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Fulltext Comparison of ASE with in-cell cleanup and the QuEChERS sample preparation methods for the analysis of pesticide residues in tea

Abdul Kadir, H., Abas, F., Mediani, A., Ismail, I.S., Lajis, N.H.

International Food Research Journal, 2017;24(1):26-267.
MyJurnal

The aim of the present work was to compare and choose the best method to extract incurred
pesticide residues from green tea. Accelerated solvent extraction (ASE) with in-cell cleanup
and the quick, easy, cheap, effective rugged and safe (QuEChERS) methods were tested on
green tea samples with incurred beta-endosulfan pesticide. The extracts were analyzed by
GC-MS/MS and the recovery and the precision of both methods were compared. The average
recovery using ASE with the in-cell cleanup method was in the range of 89 to 92% which is
better than that obtained using a QuEChERS method. Both the ASE with in-cell cleanup and
the QuEChERS methods provided good precision with RSDs in the range of 12 to 15% and
17 to 18%, respectively. This finding indicates that the ASE method with the in-cell cleanup is
more suitable for the accurate determination of pesticides incurred in tea.
Bromophenol blue binding to mammalian albumins and displacement of albumin-bound bilirubin

Kim BB, Abdul Kadir H, Tayyab S

Pak J Biol Sci, 2008 Oct 15;11(20):2418-22.
PMID: 19137852

Interaction of bromophenol blue (BPB) with serum albumins from different mammalian species, namely, human (HSA), bovine (BSA), goat (GSA), sheep (SSA), rabbit (RbSA), porcine (PSA) and dog (DSA) was studied using absorption and absorption difference spectroscopy. BPB-albumin complexes showed significant differences in the spectral characteristics, i.e., extent of bathochromic shift and hypochromism relative to the spectral features of free BPB. Absorption difference spectra of these complexes also showed variations in the position of maxima and absorption difference (deltaAbs.) values. Absorption difference spectra of different bilirubin (BR)-albumin complexes showed a significant blue shift accompanied by decrease in deltaAbs. values in presence of BPB which were indicative of the displacement of bound BR from its binding site in BR-albumin complexes. These changes in the difference spectral characteristics of BR-albumin complexes were more marked at higher BPB concentration. However, the extent of these changes was different for different BR-albumin complexes. Taken together, all these results suggest that BPB partially shares BR binding site on albumin and different mammalian albumins show differences in the microenvironment of the BR/BPB binding site.
Fulltext Induction of Mitochondria-Mediated Apoptosis in Ca Ski Human Cervical Cancer Cells Triggered by Mollic Acid Arabinoside Isolated from Leea indica

Wong YH, Abdul Kadir H

Evid Based Complement Alternat Med, 2012;2012:684740.
PMID: 22203877 DOI: 10.1155/2012/684740

Leea indica is a medicinal plant traditionally used to treat cancer. Through bioassay-guided approach, we isolated mollic acid arabinoside (MAA), for the first time from Leea indica. Here, we present the apoptosis-inducing effect of MAA on Ca Ski cervical cancer cells. Based on DAPI staining, MAA-treated cells manifested nuclear shrinkage, condensation, and fragmentation. We further confirmed the fragmentation of DNA using TUNEL assay. During early apoptosis, MAA caused the perturbation of plasma membrane through externalization of PS, followed by the formation of apoptotic blebs. Prior to these events, MAA triggered rapid dissipation of the mitochondrial membrane potential. In the upstream, MAA increased the expression of Bax, decreased the expression of Bcl-2, and augmented the Bax/Bcl-2 ratio. These findings suggested that MAA induced mitochondrial-mediated apoptosis in Ca Ski cells and thus provide the scientific explanation for the traditional application of this herbal medicine in cancer treatment.
Fulltext Leea indica Ethyl Acetate Fraction Induces Growth-Inhibitory Effect in Various Cancer Cell Lines and Apoptosis in Ca Ski Human Cervical Epidermoid Carcinoma Cells

Yau Hsiung W, Abdul Kadir H

Evid Based Complement Alternat Med, 2011;2011:293060.
PMID: 21423690 DOI: 10.1155/2011/293060

