Anti-Hyperlipidemic Components of the Leaves of Synsepalum dulcificum (Miracle Fruit).

Synsepalum dulcificum (Miracle fruit) is a tropical plant in West and Central Africa, which has been historically used for treating diarrhea in humans and animals. Pharmacological research has shown that the leaves of the plant possess anti-hyperlipidemia activity. However, its anti-hyperlipidemic components have not been reported. In this study, the leaves of S. dulcificum were extracted using 95% ethanol and the extract was fractionated using different polar solvents. The anti-hyperlipidemia activity of the extract and fractions were evaluated using the zebrafish model. The results showed that the ethyl acetate (EA) fraction displayed the best anti-hyperlipidemic effect. A comparison of the high-performance liquid chromatography equipped with diode array detector (HPLC-DAD) profiles of the ethanol extract and different fractions at 350 nm indicated that a peak at 37.4 min has the highest intensity in the EA part, relatively. Then the chemical constituents of the extract and the active fraction were extensively identified using UPLC-Q-Exactive-Orbitrap-MS/MS, showing the main peak was quercitrin and other components in the EA part mainly included quercitrin analogs. Furthermore, the quercitrin was isolated from the plant and its contents in the extract and fractions were determined using high-performance liquid chromatography with ultraviolet detector (HPLC-UV) method. The quantitative results showed that the content of quercitrin in the EA fraction was 10.04% (w/w). Further pharmacological study indicated that quercitrin also possessed potent anti-hyperlipidemia activity (improvement rates of liver fat and total cholesterol were 75.6% and 92.5% at 40 µg/mL, respectively). Besides, quercitrin showed little toxicity to zebrafish embryos.

Exploring the diversity, bioactivity of endophytes, and metabolome in Synsepalum dulcificum.

Synsepalum dulcificum exhibits high edible and medicinal value; however, there have been no reports on the exploration of its endophyte resources. Here, we conducted analyses encompassing plant metabolomics, microbial diversity, and the biological activities of endophytic metabolites in S. dulcificum. High-throughput sequencing identified 4,913 endophytic fungal amplicon sequence variants (ASVs) and 1,703 endophytic bacterial ASVs from the roots, stems, leaves, flowers, and fruits of S. dulcificum. Fungi were classified into 5 phyla, 24 classes, 75 orders, 170 families, and 313 genera, while bacteria belonged to 21 phyla, 47 classes, 93 orders, 145 families, and 232 genera. Furthermore, there were significant differences in the composition and content of metabolites in different tissues of S. dulcificum. Spearman's correlation analysis of the differential metabolites and endophytes revealed that the community composition of the endophytes correlated with plant-rich metabolites. The internal transcribed spacer sequences of 105 isolates were determined, and phylogenetic analyses revealed that these fungi were distributed into three phyla (Ascomycota, Basidiomycota, and Mucoromycota) and 20 genera. Moreover, 16S rDNA sequencing of 46 bacteria revealed they were distributed in 16 genera in three phyla: Actinobacteria, Proteobacteria, and Firmicutes. The antimicrobial activities (filter paper method) and antioxidant activity (DPPH and ABTS assays) of crude extracts obtained from 68 fungal and 20 bacterial strains cultured in different media were evaluated. Additionally, the α-glucosidase inhibitory activity of the fungal extracts was examined. The results showed that 88.6% of the strains exhibited antimicrobial activity, 55.7% exhibited antioxidant activity, and 85% of the fungi exhibited α-glucosidase inhibitory activity. The research suggested that the endophytes of S. dulcificum are highly diverse and have the potential to produce bioactive metabolites, providing abundant species resources for developing antibiotics, antioxidants and hypoglycemic drugs.

Safety of dried fruits of Synsepalum dulcificum as a novel food pursuant to Regulation (EU) 2015/2283.

