Elucidating the chemical profile and biological studies of Verbascum diversifolium Hochst. extracts.

The present study was designed to evaluate the chemical composition, antioxidant, enzyme inhibition and cytotoxic properties of different extracts from aerial parts of V. diversifolium (family Scrophulariaceae), a plant that is native to Lebanon, Syria and Turkey. Six extracts, namely, hexane, dichloromethane (DCM), ethyl acetate (EtOAc), ethanol (EtOH), 70% EtOH, and water (aqueous) were prepared by maceration. The EtOH extract was predominated by the presence of rutin (4280.20 µg g-1) and p-coumaric acid (3044.01 µg g-1) while the highest accumulation of kaempferol-3-glucoside (1537.38 µg g-1), caffeic acid (130.13 µg g-1) and 4-hydroxy benzoic acid (465.93 µg g-1) was recorded in the 70% EtOH, aqueous, and EtOAc extracts, respectively. The EtOH (46.86 mg TE/g) and 70% EtOH (46.33 mg TE/g) extracts displayed the highest DPPH radical scavenging result. Both these extracts, along with the aqueous one, exerted the highest ABTS radical scavenging result (73.03-73.56 mg TE/g). The EtOH and 70% EtOH extracts revealed the most potent anti-AChE (2.66 and 2.64 mg GALAE/g) and anti-glucosidase (1.07 and 1.09 mmol ACAE/g) activities. The aqueous extract was the most efficacious in inhibiting the proliferation of prostate cancer (DU-145) cells with an IC50 of 8.71 µg/mL and a Selectivity Index of 3.7. In conclusion, this study appraised the use of V. diversifolium aerial parts as a potential therapeutic source for future development of phytopharmaceuticals that target specific oxidative stress-linked diseases including diabetes, cancer, cardiovascular disease, and Alzheimer's disease among others.

Ethnobotanical study on medicinal plants in Melit area (North Darfur), Western Sudan.

BACKGROUND: The documentation of ethnobotanical knowledge in Sudan is restricted to specific regions, and there is a far-reaching lack of written information on the traditional use of medicinal plants in other places like Darfur State, in western Sudan. The present study was designed to document the medicinal plants used in traditional medicine of Melit area in North Darfur State. METHOD: Ethnomedicinal information was collected from 135 local informants through semi-structured questionnaires. Data were analysed for use value (UV), informant consensus factor (ICF) and fidelity level. RESULTS: A total of 59 medicinal plants, belonging to 32 families and 55 genera, were recorded for their traditional uses in Melit area. Fabaceae were represented by highest number of species (13) followed by Asteraceae and Malvaceae (4 each) and Poaceae (3). Herbs comprise the main sources (50.8%) of traditional remedies. Fruits and stem bark (17.9% each) were the major plant parts used. Decoction (36.5%) is the most mode of preparation used. Geigeria alata was most commonly used species with UV of 2.37. The highest ICF values were recorded for swellings (ICF = 1.00) and respiratory system (ICF = 0.95) categories. Ten plants, namely Carica papaya, Corchorus trilocularis, Eragrostis cilianensis, Heliotropium sudanicum, Mollugo cerviana, Psiadia punctulate, Rhynchosia minima, Solanum coagulans, Solanum forskalii and Tephrosia purpurea, were cited for the first time as medicinal plants used in Sudan traditional medicine. Resins of Boswellia papyrifera, seeds of Nigella sativa, pods of Vachellia nilotica (syn. Acacia nilotica) and clove of Syzygium aromticum were used to make different preparations for the treatment of the corona virus. CONCLUSION: This is the first ethnobotanical survey conducted in this region which is always suffering from security issues, and results indicated that Melit area harbours high diversity of plants used traditionally to cure different health conditions. The present study aids in conserving such rich heritage, and it is recommended that the newly reported species worth further studying over their phytochemical and biological properties.

HPLC-MS/MS Profiles, Antioxidant, Neuroprotective, Antidiabetic and Skin Protective Effects of Different Extracts of Vicia peregrina L. Collected from the Eastern Region of Turkey.

