Metabolomics and in-vitro bioactivities studies of fermented Musa paradisiaca pulp: A potential alpha-amylase inhibitor.

The in-vitro synthesis of bio-compounds via fermentation is a promising route for bioactive molecules intended for disease control and management. Therefore, this study evaluated the effect of fermentation on the antioxidants, antihyperglycemic and anti-inflammatory properties and the resultant chemometric phytochemical profiles of unripe plantain fruits. The results revealed that Escherichia coli and Propionibacterium spp. are suspected as the key fermenters. The E coli showed negative results to the pathogenicity test; Propionibacterium appeared to be opportunistic. A significant increase in the total polyphenols and protein and decreased flavonoids was recorded in the phytochemical profile of the methanolic extract of the fermented unripe plantain pulp; however, the ascorbic acid content was not significantly altered. The 1H NMR fingerprint showed that there is a closely related chemical shift among the shorter fermentation time (days 2-6) and the unfermented, while the more extended fermentation periods (days 7-12) with enhanced bioactivities were closely related based on the chemometrics analyses. Furthermore, the UPLC-QTOF-MS analysis annotated the presence of bioactive compounds in the day-9 fermented sample: polyhydroxy glucose conjugates (3-Methoxy-4-hydroxyphenyl 6-O-(3,4,5-trihydroxybenzoyl)-beta-D-glucopyranoside), short chain peptide (leucyl-glycyl-glycine), amino acid derivatives (4-Aminophenylalanine, and N-Acetylhistidine), linear and cyclic fatty acid derivatives (palmitoyl putrescine, ricinoleic acid, phytosphingosine, gabalid, rubrenoic acid, 2-aminocyclopentanecarboxylic and cystodienioc acid). The synergistic effect of these newly formed compounds and the increase in the phenolic content of the day-9 fermented unripe plantain may account for its more potent antioxidant, anti-inflammatory and antihyperglycemic activity. Therefore, the products obtained from the day 9 fermentation of unripe plantain pulp may serve as potential nutraceutical agents against gastro-enteric sugar digestion and absorption and sugar-induced oxidative stress, inflammation and metabolic disease.

A New Proposed Symbiotic Plant-Herbivore Relationship between Burkea africana Trees, Cirina forda Caterpillars and Their Associated Fungi Pleurostomophora richardsiae and Aspergillus nomius.

Burkea africana is a tree found in savannah and woodland in southern Africa, as well as northwards into tropical African regions as far as Nigeria and Ethiopia. It is used as fuel wood, medicinally to treat various conditions, such as toothache, headache, migraine, pain, inflammation, and sexually transmitted diseases, such as gonorrhoea, but also an ornamental tree. The current study investigated the possible symbiotic relationship between B. africana trees and the C. forda caterpillars and the mutual role played in ensuring the survival of B. africana trees/seedlings in harsh natural conditions and low-nutrient soils. Deoxyribonucleic acid isolation and sequencing results revealed that the fungal species Pleurostomophora richardsiae was highly predominant in the leaves of B. africana trees and present in the caterpillars. The second most prominent fungal species in the caterpillars was Aspergillus nomius. The latter is known to be related to a Penicillium sp. which was found to be highly prevalent in the soil where B. africana trees grow and is suggested to play a role in enhancing the effective growth of B. africana trees in their natural habitat. To support this, a phylogenetic analysis was conducted, and a tree was constructed, which shows a high percentage similarity between Aspergillus and Penicillium sp. The findings of the study revealed that B. africana trees not only serve as a source of feed for the C. forda caterpillar but benefit from C. forda caterpillars which, after dropping onto the soil, is proposed to inoculate the soil surrounding the trees with the fungus A. nomius which suggests a symbiotic and/or synergistic relationship between B. africana trees and C. forda caterpillars.

The Use of Gonimbrasia belina (Westwood, 1849) and Cirina forda (Westwood, 1849) Caterpillars (Lepidoptera: Sarturniidae) as Food Sources and Income Generators in Africa.

Gonimbrasia belina (mopane worm) and Cirina forda caterpillars (Lepidoptera: Saturniidae) are mostly found in shrubs and trees, from where they are collected as larvae and are widely consumed across southern Africa by rural and increasingly urban populations. These caterpillars are among the most prominent, traded, and economically beneficial edible insects found in Western African countries, but also in South Africa, Zimbabwe, Botswana, and the Democratic Republic of the Congo. Over the years, these caterpillars have evolved from being part of the diet in various communities to playing a vital role in income generation. In addition, consumption of G. belina and C. forda caterpillars as potential food sources has gained momentum due to their potential for contributing to livelihoods and mitigating food security challenges across Africa while providing significant benefits to developing countries on a socio-economic and ecological level. Edible caterpillars serve as a good source of rich nutrients such as proteins, fatty acids, and micronutrients and can be used in formulating nutrient-dense complementary foods. However, limited information is available, specifically on different trees that serve as hosts to these caterpillars, as they depend on the leaves as their only source of food. In addition, the review aims to critique and document knowledge on the nutritional benefits, acceptance of the use of these caterpillars as food security, commercial value, and level of acceptance towards the utilization of caterpillars as food sources.

