Thymoquinone rich fraction from Nigella sativa and thymoquinone are cytotoxic towards colon and leukemic carcinoma cell lines.

Nigella sativa has been used for centuries in Asia, Middle East and Africa to promote health and fight diseases. In this study, the anti-cancer effects of thymoquinone rich fraction (TQRF) extracted from N. sativa seeds using supercritical fluid extraction (SFE) system and commercially available thymoquinone (TQ) on colon cancer (HT29), lymphoblastic leukemia (CEMSS) and promyelocytic leukemia (HL60) cells lines were investigated. The concentration that gave 50% inhibition of cell viability (IC50) of HT29, CEMSS and HL60 cells treated with TQRF were 400, 350 and 250 µg/ml, respectively. Meanwhile, the IC50 of TQ was 8, 5 and 3 µg/ml, respectively. Cell cycle analysis shows the increment of apoptosis in a time-dependent manner. However, both TQRF and TQ were not able to arrest the cell cycle phases of the cells. Apoptosis was the main mode of HT29, CEMSS and HL60 cells death induced by both TQRF and TQ. Our findings support the potential use of TQRF and TQ for the treatment of colon cancer and leukemia

Antioxidant activity and phenolic content of phenolic rich fractions obtained from black cumin (Nigella sativa) seedcake.

The antioxidant activities of crude methanolic extract (CME) and its fractions using ethyl acetate (EAF), hexane (HF) and water (WF) of black cumin seedcake were investigated. DPPH radical scavenging activity, β-carotene-linoleate bleaching, and inhibition of corn oil oxidation were used to evaluate the antioxidant capacity. The total phenolics were found to be 78.8, 27.8, 32.1 and 12.1 mg gallic acid equivalents (GAE)/g in EAF, CME, WF and HF, respectively. The CME and EAF exhibited the highest DPPH followed by WF and HF. The extract/fractions showed high effect on reducing the oxidation of β-carotene. The effect of extract/fractions on the oxidative stability of corn oil at 70 °C was tested in the dark and compared with butylated hydroxyanisole (BHA). The oil peroxide and anisidine values were generally lower with addition of PRFs in comparison to a control. The predominant phenolic compounds identified by HPLC-DAD in CME and WF of black cumin seedcake were hydroxybenzoic, syringic and p-cumaric acids

Antioxidant activities of phenolic rich fractions (PRFs) obtained from black mahlab (Monechma ciliatum) and white mahlab (Prunus mahaleb) seedcakes

The antioxidant activities of phenolic rich fractions (PRFs) from crude methanolic extract (CME), and its fractions using ethyl acetate (EAF), hexane (HF) and water (WF) of black mahlab (Monechma ciliatum) and white mahlab (Prunus mahaleb) seedcakes were investigated. The total phenolic compounds were found to be higher in white mahlab than black mahlab seedcakes. The antioxidant activity determined by the DPPH method revealed that black mahlab PRFs had the highest antioxidant activity, compared to white mahlab fractions. The presence of antioxidants in the two mahlab PRFs reduced the oxidation of β-carotene by hydroperoxides from these extracts/fractions. The effect of the two mahlab PRFs on the oxidative stability of corn oil at 70 °C was tested in the dark and compared with butylated hydroxyanisole (BHA). The CME performed better antioxidant activity in inhibiting the formation of both primary and secondary oxidation products. The qualitative and quantitative characterisation of phenolic compounds was carried out by HPLC/DAD

Ethyl acetate extract of germinated brown rice attenuates hydrogen peroxide-induced oxidative stress in human SH-SY5Y neuroblastoma cells: role of anti-apoptotic, pro-survival and antioxidant genes

Background There are reports of improved metabolic outcomes due to consumption of germinated brown rice (GBR). Many of the functional effects of GBR can be linked to its high amounts of antioxidants. Interestingly, dietary components with high antioxidants have shown promise in the prevention of neurodegenerative diseases like Alzheimer’s disease (AD). This effect of dietary components is mostly based on their ability to prevent apoptosis, which is believed to link oxidative damage to pathological changes in AD. In view of the rich antioxidant content of GBR, we studied its potential to modulate processes leading up to AD. Methods The total phenolic content and antioxidant capacity of the ethyl acetate extract of GBR were compared to that of brown rice (BR), and the cytotoxicity of both extracts were determined on human SH-SY5Y neuronal cells using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) Assay. Based on its higher antioxidant potentials, the effect of the GBR extract on morphological changes due to hydrogen peroxide (H2O2)-induced oxidative damage in human SH-SY5Y neuronal cells was examined using inverted light microscope and fluorescence microscope by means of acridine orange-propidium iodide (AO/PI) staining. Also, evaluation of the transcriptional regulation of antioxidant and apoptotic genes was carried out using Multiplex Gene Expression System. Results The ethyl acetate extract of GBR had higher total phenolic content and antioxidant capacity compared to BR. The cytotoxicity results showed that GBR extract did not cause any damage to the human SH-SY5Y neuronal cells at concentrations of up to 20 ppm, and the morphological analyses showed that the GBR extract (up to 10 ppm) prevented H2O2-induced apoptotic changes in the cells. Furthermore, multiplex gene expression analyses showed that the protection of the cells by the GBR extract was linked to its ability to induce transcriptional changes in antioxidant (SOD 1, SOD 2 and catalase) and apoptotic (AKT, NF-Kβ, ERK1/2, JNK, p53 and p38 MAPK) genes that tended towards survival. Conclusions Taken together, the results of our study showed that the ethyl acetate extract of GBR, with high antioxidant potentials, could prevent H2O2-induced oxidative damage in SH-SY5Y cells. The potential of GBR and its neuroprotective mechanism in ameliorating oxidative stress-related cytotoxicity is therefore worth exploring further

