Three experimental animal parasite models for assessing anthelmintic efficacy and immunopotentiating properties of natural products: protocols and applications

Introduction: Helminthic infections, including soil-transmitted helminths (STHs) and intestinal tapeworms, pose a major global health challenge, classified as neglected tropical diseases (NTDs). Objective: Current control efforts are hampered by expensive treatments, emerging drug resistance, and high reinfection rates. This necessitates the urgent discovery of new anthelmintic drugs and immunomodulatory compounds. Natural products are a promising source for these new therapies. Materials and methods: This paper introduces three experimental animal parasite models specifically designed to evaluate the effectiveness and immune-boosting properties of natural products, namely Hymenolepis diminuta (tapeworm) in mice, Ascaridia galli (nematode) in chickens, and Raillietina cesticillus (tapeworm) in chickens. Rationale: These models provide controlled environments for assessing potential therapeutic compounds and offer systematic methodologies for identifying effective and immunomodulatory natural products. Conclusions: A significant challenge in treating helminth infections is targeting larval stages embedded in host tissues, such as those seen with Hymenolepis nana in humans and hypobiotic trichostrongylid nematode larvae in ruminants. Therefore, demonstrating a natural product\u27s effectiveness against A. galli larvae embedded in the intestinal mucosa is particularly important. Such efficacy would indicate that the product possesses valuable physicochemical and pharmacological properties, paving the way for developing novel treatments against hidden, tissue-dwelling parasites in both humans and animals

Antimicrobial Activities, phytochemical composition and thin layer chromatography profile of Gossypium hirsutum and Abelmoschus esculentus leaves extracts

Background: This study evaluates the antimicrobial activities, phytochemical composition, and thin layer chromatography (TLC) profile of ethanolic and aqueous leaf extracts of Gossypium hirsutum L. and Abelmoschus esculentus (L.) Moench. Aims: The objective is to investigate the phytochemical constituents and assess the antimicrobial efficacy of these plant extracts against various pathogenic microorganisms. Methods: Leaves were collected, identified, air-dried, and powdered. Ethanolic and aqueous extracts were prepared and subjected to TLC analysis using silica gel plates and a mobile phase of ethanol, methanol, and ethyl acetate (5:3:2). Phytochemical screening was conducted using standard protocols. Antimicrobial activity was evaluated using the agar well diffusion method against pathogens including Escherichia coli, Staphylococcus aureus, Klebsiella pneumoniae, Salmonella typhi, Proteus, Streptococcus pneumoniae and Candida spp. Results: TLC analysis revealed seven compounds in G. hirsutum and six in Abelmoschus esculentus with distinct Rf values. Phytochemical screening identified phenols/tannins, saponins, flavonoids, reducing sugars, terpenoids, and anthraquinones in G. hirsutum, while A. esculentus contained all tested phytochemicals except phytosterol and essential oil. The antimicrobial assay indicated that ethanolic extracts of both plants exhibited significant antibacterial activity, with G. hirsutum showing higher efficacy. Ethanolic extracts of G. hirsutum inhibited S. aureus and S. typhi effectively, while aqueous extracts showed limited activity. Extracts of A. esculentus leaves inhibited all tested organisms except Proteus, with ethanol extracts also ineffective against S. pneumoniae. Conclusion: The study demonstrates that ethanolic extracts of G. hirsutum and A. esculentus possess antimicrobial properties. These findings support the potential use of these plant extracts in developing natural antimicrobial agents

PHYTOCHEMICAL, ELEMENTAL ANALYSIS AND ANTIFUNGAL ACTIVITY OF ERIGERON FLORIBUNDUS (BILBAO FLEABANE) LEAF EXTRACT AGAINST MALASSEZIA FURFUR