Influence of seasonal and geographic variation on the anti-HSV-1 properties and chlorogenic acids content of Helichrysum aureonitens Sch. Bip.

Pharmacological studies conducted in the past revealed the potential source of medicinal plants in the development of novel medicines. The phenolic contents of medicinal plants containing chlorogenic acids (CGA) have been linked to a variety of therapeutic effects, especially antiviral activity. Helichrysum aureonitens is a medicinal plant which has been reported to contain chlorogenic acids compounds and has also shown antiviral activities against a number of virus species including Herpes Simplex Virus-1 (HSV-1). In this study, the aim was to determine both the influence of seasonal variation and locality on the antiviral properties of H. aureonitens. Since chlorogenic acids have been reported as potent antiviral compounds, these compounds were targeted to determine the effects of locality and seasonal change on the chlorogenic acid profile, and subsequent antiviral activity. The ultra-performance liquid chromatography-quadrupole time-of-flight mass spectroscopy (UPLC-qTOF-MS) was employed to determine the metabolic profile variations of three derivatives of chlorogenic acids-caffeoylquinic acid (CQA), dicaffeoylquinic acid (DCQA) and tricaffeoylquinic acid (TCQA) in the harvested plants growing in two diverse geographical climates and two different seasons (spring and autumn). Using the cytopathic effect (CPE) reduction approach, twenty-six samples of the plants' leaves and stems collected during spring and autumn at Telperion nature reserve in Mpumalanga and Wakefield farm, Midlands in KwaZulu-Natal region of South Africa were evaluated for anti-HSV activity. The MTT assay was used for the cytotoxicity evaluation of the extracts prior to antiviral determination. Seventeen (mostly spring collections) of the twenty-six extracts examined were found to have considerable anti-HSV activity as measured by a reduction in tissue culture infectious dose (TCID50) of less than 105. The UPLC-qTOF-MS result revealed that dicaffeoylquinic acid (DCQA) is the most abundant, with higher concentrations in both regions and seasons. 3-CQA was also shown to be the most abundant isomer of caffeoylquinic acid in this investigation.

NADES Compounds Identified in Hypoxis hemerocallidea Corms during Dormancy.

Soaking Hypoxis hemerocallidea corms in distilled water improved the propagation and development of cormlets, suggesting the potential leaching-out of inhibitory chemical compounds. To investigate the presence of inhibitory compounds, nuclear magnetic resonance (NMR) spectral data of the leachate from dormant H. hemerocallidea corms were obtained using a 600 MHz 1H-NMR spectrometer. The 1H-NMR analysis led to the identification of choline, succinate, propylene glycol, and lactose, as inhibitory compounds. These four chemical compounds are part of the "Natural Deep Eutectic Solvents" (NADES) that protect plant cells during stress periods, each of which has the potential to inhibit bud growth and development. These compounds are supposedly leached out of the corms during the first rain under natural conditions, possibly accompanied by changes in the ratios of dormancy-breaking phytohormones and inhibitory compounds, to release bud dormancy. The identified chemical compounds heralded a novel frontier in the vegetative propagation of H. hemerocallidea as a medicinal plant, and for its enhanced sustainable uses.

Effect of Different Climatic Regions and Seasonal Variation on the Antibacterial and Antifungal Activity, and Chemical Profile of Helichrysum aureonitens Sch. Bip.