Many Vicia species (Fabaceae) were proven to possess bioactive compounds with potential health beneficial properties. The present study was designed to determine the phenolic constituents, antioxidant and enzyme inhibition activities of aerial parts and seed of V. peregrina. Hexane, ethyl acetate and methanol extracts were prepared by maceration and aqueous extract by infusion. The chemical compositions of the extracts were determined using HPLC-MS/MS technology. The antioxidant activities were examined using various assays including free radical scavenging (ABTS and DPPH), reducing ability (CUPRAC and FRAP), metal chelation, and phosphomolybdenum. The enzyme inhibitory effects were investigated against cholinesterase, tyrosinase, amylase and glucosidase. The highest total phenolics and flavonoids contents were recorded in the methanol extracts of the seed (45.42 mg GAE/g) and aerial parts (40.33 mg RE/g) respectively. The aerial parts were characterized by higher accumulation of chlorogenic acid (9893.86 µg g-1 ), isoquercitrin (9400.33 µg g-1 ), delphindin 3,5 diglucoside (9113.28 µg g-1 ), hyperoside (6337.09 µg g-1 ), rutin (3489.83 µg g-1 ) and kaempferol-3-glucoside (2872.84 µg g-1 ). Generally, the methanol and aqueous extracts of the two studied parts exerted the best antioxidant activity with highest anti-DPPH (61.99 mg TE/g), anti-ABTS (101.80 mg TE/g) and Cu++ (16169 mg TE/g) and Fe+++ (172,36 mg TE/g) reducing capacity were recorded from the seed methanol extract. Methanol extract of the seed showed the best anti-tyrosinase activity (75.86 mg KAE/g). These results indicated that V. peregrina is rich with bioactive phenolics suggesting their use in different health promoting applications.

LC-MS/MS and GC-MS profiling, antioxidant, enzyme inhibition, and antiproliferative activities of Thymus leucostomus H ausskn. & V elen. extracts.

The chemical composition as well as antioxidant, antiproliferative, and enzyme inhibition activities of extracts from aerial parts of Thymus leucostomus H ausskn. & V elen. obtained with hexane, methanol, and water were evaluated. Results showed that the methanol extract had significantly (p < 0.05) the highest total phenolic content (TPC; 107.80 mg GAE/g) and total flavonoids content (TFC; 25.21 mg RE/g) followed by the aqueous extract (102.72 mg GAE/g and 20.88 mg RE/g, respectively). LC-MS/MS-guided profiling of the three extracts revealed that rosmarinic acid (34.8%), hesperetin (42.9%), and linoleic acid (18%) were the dominant compounds in the methanol, aqueous and hexane extracts, respectively. GC-MS analysis of the hexane extract showed that É£-sitosterol (29.9%) was the major constituent. The methanol extract displayed significantly (p < 0.05) the highest Cu++ , Fe+++ , and Mo(VI) ions scavenging and reducing properties while the aqueous extract exerted significantly (p < 0.05) the highest metal chelating power (42.51 mg EDTAE/g). Both the hexane and methanol extracts effectively inhibited the acetylcholinesterase enzyme (2.63 and 2.65 mg GALAE/g, respectively) while the former extract exerted significantly (p < 0.05) the highest butyrylcholinesterase (2.32 mg GALAE/g), tyrosinase (19.73 mg KAE/g), and amylase (1.16 mmol ACAE/g) inhibition capacity. The aqueous extract exhibited the best glucosidase inhibition property (0.49 mmol ACAE/g). The methanol and hexane extracts exerted a higher cytotoxic effect on HT-29 (IC50 : 8.12 µg/mL) and HeLa (IC50 = 8.08 µg/mL) cells, respectively. In conclusion, these results provide valuable insight into the potential use of T. leucostomus bioactive extracts in different pharmaceutical applications.

Understanding the Chemical Composition and Biological Activities of Different Extracts of Secamone afzelii Leaves: A Potential Source of Bioactive Compounds for the Food Industry.

Secamone afzelii (Roem. & Schult.) K. Schum (family Asclepiadaceae) is a creeping woody climber used to treat ailments in many traditional medicine systems. The present study aims to examine the antioxidant and enzyme inhibition activities of S. afzelii leaf using different compositions of methanol-water mixture as an extraction solvent. The extracts were characterized by HPLC-ESI-MSn in terms of chemical compounds. The in silico results show that compound 23 (quercitrin) has the higher docking scores among the selected substances and the MD simulation revealed that the interactions with the enzymatic pocket are stable over the simulation time and strongly involve the tyrosinase catalytic Cu atoms. All together the results showed that both 80% and 100% methanolic extracts contained significantly (p < 0.05) the highest total phenolics content while the highest content of total flavonoids was significantly (p < 0.05) extracted by 100% methanol. About 26 compounds were tentatively identified by HPLC-ESI-MSn and 6 of them were quantified using standards. Results showed that the extracts were rich in flavonoids with a relatively high abundance of two kaempferol glycosides comprising 60% of quantified compounds. The 100% and 80% methanol extracts recorded significantly (p < 0.05) the highest total antioxidant, DPPH and ABTS activity as well as tyrosinase and ⍺-amylase inhibitory activities. The best significant (p < 0.05) cholinesterase inhibitory activity and reducing capacity of Fe+++ and Cu++ was recorded from the 80% methanolic extract while 100% ethanolic extract gave the highest significant (p < 0.05) butyrylcholinesterase inhibitory activity. The best glucosidase activity was observed in the 50% and 80% methanolic extracts. Although the water extract displayed the least total phenolics and flavonoids content and consequently the lowest antioxidant and enzyme inhibition activity, it displayed significantly (p < 0.05) the highest chelating power. In conclusion, these results demonstrated the richness of S. afzelii leaf as a potential source of bioactive compounds for the food industry, for the preparation of food supplements and functional foods.

