Alcohol promotes liver fibrosis in high fat diet induced diabetic rats.

OBJECTIVES: Type 2 diabetes (T2DM) and alcoholism are considered to be lifestyle-associated independent risk factors in fatty liver diseases (FLD) mediated cirrhosis and hepatocellular carcinoma (HCC). A combined effect of both these conditions may exacerbate the pathological changes and a pre-clinical exploration of this is expected to provide a mechanical detail of the pathophysiology. The present study aims to understand the effect of alcohol on pre- diabetic and type 2 diabetic female Wistar rats. METHODS: In this experimental study, 12 Wistar rats (180-220 g) were randomly assigned into three groups: Normal (fed normal rat chow), alcohol (20 %) fed diabetic (HFD + STZ), and pre-diabetic rats (HFD alone). After, two months of the experimental period, blood and liver tissues were collected lipid metabolic alteration, liver injury, and fibrosis were determined following biochemical and histological methods. Data were analyzed using one-way ANOVA and Dunnett's Post Hoc test. RESULTS: Significant dyslipidemia was observed in the liver tissues of diabetic and pre-diabetic rats following alcohol ingestion. A significant (p<0.05) increase in lipid peroxidation status, and hepatic marker enzyme activities (p<0.0001) were observed in diabetic animals. In corroborating with these observations, hematoxylin and eosin staining of hepatic tissue revealed the presence of sinusoidal dilation along with heavily damaged hepatocytes and inflammatory cell infiltration. Further, significantly (p<0.001) increased hepatic hydroxyproline content and extended picrosirius red stained areas of collagen in liver tissue indicated initiation of fibrosis in alcohol-fed diabetic rats. CONCLUSIONS: Overall, the results indicate that alcohol consumption in T2DM conditions is more deleterious than pre diabetic conditions in progressing to hepatic fibrosis.

Virgin Coconut Oil Alleviates Dextran Sulphate-Induced Inflammatory Bowel Disease and Modulates Inflammation and Immune Response in Mice.

OBJECTIVE: Virgin coconut oil (VCNO), an unrefined kernel oil from Cocos nucifera L., has considerable medicinal and nutritive value. Experimental evidence suggests its antioxidant, anti-inflammatory, chemoprotective, analgesic, and hypolipidemic effects. Presently, the effect of VCNO on ameliorating dextran sodium sulfate (DSS)-induced inflammatory bowel disease and cyclophosphamide (CTX)-induced immunosuppression in experimental animals was analyzed. METHOD: DSS (4%) was administered to BALB/c mice through drinking water for 12 days to induce inflammatory bowel disease, and VCNO (500, 750, and 1000 mg/kg bwt) was supplemented orally for 12 days. For anti-inflammatory studies, lipopolysaccharide (LPS, 250 µg/animal) was injected into the intraperitoneal cavity of Swiss albino mice followed by 7 days' pretreatment of VCNO (500, 750, and 1000 mg/kg bwt). To understand the mechanism of action, serum from all animals was collected after 6 hours of LPS challenge and levels of proinflammatory cytokines were analyzed using enzyme-inked immunosorbent assay. In addition to this, immunosuppression was induced by CTX (50 mg/kg bwt, po) in Swiss albino mice. RESULTS: Oral administration of VCNO effectively reversed the pathologies associated with inflammatory bowel disease induced by DSS, including loss of body weight, increased disease activity index, shortening of colon length, diarrhea, and rectal bleeding. Histopathological examination showed that VCNO restored the damage in colon tissue induced by DSS. Similar trends were noticed in levels of myeloperoxidase and mRNA expression of proinflammatory cytokines in colon tissue. In addition to this, supplementation of VCNO markedly reduced the hike in the level of serum proinflammatory cytokines in LPS-challenged mice. Further, administration of VCNO effectively increased spleen and thymus indexes and stimulated the production of interferon-γ in serum. CONCLUSIONS: Overall, this study revealed that VCNO alleviates inflammatory bowel disease and inflammation; concurrently, it can revert immunosuppression.