The anticancer potential of Leea indica, a Chinese medicinal plant was investigated for the first time. The crude ethanol extract and fractions (ethyl acetate, hexane, and water) of Leea indica were evaluated their cytotoxicity on various cell lines (Ca Ski, MCF 7, MDA-MB-435, KB, HEP G2, WRL 68, and Vero) by MTT assay. Leea indica ethyl acetate fraction (LIEAF) was found showing the greatest cytotoxic effect against Ca Ski cervical cancer cells. Typical apoptotic morphological changes such as DNA fragmentation and chromatin condensation were observed in LIEAF-treated cells. Early signs of apoptosis such as externalization of phosphatidylserine and disruption of mitochondrial membrane potential indicated apoptosis induction. This was further substantiated by dose- and time-dependent accumulation of sub-G(1) cells, depletion of intracellular glutathione, and activation of caspase-3. In conclusion, these results suggested that LIEAF inhibited cervical cancer cells growth by inducing apoptosis and could be developed as potential anticancer drugs.
Fulltext Honey-induced protein stabilization as studied by fluorescein isothiocyanate fluorescence

Wong YH, Abdul Kadir H, Tayyab S

ScientificWorldJournal, 2013;2013:981902.
PMID: 24222758 DOI: 10.1155/2013/981902

Protein stabilizing potential of honey was studied on a model protein, bovine serum albumin (BSA), using extrinsic fluorescence of fluorescein isothiocyanate (FITC) as the probe. BSA was labelled with FITC using chemical coupling, and urea and thermal denaturation studies were performed on FITC-labelled BSA (FITC-BSA) both in the absence and presence of 10% and 20% (w/v) honey using FITC fluorescence at 522 nm upon excitation at 495 nm. There was an increase in the FITC fluorescence intensity upon increasing urea concentration or temperature, suggesting protein denaturation. The results from urea and thermal denaturation studies showed increased stability of protein in the presence of honey as reflected from the shift in the transition curve along with the start point and the midpoint of the transition towards higher urea concentration/temperature. Furthermore, the increase in ΔG D (H2O) and ΔG D (25°C) in presence of honey also suggested protein stabilization.
Fulltext Stabilizing effect of various polyols on the native and the denatured states of glucoamylase

Zaroog MS, Abdul Kadir H, Tayyab S

ScientificWorldJournal, 2013;2013:570859.
PMID: 24163624 DOI: 10.1155/2013/570859

Different spectral probes were employed to study the stabilizing effect of various polyols, such as, ethylene glycol (EG), glycerol (GLY), glucose (GLC) and trehalose (TRE) on the native (N), the acid-denatured (AD) and the thermal-denatured (TD) states of Aspergillus niger glucoamylase (GA). Polyols induced both secondary and tertiary structural changes in the AD state of enzyme as reflected from altered circular dichroism (CD), tryptophan (Trp), and 1-anilinonaphthalene-8-sulfonic acid (ANS) fluorescence characteristics. Thermodynamic analysis of the thermal denaturation curve of native GA suggested significant increase in enzyme stability in the presence of GLC, TRE, and GLY (in decreasing order) while EG destabilized it. Furthermore, CD and fluorescence characteristics of the TD state at 71°C in the presence of polyols showed greater effectiveness of both GLC and TRE in inducing native-like secondary and tertiary structures compared to GLY and EG.
Spectroscopic studies on the binding of bromocresol purple to different serum albumins and its bilirubin displacing action

