Effects of Phytic Acid Extracted From Rice Bran on Azoxymethane-Induced Colon Carcinogenesis in Rats

This research is carried out to study the potential of phytic acid extracted from rice bran in the suppression of colon carcinogenesis in rats. In the optimization of phytic acid extraction, results showed 5% H2S04 in pH 0.6 and 30 minutes of extraction time gave the highest amount of phytic acid. In animal study, 72 male Sprague-Dawley rats were divided into 6 groups with 12 rats in each group; Group 1: ADM alone, Group 2: AOM + 0.2% (w/v) Commercial Phytic Acid (CPA), G roup 3: ADM + 0.5% (w/v) Commercial Phytic Acid (CPA), Group 4: ADM + 0.2% (w/v) Extract P hytic Acid (EPA) , G roup 5: AOM + 0.5% (w/v) Extract Phytic Acid (EPA). Rats received two subcutaneous injections of azoxymethane (ADM) in saline at (15mg/kg bodyweight)" over a 2-weeks period to induce colon cancer. The treatments were given in two different concentrations of phytic acid; 0.2% (w/v) and 0.5% (w/v) during post initiation of carcinogenesis phase via drinking water. The colons of the animals were analyzed for detection and quantification of aberrant crypt foci (ACF) after 8 weeks of treatment. The finding showed treatment with 0.2% (w/v) EPA gave the greatest reduction in the formation of ACF. In addition, phytic acid significantly suppressed the number of ACF in the distal, middle and proximal colon as compared to AOM a lone (p<0.05). For the histological classification of ACF, treatment with 0.5% (w/v) CPA had the highest percentage (71%) of non-dysplastic ACF followed by treatment with 0.2% (w/v) EPA (61%). After 20 weeks of treatment, colons of the rats were excised and analyzed for tumor incidence. Results showed that administration of phytic acid reduced the incidence and multiplicity of total tumors and adenocarcinomas even though there were no significant differences between groups. For the immunohistochemical analyses, proliferating cell using Ki-67 and modulating of B-catenin and COX-2 expression were assessed as those have been shown to play a role in tumor progression . In Ki-67, there was a statistically significance difference in lowering the proliferating index between treatment groups as compared to AOM alone (p<0.05). For B-catenin a nd COX-2 expression, there was a significant difference between g roups as (p=O.OOO) and (p=0.030). In the correlation test, the results showed that there was a significant positive correlation (p=0.010) between proliferation of Ki67 and COX-2 expression. A positive linear relationship was found between total Ki67 and Bcatenin but these relationships were not statistically significant. Total B-catenin had a significant positive linear relationship with total COX-2 (p=0.044). As a conclusion, this study found the potential value of phytic acid extracted from rice bran in reducing colon cancer risk in rats. Besides identification of cancer reduction strategies based on dietary modification including looking at natural sources that may have anticancer properties, an alternative compound from local sources has been developed. Therefore, rice bran that is normally discarded as by-product of rice production will increase in value due to phytic acid potential as a nutraceutical compound in the prevention of colon cancer progression

The application of adat perpatih with regards to customary land ownership in Negeri Sembilan / Nora Ardilla Nordin, Norazalina Razali and Norzaliza Tesmin

Adat Perpatih acts as a mirror which reflects the connection between the system of Adat and the Minangkabau community in Negeri Sembilan. Adat Perpatih was considered as a unique system of social since it contained the rules and regulations which act as a yardstick for the justification and legitimacy of certain conducts of society. Kuala Pilah and Rembau are the two districts in Negeri Sembilan that are still applying and practising Adat Perpatih. As mentioned earlier, this study looks into the application of Adat Perpatih in the land administration and ownership, and also to determine whether Adat Perpatih's land ysytem is still effective to cater the needs of modern society in Kuala Pilah and Rembau. Sometimes, the problems that arise with regards to Adat Perpatih actually come from the attitude of the society themselves who had failed to implement the rules of Adat Perpatih within their daily lives and dealings such as transfer, lease and charge. In fact, many of them have turned to a conventional system of dealings and administration, especially in land matters since customary rules was seen as obsolete and not suitable in our modern world. But, with the coming into force of Negeri Sembilan Land Enactment (Cap 215), customary land is put under the exclusive jurisdiction of the Negeri Sembilan State Council and also the Adat Chiefs such as Undang and Lembaga