BRCA1/TP53 tumor proteins inhibited by novel analogues of curcumin - Insight from computational modelling, dynamic simulation and experimental validation.

The current study aimed to design novel curcumin analogue inhibitors with antiproliferative and antitumor activity towards BRCA1 and TP53 tumor proteins and to study their therapeutic potential by computer-aided molecular designing and experimental investigations. Four curcumin analogues were computationally designed and their drug-likeness and pharmacokinetic properties were predicted. The binding of these analogues against six protein targets belonging to BRCA1 and TP53 tumor proteins were modelled by molecular docking and their binding energies were compared with that of curcumin and the standard drug cyclophosphamide and its validated target. The stabilities of selected docked complexes were confirmed by molecular dynamic simulation (MDS) and MMGBSA calculations. The best-docked analogue was chemically synthesized, characterized, and used for in vitro cytotoxic screening using DLA, EAC, and C127I cell lines. In vivo antitumor studies were carried out in Swiss Albino Mice. The study revealed that the designed analogues satisfied drug-likeness and pharmacokinetic properties and demonstrated better binding affinity to the selected targets than curcumin. Among the analogues, NLH demonstrated significant interaction with the BRCA1-BRCT-c domain (TG3; binding energy -8.3 kcal/mol) when compared to the interaction of curcumin (binding energy -6.19 kcal) and cyclophosphamide (binding energy -3.8 kcal/mol) and its usual substrate (TG7). The MDS and MM/GBSA studies revealed that the binding free energy of the NLH-TG3 complex (-61.24 kcal/mol) was better when compared to that of the cyclophosphamide-TG7 complex (-21.67 kcal/mol). In vitro, cytotoxic studies showed that NLH demonstrated significant antiproliferative activities against tumor cell lines. The in vivo study depicted NLH possesses the potential for tumor inhibition. Thus, the newly synthesized curcumin analogue is probably used to develop novel therapeutic agents against breast cancer.

Normolipidic diet containing deep-fried saturated and unsaturated fatty acids rich edible oils promotes metabolic dysregulation and inflammatory microenvironment in Wistar rats.

Deleterious health impacts of high dietary intake of deep-fried edible oils have been reported however, their health impacts at normal dietary levels are yet to be evaluated. This study investigated the influence of prolonged consumption of thermally oxidised long-chain saturated and unsaturated edible oils on metabolic dysregulation and inflammation. The thermally oxidised oils used in the study possess higher p-anisidine values and free fatty acid contents compared to unoxidised oils. Fourier transform infrared spectroscopy and fatty acid methyl ester analysis confirmed the presence of free fatty acids, hydroperoxides, and aldehydes, formed during thermal oxidation. The study analysed the effects of dietary intake of 5% sunflower oil, palm oil, and their thermally-oxidised forms in male Wistar rats for six months. Unoxidised and thermally oxidised palm oil-fed animals experienced metabolic syndrome with obesity-associated inflammatory changes. However, sunflower oil-fed animals exhibited increased inflammation, as evidenced by enhanced C-reactive protein, IL-6, and lipoprotein-associated phospholipase A2 activity and hepatosteatosis condition. This study indicates that prolonged consumption of thermally oxidised oil leads to the dysregulation of carbohydrate and lipid metabolism andpromotes inflammatory microenvironment in Wistar rats that may contribute to metabolic syndrome associated with obesity and hepatic steatosis.

Structural and functional validation of a cloned parasporin from Bacillus thuringiensis isolate KAU 41 native to Western Ghats of India.