Faizul FM, Abdul Kadir H, Tayyab S

J. Photochem. Photobiol. B, Biol., 2008 Jan 30;90(1):1-7.
PMID: 18024146

The interaction between bromocresol purple (BCP) and bovine serum albumin (BSA)/porcine serum albumin (PSA) was investigated both in the absence and presence of bilirubin (BR) using absorption/absorption difference spectroscopy. A significant red shift in the absorption maxima of BCP accompanied by a decrease in absorbance was indicative of BCP binding to albumin. The titration of BSA and PSA with BCP using absorption difference spectroscopy and analysis of results by Benesi-Hildebrand equation yielded the values of association constant, K as 9.9+/-0.9x10(4)Lmol(-1) and 4.1+/-0.3x10(4)Lmol(-1) for BSA and PSA, respectively. The differential extinction coefficient (Deltaepsilon) of 34,484M(-1)cm(-1) at 615nm and 41,870M(-1)cm(-1) at 619nm were estimated for BSA and PSA, respectively. Decrease in (DeltaAbs.)(615nm) of BCP-BSA complex with the increase in ionic strength suggested the role of hydrophobic interactions in the binding phenomenon. A significant blue shift in the absorption maxima and change in (DeltaAbs)(lambdamax) values of BR-albumin complexes upon addition of increasing concentrations of BCP revealed the BR displacing action of BCP on albumin molecule.
Fulltext Bioassay-Guided Isolation of Cytotoxic Cycloartane Triterpenoid Glycosides from the Traditionally Used Medicinal Plant Leea indica

Wong YH, Abdul Kadir H, Ling SK

Evid Based Complement Alternat Med, 2012;2012:164689.
PMID: 22203865 DOI: 10.1155/2012/164689

Leea indica is a medicinal plant used traditionally to cure cancer. In this study, the cytotoxic compounds of L. indica were isolated using bioassay-guided approach. Two cycloartane triterpenoid glycosides, mollic acid arabinoside (MAA) and mollic acid xyloside (MAX), were firstly isolated from L. indica. They inhibited the growth of Ca Ski cervical cancer cells with IC(50) of 19.21 μM (MAA) and 33.33 μM (MAX). MRC5 normal cell line was used to calculate selectivity index. MAA and MAX were about 8 and 4 times more cytotoxic to Ca Ski cells compared to MRC5. The cytotoxicity of MAA was characterized by both cytostatic and cytocidal effects. MAA decreased the expression of proliferative cell nuclear antigen, increased sub-G1 cells, and arrested cells in S and G2/M phases. This study provides the evidence for the ethnomedicinal use of L. indica and paves the way for future mechanism studies on the anticancer effects of MAA.
Swietenia macrophylla King induces mitochondrial-mediated apoptosis through p53 upregulation in HCT116 colorectal carcinoma cells

Goh BH, Chan CK, Kamarudin MN, Abdul Kadir H

J Ethnopharmacol, 2014 Apr 28;153(2):375-85.
PMID: 24613274 DOI: 10.1016/j.jep.2014.02.036

Swietenia macrophylla King is a traditional herb used to treat various diseases including hypertension, diabetes and cancer. Previous study demonstrated its anti-tumor effect but the potential mechanisms have not been clearly defined. The current study was to further investigate the underlying mechanism of ethyl acetate fraction of Swietenia macrophylla (SMEAF)-induced anti-proliferative effect and apoptosis in HCT116 colorectal carcinoma cell.
Fulltext Deoxyelephantopin from Elephantopus scaber Inhibits HCT116 Human Colorectal Carcinoma Cell Growth through Apoptosis and Cell Cycle Arrest

Chan CK, Chan G, Awang K, Abdul Kadir H

Molecules, 2016 Mar 21;21(3):385.
PMID: 27007366 DOI: 10.3390/molecules21030385

Deoxyelephantopin (DET), one of the major sesquiterpene lactones derived from Elephantopus scaber was reported to possess numerous pharmacological functions. This study aimed to assess the apoptosis inducing effects and cell cycle arrest by DET followed by elucidation of the mechanisms underlying cell death in HCT116 cells. The anticancer activity of DET was evaluated by a MTT assay. Morphological and biochemical changes were detected by Hoescht 33342/PI and Annexin V/PI staining. The results revealed that DET and isodeoxyelephantopin (isoDET) could be isolated from the ethyl acetate fraction of E. scaber leaves via a bioassay-guided approach. DET induced significant dose- and time-dependent growth inhibition of HCT116 cells. Characteristics of apoptosis including nuclear morphological changes and externalization of phosphatidylserine were observed. DET also significantly resulted in the activation of caspase-3 and PARP cleavage. Additionally, DET induced cell cycle arrest at the S phase along with dose-dependent upregulation of p21 and phosphorylated p53 protein expression. DET dose-dependently downregulated cyclin D1, A2, B1, E2, CDK4 and CDK2 protein expression. In conclusion, our data showed that DET induced apoptosis and cell cycle arrest in HCT116 colorectal carcinoma, suggesting that DET has potential as an anticancer agent for colorectal carcinoma.
Dioscorea bulbifera induced apoptosis through inhibition of ERK 1/2 and activation of JNK signaling pathways in HCT116 human colorectal carcinoma cells

