A mechanistic insight into the potential anti-cancerous property of Nigella sativa on breast cancer through micro-RNA regulation: An in vitro & in vivo study.

Cancer continues to threat mortal alongside scientific community with burgeoning grasp. Most efforts directed to tame Cancer such as radiotherapy or chemotherapy, all came at a cost of severe side effects. The plant derived bioactive compounds on the other hand carries an inevitable advantage of being safer, bioavailable & less toxic compared to contemporary chemotherapeutics. Our strategic approach employed solvent extraction of Black Seed Oil (BSO) to highlight the orchestrated use of its oil soluble phytochemicals - Thymoquinone, Carvacrol & Trans-Anethole when used in cohort. These anti-cancer agents in unbelievably modest amounts present in BSO shows better potential to delineate migratory properties in breast cancer cells as compared to when treated individually. BSO was also observed to have apoptotic calibre when investigated in MDA-MB-231 and MCF-7 cell lines. We performed chemical characterization of the individual phytochemical as well as the oil in-whole to demonstrate the bioactive oil-soluble entities present in whole extract. BSO was observed to have significant anti-cancerous properties in cumulative proportion that is reportedly higher than the individual three components. Besides, this study also reports micro-RNA regulation on BSO administration, thereby playing a pivotal role in breast cancer alleviation. Thus, synergistic action of the integrants serves better combat force against breast cancer in the form of whole extract, hence aiming at a more lucrative paradigm while significantly regulating microRNAs associated with breast cancer migration and apoptosis.

Kaempferol with Verapamil impeded panoramic chemoevasion pathways in breast cancer through ROS overproduction and disruption of lysosomal biogenesis.

BACKGROUND: Reactive oxygen species (ROS) at low level promotes cell survival through lysosome induced autophagy induction. Glucose stress induced acidosis, hypoxia, ROS, upregulates markers related to cancer stemness and multidrug resistance. Also, lysosomal upregulation is proposed to be one of the important indicators of cell survival under ROS induced stress. Studies supported that, stimulation of Lysosome-TFEB-Ca2+ cascade has important role in induction of chemoresistance and survival of cancerous cells. PURPOSE: To observe the effect of synergistic drug combination, Kaempferol and Verapamil on markers regulating chemoevasion, tumor stemness & acidosis as well as lysosome upregulation pathways, under low as well as high glucose conditions. HYPOTHESIS: Based on our earlier observation as well as previous reports, we hypothesized, our drug combination Kaempferol with Verapamil could attenuate markers related to chemoevasion, tumor stemness & acidosis as well as lysosome-TFEB-Ca2+ pathway, all of which have indispensable association and role in chemoresistance. METHODS: RNA and protein expression of candidate genes, along with ROS production and Ca2+ concentrations were measured in ex vivo models in altered glucose conditions upon treatment with KV. Also, computational approaches were utilized to hypothesize the mechanism of action of the drug combination. PCR, IHC, western blotting and molecular docking approaches were used in this study. RESULTS: The overproduction of ROS by our candidate drugs KV, downregulated the chemoresistance and tumor acidosis markers along with ATP1B1 and resulted in lysosomal disruption with reduction of Ca2+ release, diminishing TFEB expression under low glucose condition. An anomalous outcome was observed in high glucose conditions. We also observed KV promoted the overproduction of ROS levels thereby inducing autophagy-mediated cell death through the upregulation of LC3-II and p62 in low glucose conditions. The ex vivo studies also corroborate with in silico study that exhibited the parallel outcome. CONCLUSION: Our ex-vivo and in-silico studies revealed that our candidate drug combination KV, could effectively target several pathways regulating chemoresistance, that were not hitherto studied in the same experimental setup and thus may be endorsed for therapeutic purposes.

A novel nanoformulation of α-eleostearic acid restores molecular pathogenesis of hypersensitivity.