Parasporins of Bacillus thuringiensis (Bt) exhibit specific toxicity to cancer cells. PCR based mining has identified apoptosis inducing parasporin in KAU41 Bt isolate from the Western Ghats of India. The study aimed to clone and overexpress the parasporin of native KAU41 Bt isolate for determining structural and functional characteristics of the protein. Parasporin gene was cloned in pGEM-T, sequenced, sub-cloned in pET30+ and overexpressed in Escherichia coli. The expressed protein was characterized by SDS-PAGE and in silico methods. Cytotoxicity of cleaved peptide was assessed by MTT assay. SDS-PAGE displayed a 31 kDa protein (rp-KAU41) overexpressed. Upon proteinase K digestion, the protein was cleaved into 29 kDa peptide which was found to be cytotoxic to HeLa cells. The protein has a deduced sequence of 267 amino acids with ß-strands folding pattern of crystal protein. Even though rp-KAU41 shared a 99.15% identity to chain-A of non-toxic crystal protein, it only showed a less similarity to the existing parasporins like PS4 (38%) and PS5 (24%) in UPGMA analysis, emphasizing the novelty of rp-KAU41. The protein is predicted to have more similarity to the pore forming toxins of Aerolysin superfamily and an additional loop in rp-KAU41 may be contributing towards its cytotoxicity. The molecular docking with caspase 3 resulted in higher Z dock and Z rank score substantiating its role in the activation of intrinsic pathway of apoptosis. The recombinant parasporin protein, rp-KAU41 is presumed to belong to the Aerolysin superfamily. An interaction with caspase 3 substantiates its role in activating the intrinsic pathway of apoptosis in cancer cells.

A High-Fructose Diet Formulated with Thermally Oxidized Monounsaturated Fat Aggravates Metabolic Dysregulation in Colon Epithelial Tissues of Rats.

BACKGROUND: Various epidemiological and clinical studies have indicated a positive association of colon cancer with high sugar and thermally oxidized fats consumption. The present study evaluated the effects of fresh and thermally oxidized coconut (CO) and mustard oils (MO) along with a high-sugar diet in the rat colon mucosa. METHODS: The animals were fed with a modified diet containing high-fructose and different edible oils as fatty acids sources over a period of 30 weeks. Further, the development of insulin resistance and hyperglycemia were estimated biochemically. The changes in the redox status were estimated in terms of reduced glutathione (GSH), antioxidant enzymes and thiobarbituric acid reactive substances (TBARS). Changes in the expression of genes associated with inflammation and cell proliferation were evaluated by qPCR. RESULTS: The animals fed with high-fructose developed hyperglycemia and insulin resistance over 30 weeks. These animals had diminished GSH level, SOD activity and a concomitant increase in the TBARS level in the colon epithelial tissues. In addition, the expression of pro-inflammatory cytokines (IL-6 and TNF-α) was elevated while P53 and PPARγ were down-regulated. This heightened body metabolic dysregulation and associated oxidative damage and inflammation in the colon were exacerbated by thermally oxidized edible oils incorporated in the diet, with a more prominent effect was observed with TMO. CONCLUSION: Feeding high-fructose diet with TMO increases the oxidative and inflammatory damages in the colon epithelium of Wistar rats. Therefore, the study cautions the prolonged consumption of thermally oxidized monounsaturated fat-rich edible oils, especially by individuals with type 2 diabetes.

Hot-processed virgin coconut oil abrogates cisplatin-induced nephrotoxicity by restoring redox balance in rats compared to fermentation-processed virgin coconut oil.