Native South Africans make use of Helichrysum aureonitens Sch. Bip. extracts for the treatment of a variety of infections and they are important in traditional medicinal preparations. This study investigated the effect of seasonal variation and geographical location on the antibacterial and antifungal activities of H. aureonitens. Material was collected in two different seasons: early spring, with high rainfall and high temperatures (October), and late autumn, with low rainfall and lower temperatures (May). Further analysis was carried out using 1H-NMR based metabolomics to analyse and compare the chemical profiles of the plants in both seasons and locations. Plant materials were collected from two sites for each season, at Wakefield farm (KwaZulu-Natal), representing a colder, wetter environment, and Telperion (Mpumalanga), representing a drier and warmer environment. Leaves of H. aureonitens were tested against bacteria (Proteus vulgaris (P. vulgaris) and Bacillus subtilis (B. subtilis)) as well as fungi (Aspergillus flavus (A. flavus), Aspergillus nomius (A. nomius), Cladosporium cladosporioides (C. cladosporioides), Fusarium oxysporum (F. oxysporum) and Penicillum halotolerans (P. halotolerans)). Extracts from the October harvest showed significant activities against the Gram-negative bacterium P. vulgaris compared to the May harvest, with an MIC value of 62.5 µg/mL. Similar activity was observed between the extracts from the wet season across the two geographically different locations. There was generally very good antifungal activity observed for all the species, with the exception of A. nomius, which had MIC values ranging from 0.39-1.56 µg/mL. Extracts of plant materials harvested in the wetter region had a significantly higher activity against A. flavus and F. oxysporum in both seasons than those from plants harvested in the drier region. Telperion-harvested plants exhibited better activity against F. oxysporum in the autumn. Hydrogen-1 NMR metabolomic analysis confirmed the significant effects of the seasons and the peculiar climates of different localities on the secondary metabolite profile of H. aureonitens.

Identification of Anti-HIV Biomarkers of Helichrysum Species by NMR-Based Metabolomic Analysis.

Several species of the Helichrysum genus have been used ethnobotanically to treat conditions that we today know have been caused by viral infections. Since HIV is a modern disease with no ethnobotanical history, we commenced with a study on the anti-HIV activity of several Helichrysum species. Drug discovery of small molecules from natural resources that is based on the integration of chemical and biological activity by means of metabolomical analyses, enables faster and a more cost-effective path to identify active compounds without the need for a long process of bioassay-guided fractionation. This study used metabolomics to identify anti-HIV compounds as biomarkers from 57 Helichrysum species in a combined study of the chemical and biological data of two previous studies. In the OPLS-DA and hierarchical cluster analyses, anti-HIV activity data was included as a secondary observation, which assisted in the correlation of the phytochemical composition and biological activity of the samples. Clear grouping revealed similarity in chemical composition and bioactivity of the samples. Based on the biological activity of polar extracts, there was a distinct phytochemical difference between active and non-active groups of extracts. This NMR-based metabolomic investigation showed that the chlorogenic acids, compounds with cinnamoyl functional groups, and quinic acid were the most prominent compounds in the Helichrysum species with anti-HIV activity. This study further revealed that the chlorogenic acid type compounds and quinic acid are biomarkers for anti-HIV activity.

New Alk(en)ylhydroxycyclohexanes with Tyrosinase Inhibition Potential from Harpephyllum caffrum Bernh. Gum Exudate.

This work presents the first report on the phytochemical investigation of Harpephyllum caffrum Bernh. gum exudate. A known cardanol, 3-heptadec-12'-Z-enyl phenol (1) and three new alk(en)ylhydroxycyclohexanes, namely, (1R,3R)-1,3-dihydroxy-3-[heptadec-12'(Z)-enyl]cyclohexane (2) (1S,2S,3S,4S,5R)-1,2,3,4,5-pentahydroxy-5-[octadec-13'(Z)-enyl]cyclohexane (3) and (1R,2S,4R)-1,2,4-trihydroxy-4-[heptadec-12'(Z)-enyl]cyclohexane (4) were isolated from the gum. The structures of the compounds were determined by extensive 1D and 2D NMR spectroscopy and HR-ESI-MS data. The ethanolic extract of the gum was found to be the most potent tyrosinase inhibitor with IC50 of 11.32 µg/mL while compounds 2 and 3, with IC50 values of 24.90 and 26.99 µg/mL, respectively, were found to be potential anti-tyrosinase candidates from the gum. Gum exudate may be a potential source for non-destructive harvesting of selective pharmacologically active compounds from plants. The results also provide evidence that H. caffrum gum may find application in cosmetics as a potential anti-tyrosinase agent.

Metabolomic profile of medicinal plants with anti-RVFV activity.