Following a request from the European Commission, the EFSA Panel on Nutrition, Novel Foods and Food Allergens (NDA) was asked to deliver an opinion on dried fruits of Synsepalum dulcificum as a novel food (NF) pursuant to Regulation (EU) 2015/2283. The NF is pitted and dried (by lyophilisation) fruits of S. dulcificum. The NF contains the glycoprotein miraculin (≤ 2.5%) which causes sour and acidic foods to taste sweet. The fruits have a documented history of consumption in Africa and products thereof can be found in different markets worldwide. Information on the production process and the composition of the NF is sufficient and does not raise safety concerns. The applicant proposes to use the NF as or in food supplements for the adult population, excluding pregnant and lactating women, at a maximum daily amount of 0.9 g. Taking into account these conditions of use, the Panel considers that the consumption of the NF is not nutritionally disadvantageous. The provided genotoxicity studies do not raise concerns for genotoxicity of the NF. The Panel concludes that the only dose tested in a 90-day oral toxicity study of 2,000 mg/kg body weight (bw) per day was not associated with adverse effects. By applying an uncertainty factor of 200, the Panel concludes that the NF is safe at an intake level of 10 mg/kg bw per day, corresponding to a maximum daily intake of 0.7 g of the NF for the target population, rather than 0.9 g/day as proposed by the applicant.

Quantification of lectins in Synsepalum dulcificum and comparison with reference foods.

A healthy life means a balance between physical activity and a diet rich in fruits and vegetables, however, some plant-based foods can have certain adverse effects due to the presence of anti-nutritional factors, such as lectins, capable of binding molecules and preventing their normal assimilation. The level of lectins in Synsepalum dulcificum fruit was determined by hemagglutination assays in human blood, and its comparison with foods characterized as having high and low lectin content. The relative hemagglutinating activity of berries from Synsepalum dulcificum compared to our positive high lectin content food reference (Pinto bean) corresponds to 3.13-6.25%, representing safe levels for nutritional food.

The Chromosome-Level Genome of Miracle Fruit (Synsepalum dulcificum) Provides New Insights Into the Evolution and Function of Miraculin.

Miracle fruit (Synsepalum dulcificum) is a rare valuable tropical plant famous for a miraculous sweetening glycoprotein, miraculin, which can modify sour flavors to sweet flavors tasted by humans. Here, we present a chromosome-level high-quality genome of S. dulcificum with an assembly genome size of ∼550 Mb, contig N50 of ∼14.14 Mb, and 37,911 annotated protein-coding genes. Phylogenetic analysis revealed that S. dulcificum was most closely related to Camellia sinensis and Diospyros oleifera, and that S. dulcificum diverged from the Diospyros genus ∼75.8 million years ago (MYA), and that C. sinensis diverged from Synsepalum ∼63.5 MYA. Ks assessment and collinearity analysis with S. dulcificum and other species suggested that a whole-genome duplication (WGD) event occurred in S. dulcificum and that there was good collinearity between S. dulcificum and Vitis vinifera. On the other hand, transcriptome and metabolism analysis with six tissues containing three developmental stages of fleshes and seeds of miracle fruit revealed that Gene Ontology (GO) terms and metabolic pathways of "cellular response to chitin," "plant-pathogen interaction," and "plant hormone signal transduction" were significantly enriched during fruit development. Interestingly, the expression of miraculin (Chr10G0299340) progressively increased from vegetative organs to reproductive organs and reached an incredible level in mature fruit flesh, with an fragments per kilobase of transcript per million (FPKM) value of ∼113,515, which was the most highly expressed gene among all detected genes. Combining the unique signal peptide and the presence of the histidine-30 residue together composed the main potential factors impacting miraculin's unique properties in S. dulcificum. Furthermore, integrated analysis of weighted gene coexpression network analysis (WGCNA), enrichment and metabolite correlation suggested that miraculin plays potential roles in regulating plant growth, seed germination and maturation, resisting pathogen infection, and environmental pressure. In summary, valuable genomic, transcriptomic, and metabolic resources provided in this study will promote the utilization of S. dulcificum and in-depth research on species in the Sapotaceae family.