Ahmad Hidayat AF, Chan CK, Mohamad J, Abdul Kadir H

Biomed Pharmacother, 2018 Aug;104:806-816.
PMID: 29860114 DOI: 10.1016/j.biopha.2018.05.073

Dioscorea bulbifera, also known as air potato, has been cultivated as food crop mainly in tropical countries in Asia and Australia. The tubers are edible and have often been used in Traditional Chinese Medicine (TCM) and Ayurvedic medicine to treat cancer, diabetes, thyroid disease, and inflammation. This study aimed to investigate the effects of D. bulbifera on HCT116 human colorectal carcinoma cells and to unravel the plausible mechanisms underlying its apoptotic effects. The ethanol crude and fractions (hexane, ethyl acetate and water) of D. bulbifera were subjected to cell viability MTT assay against various cancer cell lines. The lowest IC50 of the extract and fractions on selected cancer cells were selected for further apoptosis assay and western blot analysis. HCT116 cancer cells were treated with D. bulbifera and stained with Annexin/PI or Hoechst 33342/PI for preliminary confirmation of apoptosis. The dissipation of mitochondria membrane potential (MMP) was determined by flow cytometry. The protein expressions of apoptosis-related proteins such as Bcl-2 family, caspases, Fas, PARP, ERK1/2 and JNK were detected by western blot analysis. Moreover, the HCT116 cells were treated with UO126 and SP600125 inhibitors to verify the involvement of ERK1/2 and JNK protein expressions in inducing apoptotic cell death. Based on the result, D. bulbifera ethyl acetate fraction (DBEAF) exhibited the most compelling cytotoxicity on HCT116 cells with an IC50 of 37.91 ± 1.30 µg/mL. The induction of apoptosis was confirmed by phosphatidylserine externalization and chromatin condensation. Depolarization of MMP further conferred the induction of apoptosis was through the regulation of Bcl-2 family proteins. Activation of caspase cascades (caspase-3, -9, -8 and -10) was elicited followed by the observation of cleaved PARP accumulation in DBEAF-treated cells. Furthermore, death receptor, Fas was activated upon exposure to DBEAF. Collective apoptotic evidences suggested the involvement of intrinsic and extrinsic pathways by DBEAF in HCT116 cells. Interestingly, the attenuation of ERK1/2 phosphorylation accompanied by the activation of JNK was detected in DBEAF-treated cells. In conclusion, the findings revealed that DBEAF induced apoptosis through intrinsic and extrinsic pathways involving ERK1/2 and JNK.
Fulltext Health related quality of life (HRQOL) among low socioeconomic population in Malaysia

Wan Puteh SE, Siwar C, Zaidi MAS, Abdul Kadir H

BMC Public Health, 2019 Jun 13;19(Suppl 4):551.
PMID: 31196024 DOI: 10.1186/s12889-019-6853-7