Suppression of β-catenin and cyclooxygenase-2 expression and cell proliferation in azoxymethane-induced colonic cancer in rats by rice bran phytic acid (PA)

Background: Phytic acid (PA) is a polyphosphorylated carbohydrate that can be found in high amounts in mostcereals, legumes, nut oil, seeds and soy beans. It has been suggested to play a significant role in inhibition of colorectal cancer. This study was conducted to investigate expression changes of β-catenin and cyclooxygenase-2 COX-2) and cell proliferation in the adenoma-carcinoma sequence after treatment with rice bran PA by immunocytochemistry. Materials and Methods: Seventy-two male Sprague-Dawley rats were divided into 6 equal groups with 12rats in each group. For cancer induction two intraperitoneal injections of azoxymethane AOM) were given at 15 mg/kg bodyweight over a 2-weeks period. During the post initiation phase, two different concentrations of PA, 0.2%(w/v) and 0.5% (w/v) were administered in the diet. Results: Results of ß-catenin, COX-2 expressions and cell proliferation of Ki-67 showed a significant contribution in colonic cancer progression. For ß-catenin and COX-2 expression, there was a significant difference between groups at p<0.05. With Ki-67, there was a statistically significant lowering the proliferating index as compared to AOM alone (p<0.05). A significant positive correlation (p=0.01) was noted between COX-2 expression and proliferation. Total ß-catenin also demonstrated a significant positive linear relationship with total COX-2 (p=0.044). Conclusions: This study indicated potential value of PA extracted from om rice bran in reducing colonic cancer risk in rats

Dissecting the mechanism of growth inhibition and tumour reversion mediated by oocyte extract in human breast cancer cells

Axolotl oocyte extract (AOE) has the ability to reprogram the breast cancer cell epigenome by modifying DNA methylation and histone modifications at the promoter region of silenced tumour suppressor genes, resulting in loss of cellular tumourigenicity (Allegrucci et al., 2011). The present study aimed to determine the signalling pathways and associated chromatin modifications involved in tumour reversion mediated by AOE in an attempt to identify specific oocyte molecules responsible for the reprogramming of breast cancer cells. Microarray analysis of reprogrammed tumours revealed a cell cycle block that was confirmed by BrdU labelling of xenograft tumours. The phenotype was recapitulated in vitro cultures up to 4 days after reprogramming, with AOE-treated cells showing a significant decreased entry in S phase of the cell cycle. Reduced expression of JUN and up-regulation of CDKN1B (p27) were measured, together with a reduction of phosphorylation of p44/42 MAPK signalling pathway. CDKN1B protein also showed a nuclear localisation in reprogrammed tumour xenografts compared to untreated controls. The reprogramming of breast cancer cells at genome-wide level was associated with a decrease in H3K9me2, H3K9me3 and H4K20me1 and in increase in H3K4me3, H3K27me3, H3K9Ac, H4K20me2 and H4K20me3 immediately after reprogramming (6 hours) and the pattern of changes was generally maintained at the time point of injection into xenografts (12 hours after reprogramming). Specific histone modifiers inhibitors were used during AOE treatment to dissect histone modification activities during cells reprogramming. Altogether, the results obtained suggest that axolotl oocytes contain complex activities that may cooperate to remodel the epigenetic and signalling landscape of breast cancer cells leading to reversion of malignancy by inducing cell cycle arrest and a state of quiescence. Therefore, this work has the potential to develop new therapeutic approaches based on oocyte molecules that could be tested in different types of breast cancer, especially those with poor prognosis and patient outcome

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

Enhancement of phenolics and antioxidant activity via heat assisted extraction from Moringa oleifera using response surface methodology and its potential bioactive constituents