Chemical constituents and their antioxidant activities from the leaves of Synsepalum dulcificum.

Synsepalum dulcifificum is an evergreen shrub native of tropical West Africa. It is very effective against a variety of diseases. However, the phytochemical investigation on this plant is limited till now. In our current study, seventeen compounds were isolated and identified from the ethyl acetate extract of the leaves. All of the compounds were obtained from S. dulcifificum for the first time. Compounds 4 and 9 were isolated from natural source for the second time. Moreover, the antioxidant activities of compounds 1, 4, 5, 9-11, and 15 were evaluated firstly. Compounds 6, 7, 13, and 14 exhibited significant antioxidant activity in DPPH and ABTS + assays, suggesting their potential application for an antioxidant drug.

Antihyperglycemic and hepatoprotective properties of miracle fruit (Synsepalum dulcificum) compared to aspartame in alloxan-induced diabetic mice.

OBJECTIVE: This study was undertaken to investigate the antihyperglycemic potential of miracle fruit (MF) as well as its hepatic safety as compared to aspartame in alloxan-induced diabetic mice. METHODS: MF extracts were prepared and screened for their phytochemical composition using high-performance liquid chromatography (HPLC). Total phenolic, flavonoid and tannin contents and antioxidant potential were also determined. Additionally, MF was evaluated for its sensory attributes. For in vivo work, MF ethanol extract at high (MFH: 500 mg/kg body weight [BW]) and low (MFL: 250 mg/kg BW) doses as well as aspartame were injected intraperitoneally into alloxan-induced diabetic mice. Blood glucose levels were determined following acute and subchronic treatment. At the end of the study, animals were sacrificed, serum was collected for biochemical analysis and liver tissues were obtained for histopathological examination. RESULTS: MF ethanol extract contained more flavonoids and tannins, and had higher 1,1-diphenyl-1-picrylhydrazyl radical-scavenging activity (79.61%) compared to MF aqueous extract (P < 0.05). HPLC analysis of MF ethanol extract also revealed the presence of 10 antioxidants with quercetin comprising the major polyphenol. Additionally, sensory analysis of MF showed that its intake is effective in masking undesirable sourness. Subchronic administration of MFH proved amelioration of hyperglycemia in mice as compared to aspartame. Moreover, aspartame treatment significantly elevated (P < 0.05) the level of alanine aminotransferase and had destructive effects on the liver histopathology; however, hepatic architecture was restored by low and high doses of MF. CONCLUSION: MF is an effective antihyperglycemic with hepatoprotective properties that can be used as a healthier alternative sweetening agent in place of aspartame for sour beverages.

Positive improvement in palatability of metabolic formula with the use of miraculin protein in patients with inborn errors of metabolism and healthy adults / Cambio positivo en la palatabilidad de la fórmula metabólica con el uso de la proteína miraculina en pacientes con errores innatos del metabolismo y adultos sanos

ABSTRACT As palatability of medical formulas has been documented as unpleasant, new options are required to improve acceptance and adherence in people with inborn errors of metabolism (IEM). Miracle fruit (Synsepalum dulcificum) has a glycoprotein named miraculin that transforms a sour, bitter taste such as the one found in metabolic formula, into a sweet perception. The objective of this work is to analyze the response in the taste perception of metabolic formula with the use of the miraculin tablets in patients with IEM and healthy adults. To test this hypothesis a prospective, longitudinal, quasi-experimental, analytical study was performed. Patients with IEM and healthy adults were recruited. All participants assessed 3 different liquids (lemon, apple cider vinegar and metabolic formula) before and after the administration of miraculin tablets and completed a questionnaire. The sensory responses were evaluated using hedonic scales, analyzed with nonparametric tests for paired data. Seven patients with IEM and 14 healthy subjects were included. After miraculin intake 57% of patients (Z ≤ -1.89 p= 0.059) and healthy adults (Z≤ -2.31 p= 0.021) had a positive change in their taste perception. The absolute frequency of patients who did not like the metabolic formula decreased from 4 to 1, and in patients who liked it or loved, it increased from 0 to 2 and from 0 to 1 respectively; the frequency of patients who perceived the metabolic formula as indifferent or hated it, did not change. Response in taste perception had a positive change of 57% in both groups. The use of miraculin tablets may improve palatability of metabolic formula.