BACKGROUND: The rapid growth of economy and increasing cost of living in Malaysia have given significant impact especially to the lowest household income population. The main objective of this study was to determine risk factors for low quality of life (QOL) and poor health status of this population.
METHODS: This was a cross sectional study design. A total of 347 respondents from low household income groups, including persons with disability and Orang Asli were recruited from E-kasih. A semi-guided self-administered questionnaire was used. QOL measured by EQ. 5D utility value and health status measured by visual analogue score (VAS). Descriptive statistic, bivariate Chi-square analysis and binary logistic regression were conducted to determine factors influencing low QOL and poor health status.
RESULTS: Majority of the respondents were Malay, female (61%), 63% were married, 60% were employed and 46% with total household income of less than 1 thousand Ringgit Malaysia. 70% of them were not having any chronic medical problems. Factors that associated with low QOL were male, single, low household income, and present chronic medical illness, while poor health status associated with female, lower education level and present chronic medical illness. Logistic regression analysis has showed that determinants of low QOL was present chronic illness [AOR 4.15 95%CI (2.42, 7.13)], while determinants for poor health status were; female [AOR 1.94 95%CI (1.09,3.44)], lower education [AOR 3.07 95%CI (1.28,7.34)] and present chronic illness [AOR 2.53 95%CI (1.39,4.61)].
CONCLUSION: Low socioeconomic population defined as low total household income in this study. Low QOL of this population determined by present chronic illness, while poor health status determined by gender, education level and chronic medical illness.
Fulltext Cytotoxic Effects and Anti-Angiogenesis Potential of Pistachio (Pistacia vera L.) Hulls against MCF-7 Human Breast Cancer Cells

Seifaddinipour M, Farghadani R, Namvar F, Mohamad J, Abdul Kadir H

Molecules, 2018 Jan 05;23(1).
PMID: 29303970 DOI: 10.3390/molecules23010110

Pistachio (Pistacia vera L.) hulls (PVLH) represents a significant by-product of industrial pistachio processing that contains high amounta of phenolic and flavonoid compounds known to act as antioxidants. The current study was designed to evaluate the anti-tumor and anti-angiogenic potentials of PVLH extracts. The cytotoxic effects of hexane, ethyl acetate, methanol, and water PVLH extracts toward human colon cancer (HT-29 and HCT-116), breast adenocarcinoma (MCF-7), lung adenocarcinoma (H23), liver hepatocellular carcinoma (HepG2), cervical cancer (Ca Ski), and normal fibroblast (BJ-5ta) cells were assessed using a MTT cell viability assay. Apoptosis induction was evaluated through the different nuclear staining assays and confirmed by flow cytometry analysis. Anti-angiogenic activities were also determined using chorioallantoic membrane (CAM) assay. PVLH ethyl acetate extracts (PVLH-EAE) demonstrated a suppressive effect with an IC50 value of 21.20 ± 1.35, 23.00 ± 1.2 and 25.15 ± 1.85 µg/mL against MCF-7, HT-29 and HCT-116, respectively, after 72 h of treatment. Morphological assessment and flow cytometry analysis showed the potential of PVLH-EAE to induce apoptosis. PVLH-EAE at the highest concentration demonstrated significant inhibition of angiogenesis as comparing with control group. Also the expression of Bax increased and the expression of Bcl-2 decreased in treated MCF-7 cells. Thus, the apoptosis induction and angiogenesis potential of PVLH-EAE make it to be the most suitable for further cancer research study to deal with selective antitumor active substances to human cancers especially breast cancer.
Fulltext Loranthus micranthus Linn.: Biological Activities and Phytochemistry

Zorofchian Moghadamtousi S, Hajrezaei M, Abdul Kadir H, Zandi K

Evid Based Complement Alternat Med, 2013;2013:273712.
PMID: 24109490 DOI: 10.1155/2013/273712

Loranthus micranthus Linn. is a medicinal plant from the Loranthaceae family commonly known as an eastern Nigeria species of the African mistletoe and is widely used in folkloric medicine to cure various ailments and diseases. It is semiparasitic plant because of growing on various host trees and shrubs and absorbing mineral nutrition and water from respective host. Hence, the phytochemicals and biological activities of L. micranthus demonstrated strong host and harvesting period dependency. The leaves have been proved to possess immunomodulatory, antidiabetic, antimicrobial, antihypertensive, antioxidant, antidiarrhoeal, and hypolipidemic activities. This review summarizes the information and findings concerning the current knowledge on the biological activities, pharmacological properties, toxicity, and chemical constituents of Loranthus micranthus.
Annona muricata leaves induce G₁ cell cycle arrest and apoptosis through mitochondria-mediated pathway in human HCT-116 and HT-29 colon cancer cells

Zorofchian Moghadamtousi S, Karimian H, Rouhollahi E, Paydar M, Fadaeinasab M, Abdul Kadir H