Twenty medicinal plants with previously established anti-viral activity against a wild-type RVFV were further investigated using bio-chemometric and analytical techniques. The aim being to identify compounds common in plants with anti-RVFV activity, potentially being the major contributors to the anti-viral effect. Proton nuclear magnetic resonance (1H NMR) spectroscopy coupled with multivariate data analysis (MVDA) was applied to characterize metabolite profiles of twenty antiviral medicinal plants. Discrimination and prediction of metabolome data of active anti-RVFV from the less-active samples was assessed using the multivariate statistical models by constructing a robust principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) regression model. Annotation of metabolites in the samples with higher activity were performed by Chenomx software and the compounds confirmed using Ultra-High-Performance Liquid Chromatography-Quadrupole Time-of-Flight Mass Spectrometry (UHPLC-qTOF-MS). Both the PCA and OPLS-DA score plots showed clustering of samples; however, the OPLS-DA plot indicated a clear separation among active and less-active samples. Metabolic biomarkers were screened by p-value < 0.05 and variable importance in the projection (VIP) value >1 and S-plot. Among active samples, the most prominent metabolites putatively identified by NMR include trigonelline, vanillic acid, fumarate, chlorogenic acid, ferulate, and formate. The presence of the compounds were confirmed by UHPLC-qTOF-MS, and two hydroxylated fatty acids were additionally detected indicated by peaks at m/z 293.2116 and m/z 295.2274 13S-Hydroxy-9Z,11E,15Z-octadecatrienoic acid and 13-Hydroxy-9Z,11E-octadecadienoic acid were annotated for the first time in all the antiviral active samples and are considered potential metabolites responsible for the antiviral activity. The study provides a metabolomic profile of anti-RVFV plant extracts and report for the first time the presence of hydroxylated fatty acids 13S-Hydroxy-9Z,11E,15Z-octadecatrienoic acid and 13-Hydroxy-9Z,11E-octadecadienoic acid, present in all the tested medicinal plants with high anti-RVFV activity and is a potential target for the future development of antiviral therapeutic agents.

Anti-Pseudomonas aeruginosa activity of a C16-terpene dilactone isolated from the endophytic fungus Neofusicoccum luteum of Kigelia africana (Lam.).

Fungal endophytes have the capacity to biosynthesize secondary metabolites that are produced by their host plants. In this study, a dilactone terpenoid of C16 architecture was isolated from the fungal endophytes of Kigelia africana, in our attempt to identify anti-Pseudomonas aeruginosa metabolites. Thirty-eight fungal isolates were cultured for biomolecule production over a period of thirty days. Extracts from three (ZF 34, ZF 52 and ZF 91) of the fungi showed good anti-P. aeruginosa activity, with ZF 52 presenting the best MIC of 19.53 µg/mL and was accordingly subjected to chromatographic separation. Based on nuclear magnetic resonance (NMR) spectroscopy, high resolution mass spectrometry and single crystal X-ray diffraction (XRD) analyses, the isolated compound was identified as a C16-terpene dilactone, with a structure consistent with that of the known diterpene, CJ-14445. The isolated dilactone showed anti-P. aeruginosa activity with MIC of 0.61 µg/mL, signifying the antibacterial potential of the biomolecule. The bioactive fungal isolate (ZF 52) was identified as Neofusicoccum luteum based on genomic DNA sequencing. This is the first report of the endophyte N. luteum from K. africana and the first reported occurrence of CJ-14445 in the fungus.

Terpene-rich fractions of Ficus mucoso (Welw) modulate lipopolysaccharide-induced inflammatory mediators and aberrant permeability of the inner mitochondrial membrane in murine animal model.

Ficus mucoso is traditionally used to treat bronchial infections. This study compared the efficacy of terpene-rich fractions of F. mucoso root bark on lipopolysaccharide(LPS)-induced inflammation, liver mitochondrial permeability transition (mPT), an index of mitochondrial health, and associated pathological alterations. Terpene-Rich Fractions of Dichloromethane (TRDF) and Ethylacetate Fractions of F. mucoso (TREF) were obtained according to standard procedures. To induce systemic inflammation, a single intraperitoneal injection of 1mgLPS/kgbw was given to mice. Spectrophotometric techniques were used to evaluate the effects of the oral administration of TRDF and TREF (3 days) on levels of pro-inflammatory mediators (TNF-α, IL-1ß, IL-6) using ELSA techniques as well as antioxidant indices in normal and LPS-treated mice. The mPT pore opening, mitochondrial ATPase activity and lipid peroxidation were monitored spectrophotometrically. Our results revealed that treatment with LPS caused significant elevation in serum cytokine levels while administration of 50 and 100 mg/kg TRDF and TREF significantly reduced elevated serum levels of cytokines (TNF-α, IL-1ß, IL-6) in LPS-challenged mice. In addition, activitities of superoxide dismutase, catalase and liver marker enzymes (ALT and AST) as well as levels of mitochondrial lipid peroxides were significantly reduced in mice treated with TRDF and TREF relative to LPS-fed mice. Furthermore, LPS caused induction of opening of the liver mPT pore which was significantly inhibited by TRDF at 100 and 200 mg/kg bw by 71% and 88%, respectively, but only at 100 mg/kg TREF. Furthermore, mitochondrial ATPase activity was inhibited largely by TRDF. UPLC-ESI-MS analysis revealed the presence of terpenoid derivatives and a few aromatic metabolites in TRDF. The terpene dominance of TRDF metabolites was further justified on the 1H NMR fingerprint. Overall, TRDF is more effective as a cocktail of anti-inflammatory compounds than TREF against LPS-induced acute systemic inflammation.

