Antiproliferative mechanism of cervical cancer cells (hela) treated with biological-active subfraction from quercus infectoria extract

Cervical cancer is placed at the fourth most frequent cancer among women worldwide, which was estimated approximately 530, 000 new cases in 2012. This type of cancer has become one of the leading causes of cancer death among women worldwide. Conventional and modern cancer treatment nowadays comes with negative and adverse side effects to the patients. Therefore, lot of studies have been conducted to search for new alternative treatment which is more safe and effective such as utilisation of natural product based medications. Previous studied revealed various pharmacological activities of Quercus Infectoria (QI) galls or manjakani including anticancer activity. However, the mechanism of action lay behind was not well explained. Therefore, in this present study, QI gall was selected for the evaluation of cytotoxic activity and cell death mode of action. In this study, QI was extracted using different types of organic solvents before being tested for cytotoxicity activity and mode of cell death. MTT assay was used to determine cytotoxic activity for n-hexane (QJH), ethyl acetate (QIEA) and methanol (QIM) of QI galls extracts against cervical cancer (Hela). Cytotoxic activity of QI extracts also tested against normal fibroblast (L929) cell line to determine cytotoxic effects and cytoselective properties. Meanwhile, DMSO-treated cells served as negative control while cisplatin-treated cells served as positive control. After that, dose-response graph was plotted to determine IC50 values. Observation on nuclear morphology, apoptotic percentage and detection of apoptotic protein expressions were also conducted. Moreover, phytochemical screening analysis had been carried out to determine bioactive groups that present in the most potent extract. From the findings, QIEA extract exhibited better cytotoxic activity with best growth inhibition against Hela cells (IC50 value= 6.33 ± 0.33 µg/ml) compared to other extracts and showed cytoselective property as no cytoxicity observed in the treated normal fibroblast (L929) cells. As for the phytochemical analysis of QIEA extract, it was revealed the presence of tannin, alkaloids, glycosides, saponins, terpenoids, flavonoids and phenolic compounds The QIEA extract was then purified and resulted two compounds 4-0 methylgallic acid (MGA) and gallic acid (GA). The most potent antiproliferative activity was exhibited by MGA sub fraction with IC50 II ±0.58 Jlg/ml. The current study also revealed that Hela cells treated with MGA subfraction has undergone apoptosis as exerted by the alteration of nuclear morphology and the presence of apoptotic bodies as well as the increment of apoptosis rate in the treated cells. Furthermore, MGA subfraction triggered apoptosis through upregulation of tumor suppression protein p53 , with down regulation of Bcl-2 expression and facilitated the apoptosis execution through caspase-3 activation . In conclusion, the purification of QIEA has resulted two biologicaly active subfraction namely MGA and GA. It was cleared that MGA subfraction reduced Hela cell growth through induction of apoptosis

Re-evaluation of malnutrition risk screening tool-hospital (MRST-H) for geriatric patients: a multicentre study in Peninsular Malaysia

A local Malnutrition Risk Screening Tool-Hospital (MRST-H) has been developed to identify the risk of malnutrition among hospitalized geriatric patients in Malaysia. The aims of this multicenter study were to evaluate the criterion validity of the MRST-H against the reference standard Subjective Global Assessment (SGA) and revise its scoring criteria among Malaysian geriatric patients. A cross-sectional study was conducted among 542 geriatric patients at eight general hospitals in Peninsular Malaysia from January 2011 to February 2013. The Malay version MRST-H and SGA were administered to all participants through face-to-face interviews. Sensitivity and specificity of MRST-H were established using the Receiver Operating Characteristic (ROC) curves and the optimal cut-off scores were determined. The MRST-H had area under the ROC curve (AUC) values of 0.84 and 0.88 when validated against the SGA-determined malnutrition (SGA B+C) and severe malnutrition (SGA C) status. These high AUC values indicated that the MRST-H has very good overall diagnostic accuracy. However, the original cut-off score of five points for MRST-H has undesirable sensitivity in identifying the malnutrition (sensitivity = 0.12) and severely malnutrition (sensitivity = 0.35) status. The optimal cut-off score of MRST-H in identifying malnourished and severely malnourished participants were both established at the cut-off score of two points. The sensitivity of MRST-H increased substantially at this point without compromising its specificity. Therefore, the established cut-off score of two points with optimal sensitivity and specificity was selected to replace to original cut-off score for screening of risk of malnutrition among hospitalized geriatric patients

A natural compound from Hydnophytum formicarium induces apoptosis of MCF-7 cells via up-regulation of Bax