J Ethnopharmacol, 2014 Oct 28;156:277-89.
PMID: 25195082 DOI: 10.1016/j.jep.2014.08.011

ETHNOPHARMACOLOGICAL RELEVANCE: Annona muricata known as "the cancer killer" has been widely used in the traditional medicine for the treatment of cancer and tumors. The purpose of this study is to investigate the anticancer properties of ethyl acetate extract of Annona muricata leaves (EEAM) on HT-29 and HCT-116 colon cancer cells and the underlying mechanisms.
MATERIALS AND METHODS: The effect of EEAM on the cell proliferation of HT-29 and HCT-116 cells was analyzed by the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium) assay. High content screening system (HCS) was applied to investigate the cell membrane permeability, mitochondrial membrane potential (MMP), nuclear condensation and cytochrome c translocation from mitochondria to cytosol. Reactive oxygen species (ROS) formation, lactate dehydrogenase (LDH) release and activation of caspase-3/7, -8 and -9 were measured while treatment. Flow cytometric analysis was used to determine the cell cycle distribution and phosphatidylserine externalization. The protein expression of Bax and Bcl-2 was determined using immunofluorescence analysis. In addition, the potential of EEAM to suppress the migration and invasion of colon cancer cells was also examined.
RESULTS: EEAM exerted significant cytotoxic effects on HCT-116 and HT-29 cells as determined by MTT and LDH assays. After 24 h treatment, EEAM exhibited the IC₅₀ value of 11.43 ± 1.87 µg/ml and 8.98 ± 1.24 µg/ml against HT-29 and HCT-116 cells, respectively. Flow cytometric analysis demonstrated the cell cycle arrest at G1 phase and phosphatidylserine externalization confirming the induction of apoptosis. EEAM treatment caused excessive accumulation of ROS followed by disruption of MMP, cytochrome c leakage and activation of the initiator and executioner caspases in both colon cancer cells. Immunofluorescence analysis depicted the up-regulation of Bax and down-regulation of Bcl-2 proteins while treated with EEAM. Furthermore, EEAM conspicuously blocked the migration and invasion of HT-29 and HCT-116 cells.
CONCLUSIONS: These findings provide a scientific basis for the use of A. muricata leaves in the treatment of cancer, although further in vivo studies are still required.
Fulltext PIK3CA gene amplification and PI3K p110α protein expression in breast carcinoma

Firoozinia M, Zareian Jahromi M, Moghadamtousi SZ, Nikzad S, Abdul Kadir H

Int J Med Sci, 2014;11(6):620-5.
PMID: 24782652 DOI: 10.7150/ijms.8251

A family of PI3Ks is the lipid kinases, which enhance intracellular pools of phosphatidyl inositol 3,4,5-tri-phosphate (PIP3) through phosphorylating its precursor. Amplifications and deletions of genes, as well as somatic missense of the PIK3CA gene have been described in many human cancer varieties, including of the brain, colon, liver, lung and stomach. Immunohistochemistry and Real-time quantitative PCR tests were used to determine the PIK3CA gene amplification (gene copy number) and to detect protein expression, respectively. The results obtained were analysed and the ratio of PIK3CA to β-actin gene copy number was calculated. Positive gene amplification of PIK3CA was appointed as a copy number of ≥4. Also, PI3K p110α protein expression was scored from 0 to 3+ and the scores of 2+ and 3+ were considered as positive for PI3K p110α protein expression. We studied 50 breast carcinoma samples for PI3K p110α protein expression and PIK3CA gene copy numbers. In general, 36 out of 50 (72%) breast carcinoma samples showed a significant increase in PIK3CA gene amplification. 12 out of 50 (24%) showed positive staining, and 38 out of 50 (76%) showed negative staining for PI3K p110α expression. We have identified no significant relationship between PIK3CA amplification, race (p= 0.630) and histological type (p=0. 731) in breast carcinoma, but correlation of PIK3CA amplification and age showed a significant relationship (p=0. 003) between them. No significant relationship has been identified in correlation of PI3K p110α protein expression compared to age (p=0. 284), race (p=0. 546) and histological type (p=0. 285). Amplification of PIK3CA was frequent in breast carcinoma and occurs in stages of breast carcinoma. Our result shows that there is a relationship between gene amplification and age in breast carcinoma. We suggest that PIK3CA is significant in breast tumorigenesis serve as a prevalent mechanism contributes to the oncogenic activation pathway of PIK3CA in breast cancer.
Fulltext The chemopotential effect of Annona muricata leaves against azoxymethane-induced colonic aberrant crypt foci in rats and the apoptotic effect of Acetogenin Annomuricin E in HT-29 cells: a bioassay-guided approach