Virgin coconut oil (VCO) is a functional food oil prepared from fresh coconut kernel either by hot-processed (HPVCO) or fermentation-processed (FPVCO). The FPVCO has been widely explored for its pharmacological efficacy; while HPVCO, which has traditional uses, is less explored. The present study compared the phenolic content and nephroprotective effect of both these oils in male Wistar rats. In vitro antioxidant activity was estimated in terms of 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, ferric reducing antioxidant power and ex vivo lipid peroxidation inhibition. In in vivo models, the rats were pretreated orally with of FPVCO or HPVCO (doses 2 and 4 mL/kg) for seven days and nephrotoxicity was induced by the single intraperitoneal injection of cisplatin (10 mg/kg). The results indicated significantly higher polyphenol content in HPVCO (400.3 ± 5.8 µg/mL) than that of FPVCO (255.5 ± 5.8 µg/mL). Corroborating with the increased levels of polyphenols, the in vitro antioxidant potential was significantly higher in the HPVCO. Further, pretreatment with these VCO preparations protected the rats against the cisplatin-induced nephrotoxicity, with higher extent by HPVCO. The renal function markers like urea, creatinine and total bilirubin were significantly reduced (p < 0.05) with HPVCO pretreatment. Apart from the nephroprotective effects, HPVCO also abrogated the cisplatin-induced myelosuppression and hepatotoxicity. The restoration of hepato-renal function by the pretreatment of HPVCO was well corroborated with the improvement in functional antioxidants and subsequent reduction in renal lipid peroxidation. Supporting these observations, renal histology revealed reduced glomerular/tubular congestion and necrosis. Thus, the study concludes that HPVCO may be better functional food than FPVCO.

Virgin coconut oil complements with its polyphenol components mitigate sodium fluoride toxicity in vitro and in vivo.

Virgin coconut oil (VCO), prepared from fresh coconut kernel without any chemical refining, is an emerging functional food. The pharmacological benefits of VCO are believed to be due to the natural combination of phenolics. Although cell culture studies have demonstrated the antioxidant activity of VCO under different oxidative stress conditions, a valid in vivo demonstration of the antioxidant activity of VCO is yet to come. Sodium fluoride (NaF), an environmental pollutant, is widely used to induce oxidative stress in cell culture models and rodents to test the antioxidant potential of several compounds. Herein, VCO and its polyphenolic (VCOP) and non-phenolic oil fraction (VCOF) were individually tested in fluoride-exposed normal intestinal cells (IEC-6) and mice to address their contribution to the documented antioxidant potential. It was found that pretreatment of VCOP (40 µg/mL) was effective in mitigating the fluoride-induced cell death when compared to VCO (200 µg/mL) and VCOF (160 µg/mL). Further, exposure to fluoride (10 mM), increased the intracellular ROS measured based on the dichlorofluorescein (DCF) fluorescence, and this, in turn, was significantly reduced when the cells were supplemented with VCOP. Oral administration of VCO (2 mL/kg bwt) reversed the drop in the hepatic catalase and SOD activities to near normal with a minimal level of lipid peroxidation in fluoride intoxicated mice. However, VCOP and VCOF were less effective in lowering the fluoride-induced increase in hepatic oxidative stress markers. It is reasoned that the oil components of VCO complement the natural antioxidant molecules resulting in an overall increase in their bioavailability.

Amelioration of sodium fluoride induced oxidative stress by Cynometra travancorica Bedd in mice.

OBJECTIVES: Cynometra travancorica, endemic to Western Ghats of India is pharmacologically similar to Saraca asoca and occasionally used as substitute in a well-known Ayurvedic uterine tonic Asokarishta. S. asoca possess various biological properties, but there are no reports on C. travancorica. The present study evaluated the pharmacological properties of C. travancorica and its efficacy in attenuating the sodium fluoride (NaF) induced oxidative stress in mice. METHODS: Antioxidant potential of methanolic extract of C. travancorica (CTE) stem bark was evaluated using DPPH, superoxide radical scavenging and total antioxidant assays. The effect of CTE on mitigating NaF deteriorated redox status in the liver tissue of mice was evaluated. Functional groups in CTE were analyzed by FTIR analysis. RESULTS: CTE effectively scavenged the free radicals in in vitro condition. CTE could augment catalase (46.6%), superoxide dismutase (53.8%) activities and GSH level (48.1%) against NaF induced decline in the liver tissue of mice. The peroxidation of lipids was found to be decreased by 44.9% and tissue damage abated as inferred by histopathology. FTIR analysis revealed the presence of biologically active functional groups in CTE. CONCLUSIONS: The study revealed the ameliorative effect of C. travancorica against NaF induced deleterious effect in experimental animals by its potent antioxidant potential.