RESUMEN La palatabilidad de las fórmulas médicas se ha reportado como desagradable, se requieren nuevas opciones para mejorar la aceptación en personas con errores innatos del metabolismo (EIM). La fruta milagrosa (Synsepalum dulcificum) contiene una glucoproteína llamada miraculina que transforma el sabor agrio y amargo en dulce. El objetivo fue analizar la respuesta en la percepción del sabor de la fórmula metabólica con el uso de las tabletas de miraculina en pacientes con EIM y adultos sanos. Se realizó un estudio analítico prospectivo, longitudinal, cuasi-experimental. Los participantes evaluaron la percepción de 3 líquidos (limón, vinagre de manzana y fórmula metabólica) antes y después de la administración de tabletas de miraculina y completaron un cuestionario. Las respuestas sensoriales se evaluaron mediante escalas hedónicas, analizadas con pruebas no paramétricas para datos pareados. Se incluyeron 7 pacientes con EIM y 14 adultos sanos. Después de la miraculina el 57% de los pacientes (Z ≤ -1,89 p= 0,059) y adultos sanos (Z≤ -2,31 p= 0,021) tuvieron un cambio positivo en su percepción del sabor. La frecuencia absoluta de pacientes a los que no les gustó la fórmula disminuyó de 4 a 1, y en quienes les gustó o les encantó, aumentó de 0 a 2 y de 0 a 1 respectivamente; la frecuencia de los pacientes que percibieron la fórmula como indiferente u odiada, no cambió. La respuesta en la percepción del sabor cambió positivamente en el 57% en ambos grupos. El uso de miraculina puede mejorar la palatabilidad de la fórmula metabólica.

The cholesterol-lowering activity of miracle fruit (Synsepalum dulcificum).

Miracle fruit (Synsepalum dulcificum) is famous for its uniqueness of modifying sour taste to sweetness. However, its cholesterol-lowering activity has not been reported. This study investigated the effect of S. dulcificum on the compositional changes of plasma lipids in hamsters fed a high-cholesterol control diet. Six groups of hamsters were fed either a control diet or one of the five experimental diets containing 2% ethanol extract of leaves, 2% water extract of leaves, 2% ethanolic extract of seeds (ES), 2% water extract of seeds, or 2% dry pulp. Results showed that ES decreased the plasma total cholesterol (TC). Two triterpenoids (lupeol acetate and ß-amyrin acetate) were isolated from the ES and they added to a diet could decrease TC by 15%-20% in hamsters. It was concluded that ES showed potent TC-lowering activity and triterpenoid was one of the active components of ES. PRACTICAL APPLICATIONS: In recent years, people are more interested in phytochemicals from functional foods treated for hyperlipidemia because they possessed fewer side effects than the synthetic drugs. The triterpenoids isolated from the miracle fruit may be promising candidates for the development of cholesterol-lowering agent, especially for patients whose blood cholesterol level and body weight are high. Meanwhile, the miracle fruit have a good potential as cholesterol-lowering functional food or a natural source of cholesterol-lowering agent.

Complete chloroplast genome of Synsepalum dulcificum D.: a magical plant that modifies sour flavors to sweet.

