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This study assessed the optimum roasting conditions on the phytochemical properties of three varieties of Dioclea reflexa seeds using response surface methodology. Roasting conditions were varied using temperature (110°C~200°C) and time (10~40 min). Phytochemical components (phenolics, tannin, flavonoids, cardiac glycoside, and steroids) of the seeds were screened and estimated. The study showed that availability of phytochemical activities was heat-dependent. An increase in roasting temperature beyond 110°C for 10 min resulted in a decrease in total phenolic (TP) and flavonoid (TF) contents. However, prolonged durations of roasting favored increased amounts of TP and TF in dark and light varieties. Total sterol, tannin, and cardiac glycoside contents increased with increasing roasting temperature and time. The desirability of the models were 0.76, 0.74, and 0.72 for black, dark brown, and light brown, respectively. The coefficients of regression (R2), ranged from 0.66 to 0.98 signifying accuracy of the model. The following models (cubic, quadratic, and 2 factor interaction) were significant (P≤0.05). We found that roasting time influenced the phytochemical properties of D. reflexa to a greater extent than temperature. The optimum roasting temperature and time was found to beat 110°C, 35 min, 40 min, and 32 min in black, dark brown, and light brown varieties, respectively. Roasting conditions significantly affects the phytochemical contents of three varieties of D. reflexa seed flour (P

Dioclea reflexa bioactive compounds have been shown to contain antioxidant properties. The extracts from the same plant are used in traditional medical practices to treat various diseases with impressive outcomes. In this study, ionic mobility in Saccharomyces cerevisiae cells in the presence of D. reflexa seed extracts was monitored using electrochemical detection methods to link cell death to ionic imbalance. Cells treated with ethanol, methanol, and water extracts were studied using cyclic voltammetry and cell counting to correlate electrochemical behavior and cell viability, respectively. The results were compared with cells treated with pore-forming Amphotericin b (Amp b), as well as Fluconazole (Flu) and the antimicrobial drug Rifampicin (Rif). The D. reflexa seed water extract (SWE) revealed higher anodic peak current with 58% cell death. Seed methanol extract (SME) and seed ethanol extract (SEE) recorded 31% and 22% cell death, respectively. Among the three control drugs, Flu revealed the highest cell death of about 64%, whereas Amp b and Rif exhibited cell deaths of 35% and 16%, respectively, after 8 h of cell growth. It was observed that similar to SWE, there was an increase in the anodic peak current in the presence of different concentrations of Amp b, which also correlated with enhanced cell death. It was concluded from this observation that Amp b and SWE might follow similar mechanisms to inhibit cell growth. Thus, the individual bioactive compounds from the water extracts of D. reflexa seeds could further be purified and tested to validate their potential therapeutic application. The strategy to link electrochemical behavior to biochemical responses could be a simple, fast, and robust screening technique for new drug targets and to understand the mechanism of action of such drugs against disease models. Published version

Objective: To analyze the phytochemicals that are present in two medicinal plants which are used for the treatment of respiratory tract infections by gas chromatography-mass spectrometer. Methods: The plant leaves were extracted with n-hexane and methanol separately. Both extracts were analyzed for present phytochemicals using the method described by Harborne, 1985 while only methanol extracts were subjected to gas chromatography-mass spectrometer analysis. Results: Phytochemical screening of the methanolic extracts of Terminalia macroptera (T. macroptera) revealed the presence of glycosides, tannins, flavonoids, saponins and steroids while that of Dioclea reflexa (D. reflexa) showed the presence of flavonoids, saponins and steroids. The n-hexane extracts were devoid of the screened phytochemicals. Twelve and twenty-five compounds were identified in the leaves of T. macroptera and D. reflexa respectively. These compounds were fatty acids, fatty acid esters, other esters, heterocyclics and phenolics. The most abundant compound in T. macroptera was benzenetriol (53.30%) while the predominant compounds in D. reflexa were dodecanoic acid, methyl ester (15.31%), 5, 5, 8a-trimethyl-3, 5, 6, 7, 8, 8a-hexahydro-2H-chromene (9.73%), 10-octadecenoic acid, methyl ester and 2-hexadecanoic acid, methyl ester (8.95%). Benzofuran, 2, 3-dihydro, 3, 7, 11, 15- tetramethyl-2-hexadecen-1-ol and hexadecanoic acid, methyl ester were common in both plant extracts. The antimicrobial properties of the leaves of these plants could be responsible for their use in the treatment of respiratory tract infections. Conclusions: Some of the identified phytochemicals in the plant leaves are responsible for its use in the treatment of respiratory tract infections.