Toxicology study of vanillin on rats via oral and intra-peritoneal administration.

Vanillin is useful as anti-sickle cell anemia, anti-mutagen and anti-bacteria agent. However, vanillin must be administered at high concentration and cannot be oxidized by the upper gastrointestinal track of patients to be medically effective. In this study, we assessed the toxic effect of vanillin when administered in an un-oxidized form at high concentrations (150 and 300 mg/kg) via oral and intra-peritoneal injection. It was found that 300 mg/kg vanillin injection caused the rats to be unconscious without exerting any toxic effect on blood cells, kidney and liver. Besides, it showed blood protective property. Further analysis with GenomeLab GeXP genetic system on brain tissues showed that the expression of most xenobiotic metabolism, cell progression, tumor suppressor, DNA damage and inflammation genes were maintained at normal level. However, the expression of a few xenobiotic metabolism, cell cycle arrest and apoptosis genes were up-regulated by 5% ethanol injection. Nevertheless, when 5% ethanol was injected with the presence of vanillin, the expression was back to normal level. It is postulated that vanillin might have neuro-protective property. In conclusion, vanillin is not toxic at high concentration in both oral and intra-peritoneal injection and could provide blood and brain protective properties

Polyphenol-rich ethyl acetate fraction of Molineria latifolia rhizome restores oxidant-antioxidant balance by possible engagement of KEAP1-NRF2 and PKC/NF-κB signalling pathways

The present study examined the potential of ethyl acetate fraction (EAF) isolated from Molineria latifolia rhizome to modulate the oxidant-antioxidant balance in high fat diet and nicotinamide/streptozotocin-induced diabetic rats. Administration with EAF ameliorated systemic antioxidant status- and oxidative stress-related parameters without affecting the renal and liver functions in the diabetic rats. Further analyses on adipose, muscle and liver tissues demonstrated differing ability to scavenge free radicals and protection against lipid peroxidation. Transcriptional changes proposed concerted modulation of both KEAP1-NRF2 and PKC/NF-κB signallings in tissue-specific manner. Qualitative profiling of compounds present in EAF was analysed by non-targeted HPLC-QTOF mass spectrometry. A total of 23 unique mass signals were detected in EAF. Putative identification revealed a mixture of naturally occurring polyphenols ranging from cinnamic acid-, benzoic acid- and flavonoid-derived groups. Overall, the study demonstrated potential application of EAF to reinstate diabetes-induced oxidant-antioxidant imbalance by potentially modulating the NRF2–NF-κB signalling axis

Apoptosis and cell cycle arrest of human colorectal cancer cell line HT-29 induced by vanillin.

Background: Vanillin is responsible for the flavor and smell of vanilla, a widely used flavoring agent. Previous studies showed that vanillin could enhance the repair of mutations and thus function as an anti-mutagen. However, its role in cancer, a disease that is closely related to mutation has not yet been fully elucidated. Methods: Hence, this study investigated the cytolytic and cytostatic properties of vanillin against HT-29, a human colorectal cancer cell line. Methods used including cell viability assay, acridine orange (AO)–ethidium bromide (EB) double staining cell morphological analysis, Cell cycle analysis, annexin V–propidium iodide apoptosis test and 5-bromo-2-deoxyuridine (BrdU)-labeling cell proliferation assay. Results: Results showed that apoptosis was induced by vanillin and the IC50 for HT-29 and NIH/3T3 normal cell lines were 400 μg/ml and 1000 μg/ml, respectively. Different concentrations of vanillin arrest cell cycle at different checkpoints. 5-Bromo-2-deoxyuridine-labeling cell proliferation assay showed that G0/G1 arrest was achieved at lower concentration of vanillin (200 μg/ml) while cell cycle analysis by flow cytometer showed that G2/M arrest occurs at higher concentration of vanillin (1000 μg/ml). Conclusion: Cytolytic and cytostatic effects shown by vanillin showed that it could be a useful colorectal cancer preventive agent. Further in vivo study should be carried out to confirm that similar effects could happen in animals

