Dr. Engy Mahrous

Green beans (Phaseolus vulgaris L.) are consumed as pods or mature seeds (common beans). The pods were extracted with 95% ethanol and processed to prepare non-polar and polar fractions. Comparing the antihyperglycemic activity of both fractions, non-polar fraction (NPF, 200 mg kg  day ) lowered blood glucose in streptozotocin diabetic rats by 65% compared to 57% for the polar fraction at the same dose. When NPF treatment was combined with injection of mesenchymal stem cells (MSC) a 4.4-fold increase in serum insulin and a 73.6% reduction in blood glucose were observed compared to untreated control. Additionally, a significant decrease in malondialdehyde (76.2%), nitric oxide (68.2%), cholesterol (76.1%), and triglycerides (69.5%) and a 1.75-fold increase in HDL concentrations were observed in the group treated with this combination compared to diabetic animals. Interestingly, NPF increased homing of MSC in pancreas potentiating their antidiabetic activity. Finally, 26 compounds were identified in NPF using LC/MS analysis and four were isolated in pure form. The isolated compounds namely calotroproceryl acetate, fridelin, calotroproceryl A, and stigmasterol showed good inhibitory activity against pancreatic lipase with IC at 1.93, 1.07, 1.34 and 1.44-1 μg/ml, respectively. Additionally, these compounds inhibited α-amylase, albeit at higher concentration, with IC at 248, 212, 254, and 155 μg/ml for calotroproceryl acetate, fridelin, calotroproceryl A, and stigmasterol, respectively. Our results suggest that green beans extract can potentiate effect of MSC in diabetes directly due to its own antidiabetic effect and indirectly by increasing MSC homing in pancreatic tissues. PRACTICAL APPLICATIONS: It has been suggested in this study that green beans can improve hyperglycemia, oxidative balance in diabetes, so green beans can be promoted as a healthy nutrient for diabetic patients. Green beans also can enhance homing and differentiation of mesnchymal stem cells in the pancreas for future stem cell therapy of type I diabetes.

Jacq., family: Euphorbiaceae, is used in India and subtropical Africa to treat different skin conditions. In this study we evaluated the anti-inflammatory activity of leaves extract (JILE) using rat paw edema model. The extract was administered orally (200 and 400 mg/kg) or applied topically as creams at 2.5, 5, and 10% strength. Four hours post-treatment, maximum reduction of edema volume by 63.09% was observed after oral administration of JILE (400 mg/kg) as compared to indomethacin with 60.43%. The extract anti-inflammatory effect was accompanied by a decrease in NO, prostaglandin PGE2, TNF-α and PKC levels by 19, 29.35, 16.9, and 47.83%, respectively. Additionally, topical applications of JILE showed dose dependent reduction in paw edema and resulted in normalized levels of PGE2, TNF-α, and PKC when used as 10% cream. Signs of inflammations were reduced or absent from paw tissue of animals receiving JILE either orally or topically. Finally, liquid chromatography/mass spectrometry analysis of JILE resulted in the annotation of 133 metabolites including 24 diterpenoids, 19 flavonoids, 10 phenolic acid conjugates, 8 cyclic peptides, 6 phytosterols, 4 sesquiterpenes, and 4 coumarins. Several of the annotated metabolites have known anti-inflammatory activity including vitexin, isovitexin, fraxitin, scopeltin, stigmasterol, and many diterpenoidal derivatives.