Metabolomic Profiling of Antioxidant Compounds in Five Vachellia Species.

The genus Vachellia, previously known as Acacia, belongs to the family Fabaceae, subfamily Leguminosae, which are flowering plants, commonly known as thorn trees. They are traditionally used medicinally in various countries including South Africa for the treatment of ailments such as fever, sore throat, Tuberculosis, convulsions and as sedatives. The aim of this study was to determine biochemical variations in five Vachellia species and correlate their metabolite profiles to antioxidant activity using a chemometric approach. The antioxidant activity of five Vachellia aqueous-methanolic extracts were analyzed using three methods: 2,2-di-phenyl-1-picrylhydrazyl (DPPH) radical scavenging assay, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS+) analysis and the ferric reducing antioxidant power (FRAP) assay by means of serial dilution and bioautography with the thin-layer chromatography (TLC) method. Amongst the Vachellia extracts tested, V. karroo, V. kosiensis and V. xanthophloea demonstrated the highest DPPH, ABTS+ and FRAP inhibitory activity. The antioxidant activities of DPPH were higher than those obtained by ABTS+, although these values varied among the Vachellia species. Proton nuclear magnetic resonance (1H NMR), coupled with multivariate statistical modeling tools such as principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA), were performed to profile metabolites responsible for the observed activity. The OPLS-DA categorized the five Vachellia species, separating them into two groups, with V. karroo, V. kosiensis and V. xanthophloea demonstrating significantly higher radical scavenging activity than V. tortilis and V. sieberiana, which clustered together to form another group with lower radical scavenging activity. Annotation of metabolites was carried out using the ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-qTOF-MS), and it tentatively identified 23 metabolites of significance, including epigallocatechin (m/z = 305.0659), methyl gallate (m/z = 183.0294) and quercetin (m/z = 301.0358), amongst others. These results elucidated the metabolites that separated the Vachellia species from each other and demonstrated their possible free radical scavenging activities.

Anti-Inflammatory Potentials of the n-Hexane Fraction of Alstonia boonei Stem Bark in Lipopolysaccharide-Induced Inflammation in Wistar Rats.

BACKGROUND: Inflammation is a protective response of the host to infections and tissue damage and medicinal plants have been used to regulate inflammatory response. The phytochemical contents of the n-hexane fraction of Alstonia boonei and their anti-inflammatory potentials in lipopolysaccharide-induced inflammation were investigated in rat liver. MATERIALS AND METHODS: A quantity of 5 mg/kg lipopolysaccharide (LPS) was used to induce inflammation in twenty-five male Wistar rats, grouped (n = 5) and treated as follows: negative control (10 mL/kg saline), positive control (1 mg/kg ibuprofen); 50, 100 and 20 mg/kg of the n-hexane fraction of Alstonia boonei were administered to test groups. In another experiment, twenty rats (n = 5, without LPS) were administered the same doses of the n-hexane fraction of A. boonei and ibuprofen for seven days. At the end of the experiment, animals were sacrificed, serum was obtained from blood and liver mitochondria isolated in a refrigerated centrifuge. Mitochondrial permeability transition (mPT) pore opening and mitochondrial F0F1 ATPase (mATPase) were determined spectrophotometrically. Serum interleukins 1ß, 6 (IL-1ß, IL-6), tumour necrosis factor alpha (TNF-α), C-reactive protein (CRP) and creatine kinase (CK), gamma glutamyl transferase (GGT), aspartate and alanine aminotransferases (AST and ALT,) of the animals in which inflammation was induced using LPS but treated with graded doses of n-hexane fraction of A. boonei were determined using the ELISA technique. The phytochemical contents of the n-hexane fraction of A. boonei were determined using ultra performance liquid chromatography-tandem mass spectrometer (UHPLC-MS). RESULTS: Calcium induced mPT in 8 fold and LPS induced mPT 14 fold in the negative control while the n-hexane fraction reversed mPT in the treated groups (50, 100 and 200 mg/kg) to 2, 4, 4 folds, respectively. LPS treatment of the negative group enhanced F0F1 mATPase activity, increased CRP, TNF-α, IL-1ß, IL-6 levels as well as CK, AST, ALT and GGT activities. These values were significantly reduced by 100 and 200 mg/kg of the n-hexane fraction. UHPLC-MS analysis of the fraction revealed the presence of terpenoids, phenolics and sphingolipids. CONCLUSION: These results showed that bioactive phytochemicals present in the n-hexane fraction of A. boonei were not toxic, have an anti-inflammatory effect and could be used for the treatment of inflammatory diseases.