Synsepalum dulcificum D. belongs to the Sapotaceae family, which is an evergreen shrub native to tropical West Africa. It is a kind of magical plant that has the unique characteristic of modifying sour flavors to sweet. In this study, the chloroplast genome of S. dulcificum was sequenced, assembled, and annotated firstly. Chloroplast genome size of S. dulcificum is 158,463 bp, the circular chloroplast genome consists of four regions: a large single-copy region of 88,256 bp, two inverted repeat regions of 25,958 bp, and a small single-copy region of 18,669 bp, with the GC content of 36.87%. A total of 133 genes were annotated in the S. dulcificum chloroplast genome, of which 88 were protein-coding genes (PCGs), 37 were transfer RNA (tRNA) genes, and eight were ribosomal RNA (rRNA) genes. Phylogenetic analysis showed that Pouteria campechiana was most closely related to S. dulcificum. The study provides important genomic data for further utilization and breeding of S. dulcificum.

Antihyperglycemic and hepatoprotective properties of miracle fruit (Synsepalum dulcificum) compared to aspartame in alloxan-induced diabetic mice / 中西医结合学报

OBJECTIVE@#This study was undertaken to investigate the antihyperglycemic potential of miracle fruit (MF) as well as its hepatic safety as compared to aspartame in alloxan-induced diabetic mice.@*METHODS@#MF extracts were prepared and screened for their phytochemical composition using high-performance liquid chromatography (HPLC). Total phenolic, flavonoid and tannin contents and antioxidant potential were also determined. Additionally, MF was evaluated for its sensory attributes. For in vivo work, MF ethanol extract at high (MFH: 500 mg/kg body weight [BW]) and low (MFL: 250 mg/kg BW) doses as well as aspartame were injected intraperitoneally into alloxan-induced diabetic mice. Blood glucose levels were determined following acute and subchronic treatment. At the end of the study, animals were sacrificed, serum was collected for biochemical analysis and liver tissues were obtained for histopathological examination.@*RESULTS@#MF ethanol extract contained more flavonoids and tannins, and had higher 1,1-diphenyl-1-picrylhydrazyl radical-scavenging activity (79.61%) compared to MF aqueous extract (P < 0.05). HPLC analysis of MF ethanol extract also revealed the presence of 10 antioxidants with quercetin comprising the major polyphenol. Additionally, sensory analysis of MF showed that its intake is effective in masking undesirable sourness. Subchronic administration of MFH proved amelioration of hyperglycemia in mice as compared to aspartame. Moreover, aspartame treatment significantly elevated (P < 0.05) the level of alanine aminotransferase and had destructive effects on the liver histopathology; however, hepatic architecture was restored by low and high doses of MF.@*CONCLUSION@#MF is an effective antihyperglycemic with hepatoprotective properties that can be used as a healthier alternative sweetening agent in place of aspartame for sour beverages.

Antibacterial Property of Synsepalum dulcificum Leaves Aqueous Extract against Oral Pathogens and its Chemical Compounds

@#Synsepalum dulcificum (S. dulcificum) commonly known as “miracle fruit” because its berries have the capability to modify sour taste to the sweet taste when eaten. Beside the berries, S. dulcificum leaves were also known to possess biological properties such as antioxidant, antimutagenic and antidiabetic activities. However, the study of its antimicrobial activity against oral pathogen is still lacking. Thus, this study aimed to evaluate the antibacterial activity of its leaves against cariogenic bacteria and to analyse its phytochemical compounds. The samples of S. dulcificum leaves were collected in Kelantan, the east coast region of Peninsular Malaysia and extracted with distilled water using a Soxhlet technique. The antibacterial activity of the S. dulcificum leaves aqueous extract against Streptococcus mutans (S. mutans), Streptococcus sobrinus (S. sobrinus) and Lactobacillus salivarius (L. salivarius) was evaluated using the broth microdilution assay. The identification of the phytochemical compounds was performed using gas chromatography-mass spectrometry (GC-MS). The antibacterial study showed the minimum inhibitory concentration of S. dulcificum leaves aqueous extract against S. mutans and S. sobrinus were 16 mg/mL and 8 mg/mL, respectively. Interestingly, there was no inhibitory effect of S. dulcificum leaves aqueous extract against L. salivarius. A total of 42 chemical compounds were identified and major identified bioactive compounds groups were heterocyclic and phenolic compounds. Our results suggested S. dulcificum leaves aqueous extract has antimicrobial properties against S. mutans and S. sobrinus, but no inhibitory activity against oral normal flora, with the presence of bioactive compounds has potential in oral care products application.