Background: Elemental composition and phytochemical content of plants viz; saponins, flavonoids, alkaloids, tannins, polyphenols, terpenoids and their concentrations are potentials for their antifungal or antimicrobial activity. Aim: This study is aimed at evaluating the phytochemical, elemental composition and antifungal activity of the essential oil and dichloromethane extract of E. floribundus, against Malassezia furfur - the fungi that causes Pityriasis versicolor (eczema). Method: Assays for the antifungal activity of were performed using the agar dilution method at serial concentrations ranging from 2 to 0.06 mg/ml. Phytochemicals were determined using the suitable Official Analytical (AOAC) methods. Results: The essential oil of E. floribundus was the most potent, inhibiting the fungi at 0.25 mg/ml (IZD = 27 mm), while the dichloromethane extract inhibited the organism at 0.5 mg/ml (IZD = 25 mm), showing a better activity than the control antifungal, clotrimazole which showed an MIC of 1 mg/ml and an inhibition zone diameter (IZD) of 18 mm. Saponins showed the highest concentration of 900 mg per 5 g in the phytochemicals analyzed; with tannins showing the lowest of 56.6 mg per 5 g.  The concentrations of carbon, nitrogen, magnesium, sodium and Sulphur were 68.7 mg per 100 g, 49.4 mg per 100 g, 324.5 ppm, 78.50 ppm and nil respectively. Conclusion: The essential oil and dichloromethane extract of E. floribundus revealed a good potential as an antifungal agent against M. furfur based on their minimum inhibition concentration, which is better than the conventional antifungal agent used as control in this study. Concentrations of phytochemicals in the plant also suggest that it has potentials for the traditional treatment of eczema/skin disorders

APOPTOTIC ACTIVITY OF BOSWELLIA CARTERII EXTRACT ON HUMAN NEUTROPHILS.

Neutrophil activation is required for the initiation of the defence mechanisms which include phagocytosis. Paradoxically, neutrophils also represent one of the main mediators of tissue injury in various human diseases. The resolution of inflammation requires getting rid of excess inflammatory cells through natural cell death and phagocyte clearance. Frankincense, an oleogum resin of different species of the genus Boswellia (Burseraceae), has long been used in eastern countries\u27 traditional medicine to alleviate pain and inflammation. Although it was demonstrated that boswellic acids are potent activators of polymorphonuclear cells, little is known about the effects of the total extract on the human phagocytes’ apoptosis. We here undertake a characterization of the Boswellia carteri resin extract (BCE) effects on human neutrophils activity and viability in vitro. Oxidative burst after stimulation with BCE was evaluated by reduction of nitroblue tetrazolium (NBT) colorimetric method for superoxide anion radical in the the presence of different compounds (N-ethyl maleimide, diltiazem, chelerythrine and wortmannin). Neutrophils viability was assessed by MTT. Flow analyses were performed with propidium iodide (PI) and annexin V-FITC. Our results show that BCE induces the release reactive species of oxygen in human neutrophils in a dose dependent manner. The superoxide anion radical is principally produced via NADPH oxidase since inhibitors of the enzyme may prevent it. Ca+2 depletion reduce the magnitude of activation and PKC and PI3-K are also apparently involved in the process. The BCE has also cytotoxic activity revealed by the MTT assay. This effect seems to be produced by an apoptotic process as shown by the exclusion of the dye propidium iodide and the annexin V-FITC binding. The capacity of the Boswellia carterii extract to accelerate the cellular death by an apoptotic process suggest that either the extract or its active compounds could have applications for the resolution of some inflammatory conditions

IN VITRO EVALUATION OF THE ANTIMICROBIAL, TOXIC, ANTI-ARTHRITIC AND THROMBOLYTIC PROPERTIES OF DIFFERENT PARTS OF PLANT ORCHID RHYNCHOSTYLIS RETUSA (L.) BLUME.