AIM: The present work provides first-time empirical and molecular interaction evidence to establish the higher biofunctionality of a therapeutic lipid, α-eleostearic acid (ESA), encapsulated in a novel and thoroughly characterized biocompatible nanoemulsion (NE) system (particle size <200 nm). MATERIALS & METHODS: A novel methodology was employed to fabricate novel formulations of ESA. Molecular biological tools and assays were used to arrive at definite conclusions. RESULTS: The proinflammatory profile was found to be significantly mitigated in the hypersensitized rats administered with the ESA-NE formulation more emphatically as compared with ESA-conventional emulsion in both in vivo and ex vivo models. CONCLUSION: The novel ESA-NE formulation shows a lot of palpable promise for clinical applications.

Effects of nano-sizing on lipid bioaccessibility and ex vivo bioavailability from EPA-DHA rich oil in water nanoemulsion.

The physiological efficacy of nutraceuticals is dependent on their physicochemical nature and bioavailability across biological barriers. In the present work, effects of nano-sizing of emulsion-based delivery vehicle on the bioavailability of polyunsaturated fatty acids rich fish oil have been investigated via three-step experimental design; ex vivo rat everted intestinal sac model, cellular lipid uptake and the bioactivity in rat PBMCs. Nanoemulsion in comparison to the conventional emulsion has shown significant higher rate of uptake of polyunsaturated fatty acids in three segments of small intestine. The time-kinetics of such uptake was correlated with appearance of short-chain fatty acids in basal side of the everted sac. The bioavailability of the formulated fish oil and its inhibitory response against lipopolysaccharide-induced nitric oxide production in rat PBMCs were positively correlated. This formulation with nano-sized droplets can be utilized as smart delivery vehicles for designing oral therapies in future.

Indian freshwater edible snail Bellamya bengalensis lipid extract prevents T cell mediated hypersensitivity and inhibits LPS induced macrophage activation.

ETHNOPHARMACOLOGICAL RELEVANCE: Soup prepared from the foot of fresh water edible snail, Bellamya bengalensis, is traditionally consumed by the tribes of Jharkhand against rheumatism like bone and joint inflammation. As rheumatism has underlying involvement of cell mediated hypersensitivity, in vivo delayed-type hypersensitivity (DTH) model and in vitro LPS-induced macrophage signaling were studied to delineate the mechanism by which Bellamya bengalensis exerts its ethnomedicinal function. Since the whole meat is consumed, the lipid of Bellamya bengalensis (BBL) was hypothesized to be the active part. METHODS AND MATERIALS: BBL isolated from the foot part of this species, was characterized and given by gavage daily (10mg BBL/kg; 20mg BBL/kg) to mice for 3 weeks prior to initiating development of DTH. Effects of DTH induced changes in paw diameter, serum nitric oxide (NO), serum tumor necrosis factor (TNF)-α level, CINC1 level, splenic CD4(+)/CD8(+) cell ratios, and level of splenic Treg cells were then compared with values in untreated control mice. In vitro effect of BBL on LPS-stimulated macrophage, the immune cell that is active in DTH, was assessed by NF-kB p65 nuclear translocation, reactive oxygen species (ROS), TNFα, and NO production. RESULTS: BBL was characterized, and its supplementation in situ led to significant decrease in paw edema, tissue myeloperoxidase activity, NO level, serum TNFα level and CINC 1 level as well as decrease in splenic CD4(+)/CD8(+) ratios and increase in level of Treg cells. BBL was shown to inhibit ROS, NO, and TNFα production along with NF-kB p65 nuclear translocation in LPS stimulated macrophage. CONCLUSION: Bellamya bengalensis, traditionally used against diseases with underlying etiology of cell mediated immunity as in rheumatism, which acts through inhibition of overexpressed cell mediated immunity. The factor exerting this activity probably is the oleic acid and cyclopropane fatty acid rich lipid, isolated after the ethnomedicinal clue, from the foot of this species.

Oxalis corniculata (Oxalidaceae) leaf extract exerts in vitro antimicrobial and in vivo anticolonizing activities against Shigella dysenteriae 1 (NT4907) and Shigella flexneri 2a (2457T) in induced diarrhea in suckling mice.