Phenolic Profile, Antioxidant and Enzyme Inhibitory Activities of Leaves from Two Cassia and Two Senna Species.

Several species within the genera Cassia or Senna have a treasure of traditional medicines worldwide and can be a promising source of bioactive molecules. The objective of the present study was to evaluate the phenolic content and antioxidant and enzyme inhibition activities of leaf methanolic extracts of C. fistula L., C. grandis L., S. alexandrina Mill., and S. italica Mill. The two Cassia spp. contained higher total polyphenolic content (42.23-49.75 mg GAE/g) than the two Senna spp., and C. fistula had significantly (p ˂ 0.05) the highest concentration. On the other hand, the Senna spp. showed higher total flavonoid content (41.47-59.24 mg rutin equivalent per g of extract) than that found in the two Cassia spp., and S. alexandrina significantly (p ˂ 0.05) accumulated the highest amount. HPLC-MS/MS analysis of 38 selected bioactive compounds showed that the majority of compounds were identified in the four species, but with sharp variations in their concentrations. C. fistula was dominated by epicatechin (8928.75 µg/g), C. grandis by kaempferol-3-glucoside (47,360.04 µg/g), while rutin was the major compound in S. italica (17,285.02 µg/g) and S. alexandrina (6381.85). The methanolic extracts of the two Cassia species exerted significantly (p ˂ 0.05) higher antiradical activity, metal reducing capacity, and total antioxidant activity than that recorded from the two Senna species' methanolic extracts, and C. fistula displayed significantly (p ˂ 0.05) the highest values. C. grandis significantly (p ˂ 0.05) exhibited the highest metal chelating power. The results of the enzyme inhibition activity showed that the four species possessed anti-AChE activity, and the highest value, but not significantly (p ≥ 0.05) different from those obtained by the two Cassia spp., was exerted by S. alexandrina. The Cassia spp. exhibited significantly (p ˂ 0.05) higher anti-BChE and anti-Tyr properties than the Senna spp., and C. grandise revealed significantly (p ˂ 0.05) the highest values. C. grandise revealed significantly (p ˂ 0.05) the highest α- amylase inhibition, while the four species had more or less the same effect against the α-glucosidase enzyme. Multivariate analysis and in silico studies showed that many of the identified phenols may play key roles as antioxidant and enzyme inhibitory properties. Thus, these Cassia and Senna species could be a promising source of natural bioactive agents with beneficial effects for human health.

Chemical Profile, Antioxidant and Enzyme Inhibition Activities of Natural Saudi Sidr and Talh Honeys.

Honey is used since ancient time as a food and to cure many diseases. The present study investigated the chemical constituents, antioxidant and enzyme inhibition activities of natural Saudi Sidr (SH) and Talh (TH) honeys. Beside entire honey samples, ethyl acetate, ethanol and water extracts were prepared. The total polyphenolic content of SH, TH and their extracts was in the range of 2.86-7.21 and 3.80-17.33 mg gallic acid equivalents/g, respectively and the total flavonoids content was in the range of 0.05-1.17 and 0.18-2.38 mg rutin equivalents/g, respectively. Out of the 53 standards analyzed by HPLC, 27 compounds were detected with highest number of compounds identified in the ethyl acetate extract of TH (45 %, 24/53) and SH (26 %, 14/53), respectively. Quinic acid was dominant compound identified in all honey samples with the highest content determined in TH ethanol extract (4454 µg/g). The majority of tested samples possessed considerable anti-radicals and reducing ions capacity with the ethyl acetate extract from TH exerted significantly (p<0.05) the highest activity. All honey samples did not show chelating iron metal property. Honey samples revealed variable enzyme inhibition activity with TH (entire and/or ethyl acetate extract) showed significantly (p<0.05) the highest acetylcholinesterase, butyrylcholinesterase, tyrosinase and α-amylase inhibition activity. In conclusion, ethyl acetate is the best solvent for extraction of bioactive molecules from the two honey types. Moreover, the dark-colored TH contained the highest number of molecules and consequently exerted the best antioxidant and enzyme inhibition activities in most assays.