Introduction: Moringa oleifera Lam. is a miracle tree that has been widely utilised in folklore medicine due to its immense amount of phenolic constituents that could treat various ailments. Different techniques have been implemented to extract the phenolic but the parameters may not be optimised to further enhance the amount of phenolic extracted. Thus, the work aimed to enhance phenolic content and antioxidant activity of M. oleifera through RSM methodology, which is rapid and convenience. Methods: At first, antioxidant activity of different parts of M. oleifera (leaves, stem, pod and seed) were investigated. The plant part with the highest antioxidant activity was selected for the optimisation of extraction condition using RSM. In RSM, temperature (XA), extraction time (XB) and solid-liquid ratio (XC) were employed to study the effects on yield, total phenolics, flavonoids and antioxidant activity. Then, the optimum extraction condition obtained via RSM was utilised in LC-MS and HPLC analysis to determine the potential bioactive constituents. Results: The leaves of M. oleifera displayed the highest antioxidant activity as compared to other plant parts. The optimum extraction condition obtained for the leaves extract was: temperature (XA): 82°C, extraction time (XB): 48 min and solid-liquid ratio (XC): 1:30 g/mL (w/v). Meanwhile, LC-MS revealed the presence of gallic acid, chlorogenic acid, quercetin, kaempferol and 3-O-glucoside kaempferol. HPLC analysis detected six compounds; gallic acid, epicatechin gallate, chlorogenic acid, myricetin, quercetin and kaempferol. Conclusion: The optimisation are promising to improve yield and antioxidant activity in M. oleifera as compared to non-conventional extractions

Antioxidant properties and antiproliferative effect of brewers' rice extract (temukut) on selected cancer cell lines

Temukut, or brewers’ rice, is a mixture of broken rice, rice bran, and rice germ. Extensive studies have been conducted on rice bran, which possesses various health benefits. Temukut, however has been less well studied. The present study aimed to investigate the antioxidant and growth inhibition properties of temukut extract using colon cancer (HT-29), ovary cancer (Caov-3), and liver cancer (HepG2) cell lines. The antioxidant activity was determined by the β-carotene bleaching assay, analysis of the DPPH radical scavenging capacity, and a FRAP assay. The total phenolic compounds, oryzanol, vitamin E, and phytic acid levels in temukut were also investigated. The antiproliferative activity was evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. There was a significant difference in the cytotoxicity of two types of temukut extract (water and methanol) for HT-29 and Caov-3 cells(p < 0.05) but not for HepG2 cells. The HepG2 cell line is the least sensitive to temukut, (IC 50 = 55.30 μg/mL), whereas the highest sensitivity was observed in Caov-3 cells (IC 50=36.67 μg/ mL). No cytotoxic effect of temukut was observed on normal cells (BalBlc3T3). Although the content of the phytochemicals studied (total phenolic compounds, vitamin E, oryzanol, and phytic acid) in temukut was lower than that in rice bran, as has been previously reported, the present study demonstrated temukut’s potential to inhibit the proliferation of HT-29, Caov-3, and HepG2 cells

Optimized aqueous extraction conditions for maximal phenolics, flavonoids and antioxidant capacity from Artocarpus heterophyllus (jackfruit) leaves by response surface methodology (RSM)

Introduction: There are numerous studies on the therapeutic properties of Artocarpus heterophyllus. However, studies on the aqueous extraction of A. heterophyllus leaves are limited. This present study was conducted to optimize the extraction conditions of A. heterophyllus leaves to yield the highest phenolic, flavonoids and antioxidant contents. Methods: Response surface methodology (RSM) was employed to obtain a higher phenolic extraction parameter(s) of A. heterophyllus leaves using Central Composite Design (CCD). The antioxidant activity was then determined via ABTS (2,29-azinobis (3 ethylbenzothiazoline-6-sulfonic acid)) and DPPH (2,2-Diphenyl-1-picrylhydrazyl) assay and analysis of the individual phenolics was performed by high performance liquid chromatography (HPLC). Results: The optimum extraction conditions with higher phenolics content and antioxidant activity was achieved at 81°C, 100 min and 40 mL/g sample with a good desirability value of 0.87. Under these optimized parameters, total phenolics and flavonoids were 174.48 ± 4.05 mg GAE/g sample and 21.44 ± 0.05 mg RE/g sample, respectively. Meanwhile, antioxidant activity via ABTS and DPPH assays were 90.88% ± 0.09 and 87.22% ± 0.62, respectively. Finally, under optimal extraction conditions revealed 4 compounds identified as chlorogenic acid, quercetin, rutin and kaempferol. Conclusion: The optimisation are promising to improve phenolic yield and antioxidant activity in A. heterophyllusleaves. It also proved that A. heterophyllus leaves can be used as an alternative natural antioxidant especially in medicinal applications since all identified compound possess significant biological activities for human health