Microencapsulation of Lactococcus lactis Gh1 with Gum Arabic and Synsepalum dulcificum via Spray Drying for Potential Inclusion in Functional Yogurt.

There has been an explosion of probiotic incorporated based product. However, many reports indicated that most of the probiotics have failed to survive in high quantity, which has limited their effectiveness in most functional foods. Thus, to overcome this problem, microencapsulation is considered to be a promising process. In this study, Lactococcus lactis Gh1 was encapsulated via spray-drying with gum Arabic together with Synsepalum dulcificum or commonly known as miracle fruit. It was observed that after spray-drying, high viability (~108 CFU/mL) powders containing L. lactis in combination with S. dulcificum were developed, which was then formulated into yogurt. The tolerance of encapsulated bacterial cells in simulated gastric juice at pH 1.5 was tested in an in-vitro model and the result showed that after 2 h, cell viability remained high at 1.11 × 106 CFU/mL. Incubation of encapsulated cells in the presence of 0.6% (w/v) bile salts showed it was able to survive (~104 CFU/mL) after 2 h. Microencapsulated L. lactis retained a higher viability, at ~107 CFU/mL, when incorporated into yogurt compared to non-microencapsulated cells ~105 CFU/mL. The fortification of microencapsulated and non-microencapsulated L. lactis in yogurts influenced the viable cell counts of yogurt starter cultures, Lactobacillus delbrueckii subs. bulgaricus and Streptococcus thermophilus.

Beneficial effects of an investigational wristband containing Synsepalum dulcificum (miracle fruit) seed oil on the performance of hand and finger motor skills in healthy subjects: A randomized controlled preliminary study.

Miracle fruit (Synsepalum dulcificum) seed oil (MFSO) contains phytochemicals and nutrients reported to affect musculoskeletal performance. The purpose of this study was to assess the safety and efficacy of a compression wristband containing MFSO on its ability to measurably improve the hand and finger motor skills of participants. Healthy right-handed participants (n = 38) were randomized in this double-blind, placebo-controlled study of MFSO and vehicle wristbands. Subjects wore the wristband on their left hand 4-6 weeks and then only on their right hand 2-4 weeks; the contralateral untreated hand served as an additional control. Twelve hand/finger motor skills were measured using quantitative bio-instrumentation tests, and subject self-assessment questionnaires were conducted. With each hand, in 9/12 tests, the MFSO group showed a clinically meaningful average improvement compared with an average worsening in the vehicle group. Statistical superiority to the control treatment group was exhibited in 9/12 tests for each hand (p < .01). After discontinuing the MFSO wristband on the left hand, test values regressed toward baseline levels. Subjects favored the MFSO wristband over the control, rating it as effective in improving their motor skills. Use of the MFSO wristband may improve an individual's manual dexterity skills and ability to maintain this performance.

Synsepalum dulcificum extracts exhibit cytotoxic activity on human colorectal cancer cells and upregulate c-fos and c-jun early apoptotic gene expression