Chemical profile, anti 5-lipoxygenase and cyclooxegenase inhibitory effects of ginger (Zingiber officinale) rhizome, callus and callus treated with elicitors / Perfil químico, anti-5-lipoxigenase e efeitos inibitórios da ciclooxegenase do rizoma de gengibre (Zingiber officinale), do calo e calos tratados com elicitores

The present study investigated the chemical profiles and evaluated the inhibitory effect against 5-Lipoxygenase (5-Lox) activity for extracts of ginger rhizome, callus, and callus treated with the elicitors; yeast extract (100, 300 and 500 mg/L), glycine (100, 200 and 300 mg/L) and salicylic acid (100 and 200 mg/L). Oils and chloroform: methanol (CM) extracts were prepared by maceration in petroleum ether and CM (1:1, v/v), respectively. Chemical profiles were determined by gas chromatography/mass spectrometry (GC/MS) analysis. Oil of the callus recorded higher 5-Lox inhibitory effect (IC50 58.33±4.66 µg/mL) than the oil of rhizome (IC50168.34±15.64 µg/mL) and comparable to that of the positive control; Nordihydroguaiaretic acid (IC50 61.25±1.02 µg/mL). The chemical profile of the callus oil contained large amounts of fatty acids, mainly the unsaturated fatty acid oleic acid (31.11%) and saturated fatty acid palmitic acid (28.56%). Elicitors modified the chemical profile of the callus and ameliorated the anti-5-Lox activity of CM extract of the callus. CM extracts of callus treated with 100 and 300 mg/L yeast extract and 50 mg/L salicylic acid significantly suppressed (P ≤ 0.05) the 5-Lox activity by 33.16%, 25.46% and 16%, respectively as compared to the CM extract of untreated callus. In conclusion, ginger callus could be considered as a valuable dietary supplement in the treatment of various inflammatory disorders.

O presente estudo teve como objetivo investigar os perfis químicos e avaliar o efeito inibitório da atividade da 5-Lipoxigenase (5-Lox) em extratos de rizoma, calo e calo de gengibre tratados com os eliciadores; extrato de levedura (100, 300 e 500 mg / L), glicina (100, 200 e 300 mg / L) e ácido salicílico (100 e 200 mg / L). Extratos de óleos e clorofórmio: metanol (CM) foram preparados por maceração em éter e CM (1: 1, v / v), respectivamente. Os perfis químicos foram determinados por análise de cromatografia gasosa / espectrometria de massa (GC / MS). O óleo do calo registrou maior efeito inibitório de 5-Lox (IC50 58,33 ± 4,66 µg / mL) do que o óleo de rizoma (IC50168,34 ± 15,64 µg / mL) e comparável ao do controle positivo; Ácido nordi-hidroguaiarético (IC50 61,25 ± 1,02 µg / mL). O perfil químico do óleo de calo continha grandes quantidades de ácidos graxos, principalmente o ácido graxo insaturado ácido oleico (31,11%) e ácido graxo saturado palmítico (28,56%). Os elicitores modificaram o perfil químico do calo e melhoraram a atividade anti-5-Lox do extrato de CM do calo. Extratos de CM de calos tratados com 100 e 300 mg / L de extrato de levedura e 50 mg / L de ácido salicílico suprimiram significativamente (P ≤ 0,05) a atividade de 5-Lox em 33,16%, 25,46% e 16%, respectivamente, em comparação com o extrato de CM de calo não tratado. Em conclusão, o calo de gengibre pode ser considerado um suplemento dietético valioso no tratamento de vários distúrbios inflamatórios.

Evaluation of Antiviral, Antibacterial and Antiproliferative Activities of the Endophytic Fungus Curvularia papendorfii, and Isolation of a New Polyhydroxyacid.

An endophytic fungus isolated from Vernonia amygdalina, a medicinal plant from Sudan, was taxonomically characterized as Curvularia papendorfii. Ethyl acetate crude extract of C. papendorfii revealed an important antiviral effect against two viral pathogens, the human coronavirus HCoV 229E and a norovirus surrogate, the feline coronavirus FCV F9. For the last one, 40% of the reduction of the virus-induced cytopathogenic effect at lower multiplicity of infection (MOI) 0.0001 was observed. Selective antibacterial activity was obtained against Staphylococcus sp. (312 µg/mL), and interesting antiproliferative activity with half maximal inhibitory concentration (IC50) value of 21.5 ± 5.9 µg/mL was observed against human breast carcinoma MCF7 cell line. Therefore, C. papendorfii crude extract was further investigated and fractionated. Twenty-two metabolites were identified by gas chromatography coupled to mass spectrometry (GC-MS), and two pure compounds, mannitol and a new polyhydroxyacid, called kheiric acid, were characterized. A combination of spectroscopic methods was used to elucidate the structure of the new aliphatic carboxylic acid: kheiric acid (3,7,11,15-tetrahydroxy-18-hydroxymethyl-14,16,20,22,24-pentamethyl-hexacosa-4E,8E,12E,16,18-pentaenoic acid). Kheiric acid showed an interesting result with a minimum inhibitory concentration (MIC) value of 62.5 µg/mL against meticillin-resistant Staphylococcus aureus (MRSA). Hence, endophytes associated with medicinal plants from Sudan merit more attention, as they could be a treasure of new bioactive compounds.