<p>Abstract</p> <p>Background</p> <p><it>Hydnophytum formicarium </it>Jack is an epyphytic shrub that belongs to the family of Rubiaceae and is native to the tropical rain forests of the Asean region, which includes Malaysia. A flavanoid derivative, 7, 3', 5'-trihydroxyflavanone (3HFD), isolated from <it>H. formicarium </it>has been reported to have cytotoxic effects on the human breast carcinoma cell line MCF-7. The aim of the current study was to investigate the mode of cell death in MCF-7 cells treated with 3HFD. A DNA fragmentation assay was conducted on isolated genomic DNA, a TUNEL assay was used to determine the mode of cell death and Western blotting was used to evaluate the expression levels of Bax and Bcl-2. Immunofluorescence staining of MCF-7 cells was also performed to confirm the up-regulation of the Bax protein.</p> <p>Results</p> <p>The ladder pattern resulting from the DNA fragmentation assay was a multimer of 180 kb. The morphological changes of cells undergoing apoptosis were visualised by a TUNEL assay over time. The percentage of apoptotic cells increased as early as 6 hours post treatment compared to untreated cells. Western blotting revealed up-regulation of the pro-apoptotic protein Bax. However, 3HFD did not affect expression of the anti-apoptotic protein Bcl-2.</p> <p>Conclusions</p> <p>Our results provide evidence that plant-derived 3HFD was able to induce the apoptotic cell death of MCF-7 cells by increasing Bax expression level and makes 3HFD a promising agent for chemotherapy, which merits further study.</p

The Potential Role of Quercus Infectoria Gall Extract on Osteoblast Function and Bone Metabolism

The galls of the Quercus infectoria (QI) tree are traditionally believed to have great medicinal value. Pharmacologically the galls are claimed to have various biological activities such as astringent effect, antidiabetic, antitremorine, local anaesthetic, antipyretic, anti-inflammatory, antibacterial, antiviral and many more. These pharmacological activities of gall extracts were reported to be due to its excellent antioxidant activity with phytochemicals constituents of phenolic and flavanoid compounds. The phenolic compounds or polyphenols can act on bone metabolism by modulating os-teoblast proliferation, differentiation and mineralization, as well as osteoclastogenesis. In addition, elemental and phys-ico-chemical analysis indicated the presence of important minerals in QI, such as calcium, magnesium, phosphorus, oxygen, potassium, aluminium, carbon, zinc, iron, manganese, nickel and silica. The current review will be focusing on the potential bone health benefits of the well-known traditional herbal medicine, QI or locally known as the “manja-kani”

Thyroglossal duct carcinoma

We report a 56-year-old Malay woman with a tumour that involved the skin and caused hyoid bone erosion. There was no clinical or radiological evidence of regional lymph node involvement. A modified radical neck dissection with preservation of the accessory nerve and internal jugular vein was performed, followed by an "extended" Sistrunk operation. The surgical defect was reconstructed with a pectoralis major myocutaneous flap. Our literature review showed that this is the first reported thyroglossal duct carcinoma which involved the skin and required a pedicle flap reconstruction

Human natural killer (NK) cell activation by luteolin from Brucea javanica leaves

There is growing interest in using natural substances from plants to potentially enhance anticancer activity. A key target in this respect is natural killer (NK) cells in order to generate an anticancerimmune response. Luteolin used in this study was derived from a local plant Brucea javanica leaves extract; through a bioassay-guided fractionation. The expression of interleukin-2 (IL-2) and interferon-γ (IFN-γ) was increased after 20 h treatment of 0.1 µM luteolin in human mixed lymphocytes, which reflects the activation of NK cells. This was further confirmed in co-culture experiments. NK cellinduced K562 target cell death was increased in the presence of luteolin. These results show that luteolin activates NK cells to kill target cells indicating the potential use of luteolin as an anticancer immunostimulant

Cytotoxicity of Clinacanthus nutans and mechanism of action of its active fraction towards human cervical cancer cell line, HeLA

Traditionally, Clinacanthus nutans (CN) or locally named as ‘Belalai Gajah’ is one of the herbal plant claimed to be able to treat cancer. The aimd of this study are to extract, isolate and characterize the active anticancer compound from CN and to determine the mode of cell death induced by the compound. Bioassay guided fractionation was done on the CN extract by using column chromatography. The cytotoxicity activities of these fractions toward HeLA cells were examined by MTT assay. The nuclear morphology was examined by Hoechst 33258 staining and the cell cycle arrest was evaluated by propium iodide staining using flow cytometry. The presence of active compound in the chosen fraction was determined by Liquid Chromatography Mass Spectrometry (LCMS). Out of 16 fractions collected, Fraction 11(F11) showed the lowest IC50 value with 27 ± 2.6 μg/mL. The value of IC50 for F11 towards normal cell, NIH 3T3 cell and L929 cell, were 70 ± 4.0 μg/mL and 45 ± 1.5 μg/mL respectively. These values were higher than tamoxifen, therefore indicating that tamoxifen is more toxic towards normal cells compared to F11. Nuclear morphology of HeLA cell displayed DNA fragmentation, nuclear condensation and formation of apoptotic bodies upon treatment with F11 for 24 hours. The cell cycle distribution of HeLA cell treated with F11 was arrested at G1 phase. The active compound identified to potentially possess the anticancer property is 19-Oxo-all-trans-retinoic acid. In conclusion, 19-Oxo-all-trans-retinoic acids from F11 of the CN extract, is a potential anticancer agent for cervical cancer