Objective:To explore cytotoxicity of Synsepalum dulcificum (S.dulcificum) Daniell (Sapotaceae) on human colon cancer (HCT-116 and HT-29),human monocytic leukemia (THP-1) and normal (HDFn) cell lines,and its effect on the expression of early apoptotic genes,c-fos and c-jun.Methods:Leaf,stem and berry of S.dulcificum were separately extracted by using 2 solvents,10％ ethanol (EtOH) and 80％ methanol (MeOH).PrestoBlue(R)cell viability assay and qRT-PCR assay were conducted to examine the above objectives respectively.Results:Stem MeOH,stem EtOH,and berry EtOH extracts of S.dulcificum were cytotoxic to HCT-116 and HT-29 human colon cancer cells.For HCT-116,IC50 values of these 3 extracts were not significantly different (P＞0.05) from that of the positive control bleomycin (IC5o of 33.57 μg/mL),while for HT-29,IC5o values of these 3 extracts were significantly lower (P＜0.05) than that of bleomycin (IC50 of 25.24 μg/mL).None of the extracts were cytotoxic to the THP-1 monocytic leukemia cells and HDFn normal human dermal fibroblasts.For both HCT-116 and HT-29,these extracts significantly up-regulated (P＜0.05) the expression of c-fos and c-jun compared to the untreated negative control.Conclusions:The results of this study suggest that cytotoxicity of stem MeOH,stem EtOH,and berry EtOH extracts of S.dulcificum on HCT-116 and HT-29 colon cancer cells is due to the induced apoptosis which is caused by the up-regulation of the expression of early apoptotic genes,c-fos and c-jun.

Synsepalum dulcificum extracts exhibit cytotoxic activity on human colorectal cancer cells and upregulate c-fos and c-jun early apoptotic gene expression

Objective: To explore cytotoxicity of Synsepalum dulcificum (S. dulcificum) Daniell (Sapotaceae) on human colon cancer (HCT-116 and HT-29), human monocytic leukemia (THP-1) and normal (HDFn) cell lines, and its effect on the expression of early apoptotic genes, c-fos and c-jun. Methods: Leaf, stem and berry of S. dulcificum were separately extracted by using 2 solvents, 10% ethanol (EtOH) and 80% methanol (MeOH). PrestoBlue® cell viability assay and qRT-PCR assay were conducted to examine the above objectives respectively. Results: Stem MeOH, stem EtOH, and berry EtOH extracts of S. dulcificum were cytotoxic to HCT-116 and HT-29 human colon cancer cells. For HCT-116, IC

Antidiabetic potential of methanolic and flavonoid-rich leaf extracts of Synsepalum dulcificum in type 2 diabetic rats.

BACKGROUND: Synsepalum dulcificum is a plant indigenous to West Africa. The fruit is used to modify taste of foods to sweetness. OBJECTIVES: This study aims to investigate the antidiabetic potentials of both methanolic and flavonoid-rich leaf extracts of S. dulcificum (MSD and FSD respectively) in type 2 diabetic Wistar albino rats. MATERIALS AND METHODS: Sixty three rats were randomly distributed into nine groups of seven animals each with group 1 serving as the normal control. Groups 2 to 7 were given 10% fructose in their drinking water for 14 days, after which 40 mg/kg of streptozotocin was administered. Group 2 animals served as the diabetic control, while groups 3, 4, 5, 6 and 7 were treated with 30 mg/kg MSD, 60 mg/kg MSD, 30 mg/kg FSD, 60 mg/kg FSD and 5 mg/kg glibenclamide respectively. Groups 8 and 9, contained healthy animals, and were treated with only 60 MSD, and 60 mg/kg FSD respectively. Biochemical parameters such as liver and kidney function tests, lipid profile, as well as lipid peroxidation and antioxidant enzymes were assessed in addition to histopathology. RESULTS: It was observed that daily oral administration of MSD and FSD for 21 days significantly (p < 0.05) improved the observed pathological changes as a result of type 2 diabetes. CONCLUSION: It could be deduced from results obtained in this study that methanolic and flavonoid-rich leaf extracts of S.dulcificum have antidiabetic potential in type 2 diabetic rats.

Effect of Miracle Fruit (Synsepalum dulcificum) Seed Oil (MFSO®) on the Measurable Improvement of Hair Breakage in Women with Damaged Hair: A Randomized, Double-blind, Placebo-controlled, Eight-month Trial.