Trans-3,5-dicaffeoylquinic acid from Geigeria alata Benth. & Hook.f. ex Oliv. & Hiern with beneficial effects on experimental diabetes in animal model of essential hypertension.

Geigeria alata Benth. & Hook.f. ex Oliv. & Hiern (Asteraceae) is used in Sudanese folk medicine for treatment of diabetes. The study aimed to estimate the acute oral toxicity of trans-3,5-dicaffeoylquinic acid (3,5-diCQA) from G. alata roots and to assess its antihypeglycemic, antioxidant and antihypertensive effects on chemically-induced diabetic spontaneously hypertensive rats (SHRs). The structure of 3,5-diCQA was established by NMR and HRMS spectra. Type 2 diabetes was induced by intraperitoneal injection of streptozotocin. 3,5-diCQA was slightly toxic with LD50 = 2154 mg/kg. At 5 mg/kg 3,5-diCQA reduced significantly (p < 0.05) the blood glucose levels by 42%, decreased the blood pressure by 22% and ameliorated the oxidative stress biomarkers reduced glutathione, malondialdehyde, and serum biochemical parameters. The beneficial effect on antioxidant enzymes was evidenced by the elevated glutathione peroxidase, glutathione reductase, and glutathione S-transferase activitiy in the livers of diabetic animals. 3,5-diCQA prevents the histopathological changes related to diabetes and hypertension. 3,5-diCQA was more potent α-glucosidase inhibitor (IC50 27.24 µg/mL) than acarbose (IC50 99.77 µg/mL). The antihyperglycemic action of the compound was attributed to the α-glucosidase inhibition. The beneficial effects of 3,5-diCQA on streptozotocin-induced diabetic hypertensive rats support the traditional use of G.alata for the management of diabetes.

Cytotoxicity of cucurbitacin E from Citrullus colocynthis against multidrug-resistant cancer cells.

BACKGROUND: Cucurbitacin E (CuE) is an oxygenated tetracyclic triterpenoid isolated from the fruits of Citrullus colocynthis (L.) Schrad. PURPOSE: This study outlines CuE's cytotoxic activity against drug-resistant tumor cell lines. Three members of ABC transporters superfamily, P-glycoprotein (P-gp), breast cancer resistance protein (BCRP) and ABCB5 were investigated, whose overexpression in tumors is tightly linked to multidrug resistance. Further factors of drug resistance studied were the tumor suppressor TP53 and the epidermal growth factor receptor (EGFR). METHODS: Cytotoxicity assays (resazurin assays) were used to investigate the activity of Citrullus colocynthis and CuE towards multidrug resistant cancer cells. Molecular docking (In silico) has been carried out to explore the CuE's mode of binding to ABC transporters (P-gp, BCRP and ABCB5). The visualization of doxorubicin uptake was done by a Spinning Disc Confocal Microscope. The assessment of proteins expression was done by western blotting analysis. COMPARE and hierarchical cluster analyses were applied to identify, which genes correlate with sensitivity or resistance to cucurbitacins (CuA, CuB, CuE, CuD, CuI, and CuK). RESULTS: Multidrug-resistant cells overexpressing P-gp or BCRP were cross-resistant to CuE. By contrast, TP53 knock-out cells were sensitive to CuE. Remarkably, resistant cells transfected with oncogenic ΔEGFR or ABCB5 were hypersensitive (collateral sensitive) to CuE. In silico analyses demonstrated that CuE is a substrate for P-gp and BCRP. Immunoblot analyses highlighted that CuE targeted EGFR and silenced its downstream signaling cascades. The most striking result that emerged from the doxorubicin uptake by ABCB5 overexpressing cells is that CuE is an effective inhibitor for ABCB5 transporter when compared with verapamil. The COMPARE analyses of transcriptome-wide expression profiles of tumor cell lines of the NCI identified common genes involved in cell cycle regulation, cellular adhesion and intracellular communication for different cucurbitacins. CONCLUSION: CuE represents a potential therapeutic candidate for the treatment of certain types of refractory tumors. To best of our knowledge, this is the first time to identify CuE and verapamil as inhibitors for ABCB5 transporter.