Zorofchian Moghadamtousi S, Rouhollahi E, Karimian H, Fadaeinasab M, Firoozinia M, Ameen Abdulla M, et al.

PLoS One, 2015;10(4):e0122288.
PMID: 25860620 DOI: 10.1371/journal.pone.0122288

Annona muricata has been used in folk medicine for the treatment of cancer and tumors. This study evaluated the chemopreventive properties of an ethyl acetate extract of A. muricata leaves (EEAML) on azoxymethane-induced colonic aberrant crypt foci (ACF) in rats. Moreover, the cytotoxic compound of EEAML (Annomuricin E) was isolated, and its apoptosis-inducing effect was investigated against HT-29 colon cancer cell line using a bioassay-guided approach. This experiment was performed on five groups of rats: negative control, cancer control, EEAML (250 mg/kg), EEAML (500 mg/kg) and positive control (5-fluorouracil). Methylene blue staining of colorectal specimens showed that application of EEAML at both doses significantly reduced the colonic ACF formation compared with the cancer control group. Immunohistochemistry analysis showed the down-regulation of PCNA and Bcl-2 proteins and the up-regulation of Bax protein after administration of EEAML compared with the cancer control group. In addition, an increase in the levels of enzymatic antioxidants and a decrease in the malondialdehyde level of the colon tissue homogenates were observed, suggesting the suppression of lipid peroxidation. Annomuricin E inhibited the growth of HT-29 cells with an IC50 value of 1.62 ± 0.24 μg/ml after 48 h. The cytotoxic effect of annomuricin E was further substantiated by G1 cell cycle arrest and early apoptosis induction in HT-29 cells. Annomuricin E triggered mitochondria-initiated events, including the dissipation of the mitochondrial membrane potential and the leakage of cytochrome c from the mitochondria. Prior to these events, annomuricin E activated caspase 3/7 and caspase 9. Upstream, annomuricin E induced a time-dependent upregulation of Bax and downregulation of Bcl-2 at the mRNA and protein levels. In conclusion, these findings substantiate the usage of A. muricata leaves in ethnomedicine against cancer and highlight annomuricin E as one of the contributing compounds in the anticancer activity of A. muricata leaves.
Fulltext Anticancer and antitumor potential of fucoidan and fucoxanthin, two main metabolites isolated from brown algae

Zorofchian Moghadamtousi S, Karimian H, Khanabdali R, Razavi M, Firoozinia M, Zandi K, et al.

ScientificWorldJournal, 2014;2014:768323.
PMID: 24526922 DOI: 10.1155/2014/768323

Seaweed is one of the largest producers of biomass in marine environment and is a rich arsenal of active metabolites and functional ingredients with valuable beneficial health effects. Being a staple part of Asian cuisine, investigations on the crude extracts of Phaeophyceae or brown algae revealed marked antitumor activity, eliciting a variety of research to determine the active ingredients involved in this potential. The sulfated polysaccharide of fucoidan and carotenoid of fucoxanthin were found to be the most important active metabolites of brown algae as potential chemotherapeutic or chemopreventive agents. This review strives to provide detailed account of all current knowledge on the anticancer and antitumor activity of fucoidan and fucoxanthin as the two major metabolites isolated from brown algae.
Fulltext (R)-(+)-α-lipoic acid protected NG108-15 cells against H₂O₂-induced cell death through PI3K-Akt/GSK-3β pathway and suppression of NF-κβ-cytokines