In this study, the extract of a green leafy vegetable Oxalis corniculata (Oxalidaceae) was evaluated for its in vitro antibacterial and in vivo anti colonizing effect against common intestinal pathogenic bacteria. Methanolic extract (80%) of Oxalis corniculata (Oxalidaceae) leaf contained a polyphenol content of 910 mg gallic acid equivalent per gram of dry weight and the yield was 8%. The flavonoid content was 2.353 g quercetin equivalent per 100 g of the extract. In vitro studies indicated that the extract inhibited numerous pathogenic bacteria like Staphylococcus aureus (ATCC 25922), Escherichia coli (ATCC 25923), Shigella dysenteriae 1 (NT4907), Shigella flexneri 2a (2457T), Shigella boydii 4 (BCH612), and Shigella sonnie phase I (IDH00968). The minimum inhibitory concentration (MIC) against E. coli (ATCC 25923) was minimal (0.08 mg/mL), whereas MIC against S. flexneri 2a (2457T) was higher (0.13 mg/mL). A suckling mouse model was developed which involved challenging the mice intragastrically with S. flexneri 2a (2457T) and S. dysenteriae 1 (NT4907) to study the anticolonization activity. It was revealed that the extract was more potent against S. dysenteriae 1 (NT4907) as compared to S. flexneri 2a (2457T). It was also found that simultaneous administration of extract along with bacterial inoculums promoted good anticolonization activity. Significant activity was observed even when treated after 3 h of bacterial inoculation.

Protective effect of secondary plant metabolites from Ipomoea aquatica Forsk. against carbofuran induced damages.

Plausible interactions between food contaminants and natural constituents in vivo and protective effect of polyphenols present in I. aquatica against carbofuran toxicity in Charles Foster rats were evaluated. Determinations based on antioxidant enzyme activities showed significant alterations in glutathione, glutathione peroxidase, superoxide dismutase and catalase in tissues (liver and brain) and plasma of pesticide treated group while polyphenolic extracts from I. aquatica (IAE) attenuated their activities when given alongwith carbofuran. IAE decreased enhanced lipid peroxidation levels in plasma and erythrocyte membrane and cholesterol levels in brain and plasma. IAE also minimized histopathological degenerative changes produced by carbofuran. While single cell gel electrophoresis showed that secondary metabolites in leafy vegetables produced a combinatorial effect with pesticide at cellular level, DNA fragmentation level in bone marrow cells showed a decline in the IAE treated rats. Food safety adversely affected by various chemical contaminants can be retained by plant polyphenols and secondary plant constituents that can be found together in bolus. Therefore, the present study gives an insight into the protective role of naturally found polyphenols against pesticide toxicity.

Dietary effects of pure and diacylglycerol-rich rice bran oil on growth pattern and lipid profile of rats.

Diacylglycerol (DAG) oil is becoming quite well known for its beneficial effects on obesity and weight related disorders as they are digested and metabolized in a different way, which significantly reduces body weight. The present study was conducted to evaluate the nutritional effect of rice bran oil (RBO) and DAG-rich rice bran oil on rat model. DAG-rich RBO (46.10% DAG) was prepared in the laboratory by enzymatic (TLIM) glycerolysis process. For the feeding experiment, rats (average body weight 130 g) were divided into two groups, each consisting of eight rats. Group RBO was taken as control while DAG-rich RBO (Group DGR) was taken as experimental. Food efficiency ratio, lipid profile of plasma, liver, mesentery and erythrocyte membrane (EM) lipid profile, plasma leptin content, HMG-CoA reductase activity were measured using standard procedures. The results indicated that DAG-rich rice bran oil significantly decreased body weight of rats and content of total cholesterol and triglyceride in plasma compared to the control RBO. The long term use of DAG-rich rice bran oil might be useful for prevention of obesity and incidence of hypercholesterolemia.

Comparative evaluation of essential fatty acid composition of mothers' milk of some urban and suburban regions of West Bengal, India.