Jacq., family: Euphorbiaceae, is used in India and subtropical Africa to treat different skin conditions. In this study we evaluated the anti-inflammatory activity of leaves extract (JILE) using rat paw edema model. The extract was administered orally (200 and 400 mg/kg) or applied topically as creams at 2.5, 5, and 10% strength. Four hours post-treatment, maximum reduction of edema volume by 63.09% was observed after oral administration of JILE (400 mg/kg) as compared to indomethacin with 60.43%. The extract anti-inflammatory effect was accompanied by a decrease in NO, prostaglandin PGE2, TNF-α and PKC levels by 19, 29.35, 16.9, and 47.83%, respectively. Additionally, topical applications of JILE showed dose dependent reduction in paw edema and resulted in normalized levels of PGE2, TNF-α, and PKC when used as 10% cream. Signs of inflammations were reduced or absent from paw tissue of animals receiving JILE either orally or topically. Finally, liquid chromatography/mass spectrometry analysis of JILE resulted in the annotation of 133 metabolites including 24 diterpenoids, 19 flavonoids, 10 phenolic acid conjugates, 8 cyclic peptides, 6 phytosterols, 4 sesquiterpenes, and 4 coumarins. Several of the annotated metabolites have known anti-inflammatory activity including vitexin, isovitexin, fraxitin, scopeltin, stigmasterol, and many diterpenoidal derivatives.

Maiden & Betche is cultivated around the world for its highly valued nuts (macadamia nuts). Although the chemical composition of the edible macadamia oil has been repeatedly investigated, other plant organs have not been phytochemically or biologically assessed. In this study, ethanolic extract of leaves was phytochemically investigated which led to the isolation of 6 compounds. Two functional galactolipids, monogalactosyl diacylglycrol 36:4 (MGDG 36:4), digalactosyl monoacylglycerol 18:2 (DGMG 18:2), gallic acid and protocatechuic acid were identified in the genus for the first time, in addition to the cyanogenic glycoside dhurrin and -sitosterol. Additionally, anti-tyrosinase activity of the extract, its fractions and isolated compounds was investigated and a good tyrosinase inhibitory activity was observed for the extract, IC=85 µg/mL and its polar fractions (ethyl acetate at 60 µg/mL and -butanol at 75 µg/mL), with gallic acid showing strong anti-tyrosinase activity at IC 56 µg/mL.

Plants of the genus Pulicaria (family: Asteraceae) are widely used in central Asia and the Middle East for treatment of different human diseases. Ultra performance liquid chromatography coupled to high resolution mass spectrometry (UPLC/MS) was utilized to establish the metabolic profiles of two Pulicaria species: P. crispa and P. incisa. 122 metabolites were identified including flavonoids (37), phenolic acids (22), sesquiterpenes (17), diterpenes (7), and fatty acids (27), with enrichment in methoxylated flavonoids (20), caffeoylquinic acid conjugates (14) xanthane sesquiterpenes (9) and hydroxylated fatty acids (20) in both Pulicaria species. The metabolite profile of P. incisa was characterized by the presence of tri- and tetra-methoxylated flavonoids while xanthane sesquiterpenes were the main chemical markers of P. crispa. Additionally, a novel sesquiterpene acid (dihydropulicaric acid) was annotated in both species based on its MS fragments. Antioxidant activity for P. crispa and P. incisa methanol extracts was assessed in vitro based on DPPH and ABTS assays and further in vivo using chlorpromazine intoxicated rat model. Results revealed that P. incisa extract was more effective in inhibiting both DPPH and ABTS free radicals (IC50 0.36 and 0.52 mg/mL, respectively) than P. crispa (IC50 0.51 and 0.73 mg/mL). In the animal model, antioxidant activity of P. incisa (20 mg/kg/day) was also slightly higher causing a 55 % reduction in MDA levels and 65 % increase in GSH activity compared to untreated animals. Furthermore, both extracts showed a hepatoprotective effect as revealed by improvement in levels of serum biomarkers of liver functions: total bilirubin, alanine transaminase (ALT) and aspartate transaminase (AST) comparable to silymarin at 25 mg/kg/day. These findings were also supported by the preserved integrity of the hepatic tissues of animals receiving either extracts at a dose of 20 mg/kg b.wt. The present study reveals for the potential antioxidant and hepatoprotective effects for Pulicaria in relation to its bioactive metabolites.