Background: Hair breakage is a common unrecognized form of hair loss in women most often the result of hair weathering and traumatic grooming practices. Lipids are major determinants of the physical properties of the hair. Synsepalum dulcificum seed oil (MFSO®; Miracle Fruit Oil Co., Miami Beach, Florida), is an exotic fruit oil with physicochemical properties suited to providing a superior ability to reduce hair breakage. Objective: To assess the safety and efficacy of a hair oil containing MFSO and its effects on hair breakage rates. Methods: Healthy, long-haired women (age range: 19-63 years, mean age: 36.7 years, standard deviation: 10.77 years) with excessive hair breakage were randomized in this double-blind, placebo-controlled study to receive MFSO (n=24), vehicle (n=17), or argan oil (n=16). Measurements of hair length, hair diameter, and Hair Mass Index were performed at baseline, Month 4, and Month 8. Hair Breakage Index and the Healthy Hair Index values were calculated from the trichometer measurements, and subject self-assessment questionnaires were conducted. The primary efficacy endpoints were the percent change in Healthy Hair Index 75 and Healthy Hair Index 50 measurements from baseline to the eighth month. Results: The Healthy Hair Index calculations, expressed as percent change from baseline to Month 4 and from baseline to Month 8, revealed that the MFSO® treatment group improved by 103.6 percent and 215.7 percent for the Healthy Hair Index 75 and 133.7 and 188.3 percent for the Healthy Hair Index 50 values, respectively. When compared with the vehicle and the argan oil brand groups, the Healthy Hair Index levels were significantly higher (p < 0.001) for the MFSO® treatment group, indicating a much greater ability to increase the levels of unbroken hairs by reducing hair breakage. With respect to the mean percent improvements from baseline to Month 4 and Month 8, the MFSO® hair oil treatment group was better than each of the other two treatment groups by at least 117.6 percent and 234.9 percent for the Healthy Hair Index 75 and 316.5 percent and 312 percent for the Healthy Hair Index 50 values, respectively, thereby achieving the primary efficacy objective. Subjects favored the MFSO® hair oil treatment, rating it as safe, effective, and aesthetically pleasing. Conclusions: The MFSO hair oil product is a safe and effective option for the treatment of women suffering from hair breakage and damaged hair.

Amino acid profile and oxidizable vitamin content of Synsepalum dulcificum berry (miracle fruit) pulp.

The amino acid profile of the Synsepalum dulcificum berry was studied. Among the essential amino acid observed, leucine (2.35 g/100 g protein) was the highest while methionine (0.31 g/100 g protein) was the lowest. The nonessential amino acids were also discovered, with glutamic acid (3.43 g/100 g protein) being the highest and glycine (0.38 g/100 g protein), the lowest. The study of the oxidizable vitamins revealed that vitamin C (1.33 mg/100 g) was more abundant than vitamin A (2.54 µg) and vitamin E (0.78 mg/100 g). This information will hopefully enhance the fruits acceptability by more people and thus, generally promote its utilization and appreciation in our diets.

Optimization of conditions for the single step IMAC purification of miraculin from Synsepalum dulcificum.

In this study, the methods for extraction and purification of miraculin from Synsepalum dulcificum were investigated. For extraction, the effect of different extraction buffers (phosphate buffer saline, Tris-HCl and NaCl) on the extraction efficiency of total protein was evaluated. Immobilized metal ion affinity chromatography (IMAC) with nickel-NTA was used for the purification of the extracted protein, where the influence of binding buffer pH, crude extract pH and imidazole concentration in elution buffer upon the purification performance was explored. The total amount of protein extracted from miracle fruit was found to be 4 times higher using 0.5M NaCl as compared to Tris-HCl and phosphate buffer saline. On the other hand, the use of Tris-HCl as binding buffer gave higher purification performance than sodium phosphate and citrate-phosphate buffers in IMAC system. The optimum purification condition of miraculin using IMAC was achieved with crude extract at pH 7, Tris-HCl binding buffer at pH 7 and the use of 300 mM imidazole as elution buffer, which gave the overall yield of 80.3% and purity of 97.5%. IMAC with nickel-NTA was successfully used as a single step process for the purification of miraculin from crude extract of S. dulcificum.