This study investigated the fatty acid composition of lipid present in breast milk of mothers residing in urban and suburban regions of West Bengal with special emphasis on n-6 and n-3 polyunsaturated fatty acids, which played a crucial role in the growth and development of neonates. Milk samples collected from 135 mothers of middle income group (average monthly income around 'Rs 10,000/-') were analysed by gas liquid chromatography after extraction and transmethylation to determine fatty acid composition. Information about the dietary intake of individual mothers was obtained through food frequency questionnaire. The fractions of n-6 and n-3 polyunsaturated fatty acids available in milk of urban mothers were 13.59 ± 0.94 and 3.65 ± 0.49, respectively, and in suburban mothers 12.74 ± 0.89 and 4.36 ± 0.39, respectively. The green leafy vegetables, fishes and vegetable oils were the major sources of essential fatty acids in the diet of the experimental groups of Bengali mothers. This study revealed a relationship between the alimentary habits of mothers and the concentration of essential fatty acids in breast milk of Bengali mothers.

In vitro antioxidant activity of different cultivars of banana flower (Musa paradicicus L.) extracts available in India.

Six different cultivars of banana flowers (Musa paradicicus) (Kathali, Bichi, Shingapuri, Kacha, Champa, and Kalabou) were analyzed for the content of polyphenol expressed as gallic acid equivalent and flavonoid expressed as quercetein equivalent, and the in vitro total antioxidative activities of the flower extracts were compared with standard and expressed as trolox equivalent. The reducing power, 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) radical cation (ABTS•(+)) scavenging activities, inhibition of lipid peroxidation in a linoleic acid emulsion system, and liposome peroxidation system were measured and compared with respective standard antioxidants. Iron-mediated Fenton reaction was carried out to evaluate the protective effect of the extract of banana flower (Kacha cultivar) against H(2)O(2)-induced DNA damage. The Kacha variety contains the maximum amount of polyphenol (11.94 ± 0.03 mg of gallic acid equivalent/g of dry weight) and flavonoid (0.174 ± 0.001 g of quercetin equivalent/g of polyphenol). It also has the highest total antioxidant capacity, DPPH radical scavenging activity, and ABTS•(+) radical scavenging activity with a least EC(50) value of 0.051 mg/mL. Hepatic cell damage in iron-mediated Fenton reaction caused by free radicals is reduced by the banana flower extract. On the basis of the results obtained, the banana flowers are found to be a potential source of natural antioxidants. This is the first report on the antioxidant properties of the extracts from banana flowers. The study suggests that the flowers of M. paradicicus that are found in India and consumed as vegetable can provide valuable functional ingredients that help in the prevention of oxidative stress.

Influence of polyphenolic extracts from Enydra fluctuans on oxidative stress induced by acephate in rats.

Acephate, an organophosphorus pesticide, has been proved to play an important role in tissue damage by inducing oxidative stress through the release of free radicals. The aim of this study was to evaluate the protective effect of the plant phenolic compounds present in Enydra fluctuans against acephate toxicity based on lipid peroxidation and antioxidant enzymes profile in rats. An oral dose of acephate at 30 mg/kg of body weight has been given against the extracts containing 20mg of polyphenolic compounds (expressed as gallic acid equivalents)/kg body weight for 14 days. The results showed that under the influence of the pesticides, there was significant decrease in the activities of the antioxidant enzymes SOD, Catalase and Glutathione peroxidase (GPx) and an increase in the non-enzymatic Glutathione, with respect to the normal and the plant extract gavaged groups. Also that there was an increase in the plasma and erythrocyte membrane lipid peroxidation levels in the pesticide treated group compared to the normal or the group treated with the plant extract. The present study thus gives an insight into the ill-effects of this organophosphate and the protective role of plant polyphenols in minimizing those effects.

Lutein content and in vitro antioxidant activity of different cultivars of Indian marigold flower ( Tagetes patula L.) extracts.