Ethnobotanical investigation on medicinal plants in Algoz area (South Kordofan), Sudan.

BACKGROUND: The inhabitants of western Sudan use traditional medicine for the treatment of various ailments due to lack of medical doctors and unaffordable prices of pharmaceutical products. The present study is the first documentation of the traditional plant knowledge on medicinal uses of plants by healers in Algoz (South Kordofan), Sudan. METHOD: Ethnobotanical data were collected over a period from March to November 2015 using semi-structured interviews with 30 healers (24 male and 6 female) living in the investigated area. Quantitative indices such as use categories, use value (UV) and informant consensus factor (ICF) were intended to evaluate the importance of medicinal plant species. RESULTS: A total of 94 medicinal plants, which belong to 45 families and 81 genera, were recorded in the study area. The most represented families are Leguminosae with 20 species followed by Combretaceae (6 species), Rubiaceae (5 species) and Asteraceae (4 species). The reported species were belonging to herbs (43%), trees (28%), shrubs (22%), climbers (4%) and parasites (3%). Root and stem (21% each) were the most plant parts used. A majority of remedies are administered orally (67%) where infusion (36%) and maceration (32%) are the most used methods. The highest ICF (0.87) was reported for poisonous animal bites followed by urinary system diseases (0.89), blood system disorders (0.88) and gynaecological diseases (0.87). Anastatica hierochuntica, Ctenolepis cerasiformis, Echinops longifolius, Cleome gynandra, Maerua pseudopetalosa, Martynia annua, Oldenlandia uniflora, Opuntia ficus-indica, Solanum dubium, Sonchus cornutus, Tribulus terrestris and Drimia maritima were reported for the first time in this study. CONCLUSION: The number of medicinal plants reported in this paper reflects evidence that Algoz area had a high diversity of medicinal plants which will continue to play an important role in the healthcare system in the study area.

In Vitro Antioxidant and Cytotoxic Activities of 18 Plants from the Erkowit Region, Eastern Sudan.

We investigated the antioxidant potential and cytotoxicity towards human CCRF-CEM leukemia cells of 57 extracts obtained from 18 plants collected in the Erkowit region, eastern Sudan. The antioxidant activity was determined by measuring the radical scavenging effects against 2,2-diphenyl-1-picrylhydrazyl (DPPH) and N,N-dimethyl-p-phenylendiamine (DMPD), metal-chelation capacity, ferric-reducing (FRAP) and phosphomolibdenum-reducing antioxidant power (PRAP) methods using ELISA microtiter assays. Total phenol and flavonoid amounts of the extracts were determined spectrophotometrically. Cytotoxicity towards CCRF-CEM cells was evaluated by the resazurin reduction assay. Geranium favosum followed by Kalanchoe glaucescens, Malva parviflora, Aizoon canariense, and Coleus barbatus, respectively, possessed the highest antioxidant activity among the studied plants. Chrozophora oblongifolia and K. glaucescens exerted considerable cytotoxicity against CCRF-CEM leukemia cells. These plants may serve as source for the further development of natural antioxidant and antitumor agents.

Traditional medicinal plants used for the treatment of diabetes in the Sudan : A review

The prevalence of metabolic disorders such as diabetes among population is of increasing concern worldwide. Sudan is a developing country, where several areas frequently depend on folk medicine. Several herbal preparations have been used in folklore practice in Sudan for the management of diabetes with claims asserting their hypoglycemic effect. Basic research relating to these plants are reviewed in this paper with the intention to highlight their therapeutic potential for the treatment of diabetes and promote their regular use in Sudan. Ethnobotanical information was obtained by an assessment of the available literature in electronic data bases with publications describing the medicinal plants used for the treatment of diabetes. In this review paper, different parts of 38 plant species, are described that are used in the Sudanese traditional medicine and belong to 35 genera and 23 families. Thirty three plants have been documented in scientific literature to possess in vivo antidiabetic activity and only one was ineffective in lowering blood glucose level, namely Striga hermonthica. Many of the plants in the study review have been studied in in vitro models (such as α-amylase or α-glucosidase inhibition) in an effort to explain some of their biomedical interaction. The role of isolated bioactive compounds like trigonelline and 3, 5-dicaffeoylquinic acid in diabetes management is also evaluated in the present review. Ten plants original from Sudan have been already used in clinical trials for the treatment of type 2 diabetes. This review provides useful information on the characterization of such herbal medicines that are utilized in the Sudanese traditional medicine for the control of metabolic syndromes such as diabetes

HPLC-UV and LC-MS Analyses of Acylquinic Acids in Geigeria alata (DC) Oliv. & Hiern. and their Contribution to Antioxidant and Antimicrobial Capacity.