Desmodium gyrans dc modulates lipid trafficking in cultured macrophages and improves functional high-density lipoprotein in male wistar rats.

OBJECTIVE: High-density lipoprotein (HDL) cholesterol-mediated atherosclerotic plaque regression has gained wide therapeutic attention. The whole plant methanolic extract of the medicinal plant Desmodium gyrans Methanolic Extract (DGM) has shown to mitigate hyperlipidemia in high fat- and-cholesterol fed rats and rabbits with significant HDL enhancing property. The study aimed to assess the functionality and mechanistic basis of HDL promoting effect of DGM. MATERIALS AND METHODS: Macrophage cholesterol efflux and foam cell formation assays were performed in THP-1 macrophages. Male Wistar rats were given DGM extract over 1 month and assessed the serum HDL, Apolipoprotein A1 (Apo-A1), and paraoxonase activity. Quantitative Polymerase chain reaction was carried out to assess the expression level of Apo-A1, SR-B1 (Scavenger receptor B1), and Cholesteryl ester transfer protein (CETP) on cDNA of HepG2 cells exposed to DGM. RESULTS: Pretreatment of DGM inhibited uptake of oxidized lipids and enhanced the lipid efflux by THP-1-derived macrophages. Oral administration of DGM (100 and 250 mg/kg) progressively enhanced the serum HDL, Apo-A1 level, and associated paraoxonase activity in normal male Wistar rats. In support to this, DGM exposed HepG2 cells documented dose-dependent increase in the expression of SR-B1 and Apo-A1 mRNA, while reduced the CETP expression. CONCLUSION: Overall the results indicated that DGM modulates lipid trafficking and possesses functional HDL enhancing potential through increased Apo-A1 levels and paraoxonase activity. Further, reduced CETP expression and increased expression of SR-B1 suggest the reverse cholesterol transport promoting role of DGM.

Atherogenic oxoaldehyde of cholesterol induces innate immune response in monocytes and macrophages.

Cholesterol oxidation product, 3ß-hydroxy-5-oxo-5,6-secocholestan-6-al (cholesterol 5,6-secosterol, ChSeco or Atheronal-A), formed at inflammatory sites, has been shown to promote monocyte differentiation into macrophages and cause elevated expression of macrophage scavenger receptors. Since inflammation is a key event at all stages of atherosclerotic plaque formation, the pro-inflammatory actions of ChSeco in human THP-1 monocytes and mouse J774 macrophages were investigated in the present study by employing ELISA, qRT-PCR, and functional assays. An increase in the secretion of interleukin-8 and platelet-derived growth factor (PDGF) isoform AA and, to a limited extent, PDGF isoform BB was observed into the culture medium of THP-1 monocytes exposed to ChSeco. However, no changes were seen in the secretion of tumor necrosis factor-alpha. In J774 macrophages treated with ChSeco, there was an upregulation of gene expression of several pro-inflammatory cytokines and their receptors. Concomitantly, there was down-regulation of gene expression of interleukin-1ß, interleukin-10, and lymphotoxin-beta. An increase in the release of interleukin-18 and chemokine (C-C motif) ligand-20 from J774 macrophages (which corroborated well with their gene expression profiles) and increased binding of THP-1 monocytes to ChSeco-exposed human aortic endothelial cells were observed. The results of the present study, for the first time, demonstrate the pro-inflammatory action of ChSeco and suggest the underlying pro-atherogenic mechanisms. These could be mediated through enhanced monocyte recruitment into the sub-endothelial space and subsequent proliferation of smooth muscle cells under the influence of monocyte-derived PDGF.

Thermally Oxidized Coconut Oil as Fat Source in High-Fat Diet Induces Hepatic Fibrosis in Diabetic Rat Model.