Three different cultivars of marigold flowers ( Tagetes patula L.) (marigold orange, marigold yellow, and marigold red) were analyzed for the lutein ester contents, and the in vitro antioxidative activities of the flower extracts were compared. The total antioxidant capacity, reducing power, hydroxyl, DPPH, and ABTS(*+) radical scavenging activities, iron chelation capacity, and inhibition of lipid peroxidation in a linoleic acid emulsion system were measured. Iron-mediated Fenton reaction was carried out to evaluate the protective effect of leutin against DNA damage. The marigold orange (MGO) variety contains the maximum amount of lutein. It also had the highest DPPH radical scavenging activity and ABTS radical scavenging activity, with an EC(50) value of 0.344 mg/mL. It was also the most effective against lipid peroxidation and hydroxyl radical scavenging activities. The MGO extract has the maximum reducing power. Hepatic cell damage in iron-mediated Fenton reaction caused by free radicals was reduced by the marigold extracts. Marigold flowers of Indian variety can be effectively utilized to produce lutein ester, which can be used as a food supplement or as an accessible source of natural antioxidants.

Ameliorative influence of sesame lignans on lipid profile and lipid peroxidation in induced diabetic rats.

Sesame lignans are working as antioxidants in various physiological functions. In the present study, the antioxidative effect of sesame lignans is examined in chemically induced diabetes mellitus (DM) in rats against lipid profile and lipid peroxidations. DM was induced in four groups of rats by injection of alloxan. The control groups (non-diabetic and diabetic) received a diet containing sunflower oil while the rest of the three experimental diabetic groups received a diet containing 0.25% alpha-tocopherol (D-Toc), 0.5% sesame lignan (D-SL), and 0.25% alpha-tocopherol+0.25% sesame lignan (D-Toc-SL) in sunflower oil for 4 weeks. Lipid profile and lipid peroxidations of plasma, erythrocyte membrane (EM), and liver tissues were measured. The total cholesterol, non-HDL cholesterol, plasma lipid peroxidation, and also LDL-peroxidation decreased, and HDL cholesterol increased significantly (P<0.05) in all the experimental groups as compared to the control diabetic sunflower oil group. The triacylglycerol (TAG) level in plasma decreased significantly in the D-SL and D-Toc-SL groups as compared to control diabetic group. Significant decrease in TAG level was observed in the D-SL group as compared to the D-Toc group. LDL peroxidation also decreased significantly in the D-Toc-SL group as compared to the D-Toc group. EM lipid peroxidation and liver lipid peroxidation decreased significantly in the D-Toc, D-SL, and D-Toc-SL groups as compared to the control diabetic group. Liver TAG level decreased more significantly in the D-SL and D-Toc-SL groups than in the control diabetic group. So, sesame lignans at 0.5% level and sesame lignan + alpha-tocopherol significantly ameliorate the alteration in lipid profile and the adverse free radical generative influence of DM induced by alloxan.

Antioxidative effect of conjugated linolenic acid in diabetic and non-diabetic blood: an in vitro study.

The present study examined the in vitro antioxidant activity of conjugated octadecatrienoic fatty acid (9cis, 11 trans, 13 trans-18:3), alpha-eleostearic acid present in karela seed oil (Momordica charantia) at about 55% level. The in vitro antioxidant properties of alpha-eleostearic acid are investigated on oxidative modification of human plasma, low-density lipoprotein (LDL) and erythrocyte membrane lipid. Blood samples are collected from diabetic and non-diabetic (normal) healthy individuals. alpha-eleostearic acid is added at 0.05% and 0.1% concentrations to plasma, LDL and erythrocyte membrane isolated from the respective blood samples and peroxidations are determined against control samples. A significant increase of respective peroxidation levels has been observed in diabetic control blood than the non-diabetic control blood. alpha-eleostearic acid has decreased lipid peroxidation level against control samples in a dose dependent manner. The present findings suggest that CLnA, 9cis, 11trans, 13trans-18:3 is a potentially effective antioxidant that can protect plasma, low density lipoprotein and erythrocyte membrane from oxidation which may be effective in reducing the risk of coronary heart disease in diabetes mellitus.