NMR-Based Metabolomic Analyses to Identify the Effect of Harvesting Frequencies on the Leaf Metabolite Profile of a Moringa oleifera Cultivar Grown in an Open Hydroponic System.

Moringa oleifera Lam. is one of the world's most useful medicinal plants. Different parts of the M. oleifera tree contain a rich profile of important minerals, proteins, vitamins, and various important bioactive compounds. However, there are differences in the phytochemical composition of the medicinal plant's raw materials due to seasonal variation, cultivation practices, and post-harvest processing. The main objective of this study was therefore to determine the effect of harvesting frequencies on selected bioactive compounds of a M. oleifera cultivar (PKM1) grown in a hydroponic system under a shade net structure. Three harvesting frequency treatments were applied in the study, with the plants harvested at every 30 days (high frequency), 60 days (intermediate frequency), and 90 days (low frequency) respectively. 1H-NMR was used for data acquisition, and multivariate data analysis by means of principal component analysis (PCA), partial least square discriminatory analysis (PLS-DA), and orthogonal partial least square discriminatory analysis (OPLS-DA) were applied to determine the changes in the leaf metabolite profile, and also to identify the spectral features contributing to the separation of samples. Targeted metabolite analysis was used to match the NMR peaks of the compounds with the NMR chemical shifts of the contribution plot. The contribution plot showed that the increase in concentration of some compounds in aliphatic, sugar and aromatic regions contributed to the separation of the samples. The results revealed that intermediate and low harvesting frequencies resulted in a change in the leaf metabolite profile. Compounds such as chlorogenic acid, ferulic acid, vanillic acid, wogonin, esculetin, niazirin, and gamma-aminobutyric acid (GABA) showed an increase under intermediate and low harvesting frequencies. These results provide insight into the effect of harvesting frequencies on the metabolite profile and associated medicinal activity of M. oleifera.

1H-NMR and LC-MS Based Metabolomics Analysis of Wild and Cultivated Amaranthus spp.

Amaranthus crops are important for their use as food and nutritional sources, as well as for their medicinal properties. They are mostly harvested from the wild, and cultivation of Amaranthus species is still rare, and therefore, attempts are being made to commercialize and market this important crop. This research investigated the effect of cultivation and environment on the chemical profile of both cultivated and wild A. cruentus and A. hybridus by multivariate statistical analysis of spectral data deduced by Nuclear Magnetic Resonance (NMR). Furthermore, wild samples of A. cruentus and A. hybridus were subjected to Liquid Chromatography-Mass Spectrometry (LC-MS) for further analysis. Through NMR analysis, it was found that maltose and sucrose increased in both cultivated A. cruentus and A. hybridus. Moreover, the amino acid, proline was present in cultivated A. cruentus in high quantity whereas, proline and leucine were prominent in A. hybridus. Other compounds that were found in both wild and cultivated A. cruentus and A. hybridus are trehalose, trigonelline, lactulose, betaine, valine, alanine, fumarate, formate and kynurenine. LC-MS analysis revealed the presence of rutin, 2-phenylethenamine and amaranthussaponin I in both wild A. cruentus and A. hybridus, while chlorogenic acid was identified only in cultivated A. hybridus. On the contrary, L-tryptophan, kaempferol, phenylalanine and quercetin were detected only in wild A. cruentus. Amaranth is not only rich in macro and micronutrients, but the leaves also contain phytochemicals that vary between species and cultivated plants, and might, therefore, affect the medicinal properties of the material.

In Vitro Evaluation of Anti-Rift Valley Fever Virus, Antioxidant and Anti-Inflammatory Activity of South African Medicinal Plant Extracts.

Rift valley fever virus (RVFV) is a mosquito-borne virus endemic to sub-Saharan African countries, and the first sporadic outbreaks outside Africa were reported in the Asia-Pacific region. There are no approved therapeutic agents available for RVFV; however, finding an effective antiviral agent against RVFV is important. This study aimed to evaluate the antiviral, antioxidant and anti-inflammatory activity of medicinal plant extracts. Twenty medicinal plants were screened for their anti-RVFV activity using the cytopathic effect (CPE) reduction method. The cytotoxicity assessment of the extracts was done before antiviral screening using the MTT assay. Antioxidant and reactive oxygen/nitrogen species' (ROS/RNS) inhibitory activity by the extracts was investigated using non-cell-based and cell-based assays. Out of twenty plant extracts tested, eight showed significant potency against RVFV indicated by a decrease in tissue culture infectious dose (TCID50) < 105. The cytotoxicity of extracts showed inhibitory concentrations values (IC50) > 200 µg/mL for most of the extracts. The antioxidant activity and anti-inflammatory results revealed that extracts scavenged free radicals exhibiting an IC50 range of 4.12-20.41 µg/mL and suppressed the production of pro-inflammatory mediators by 60-80% in Vero cells. This study demonstrated the ability of the extracts to lower RVFV viral load and their potency to reduce free radicals.