Dioclea reflexa bioactive compounds have been shown to contain antioxidant properties. The extracts from the same plant are used in traditional medical practices to treat various diseases with impressive outcomes. In this study, ionic mobility in Saccharomyces cerevisiae cells in the presence of D. reflexa seed extracts was monitored using electrochemical detection methods to link cell death to ionic imbalance. Cells treated with ethanol, methanol, and water extracts were studied using cyclic voltammetry and cell counting to correlate electrochemical behavior and cell viability, respectively. The results were compared with cells treated with pore-forming Amphotericin b (Amp b), as well as Fluconazole (Flu) and the antimicrobial drug Rifampicin (Rif). The D. reflexa seed water extract (SWE) revealed higher anodic peak current with 58% cell death. Seed methanol extract (SME) and seed ethanol extract (SEE) recorded 31% and 22% cell death, respectively. Among the three control drugs, Flu revealed the highest cell death of about 64%, whereas Amp b and Rif exhibited cell deaths of 35% and 16%, respectively, after 8 h of cell growth. It was observed that similar to SWE, there was an increase in the anodic peak current in the presence of different concentrations of Amp b, which also correlated with enhanced cell death. It was concluded from this observation that Amp b and SWE might follow similar mechanisms to inhibit cell growth. Thus, the individual bioactive compounds from the water extracts of D. reflexa seeds could further be purified and tested to validate their potential therapeutic application. The strategy to link electrochemical behavior to biochemical responses could be a simple, fast, and robust screening technique for new drug targets and to understand the mechanism of action of such drugs against disease models.

Seeds of Hexalobus crispiflorus, Clitandra togolana and Dioclea reflexa were analyzed for proximate composition, cell wall carbohydrates, nutritive and non-nutritive components and then their energy values were estimated. Crop seeds contained 7.11-13.81% crude protein, 8.30-35.86% fat, 7.38-8.61% crude fibre, 3.11-4.25% total ash, 35.03-59.62% soluble carbohydrate; 7.10-9.80% cell wall carbohydrate; 3.20-4.60% cellulose, 3.90-5.20% hemicelluloses, 91.82-92.62% cellular content, 89.09-89.42% organic cellular content, 2.30-3.20% soluble ash, 0.66-1.06% acid insoluble ash, 0.80-1.20% lignin, 1507.332055.18 KJ total energy and 2.6156-9.0476 g digestible crude protein. It was concluded that all the seed samples are potentially good sources of dietary energy for animals. The need to further investigate the inorganic matter for their mineral element constituents in order to ascertain adequacy or otherwise in meeting the animals’ requirement is suggested.

The study aimed at exploring the anti-diabetic effect of the seed flour of Dioclea reflexa on alloxan-induced diabetic rats. Thirty Wistar rats of both sexes were exposed to respective doses of the seed flour and the standardized drugs (Gliclazide 6.1 mg/kd body weight and Metformin 60 mg/kg body weight) according to the group specifications. The seed flour at a dose of 2 g/kg body weight per oral once daily was found to be more effective than any of the standard drugs with which it was compared. The result showed that D. reflexa seed flour possessed potent oral hypoglycaemic property probably mediated via increased peripheral utilization of glucose. Key words: Dioclea reflexa, anti-diabetic, anti-hyperglycaemia, seed flour, hypoglycaemic, glucose utilization, alloxan-induced diabetes.

The aim of the present study is to evaluate the colon cancer chemopreventive effect of dietary inclusion of Diocleareflexaseed, a popular Nigerian soup condiment in male albino rats following ten weeks of intra-rectal instillation with N-methyl-N-nitrosourea (MNU). Male albino wistar rats, randomly divided into 6 groups of 7 rats each, were fed D. reflexa diatary inclusion at 0%, 2.5%, 5%, and 10% for ten weeks with concomitant of intra-rectal instillation with MNU at MNU at a dose of 1.2mg/rat. At the end of feeding experiment, rats were sacrificed, blood and tissues were collected for biochemical, histological and immunohistochemical analysis. The flavonoids, total polyphenols and fiber content of the experimental diets were also assessed.The level of carcino-embryonic antigen (CEA) and malondialdehyde (MDA) were significantly (p Key words: Dioclea reflexa seeds; dietary inclusion; colon carcinogenesis; cancer chemoprevention; experimental carcinogenesis; dietary supplementation Citation Format: Uju D. Iliemene, Sunday Ene-Ojo Atawodi. Chemopreventive effect of Dioclea reflexa seeds dietary inclusion in experimental colon carcinogenesis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4007.