Gallic acid and methyl gallate enhance antiproliferative effect of cisplatin on cervical cancer (HeLa) cells

Cervical cancer is the fourth most common cancer-related death affecting women. The drug resistance, toxicities and undesired side effects become the major limitation in cisplatin-based chemotherapy. Gallic acid and methyl gallate are the most abundance phenolic compounds that are widely distributed in plants. This study was conducted to evaluate the antioxidant and antiproliferative activity of gallic acid and methyl gallate and their synergistic effects in combination with cisplatin towards cervical cancer (HeLa) cells. The antioxidant activity of gallic acid and methyl gallate was measured by using 1, 1-diphenyl-2-picrylhydrazyl radical (DPPH) scavenging assay. Antiproliferative activity of gallic acid, methyl gallate and cisplatin on HeLa and NIH/ 3T3 cells was determined using MTT assay. The effect of gallic acid and methyl gallate combined with cisplatin were then determined by CompuSyn software. Gallic acid and methyl gallate showed strong antioxidant activity with EC50 value of 18.23 µM and 19.39 µM, respectively. The IC50 of gallic acid, methyl gallate and cisplatin on HeLa cells were 13.44 µg/mL, 16.55 µg/mL, and 8.04 µg/mL whereas in NIH/3T3 cells were 32.90 µg/mL, 35.70 µg/mL, and 6.57 µg/mL. Cisplatin combined with fixed concentration of gallic acid and methyl gallate could inhibit the proliferation of HeLa cells greater than cisplatin alone. Interestingly, gallic acid and methyl gallate in combination with cisplatin at the concentration of 0.51-4.02 µg/mL have shown synergistic effects. Therefore, our study suggested that gallic acid and methyl gallate in combination with cisplatin have the potential to be developed as chemotherapeutic agents for cervical cancer

Design of smart waste bin and prediction algorithm for waste management in household area

Maintaining current municipal solid waste management (MSWM) for the next ten years would not be efficient anymore as it has brought many environmental issues such as air pollution. This project has proposed Artificial Neural Network (ANN) based prediction algorithm that can forecast Solid Waste Generation (SWG) based on household size factor. Kulliyyah of Engineering (KOE) in International Islamic University Malaysia (IIUM) has been chosen as the sample size for household size factor. A smart waste bin has been developed that can measure the weight, detect the emptiness level of the waste bin, stores information and have direct communication between waste bin and collector crews. This study uses the information obtained from the smart waste bin for the waste weight while the sample size of KOE has been obtained through KOE’s department. All data will be normalized in the pre-processing stage before proceeding to the prediction using Visual Gene Developer. This project evaluated the performance using R2 value. Two hidden layers with five and ten nodes were used respectively. The result portrayed that the average rate of increment of waste weight is 2.05 percent from week one until week twenty. The limitation to this study is that the amount of smart waste bin should be replicated more so that all data for waste weight is directly collected from the smart waste bin

Aqueous polyphenol semi-purified fractions from galls of Quercus infectoria increase ALP activity and mineralisation of hFOB 1.19 cells

The galls of Quercus infectoria (QI) have been reported to possess numerous medicinal values and give a positive effect on bone metabolism. This study investigated the effects of semi-purified fractions of QI gall extract on the proliferation, alkaline phosphatase (ALP) activity, and mineralisation of human foetal osteoblast cell line (hFOB 1.19). The semi-purified fractions (fraction A and B) were prepared by column chromatography methods. MTT assay was used to measure cell proliferation activity to obtain half maximal concentration (EC50) of the cells treated with fraction A (phenolic components and contained amide), fraction B (phenolic components with the presence of alkene), and pamidronate (drug control). The most potent or lowest EC50 was further used to measure ALP activity in the treated and untreated cells at day 1, 3, 7, 10, and 14 by ELISA. Cell mineralisation was determined by von Kossa staining for phosphate depositions and Alizarin Red S staining for calcium depositions. The EC50 values for fraction A and B were 8.86 and 9.92 μg/mL, respectively, which showed a greater effect compared to the pamidronate (15.27 μg/mL). The ALP activity of both fractions in the treated cells were also greater compared to the two control groups (cells treated with pamidronate and untreated cells), starting from day 3 onwards. The calcium depositions appeared as red spots, while phosphate depositions appeared as black spots. Interestingly, the calcium depositions of cells treated with both fractions were higher than those of the two control groups. In conclusion, semi-purified fractions of QI gall extract enhanced proliferation, improved mineralisation, and increased ALP activity of hFOB 1.19 cells