Antimicrobial, Cytotoxic and Oxidative Stress Inhibitory Activities of Terpenoids and Flavonols from Senegalia nigrescens (Oliv.) P.J.H. Hurter.

Senegalia nigrescens (knob thorn) is a deciduous tree distributed in savannah regions from Tanzania to South Africa used for timber but also medicinally for the treatment of convulsions, wounds, and skin problems. In this study, the biological activities of six phytocompounds, namely: 3 ß -hydroxy-20(29)-en-lupan-30-al (1), 30-hydroxylup-20(29)-en-3 ß -ol (2), ent-kaur-15-en-18,20-diol (3), melanoxetin (4), quercetin (5) and quercetin-3-O-methyl ether (6), isolated from S. nigrescens were investigated. The compounds were screened against two bacterial (Escherichia coli and Staphylococcus aureus) and one fungal (Candida albicans) strain and were also tested for their cytotoxicity on breast cancer (MDA-MB-231) and normal murine macrophage (RAW 264.7) cell line. Effects of the compounds on attenuating the lipopolysaccharide (LPS)-induced intracellular reactive oxygen species (ROS) production in RAW 264.7 cells were quantified with the H2DCF-DA assay. This study revealed that flavonols (5 and 6) had the strongest antibacterial and antifungal effects, both having MIC values of 62.5, 31.25 and 31.25 µg/mL on E. coli, S. aureus and C. albicans, respectively. Compounds 2, 3 and 6 were the most cytotoxic against the breast cancer cells with IC50 values of 11.86, 12.62 and 14.03 µg/mL, respectively, while the least toxicity towards normal cells were observed in compounds 2, 5 and 6. All compounds (1-6) significantly lowered ROS production in RAW264.7 cells. In conclusion, tested compounds represent potential promising candidates as antimicrobial, anticancer and antidotes for LPS-induced oxidative stress. This is the first report on the antifungal, cytotoxicity and antioxidative activities of the ent-kaurene diterpenoid, ent-kaur-15-en-18,20-diol (3).

Comparative antimalarial, toxicity and mito-protective effects of Diospyros mespiliformis Hochst. ex A. DC. and Mondia whitei (Hook.f.) Skeels on Plasmodium berghei infection in mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Diospyros mespiliformis Hochst. ex A. DC. and Mondia whitei (Hook.f.) Skeels are traditionally used in Africa for the treatment of malaria. However, scientific evidence to substantiate this folkloric claim and their effects on liver mitochondria during malaria treatment have not been reported. AIM OF THE STUDY: This study investigated the efficacy of D. mespiliformis and M. whitei against chloroquine-sensitive and resistant strains of malarial parasites in mice. It also investigated the toxicity and protection against cellular organelles like mitochondria. MATERIALS AND METHODS: Male Swiss mice were infected with a chloroquine resistant (ANKA) strain of Plasmodium berghei and were treated via oral gavage with methanol extracts of D. mespiliformis and M. whitei reconstituted in diluted dimethylsulfoxide as vehicle (DMSO, 5% v/v) for five consecutive days. Percentage parasite load and clearance were assessed by microscopy. The infected control was treated with the vehicle. Hematological indices were assessed using standard procedures. Alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP) were determined using assay kits. Hepatic mitochondria were isolated via centrifugation, and their permeability transition (mPT), ATPase (mATPase) activity and lipid peroxidation (mLPO) were determined spectroscopically. Liver tissue histology was carried out by standard laboratory procedures. Phytochemical analysis of both extracts were performed using LC-MS to identify the most prominent compounds from each of the extracts. RESULTS: After treatment on day 5, D. mespiliformis and M. whitei at 400 mg/kg decreased mean values for: percentage parasitemia (5.0 ± 1.0, 2.0 ± 0.2), increased Packed Cell Volume (PCV) (36.0 ± 1.4, 36.0 ± 0.0%) and platelets (2.0 ± 1.4, 2.0 ± 2.8 × 105mm3) relative to the untreated control (20.0 ± 5.2; 30.0 ± 0.0%; 1.4 ± 1.4 × 105 mm3, respectively). At the same dose, D. mespiliformis and M. whitei decreased ALT (8.0 ± 3.8, 24.2 ± 4.0U/L), AST (6.2 ± 0.8, 8.0 ± 0.9U/L) and ALP (56.0 ± 0.7, 51.0 ± 1.0U/L) activities compared to the infected control (77.0 ± 10.9U/L, 14.0 ± 0.7U/L and 76.0 ± 6.0U/L, respectively). Both D. mespiliformis and M. whitei reversed mPT opening, decreased mATPase enhancement and mLPO, relative to the control. Histopathology of the liver showed extensive hemorrhagic lesions and severe disseminated congestion in the infected control while both D. mespiliformis and M. whitei were well tolerated at the highest dose. The LC-MS analysis of D. mespiliformis showed the presence of betulinic acid, tocopherol and kaempferol with antimalarial and antioxidant properties while the M. whitei sample contained coumarin and chlorogenic acid that have antimalarial and hepato-protective properties. CONCLUSIONS: D.mespiliformis and M. whitei show antimalarial effects against resistant Plasmodium berghei infection, enhanced cell viability, mito-protection and are not toxic in mice.