In the present study, HFD/STZ-mediated type 2 diabetic rodent model was used to comparatively evaluate coconut oil (CO) and thermally oxidized CO (TCO) as fat sources for the development of NAFLD. Female Wistar rats (six in each group; average bwt 200 g) fed HFD containing either CO or TCO for 2 months along with an intraperitoneal injection of streptozotocin (30 mg/kg bwt) at the end of 1-month feeding were found to develop fatty liver and subsequent inflammatory changes when compared to the normal laboratory diet-fed animals over 2-month period. Dyslipidemia as well as enhanced activities of serum hepatic marker enzymes (e.g., AST, ALT, and ALP) were prominent in TCO-fed animals. Further, HFD-fed animals showed alterations in their hepatic redox equilibrium. Hepatic GSH and antioxidant enzyme activities that form the part of a protective mechanism against oxidative/carbonyl stress were found to be increased in HFD-fed rats. Supporting this, CO- and TCO-containing-HFD-fed animals had enhanced lipid peroxidation (increased TBARs). Thus, fatty liver with heightened antioxidant defense, lipid peroxidation, and inflammation indicate hepatosteatosis. Histological details of the hepatic tissues corroborated sufficiently with these observations and showed an increased incidence of macrovesicles, inflammation, and hepatocyte ballooning in the TCO-fed rats than in CO-fed animals. Further, in support of this proposition, hydroxyproline, an index of collagen formation, was found to be significantly increased in TCO-fed rats than in the CO-fed group. Overall, the study shows that the formulation of HFD incorporated with TCO as a fat source, combined with STZ injection, is an efficient dietary model for developing hepatosteatosis with fibrotic stage in rats within 2 months. Administration of this modified diet for a more extended period may be a good model for cirrhotic and hepatocellular carcinoma studies, which need to be further assessed.

Variations in natural polyphenols determine the anti-inflammatory potential of virgin coconut oils.

Virgin coconut oil (VCO), an edible oil prepared from fresh coconut kernel by natural or mechanical means without undergoing chemical refining, has been in the limelight of research as functional food oil. The phenolic components in VCO have been accredited with these pharmacological benefits. The present study compared the phenolic constituents of freshly prepared fermentation processed (FVCO) and hot-pressed VCO (HVCO) and their anti-inflammatory efficacies. The biochemical analysis documented quantitative variation in the phenolic content, being higher in HVCO than FVCO (40.03 ± 5.8 µg and 25.55 ± 5.8 µg/mL of oil, respectively). In vitro studies observed nitric oxide radical scavenging efficacy (IC50 value of 14.84 ± 0.81 µg/mL) for HVCO polyphenols, which shows higher inhibition efficacy than FVCO (29.41 ± 1.7 µg/mL). In dextran and formalin mediated acute and chronic inflammation in mice, HVCO displayed more protective efficacy (40.5 and 46.4% inhibition) than FVCO (33.3 and 43.8% inhibition), which is similar to the standard diclofenac (55.6 and 59.8% inhibition). The study, thus, concludes that compared to FVCO, HVCO is a more active anti-inflammatory agent. PRACTICAL APPLICATION: Virgin coconut oil, a widely used edible oil in South Asian countries, has been shown to have health benefits possibly exerted by the natural phenolics it contains. However, different modes of preparations of VCO determine the phenolic combinations and efficacy as well. Our study compared two different VCO preparations and suggests that the VCO prepared by the traditional way (HVCO) is pharmacologically potent than that prepared by simple fermentation process (FVCO) in reducing inflammation. The efficacy is attributed to the variations in phenolic profile revealed by LC-MS analysis. Hence, the current study suggests HVCO as a potential food supplement that can reduce the incidence of degenerative diseases.

Targeting Histone Onco- Modifications Using Plant-Derived Products.