Background: Infection due to microbes, trauma, cancer, arthritis and thrombosis are the most common problems worldwide. Traditionally, a large number of herbal extracts are used for the treatment of those disorders. Aim: Ethanolic extracts from leaves, roots and stems of Rhynchostylis retusa (L.) Blume were phytochemically characterised and then tested for their antibacterial, toxicity, anti-arthritic, and thrombolytic properties in-vitro. Methods and Results: Disk diffusion inhibition tests with a panel of fourteen different microorganisms was used as the antimicrobial assays. Antimicrobial activity of the stem extract found to be higher (10-11 mm, 400 µg/disk) than that of root extract whereas leaf showed no activity with ciprofloxacin (43-46 mm, 5 µg/disc) serving as reference standard. Toxicity of the plant extracts in terms of their LC50 values against the brine shrimp (A. salina) was assessed. The toxic potential of leaf extract was 6.7 μg/ml, which is higher than that of stem and root extract (c.a. 8.5 μg/ml). Thrombolytic properties were tested in vitro using streptokinase as reference (%Clot lysis after 1.5h & 24h was 76.15±1.94 & 92.59±2.35, 30000iu). The leaf extract exhibited higher activity. Leaf extract (1000 ppm) lysed of 33.08±1.24% of the clot and 60.47±2.33 after 1.5 h and 24 h of incubation, respectively. Anti-arthritic effects measured as albumin denaturation using diclofenac sodium as reference (IC50=60.25 ppm) revealed the stem extract as more active. Conclusion: our research will help to provide evidence for some of the traditional uses of this medicinal plant

FORMULATION OF AN ANTIOXIDANT HYDROGEL FACE MASK USING AN ORGANIC, SUSTAINABLY SOURCED, STANDARDISED WATER EXTRACT OF YERBA MATE (ILEX PARAGUARIENSIS) AS ACTIVE INGREDIENT.

Background: The use of natural products in cosmetics has become increasingly popular in recent years. One such natural product that has shown promise as a cosmetic ingredient is Ilex paraguariensis, also known as yerba mate. Aim: to formulate an antioxidant hydrogel face mask using an organic, sustainably sourced, standardised water extract of yerba mate (Ilex paraguariensis) as active ingredient. Methods: Organic yerba mate was water-extracted at 40°C and 80°C. Theobromine, theophylline, caffeine, and various phenolic acids were quantified using RP-HPLC-UV analysis. Extracts were then evaluated for antioxidant activity (DPPH assay). A commercially available mask-making machines for home use was used to produce hydrogel masks based on marine collagen and these were analysed for active ingredient distribution (homogeneity). Results: The differences between 40°C and 80°C water extracts of yerba mate. The content in phytochemicals ranged 0.02-1 mg/ml. The extract equivalence in chlorogenic acid (CA) according to the DPPH assay was 0.25 mgCA / mgextract. The hydrogel facial mask was successfully formulated with 0.1 mg/mL of standardised yerba mate extract. The content in phytochemicals within the gel varied most for caffeic acid and theobromine (approx. 20%) but was <10% for theophylline, caffeine, chlorogenic acid, ferulic acid, rutin, quercitrin, and quercetin. Conclusion: Face masks “home” machines can deliver hydrogel face masks based on marine collagen can incorporate yerba mate (Ilex paraguariensis) water extracts as active ingredient without detriment of the consistency, texture and sensory characteristics with relatively homogenous concentrations of the active principles

Soxhlet extraction cycle-dependent diversity in phenolic profile and antioxidant potency of red cabbage (Brassica oleracea var. capitata F. rubra)