Salvadora persica L. (toothbrush tree, Miswak) is well recognized in most Middle Eastern and African countries for its potential role in dental care, albeit the underlying mechanism for its effectiveness is still not fully understood. A comparative MS and NMR metabolomics approach was employed to investigate the major primary and secondary metabolites composition of S. persica in context of its organ type viz., root or stem to rationalize for its use as a tooth brush. NMR metabolomics revealed its enrichment in nitrogenous compounds including proline-betaines i.e., 4-hydroxy-stachydrine and stachydrine reported for the first time in S. persica. LC/MS metabolomics identified flavonoids (8), benzylurea derivatives (5), butanediamides (3), phenolic acids (8) and 5 sulfur compounds, with 21 constituents reported for the first time in S. persica. Principal component analysis (PCA) and hierarchical cluster analysis (HCA) of either NMR or LC/MS dataset clearly separated stem from root specimens based on nitrogenous compounds abundance in roots and is justifying for its preference as toothbrush versus stems. The presence of betaines at high levels in S. persica (9−12 μg/mg dry weight) offers novel insights into its functioning as an osmoprotectant that maintains the hydration of oral mucosa. Additionally, the previously described anti-inflammatory activity of stachydrine along with the antimicrobial effects of sulfonated flavonoids, benzylisothiocynate and ellagic acid derivatives are likely contributors to S. persica oral hygiene health benefits. Among root samples, variation in sugars and organic acids levels were the main discriminatory criterion. This study provides the first standardization of S. persica extract using qNMR for further inclusion in nutraceuticals.

Estrogen signaling is crucial for breast cancer initiation and progression. Endocrine-based therapies comprising estrogen receptor (ER) modulators and aromatase inhibitors remain the mainstay of treatment. This study aimed at investigating the antitumor potential of the most potent compounds in citrus peels on breast cancer by exploring their anti-estrogenic and anti-aromatase activities. The ethanolic extract of different varieties of citrus peels along with eight isolated flavonoids were screened against estrogen-dependent breast cancer cell lines besides normal cells for evaluating their safety profile. Naringenin, naringin and quercetin demonstrated the lowest IC and were therefore selected for further assays. In silico molecular modeling against ER and aromatase was performed for the three compounds. In vivo estrogenic and anti-estrogenic assays confirmed an anti-estrogenic activity for the isolates. Moreover, naringenin, naringin and quercetin demonstrated in vitro inhibitory potential against aromatase enzyme along with anticancer potential in vivo, as evidenced by decreased tumor volumes. Reduction in aromatase levels in solid tumors was also observed in treated groups. Overall, this study suggests an antitumor potential for naringenin, naringin and quercetin isolated from citrus peels in breast cancer via possible modulation of estrogen signaling and aromatase inhibition suggesting their use in pre- and post-menopausal breast cancer patients, respectively.

Plants of the genus Pulicaria (family: Asteraceae) are widely used in central Asia and the Middle East for treatment of different human diseases. Ultra performance liquid chromatography coupled to high resolution mass spectrometry (UPLC/MS) was utilized to establish the metabolic profiles of two Pulicaria species: P. crispa and P. incisa. 122 metabolites were identified including flavonoids (37), phenolic acids (22), sesquiterpenes (17), diterpenes (7), and fatty acids (27), with enrichment in methoxylated flavonoids (20), caffeoylquinic acid conjugates (14) xanthane sesquiterpenes (9) and hydroxylated fatty acids (20) in both Pulicaria species. The metabolite profile of P. incisa was characterized by the presence of tri- and tetra-methoxylated flavonoids while xanthane sesquiterpenes were the main chemical markers of P. crispa. Additionally, a novel sesquiterpene acid (dihydropulicaric acid) was annotated in both species based on its MS fragments. Antioxidant activity for P. crispa and P. incisa methanol extracts was assessed in vitro based on DPPH and ABTS assays and further in vivo using chlorpromazine intoxicated rat model. Results revealed that P. incisa extract was more effective in inhibiting both DPPH and ABTS free radicals (IC 0.36 and 0.52 mg/mL, respectively) than P. crispa (IC 0.51 and 0.73 mg/mL). In the animal model, antioxidant activity of P. incisa (20 mg/kg/day) was also slightly higher causing a 55 % reduction in MDA levels and 65 % increase in GSH activity compared to untreated animals. Furthermore, both extracts showed a hepatoprotective effect as revealed by improvement in levels of serum biomarkers of liver functions: total bilirubin, alanine transaminase (ALT) and aspartate transaminase (AST) comparable to silymarin at 25 mg/kg/day. These findings were also supported by the preserved integrity of the hepatic tissues of animals receiving either extracts at a dose of 20 mg/kg b.wt. The present study reveals for the potential antioxidant and hepatoprotective effects for Pulicaria in relation to its bioactive metabolites.

