicidal activities

Abstract Background The emerging evolution of antibiotic and anthelmintic resistance and inefficiency of some synthetic drugs elicit the need to investigate new drug sources. In this context, Eucalyptus camaldulensis is an evergreen tree that has been widely used in traditional medicine for the treatment of various health disorders. Results Organic solvent extracts from Eucalyptus camaldulensis leaves were assessed for their antimicrobial activity. Among these extracts, ethyl acetate (EtOAc) and water extracts showed the highest antimicrobial activity against Staphylococcus aureus, Pseudomonas aeruginosa, Candida albicans, and Aspergillus niger. EtOAc extract was further subjected to vacuum liquid chromatography (VLC) technique for the isolation of its polyphenolic ingredients. VLC yielded 13 fractions (1–13) that were assayed for their antimicrobial activities. Six of these fractions, namely 4, 5, 6, 7, 8, and 9, showed antimicrobial activity against all the tested microbes except the fungus Aspergillus niger. Fractions 5 and 6 having considerably the highest antimicrobial activity with inhibition zones ranged from 5 to 14 mm. Moreover, fraction 5 was tested as a larvicidal agent against miracidia and cercariae of Schistosoma mansoni. At concentration of 200 mg/L, the mortality rates of miracidia and cercariae of Schistosoma mansoni were 30%, 20%, 50%, 40%, 80%, 20%, and 100%, 80% at 5, 10, 15, and 20 min, respectively. Chromatographic isolation of the EtOAc extract led to identification of six compounds: gallic acid (1), taxifolin (2), methyl gallate (3), quercetin (4), luteolin (5), and hesperidin (6). Conclusions Ethyl acetate extract from Eucalyptus camaldulensis leaves showed a potent antimicrobial and antischistosomal activity. This activity may be attributed to the six phenolic compounds identified through structure elucidation. Thus, these compounds can be good candidates for treatment of microbes and for the control of schistosomiasis

Artemisia campestris and Artemisia herbaalba: LC-HRESI-MS Profile Alongside Their Antioxidant and Antimicrobial Evaluation

Artemisia species have been traditionally used to treat various ailments and health problems like colds, digestive troubles, gastric ulcer, menstrual pain, and diarrhea. In our study, total phenolic and flavonoid contents of 80% aqueous methanol extracts of Artemisia campestris and Artemisia herba-alba plants were investigated. Furthermore, their in vitro antioxidant and antimicrobial activities were evaluated. Also, their phytochemical profiling was performed via using LC-HRESI-MS analysis. Both plant extracts showed strong antioxidant activity using DPPH, ABTS, and phosphomolybdenum assays. The results revealed A. herba-alba extract showed moderate antimicrobial activity against bacteria including Staphylococcus epidermidis and Staphylococcus aureus. While A. campestris extract exhibited antimicrobial activity against different microbial populations such as Pseudomonas aeruginosa and Candida albicans. Also, the results revealed that A. herba-alba extract contains high amounts of 3-O-methylquercetin, eupatilin and acacetin (ranging from 22.04 to 31.88 mg/g), while A. campestris extract contains significant amounts of 3-O-methylquercetin, rutin and chlorogenic acid (82.98, 79.44 and 29.54 mg/ g, respectively). In conclusion, the aqueous methanol extracts of both studied Artemisia species could be promising candidates for treating microbial infections and oxidative stress

Anti-Helicobacter pylori, anti-biofilm activity, and molecular docking study of citropten, bergapten, and its positional isomer isolated from Citrus sinensis L. leaves

Introduction: Citrus sinensis L. is a candidate plant with promising antimicrobial potential. In the current study, the phytochemical investigation of C. sinensis leaf extract led to the isolation of three coumarins, namely bergapten, xanthotoxin, and citropten. Methods: The chemical structures of the isolated coumarins were elucidated using NMR and ESI-MS techniques. The total aqueous ethanol leaf extract and the isolated coumarins were evaluated for their antimicrobial effects against Helicobacter pylori using the MTT-micro-well dilution method and its anti-biofilm activity using MBEC assay, as compared to clarithromycin. Results: The results showed that citropten scored the lowest MIC value at 3.9 μg/mL and completely inhibited the planktonic growth of H. pylori. In addition, it completely suppressed H. pylori biofilm at 31.25 μg/mL. These findings have been supported by molecular docking studies on the active sites of the H. pylori inosine 5′-monophosphate dehydrogenase (HpIMPDH) model and the urease enzyme, showing a strong binding affinity of citropten to HpIMPDH with seven hydrogen bonds and a binding energy of −6.9 kcal/mol. Xanthotoxin and bergapten showed good docking scores, both at −6.5 kcal/mol for HpIMPDH, with each having four hydrogen bondings. Furthermore, xanthotoxin showed many hydrophobic interactions, while bergapten formed one Pi-anion interaction. Concerning docking in the urease enzyme, the compounds showed mild to moderate binding affinities as compared to the ligand. Thus, based on docking results and good binding scores observed with the HpIMPDH active site, an in-vitro HpIMPDH inhibition assay was done for the compounds. Citropten showed the most promising inhibitory activity with an IC50 value of 2.4 μM. Conclusion: The present study demonstrates that C. sinensis L. leaves are a good source for supplying coumarins that can act as naturally effective anti-H. pylori agents