Introduction: The Brassicaceae vegetables are a rich source of secondary metabolites that exhibit several health benefits and protection against numerous degenerative diseases. Objectives: The current study was performed to investigate the effect of Soxhlet extraction cycles on the phenolic profile of red cabbage (Brassica oleracea var. capitata F. rubra) and its biological activities. Materials and Methods: The red cabbage sample was harvested from the research fields of Ayub Agricultural Research Institute Faisalabad, Pakistan. The ethanol extracts were prepared using 1-, 2-, 4-, and 8-cycles Soxhlet extraction technique. The antioxidant potential of red cabbage extracts was estimated by evaluation of total phenolic contents (TPC), total flavonoid contents (TFC), DPPH radical scavenging capacity and reducing power. The identification and quantification of polyphenols were carried out by RP-HPLC. The antibacterial activity of red cabbage extracts was determined against Escherichia coli and Staphylococcus aureus and the antiproliferative activity was carried out by MTT mitochondrial viability assay against the human A549 cancer cell line. Results: Gallic acid, p-hydroxyl benzoic acid, chlorogenic acid, and p-coumaric acid were the major phenolic acids, whereas catechin and quercetin were the major flavonoids detected in the red cabbage extracts. The extraction cycles were found to have significant (p ≤ 0.05) effects on the phenolic profile of the red cabbage extracts.  TPC of extracts ranged from 5.22-11.72 mg/g dry matter, measured as gallic acid equivalent, while the TFC ranged from 1.64-5.19 mg/g dry matter; measured as catechin equivalent. The 4-cycles extract of red cabbage exhibited the maximum TPC, TFC, and DPPH free-radical scavenging and reducing activity. Conclusion: The study concluded that the Soxhlet extraction cycles could exert a considerable effect on the yield and polyphenol composition of red cabbage extracts as well as their antioxidant potential. Antibacterial and antiproliferative activities were observed by all the extracts of red cabbage

The Proceedings of the 4th CNPD Conference: “NATURAL PRODUCTS IN COSMETICS, FOOD AND MEDICINE”

The 4th Annual CNPD (Centre for Natural Products Discovery) Conference was held on 19-22 June 2023 at the John Lennon Arts and Design Building (Liverpool John Moores University). The Organisation Committee has kindly approved the publication of all communications presented to this event in this issue of the Journal of Natural Products Discovery. This issue will replace the classic “Book of Abstracts” that was published in previous years. Therefore, authors will have a collective D.O.I. associated with their presentations instead of an ISSN. We hope that in this way both the event and splendid work of the attendants will be more publicly accessible. Please, follow this example to cite any of the abstracts: Gibbons, S. (2023). " Phytochemistry: not just for drugs and busting bugs." Journal of Natural Products Discovery 2(1): 2. Prof. Dr. Satyajit D. Sarker Editor in Chief Dr. Jose M. Prieto-Garcia Executive Editor Dr Alistair Fielding Member of the Editorial Committe

LEGALISING DRUGS OF ABUSE: THE LE CHATELIER THOUGHT EXPERIMENT

BackgroundThere is currently much debate on legalising some psychoactive materials such as Cannabis. This could stabilise policing costs, reduce crime, generate tax revenue and control sale by registration of outlets. Such outcomes are valuable, but legalisation may have unintended consequences that are deleterious to society or have no effects on crime or user’s health. Here, I propose that behaviour following changes to control of possession and supply may have outcomes that could be predicted by the equilibrium law proposed by Henry Le Chatelier. This law describes a system moving in such a way as to remove the constraints placed upon it. AimsThe purpose of this opinion paper is to follow Le Chatelier’s Principle (LCP) as a thought experiment to predict the outcomes (products) of drug consumption (reactants) when factors influencing this consumption (conditions) are changed. If this equilibrium law can be applied to human behaviour, it may have value to predict the outcomes of these conditions and assess whether they are positive or injurious. It must be noted that the views expressed here are solely those of the author.MethodsPubmed, ScienceDirect, SpringerLink and Google were used for literature searches and “drugs” will be used for drugs of abuse and distinct from medicinal drugs, apart from methadone and heroin (diamorphine). ResultsApplying LCP predicts that drug use may not necessarily decrease, but availability, poor health and criminal activity may increase use. The businesses behind drug treatment and illegal distribution have too much financial incentive to halt operations. ConclusionOrganisations recommending legislative changes to drug control should conduct thought experiments to horizon scan for unintended consequences, that may be costly and deleterious to society. Governments often focus too heavily on the evidence base as the sole driver for change. However, common sense drawn from opinion from these experiments may lead to wisdom. Keywords; War on drugs, legalisation, Le Chatelier’s Principle, psychoactive substance