The regulatory mechanisms lying over the genome that determines the differential expression of genes are termed epigenetic mechanisms. DNA methylation, acetylation, and phosphorylation of histone proteins and RNAi are typical examples. These epigenetic modifications are important determinants of normal growth and metabolism; at the same time, aberrant histone modifications play a major role in pathological conditions and are emerging as a new area of research for the last decades. Histone onco-modification is a term introduced by the scientific world to denote histone post-translational modifications that are associated with cancer development and progression. These modifications are likely to act in certain conditions as adaptive mechanisms to environmental and social factors. The enzymes that regulate DNA methylation as well as histone modifications are thus become a target for cancer therapy and chemoprevention. Since oxidative stress has been shown to modulate epigenetic changes, and phytocompounds with powerful antioxidant properties have a significant role in disease prevention. Nowadays, "nutri- epigenetics" is becoming an emerging area of research that deals with the influence of dietary compounds in epigenetics. This review aims to discuss the biological efficacy of promising phytocompounds that are able to counteract deleterious epigenetic modifications, especially histone onco- modifications.

Curcumin Enriched VCO Protects against 7,12-Dimethyl Benz[a] Anthracene-Induced Skin Papilloma in Mice.

Virgin coconut oil (VCO) and turmeric are traditionally being used in Indian cuisine systems; VCO is a natural combination of medium-chain triglyceride and polyphenols with established pharmacological potential. Curcumin isolated from turmeric is renowned for its anticancer properties, however, with limited clinical success due to poor bioavailability. Considering the lipophilic nature of VCO, curcumin added to VCO is expected to have synergistic/additive actions. In this study, the chemopreventive potential of curcumin enriched VCO (VCr) (4 and 8 mL/Kg orally) was analyzed in 7,12-dimethyl benz[a]anthracene (DMBA;470 nmoles/200 µL/week for two weeks topical)/croton oil (3% v/v in 200 µL acetone twice a week for 6 weeks topical) induced skin papilloma. In DMBA control animals, an average incidence of 13 papilloma/mice (latency period of 11.6 ± 1.5 weeks) was recorded. Pretreatment with VCrH (8 mL/kg) had a 60% inhibition of tumor index, and an increased latency period (12.5 ± 0.9 weeks). Additionally, DMBA/croton oil-induced reduction in glutathione levels and concomitant increase in thiobarbituric acid reactive substance (TBARS) in the skin microenvironment were restored by VCr. The study thus suggests that the VCr promotes antioxidant status in vivo and imparts an improved anticarcinogenic potential. However, further studies are necessary to ascertain the improvement in bioavailability of curcumin .

Natural combination of phenolic glycosides from fruits resists pro-oxidant insults to colon cells and enhances intrinsic antioxidant status in mice.

A combination of fresh fruits adequately supplying required nutrients is likely to have better health benefits by virtue of the synergistic/additive effect of its natural constituents. With this view and aiming to obtain phenolic glycosides in combination, fresh apple, grape, orange, pomegranate, and sapota fruit juices were combined and lyophilized. An aqueous extract of this fruit combination (AEFC) had polyphenols as a major constituent (47.36 µg GAE/mL) and LC-MS analysis documented the presence of cyanidin and pallidol 3-O-glucosides, phloridzin, delphinidin-3-O-rutinoside, kaempferol-3-O-pentoside, quercetin-3-O-rutinoside, trans-caffeic acid. Corroborating this, AEFC exhibited significant DPPH and superoxide radical scavenging activities (IC50values 43.63 and 49.01 µg/mL) and protected colon epithelial cells (HCT-15) against H2O2 and AAPH induced cell death by 40 and 72.62% and buthionine sulfoximine (BSO) induced GSH depletion by 52.43%. In normal Swiss albino mice, administration of AEFC for over 30 days improved hepatic and renal GPx, SOD, and catalase activities and GSH levels. The study thus suggests the combinatorial effects of natural phenolic glycosides from fruits in resisting oxidative insults and associated disease pathology.

Non-polar lipid carbonyls of thermally oxidized coconut oil induce hepatotoxicity mediated by redox imbalance.