Salvadora persica L. (toothbrush tree, Miswak) is well recognized in most Middle Eastern and African countries for its potential role in dental care, albeit the underlying mechanism for its effectiveness is still not fully understood. A comparative MS and NMR metabolomics approach was employed to investigate the major primary and secondary metabolites composition of S. persica in context of its organ type viz., root or stem to rationalize for its use as a tooth brush. NMR metabolomics revealed its enrichment in nitrogenous compounds including proline-betaines i.e., 4-hydroxy-stachydrine and stachydrine reported for the first time in S. persica. LC/MS metabolomics identified flavonoids (8), benzylurea derivatives (5), butanediamides (3), phenolic acids (8) and 5 sulfur compounds, with 21 constituents reported for the first time in S. persica. Principal component analysis (PCA) and hierarchical cluster analysis (HCA) of either NMR or LC/MS dataset clearly separated stem from root specimens based on nitrogenous compounds abundance in roots and is justifying for its preference as toothbrush versus stems. The presence of betaines at high levels in S. persica (9-12 μg/mg dry weight) offers novel insights into its functioning as an osmoprotectant that maintains the hydration of oral mucosa. Additionally, the previously described anti-inflammatory activity of stachydrine along with the antimicrobial effects of sulfonated flavonoids, benzylisothiocynate and ellagic acid derivatives are likely contributors to S. persica oral hygiene health benefits. Among root samples, variation in sugars and organic acids levels were the main discriminatory criterion. This study provides the first standardization of S. persica extract using qNMR for further inclusion in nutraceuticals.

This study was designed to investigate the effect of Morus nigra fruit extract in retarding the progression of diabetic nephropathy in streptozotocin (STZ)-induced diabetic rats.Diabetic male Wistar rats were injected with black mulberry fruit extract (BMFE) at doses of 150 and 300 mg/kg body weight. After 4 weeks, microalbuminuria was estimated in addition to serum concentrations of glucose, insulin, creatinine and albumin.The study revealed a significant amelioration of all the measured parameters in diabetic animals. In addition, MDA, lipid peroxide levels and catalase activity were also improved. The histopathological examination of kidney tissues revealed significant improvement of the pathological changes and glomerular sclerosis in diabetic rats treated with BMFE. Treated rats showed downregulation of TNF-α, vascular cell adhesion molecule-1 (VCAM-1) and fibronectin mRNA expression.The ameliorative effect of BMFE on diabetic nephropathy is not only through its potent antioxidant and hypoglycaemic effects but also through its downregulation of TNF-α, VCAM-1 and fibronectin mRNA expression in renal tissues of diabetic-treated rats. Therefore, BMFE as dietary supplement could be a promising agent in improving diabetic nephropathy.