Germinated brown rice (GBR) reduces the incidence of aberrant crypt foci with the involvement of β-catenin and COX-2 in azoxymethane-induced colon cancer in rats

Chemoprevention has become an important area in cancer research due to the failure of current therapeutic modalities. Epidemiological and preclinical studies have demonstrated that nutrition plays a vital role in the etiology of cancer. This study was conducted to determine the chemopreventive effects of germinated brown rice (GBR) in rats induced with colon cancer. GBR is brown rice that has been claimed to be richer in nutrients compared to the common white rice. The male Sprague Dawley rats (6 weeks of age) were randomly divided into 5 groups: (G1) positive control (with colon cancer, unfed with GBR), (G2) fed with 2.5 g/kg of GBR (GBR (g)/weight of rat (kg)), (G3) fed with 5 g/kg of GBR, (G4) fed with 10 g/kg of GBR and (G5) negative control (without colon cancer, unfed with GBR). GBR was administered orally once daily via gavage after injection of 15 mg/kg of body weight of azoxymethane (AOM) once a week for two weeks, intraperitonially. After 8 weeks of treatment, animals were sacrificed and colons were removed. Colonic aberrant crypt foci (ACF) were evaluated histopathologically. Total number of ACF and AC, and multicrypt of ACF, and the expression of β-catenin and COX-2 reduced significantly (p < 0.05) in all the groups treated with GBR (G2, G3 and G4) compared to the control group (G1). Spearman rank correlation test showed significant positive linear relationship between total β-catenin and COX-2 score (Spearman's rho = 0.616, p = 0.0001). It is demonstrated that GBR inhibits the development of total number of ACF and AC, and multicrypt of ACF, reduces the expression of β-catenin and COX-2, and thus can be a promising dietary supplement in prevention of colon cancer

Detection of calreticulin as a candidate prognostic biomarker in invasive breast carcinoma from a biological scaffold-based 3D co-culture system

Introduction: Breast cancer is the most common cancer in women and the world’s second leading cause of death in women, after lung cancer. Calreticulin (CRT), an endoplasmic reticulum (ER) multipurpose protein, has been proposed as a potential biomarker for breast cancer. However, reports on the correlation between CRT expression and cell invasiveness in breast cancer micro-tissues are scarce. Thus, in the current study, we analyzed the potential correlation between CRT and invasiveness of breast cancer in a biological scaffold-based 3D co-culture system. Methods: MCF7, MDA-MB-231 and MCF-10A breast cell lines were co-cultured in a 3-dimensional (3D) system with MRC-5 lung fibroblast cell line in the cell density ratio of 3:1. Thereafter, calreticulin gene and protein expression levels were determined based on quantitative reverse transcription-polymerase chain reaction (RT-qPCR) and immunohistochemistry, respectively. Moreover, via RT-qPCR analysis, the gene expression levels of calreticulin-related candidate metastasis genes in breast cancer micro-tissues were carried out. Results: The results showed occasional foci of lumen-like morphology in the non-cancerous breast micro-tissues and the formation of solid clusters for breast cancer micro-tissues. Moreover, immunohistochemistry results revealed protein expression of calreticulin in non-cancerous and cancerous breast micro-tissues with cytoplasmic and nucleic acid localizations. It was found that PCMT1 and ER-α genes were significantly downregulated (p < 0.01) in invasive breast cancer micro-tissues. Conclusion: This study suggests that CRT and CRT-related candidate metastasis genes may potentially serve as prognostic biomarkers in invasive breast carcinoma