Thermal oxidation products of edible oils including aldehydes, peroxides and polymerized triglycerides formed during the cooking process are increasingly debated as contributory to chronic degenerative diseases. Depending on the oil used for cooking, the source of fatty acids and its oxidation products may vary and would have a differential influence on the physiological process. Coconut oil (CO) is a medium chain triglyceride-rich edible oil used in South India and other Asia Pacific countries for cooking purposes. The present study evaluated the biological effects of thermally oxidized coconut oil (TCO) as well as its non- polar hexane (TCOH) and polar methanol (TCO-M) sub-fractions in male Wistar rats. Results showed an increase in the thiobarbituric acid reactive substances (TBARs) and conjugated diene levels in TCO, which was extracted to TCOH fraction. The animals consumed TCO and its hexane and methanol fractions had a considerable increase in weight gain. However, serum and hepatic triglycerides were increased only in animals with TCO and TCOH administration. In these animals, the hepatic redox balance was disturbed, with a reduction in GSH and a concomitant increase in thiobarbituric acid reactive substances (TBARs). Increased incidence of microvesicles in hepatic histological observations also supported this assumption. Together, the study shows that TCO consumption is unhealthy, where the nonpolar compounds generated during thermal oxidation may be involved in the toxic insults.

A parasporin from Bacillus thuringiensis native to Peninsular India induces apoptosis in cancer cells through intrinsic pathway.

Parasporins, a class of non-insecticidal crystal proteins of Bacillus thuringiensis (Bt) are being explored as promising anticancer agents due to their specific toxicity to cancer cells. The present study has identified 25 Bt isolates harbouring parasporin genes from Western Ghats region, the hotspot of biodiversity in India. Among these, the isolate, KAU 41 (Kerala Agricultural University isolate 41) contained non-hemolytic homogenous crystals showing specific cytotoxicity towards cancer cells. SDS-PAGE analysis of this crystal, isolated by aqueous biphasic separation, revealed a 31 kDa sized peptide. The N-terminal sequence deciphered in BLAST analysis showed homology to a hypothetical Bt protein. Upon proteolysis, a 29 kDa active peptide was generated which exhibited heterogenic cytotoxic spectrum on various cancer cells. HeLa cells were highly susceptible to this peptide with IC 50 1 lg/mL and showed characteristics of apoptosis. RT-qPCR analysis revealed the overexpression of APAF1, caspase 3 and 9 by 14.9, 8 and 7.4 fold, respectively which indicates the activation of intrinsic pathway of apoptosis. However, at higher concentrations of peptide (greater than 3 lg/mL), necrotic death was prominent. The results suggest that the 31 kDa protein from Bt isolate, KAU 41 is a parasporin that may have high therapeutic potential.

Non-enzymatic conversion of primary oxidation products of Docosahexaenoic acid into less toxic acid molecules.

Docosahexaenoic acid (DHA) is long chain omega-3 fatty acid with known health benefits and clinical significance. However, 4-hydroxy hexenal (HHE), an enzymatic oxidation product of DHA has recently been reported to have health-damaging effects. This conflict raises major concern on the long-term clinical use of these fatty acids. Even though the enzymatic and non-enzymatic conversion of HHE to nontoxic acid molecules is possible by the aldehyde detoxification systems, it has not yet studied. To address this, primary oxidation products of DHA in lipoxidase system were subjected to non-enzymatic conversion at physiological temperature over a period of 1 week. The reaction was monitored using HPLC, IR spectroscopy and biochemical assays (based on the loss of conjugated dienes, lipid peroxides aldehydes). Short term and long term cytotoxicity of the compounds generated at various time points were analyzed. IR and HPLC spectra revealed that the level of aldehydes in the primary oxidation products reduced over time, generating acids and acid derivatives within a week period. In short term and long term cytotoxicity analysis, initial decomposition products were found more toxic than the 1-week decomposition products. Further, when primary oxidation products were subjected to aldehyde dehydrogenase mediated oxidation, it generated products that are also less toxic. The study suggests the possible non-enzymatic conversion of primary oxidation products of DHA to less cytotoxic acid molecules. Exploration of the physiological roles of these acid molecules may explain the biological potential of omega-3 fatty acids.