Comprehensive chemical profiling of Bassia indica Wight. aerial parts extract using UPLC-ESI–MS/MS, and its antiparasitic activity in Trichinella spiralis infected mice: in silico supported in vivo study

Abstract Background Trichinellosis is a public health threat infected both animals and humans as a result of eating undercooked meat. It caused by Trichinella spiralis that has widespread drug resistance and even developed many sophisticated strategies for their survival, this increases the demand in searching for new anthelmintic drugs from natural source. Methods Our objectives were to test the in vitro and in vivo anthelmintic activity of Bassia indica BuOH frac., and to characterize its chemical composition using UPLC-ESI–MS/MS. Besides an in silico molecular docking study with the prediction of the PreADMET properties. Results In vitro investigation of B. indica BuOH frac., showed severe destruction of the adult worm and larvae, marked cuticle swelling, areas with vesicles, blebs and loss of annulations. This was assured via in vivo study, which revealed a significant reduction (P < 0.05) in the mean adult worm count with efficacy of 47.8% along with a significant decrease (P < 0.001) in the mean larval count per gram muscle with efficacy 80.7%. Histopathological examinations of the small intestine and muscular sections showed marked improvement. In addition, immunohistochemical findings demonstrated that B. indica BuOH frac. depressed the proinflammatory cytokines expressions of TNF-α, which was obviously upregulated by T. spiralis. Precise chemical investigation of the BuOH frac. using UPLC-ESI–MS/MS resulted in the identification of 13 oleanolic type triterpenoid saponins; oleanolic acid 3-O-6´-O-methyl-β-D-glucurono-pyranoside (1), chikusetsusaponin-IVa (2) and its methyl ester (3), chikusetsusaponin IV (4) and its methyl ester (5), momordin-Ic (6) and its methyl ester (7), betavulgaroside-I (8), -II (9) -IV (10), -X (11), licorice-saponin-C2 (12) and -J2 (13). In addition, 6 more phenolics were identified as syringaresinol (14), 3,4-di-O-caffeoylquinic acid (15), 3-O-caffeoyl-4-O-dihydrocaffeoylquinic acid (16), 3,4-di-O-caffeoylquinic acid butyl ester (17), 3,5-di-O-galloyl-4-O-digalloylquinic acid (18) and quercetin 3-O-(6´´-feruloyl)-sophoroside (19). The auspicious anthelmintic activity was further ascertained using in silico molecular docking approach that targeted certain protein receptors (β-tubulin monomer, tumor necrosis factor alpha (TNF-α), cysteine protease (Ts-CF1), calreticulin protein (Ts-CRT)), all the docked compounds (1–19) fit into the binding site of the active pocket with binding affinities noteworthy than albendazole. In addition, ADMET properties, drug score and drug likeness were predicted for all compounds

Fatty acid and amino acid composition of citrullus colocynthis seeds growing in Algeria

Citrullus colocynthis is an herbaceous perennial wild species of the Cucurbitaceae family, used in traditional medicine in the treatment of diabetes mellitus. In the present work, C. colocynthis seed oils have been assessed for their fatty acids profile and concentration by Gas Chromatography–Mass Spectrometry (GC-MS), while oil and protein extracts were investigated in respect to their amino acid compositions by using Ultra High-Performance Liquid Chromatography-Electrospray Ionization-Triple Quadrupole Mass Spectrometry (UHPLC-ESI-QqQ-MS/MS) instrumentation. Results revealed that the predominant fatty acids in C. colocynthis seeds were linoleic (70.7 %), oleic (10.9 %), palmitic (8.3 %), and stearic (7.8 %) acids. On other hand, the analysis of the free amino acids content noticed the presence of threonine (0.32 μgmL), valine (0.26 μgmL), and tryptophan (0.19 μgmL). Besides, the C. colocynthis seeds oil contained the non-essential amino acids like serine (0.29 μgmL), ethanolamine (0.18 μgmL), glycine (0.25 μgmL) and aspartic acid (0.12 μgmL). In conclusion, the results obtained revealed the valuable nutritional value of C. colocynthis and derived co-products, suggesting its suitability to be used as a nutritional ingredient and to be considered in the development of dietary supplements.The authors are grateful to the DGRSDT and the Ministry of Higher Education and Scientific Research of Algeria for PRFU Project (A16N01UN030120180002), and the team of Medicinal Chemistry Department, Theodor Bilharz Research Institute, Egyp