INTRODUCTION: Geigeria alata is a traditional plant used in Sudanese folk medicine for treatment of diabetes, cough, epilepsy and intestinal complaints. OBJECTIVE: To analyze phenolic acids in Geigeria alata roots and leaves and to evaluate their antioxidant and antimicrobial activities. METHODOLOGY: Phenolic acids in the aqueous-methanol extracts were identified by LC-MS. Major compounds were isolated using low-pressure liquid chromatography. The quantitative analysis of phenolic acids was performed by a validated HPLC-UV method with limits of detection ranging from 0.04 to 0.57 µg/mL. 2,2-Diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azinobis-(3-ethylbenzothiazine-6-sulphonic acid) (ABTS) and ferric reducing antioxidant power (FRAP) methods were used for antioxidant activity evaluation. In addition, the minimal inhibitory concentration and the minimal bactericidal concentration against a panel of pathogenic bacteria and fungi were determined by the broth microdilution test. RESULTS: For the first time protocatechuic, caffeic, p-coumaroylquinic, caffeoylsinapoylquinic, caffeoylferuloylquinic, three feruloylquinic, six caffeoylquinic acids, and a caffeic acid hexoside were detected in Geigeria alata roots by LC-MS. HPLC-UV analyses showed that 3,5-dicaffeoylquinic acid (25.96 ± 2.08 mg/g dry weight (DW)) was the most abundant phenolic acid in roots, while 4,5-dicaffeoylquinic acid (8.99 ± 0.56 mg/g DW) was the main compound present in leaves. 3,5-Dicaffeoylquinic acid demonstrated stronger radical scavenging activity and reducing power compared with the crude extracts and the positive control 5-caffeoylquinic acid. 3,4,5-Tricaffeoylquinic acid revealed the highest antibacterial potential against the penicillin sensitive and resistant Staphylococcus aureus strains, as well as methicillin-resistant S. aureus. CONCLUSION: The caffeoylquinic acids content of up to 6.22% in Geigeria alata roots establishes this species as a new source rich in these bioactive molecules. Copyright © 2016 John Wiley & Sons, Ltd.

Endophytic fungi associated with Sudanese medicinal plants show cytotoxic and antibiotic potential.

In this study, we isolated 15 endophytic fungi from five Sudanese medicinal plants. Each fungal endophytic strain was identified by sequencing of internal transcribed spacer (ITS) regions of rDNA. Ethyl acetate extracts were prepared from each endophyte cultivated in vitro and tested for their respective antibacterial activities and antiproliferative activities against human cancer cells. Antibacterial screening was carried out against two bacterial strains: Gram-negative Escherichia coli and Gram-positive methicillin-resistant Staphylococcus aureus, by the broth dilution method. Cell viability was evaluated by the MTT procedure after exposure of MCF7 breast cancer cells and HT29 or HCT116 human colon adenocarcinoma cells to each endophytic extract. Of interest, Byssochlamys spectabilis isolated from Euphorbia prostata showed cytotoxicity (IC50 = 1.51 ± 0.2 µg mL(-1)) against MCF7 cells, but had a low effect against HT29 or HCT116 cells (IC50 > 20 µg mL(-1)). Cladosporium cladosporioides 2, isolated from Vernonia amygdalina leaves, showed antiproliferative activities against MCF7 cells (IC50 = 10.5 ± 1.5 µg mL(-1)) only. On the other hand, B. spectabilis and Alternaria sp. extract had antibacterial activities against the S. aureus strain. The findings of this work revealed that endophytic fungi associated with medicinal plants from Sudan could be considered as an attractive source of new therapeutic compounds.

A pilot study of antioxidant potential of endophytic fungi from some Sudanese medicinal plants.

OBJECTIVE: To evaluate the total phenolic content and total antioxidant capacity of ethyl acetate extracts of 21 endophytic fungi isolated from five Sudanese medicinal plants: Calotropis procera, Catharanthus roseus, Euphorbia prostrate, Vernonia amygdalina and Trigonella foenum-graecum. METHODS: Crude extracts of endophytic fungi and their host plants were tested by classical Folin-Ciocalteu colorimetric method to determine the total phenolic content, also total antioxidant capacity was estimated using 1,1-diphenyl-2-picrylhydrazyl free radical scavenging in vitro method. RESULTS: Among the endophytes, endophytic fungus Aspergillus sp. from Trigonella foenum-graecum seeds demonstrated the highest both total phenolic content in term of gallic acid equivalent [(89.9 ± 7.1) mg GAE/g] and antioxidant activity for 1,1-diphenyl-2-picrylhydrazyl radical scavenging assay [IC50: (18.0 ± 0.1) µg/mL]. A high positive linear correlation (R(2) = 0.999 1) was found between total antioxidant capacity and total phenolic content of endophytic fungi isolated from Vernonia amygdalina. CONCLUSIONS: The present study revealed that some endophytic fungi from the five Sudanese medicinal plants could be a potential source of novel natural antioxidant compounds.