NMR-Based Metabolomic Analysis and Microbial Composition of Soil Supporting Burkea africana Growth.

Burkea africana is a leguminous tree used for medicinal purposes, growing in clusters, on soils impoverished from most nutrients. The study aimed to determine the factors responsible for successful reproduction and establishment of the B. africana trees in nature, as all efforts for commercial production has been proven unsuccessful. An investigation was carried out to determine the metabolomic profile, chemical composition, and microbial composition of the soils where B. africana grows (Burkea soil) versus the soil where it does not grow (non-Burkea soil). 1H-NMR metabolomic analysis showed different metabolites in the respective soils. Trehalose and betaine, as well as a choline-like and carnitine-like compound, were found to be in higher concentration in Burkea soils, whereas, acetate, lactate, and formate were concentrated in non-Burkea soils. Liquid Chromatography-Mass Spectrometry analysis revealed the presence of numerous amino acids such as aspartic acid and glutamine to be higher in Burkea soils. Since it was previously suggested that the soil microbial diversity is the major driver for establishment and survival of seedlings in nature, Deoxyribonucleic acid (DNA) was extracted and a BLAST analysis conducted for species identification. Penicillium species was found to be highly prevalent and discriminant between the two soils, associated with the Burkea soils. No differences in the bacterial composition of Burkea and non-Burkea soils were observed. The variances in fungal composition suggests that species supremacy play a role in development of B. africana trees and is responsible for creating a supporting environment for natural establishment and survival of seedlings.

Ethnobotany, phytochemistry and pharmacological significance of the genus Bulbine (Asphodelaceae).

ETHNOPHARMACOLOGICAL RELEVANCE: The genus Bulbine (Asphodelaceae) is spread across Southern Africa and Australia and has been traditionally used for various medicinal applications such as treating skin diseases, burns, diarrhoea, and sexually transmitted diseases. AIM OF THIS REVIEW: The aim is to present a critical review of the ethnomedicinally important species of the genus Bulbine with a comprehensive overview of their chemical constituents and biological activities. MATERIALS AND METHODS: This paper is an overview of literature published on the genus Bulbine in the last six decades with regards to phytochemical composition and their respective pharmacological potentials with the aid of data obtained from the search engine Google Scholar with string searches performed using keywords to obtain relevant publications from scientific databases including ACS Journals, PubMed, Science Direct, SciELO, Sci Finder, Springer, Tailor & Francis, The Plant List Database, Web of Science and Wiley. RESULTS: The literature survey reveals that only 12 species in the genus Bulbine have been reported to be used traditionally with scientific records of ethnomedicinal usage Anthraquinones appeared as the most abundant phytochemicals in the genus. Other isolated/detected metabolites include isofuranonaphthoquinones, flavonoids, and triterpenoids. Promising pharmacological activities have been reported by members of the genus with antiplasmodial, antitrypanosomal, antiviral, antioxidant, anticancer, anti-inflammatory and anti-microbial activity, potent wound healing properties as well as improved reproduction. CONCLUSIONS: This review showed the traditional uses of this genus and its preventative and curative properties in the management of the listed diseases providing support from bioassays of the tested compounds and extracts. State-of-the-art analytical techniques are required for the characterisation and quantification of the compounds within the genus. The efficacy of the therapeutic potential of the Bulbine species need to be further confirmed with pre-clinical and clinical studies.