Powerful antioxidants can delay progression of diabetes and protect vital organs for tissue damage caused by high concentration of free radicals and reactive oxygen species generated by a sustained state of hyperglycemia. Preliminary studies point to the positive effect of the alcoholic extract of Bauhinia vahlii leaves, a plant traditionally used for treatment of diabetes, in controlling oxidative stress associated with diabetes. Evidence indicates that B. vahlii exerts its effect through moderate antihyperglycemic effect and strong antioxidant activity. B. vahlii extract when given to diabetic Wister rats for 4 weeks was able to reduce fasting blood glucose and glycated hemoglobin by up to 43% and 37%, respectively, indicating moderate antihyperglycemic effect, which may be mediated by strong inhibition of α-glucosidase enzyme (IC50=30.7µg/mL). On the other hand, the antioxidant activity of B. vahlii allowed a full restoration of serum total antioxidant capacity, hepatic malonyldialdehyde, and reduced glutathione to their basal levels in normal nondiabetic animals after treatment with a daily dose of 300mg/kg b.w. Moreover, normal histological features and liver functions were observed in diabetic animals receiving the same dose of B. vahlii extract but not with the standard antidiabetic metformin.

Fava beans are considered basic food in Egypt and all over the world. They are a source of many bioactive secondary metabolites. Since there are many Egyptian cultivars of fava beans, this study addresses comparative chemical profiling of three Egyptian cultivars of fava beans namely Sakha 3, Nubaria 3, and Giza 843. The study also focuses on the effect of germination on the chemical profile of phenolic compounds and saponins. Sixty-five metabolites were characterized based on their UV spectra, accurate MS, and MS/MS data using RP-HPLC-DAD-ESI-QTOF-MS, including 37 metabolites reported for the first time in fava beans. The observed metabolites are phenolic compounds (flavonoids and phenolic acids), saponins, alkaloids, amino acids, nucleosides, organic acids, sugars, and a jasmonate. In fact, upon quantifying the area of metabolites in their chromatographic profiles, it was remarkable that germination increased the quantities of flavonoids, phenolic acids, and saponins. Moreover, the yield (%) of extracts, TPC, and DPPH increased upon sprouting. The TPC and the relative amounts of flavones, saponins, and amino acids showed a good positive correlation with DPPH. These findings elucidated the first report on the metabolic profiling of these cultivars as well as their antioxidant activities.

Vicia faba L. is a legume from the family Fabaceae. Ancient Egyptians consumed fava beans thousands of years ago and they are still one of the most popular foods in Egypt. The current study examined the anti-Parkinson effect of 80% methanolic extracts of seeds or sprouts of the fava ‘Sakha 3 ‘cultivar which has been selected based on the total phenol content among three cultivars tested. In addition, the extracts were characterized by reversed-phase high-performance liquid chromatography coupled with diode array detection and quadrupole-time-of-flight-mass spectrometry (RP-HPLC–DAD-QTOF-MS). Three doses (200, 400, and 600 mg/kg) of 80% methanol extracts of seeds or sprouts of the Sakha 3 cultivar were evaluated in rotenone–Parkinsonian mice from behavioral, biochemical, and histopathological aspects. The extract of fava sprouts (600 mg/kg dose) showed the most beneficial effect. It improved motor activity, enhanced striatal dopamine level, and decreased the striatal malondialdehyde, as well as the expression of the inflammatory markers, compared with the rotenone control group and groups receiving lower therapeutic doses of the extracts or l-Dopa. In addition, these findings were supported by a histopathological investigation which indicated that mice treated with the 600-mg/kg dose of the sprout extract showed a low number of degenerated neurons. The application of RP-HPLC–DAD-QTOF-MS and mass/mass spectroscopy enabled the metabolic profiling of the sprouts and seeds of the ‘Sakha 3′ cultivar. It is obvious that germination increased the amounts of phenolic acids, saponins, and aromatic amino acids, together with a dramatic increase in flavonoids. In conclusion, the 80% methanolic extract of sprouts of the fava “Sakha 3” cultivar may be a promising candidate for treating Parkinsonism if appropriate safety data are available.