Comparative antioxidant activity appraisal of traditional Sudanese kisra prepared from two sorghum cultivars.

The effect of fermentation and heating on the antioxidant activity of the fermented and fermented baked (kisra) dough prepared from two Sorghum cultivars (Tabat and Wad Ahmed) was evaluated. Kisra prepared from Tabat cultivar showed higher DPPH (1,1-diphenyl-2-picrylhydrazyl radical) scavenging and ferric reducing/antioxidant power (FRAP) than that of the Wad Ahmed cultivar. Baking improves the DPPH and FRAP of the kisra prepared from two cultivars. Baking caused a variable effect on the total phenol, tannins and flavonoids content across different periods of fermentation where an increase was mainly observed for samples subjected to longer periods of fermentation. A positive high correlation between the total phenol and antioxidant activity, using the DPPH and FRAP assays, was obtained for kisra prepared from both cultivars. The same observation was obtained for tannin content. In conclusion, fermentation and heating improve the antioxidant capacity of the sorghum grains from Tabat and Wad Ahmed cultivars.

Cucurbitacin L 2-O-ß-Glucoside Demonstrates Apoptogenesis in Colon Adenocarcinoma Cells (HT-29): Involvement of Reactive Oxygen and Nitrogen Species Regulation.

Emerging evidence suggests that reactive oxygen (ROS) and nitrogen (RNS) species can contribute to diverse signalling pathways of inflammatory and tumour cells. Cucurbitacins are a group of highly oxygenated triterpenes. Many plants used in folk medicine to treat cancer have been found to contain cucurbitacins displaying potentially important anti-inflammatory actions. The current study was designed to investigate the anti-ROS and -RNS effects of cucurbitacin L 2-O-ß-glucoside (CLG) and the role of these signaling factors in the apoptogenic effects of CLG on human colon cancer cells (HT-29). This natural cucurbitacin was isolated purely from Citrullus lanatus var. citroides (Cucurbitaceae). The results revealed that CLG was cytotoxic to HT-29. CLG increased significantly (P < 0.05) RNA and protein levels of caspase-3 in HT-29 cells when verified using a colorimetric assay and realtime qPCR, respectively. The results showed that lipopolysaccharide/interferon-gamma (LPS/INF-γ) increased nitrous oxide (NO) production inR AW264.7macrophages, whereas N(G)-nitro-L-argininemethyl ester (L-NAME) and CLG curtailed it. This compound did not reveal any cytotoxicity on RAW264.7 macrophages and human normal liver cells (WRL-68) when tested using the MTT assay. Findings of ferric reducing antioxidant power (FRAP) and oxygen radical absorption capacity (ORAC) assays demonstrate the antioxidant properties of CLG. The apoptogenic property of CLG on HT-29 cells is thus related to inhibition of reactive nitrogen and oxygen reactive species and the triggering of caspase-3-regulated apoptosis.

The antidiabetic effect of Geigeria alata is mediated by enhanced insulin secretion, modulation of ß-cell function, and improvement of antioxidant activity in streptozotocin-induced diabetic rats.

In Sudanese folk medicine, Geigeria alata roots have been used for the management of diabetes for a long time. However, its antidiabetic activity is unreported. In this study, G. alata methanolic extract was tested for its antidiabetic, antioxidant, and ß-cell modulatory effects in a streptozotocin-induced diabetic rat model. In this model of diabetic rats, the oral glucose tolerance test with G. alata extract at 125, 250, and 500  mg/kg doses revealed the efficacy of the 250  mg/kg dose in improving glucose tolerance comparable to the standard drug glibenclamide. Diabetic rats were treated with a 250  mg/kg dose of G. alata extract orally for 2  h (acute) or 14 days (chronic). In the case of acute treatment, the extract lowered blood glucose levels significantly at 120  min both in nondiabetic and diabetic rats. Chronic treatment of diabetic rats with 250  mg/kg of G. alata extract resulted in a significant decrease in blood glucose level closer to that of nondiabetic rats. Interestingly, increased serum insulin, improved ß-cell function, and antioxidant status were observed in G. alata-treated diabetic rats. G. alata also showed strong antioxidant and α-glucosidase inhibitory activities in in vitro assays. These data show direct evidence that G. alata has antidiabetic activity and suggest that the antidiabetic activity is due to enhanced insulin secretion, modulation of ß-cell function, and improvement of antioxidant status.