Effects of germinated brown rice on the expression of alzheimer’s disease biomarkers in high fat/cholesterol diet-induced neurodegenerative changes

Excessive amyloid-β (Aβ) accumulation has been strongly implicated in Alzheimer’s disease (AD). Various experiments utilized diet-induced animal models of atherosclerosis/hypercholesterolemia and diabetes to investigate the convergent mechanisms between metabolic disorders and sporadic AD. While saturated fats and high cholesterol levels are associated with an increased risk of AD, consumption of certain dietary components may exert a protective role. Herewith, the effects of germinated brown rice (GBR) extract in a high fat/cholesterol diet (HFCD)-induced sporadic model of AD were investigated for 24 weeks in adult Sprague-Dawley rats. Rats were divided into 7 groups (n=70): Normal control, HFCD, HFCD with Donepezil (1.5 mg/kg BW), HFCD with Simvastatin (10 mg/kg BW), HFCD with Probucol (25 mg/kg BW), HFCD with GBR-100 (100 mg/kg BW), and HFCD with GBR-200 (200 mg/kg BW). Biochemical assays, mRNA levels of inflammatory-related genes and markers of neurodegeneration in the hippocampus and frontal cortex were analyzed. GBR regulated inflammatory genes such as CRP, PPAR-γ and TNF-α, and the expressions were significantly different from HFCD group. HFCD-fed rats also exhibited increased Aβ levels and altered expressions of proteins involved in Aβ generation, degradation and clearance. In terms of these biomarkers, GBR groups exhibited significant differences when compared to HFCD group, and the effects were comparable with the normal and some of the drug control groups. Taken together, the results suggested that GBR had the potential to attenuate HFCD diet-induced neurodegenerative changes, likely due to reduced brain inflammation in addition to modulation of Aβ processing and metabolism pathway

Rice bran phytic acid (IP6) induces growth inhibition, cell cycle arrest and apoptosis on human colorectal adenocarcinoma cells

Phytic acid (inositol hexaphosphate or IP6) is one of the bioactive compound that is present in cereals, nuts and legumes. IP6 is a naturally occurring polyphosphorylated carbohydrate, recognized to posses various significant health benefits including anticancer effects. Several in vitro and in vivo studies provide convincing evidence for the anticarcinogenic properties of commercial rice IP6 whilst the underlying mechanisms by which IP6 exerts anti-tumorigenic effects are still not fully known. The purpose of this present study is to investigate the growth inhibitory effects of IP6 extracted from rice bran on human colorectal cancer cell line (HT-29). IP6 extracted from rice bran induced marked growth inhibition in HT-29 with an IC50 value of 12.0 ± 2 μg/ml, in a dose and time dependent manner. Flow cytometry was performed for the analysis of cell cycle and apoptosis. Rice bran IP6-extract induced cell cycle arrest in HT-29 cell at G0/G1 phase. Staining with Annexin V-based assay and propidium iodide confirmed that apoptosis occurred early and late in the HT-29. IP6 is expected to exert anticarcinogenic activity through disruption of cell cycle progression and induction of apoptosis. Our study further supports the function of rice bran IP6 as a chemopreventive agent for human colorectal cancer

Probiotic properties of Bacillus strains isolated from stingless bee (Heterotrigona itama) honey collected across Malaysia

This study aimed to isolate, identify, and evaluate the probiotic properties of Bacillus species from honey of the stingless bee Heterotrigona itama. Bacillus spp. were isolated from five different H. itama meliponicultures, and the isolates were characterized through Gram-staining and a catalase test. Tolerance to acidic conditions and bile salt (0.3%), hydrophobicity, and autoaggregation tests were performed to assess the probiotic properties of the selected isolates, B. amyloliquefaciens HTI-19 and B. subtilis HTI-23. Both Bacillus isolates exhibited excellent antimicrobial activity against both Gram-positive and Gram-negative bacteria and possessed significantly high survival rates in 0.3% bile solution for 3 h. Their survival rates in acidic conditions were also comparable to a commercial probiotic strain, Lactobacillus rhamnosus GG. Interestingly, the hydrophobicity and autoaggregation percentage showed no significant difference from L. rhamnosus GG, a commercial probiotic strain. The results from this study suggest that B. amyloliquefaciens HTI-19 and B. subtilis HTI-23 isolated from stingless bee honey have considerably good probiotic properties. Therefore, more studies should be done to investigate the effects of these bacteria cultures on gastrointestinal health