Lectins with lactose binding specificity isolated from seeds of phylogenetically close plant species of Diocleinae sub-tribe present different characteristics when compared to the extensively studied mannose/glucose-specific lectins. In this study, a new lactose-specific lectin from Dioclea reflexa seeds (DrfL II) was isolated by a two-step chromatography in Sephadex® G-50 and lactose-Sepharose columns. The SDS-PAGE showed that the pure lectin is composed of a single band, slightly lower than 30 kDa, presenting a different profile from ConA-like mannose/glucose specific lectins. Partial sequence of the protein was obtained by electrospray ionization mass spectrometry covering 30% of the total sequence. DrfL II presented sugar specificity to α-lactose, and high stability at wide range of pH. DrfL II showed no toxicity against Artemia nauplii and induced acute inflammation (paw edema and hipernociception) in mice with the participation of the lectin domain and nitric oxide. Thus, it is important to further determine the complete sequence and tridimensional structure of this new type of lactose-binding lectin in order to be able to compare it with mannose/glucose lectins and their biotechnological applications. Lectinas com afinidade de ligação a lactose isoladas de sementes de espécies vegetais filogeneticamente próximas da sub-tribo Diocleinae apresentam diferentes características quando comparadas à outras lectinas extensivamente estudadas com afinidade de ligação a glicose-manose. Nesse estudo, uma nova lectina específica a lactose extraída a partir de sementes de Dioclea reflexa (DrfL II) foi isolada por dois passos cromatográficos em colunas de Sephadex G-50 e Sepharose-lactose. A SDS-PAGE revelou uma lectina como uma banda única em torno de 29 kDa, diferindo do perfil já conhecido de lectinas ConA-like específicas a glicose/manose. A sequência parcial da lectina foi obtida por espectrometria de massa com fonte ionizadora eletrospray cobrindo cerca de 30% da sequência total da proteína. DrfL II apresentou especificidade à α-lactose e alta estabilidade a um vasto intervalo de pH. DrfL II mostrou ausência de toxicidade contra Artemia nauplii e induziu inflamação aguda (edema de pata e hipernocicepção) em camundongos com a participação do domínio da lectina e óxido nítrico. Visto que poucas informações têm sido publicadas a respeito de lectinas lactose específicas de Diocleinae, se faz necessário, portanto, que o estudo seja continuado visando a determinação completa da estrutura primária e tridimensional da lectina, para comparação com os dados existentes de lectinas ConA-like e lactose específicas

Reflevone ( 1 ), a new flavone, together with, mearnsetin ( 2 ), 7,4'-dihydroxyflavone ( 3 ) and two known phytosterols, β-sitosterol ( 4 ) and stigmasterol ( 5 ) were isolated from ethanol soluble part of the roots of Dioclea reflexa (Hook F.). Their structures were elucidated by analysis of spectroscopic data involving 1D and 2D NMR, MS, IR and UV experiments. The crude roots extract showed significant activity with IC 50 = 5.18 µg/mL against the prostate cancer cell line. KEY WORDS : Phytochemical analysis, Dioclea reflexa , Fabaceae, Cytotoxic activity, Flavonoid Bull. Chem. Soc. Ethiop. 2015 , 29(3), 441-448. DOI: http://dx.doi.org/10.4314/bcse.v29i3.12

Methanolic extract of Dioclea reflexa seed was investigated following acute or chronic carbon tetrachloride intoxication in male albino rats. In the acute experiment, rats were pre-treated with extract at 5 mg/kg for three days before administration of carbon tetrachloride(0.6 mL/kg) while in the chronic model, rats received extract at 2.5 mg/kg for 12 consecutive days with 72 hourly administration of carbon tetrachloride at 0.3 mL/kg. In both models, the levels of transaminases and malondialdehyde were significantly (p

Aims: Dioclea reflexa Hook F., is a woody vine widely distributed within tropical Africa and South America, in Nigeria flour prepared from the seeds is used as a soup thickener. The objectives of this study were to extract and modify the native gum from the seeds of D. reflexa, and evaluate their physicochemical and functional properties as a potential pharmaceutical excipient. Place and Duration of Study: Department of Pharmaceutical Technology and Raw Materials Development, National Institute for Pharmaceutical Research and Development, Abuja, Nigeria. Between January 2009 and October 2010. And Department of Pharmaceutics, Faculty of Pharmaceutical Sciences, University of Nigeria, Nsukka, Nigeria. Between June 2010 and October 2011. Methodology: Native D. reflexa gum (DR-gum) was extracted from the seeds of D. reflexa and modified by ionotropic gelation in calcium chloride solution. The gum’s gel clarity and swelling in buffer solutions of different pH (4, 7, and 9.2), as well as the moisture sorption characteristics at different relative humidity (RH) were determined in relation to guar gum (G-gum). Also, the Fourier transform infrared (FT-IR) and Differential scanning calorimetry (DSC) analysis as well as the powders’ flow properties were evaluated. Results: DR-gum, XDR-gum and G-gum are typical hydrogels showing considerable responsiveness in swelling and gel clarity to changes in pH. The DSC thermographs of DR-gum and G-gum were similarly characterized by a glass transition and a cold crystallization transitions peaks as compared to the glass transition and melting transition peak of XDR-gum. The moisture sorption profile indicated that DR-gum and G-gum are moderately hygroscopic while XDR-gum is slightly hydroscopic. The FT-IR of the gums showed the basic differences in the functional groups of the respective polymer chain network. The bulk particles of the three gums showed comparative good flow. Conclusion: The basic similarities and differences in the physicochemical and functional properties of DR-gum and XDR-gum, and G-gum indicate their potential as a pharmaceutical excipient.