Kamarudin MN, Mohd Raflee NA, Hussein SS, Lo JY, Supriady H, Abdul Kadir H

Drug Des Devel Ther, 2014;8:1765-80.
PMID: 25336920 DOI: 10.2147/DDDT.S67980

Alpha-lipoic acid, a potent antioxidant with multifarious pharmacological benefits has been reported to be neuroprotective in several neuronal models and used to treat neurological disorders such as Alzheimer's disease. Nonetheless, conclusive mechanisms of alpha-lipoic acid for its protective effects particularly in NG108-15 cells have never been investigated. In this study, the intricate neuroprotective molecular mechanisms by (R)-(+)-alpha-lipoic acid (R-LA) against H2O2-induced cell death in an in vitro model of neurodegeneration were elucidated. Pretreatment with R-LA (2 hours) significantly increased NG108-15 cell viability as compared to H2O2-treated cells and mitigated the induction of apoptosis as evidenced by Hoechst 33342/propidium iodide staining. R-LA (12.5-50 μM) aggrandized the reduced glutathione over glutathione disulfide ratio followed by a reduction in the intracellular reactive oxygen species level and an increase in mitochondrial membrane potential following H2O2 exposure. Moreover, pretreatment with R-LA stimulated the activation of PI3K-Akt through mTORC1 and mTORC2 components (mTOR, rictor and raptor) and production of antiinflammatory cytokine, IL-10 which led to the inactivation of glycogen synthase kinase-3β (GSK-3β) and reduction of both Bax/Bcl2 and Bax/Bcl-xL ratios, accompanied by inhibition of the cleaved caspase-3. Additionally, this observation was preceded by the suppression of NF-κβ p65 translocation and production of proinflammatory cytokines (IL-6 and TNF-α). The current findings accentuate new mechanistic insight of R-LA against apoptogenic and brain inflammatory factors in a neuronal model. These results further advocate the therapeutic potential of R-LA for the treatment of neurodegenerative diseases.
Fulltext Prevalence and risk factors of sonographically detected non alcoholic fatty liver disease in a screening centre in Klang Valley, Malaysia: an observational cross-sectional study

Khammas ASA, Hassan HA, Salih SQM, Kadir H, Ibrahim RM, Nasir NNM, et al.

Porto Biomed J, 2018 09 19;4(2):e31.
PMID: 31595260 DOI: 10.1016/j.pbj.0000000000000031

Objectives: Nonalcoholic fatty liver disease (NAFLD) is a very common liver disorder in Western countries. As of late, it has been found to be prevalent in Asia as well. It is a benign disease unless it develops into necroinflammation and fibrosis. This study was proposed to determine the prevalence and risk factors of sonography-detected NAFLD among Malaysian adults in Klang Valley, West Malaysia.
Study design: An observational cross-sectional study.
Methods: The participants were aged between 45 and 75 years who participated in a screening program at the Golden Horses Health Sanctuary in Klang Valley. Lipid profile and anthropometric measurements were collected from the subjects' medical records. Ultrasound machine and a structured self-administered questionnaire were used as instruments for recruiting data from the subjects. The subjects who consumed alcohol (>140 g/wk for men and >70 g/wk for females), had hepatitis B or C viruses, liver insults, and surgery, and taken lipid-lowering medications were excluded from the study.
Results: A total of 628 subjects were analyzed, and 235 (37.4%) subjects were diagnosed with definite NAFLD. They comprised 518 (82.5%) Chinese, 92 (14.6%) Malays, and 18 (2.9%) Indians. Peak prevalence of NAFLD was found in 53 to 60 years age group. The higher prevalence of NAFLD was among men (48.3%) than women (27.3%) and among Indians (61.1%) and Malays (51.1%) than among Chinese (34.2%). NAFLD has been found to be strongly correlated with male sex, high body mass index (≥23.0 kg/m2), hypertriglyceridemia, low high-density lipoprotein cholesterol, diabetes mellitus, and hypertension.
Conclusion: NAFLD is quite common among adults in Malaysian urban population. The prevalence of NAFLD was inordinately high among the 53 to 60 years age group, male sex, Indians, and Malays (as compared with Chinese). Age >60 years, male sex, high body mass index (≥23.0 kg/m2), hypertriglyceridemia, and diabetes mellitus were proven to be risk predictors for NAFLD.

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