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1.  Angiotensin-I Converting Enzyme (ACE) Inhibitory and Anti-Hypertensive Effect of Protein Hydrolysate from Actinopyga lecanora (Sea Cucumber) in Rats 
Marine Drugs  2016;14(10):176.
Food protein hydrolysates are known to exhibit angiotensin converting enzyme (ACE) inhibitory properties and can be used as a novel functional food for prevention of hypertension. This study evaluated the ACE inhibitory potentials of Actinopyga lecanora proteolysate (ALP) in vivo. The pre-fed rats with ALP at various doses (200, 400, 800 mg/kg body weight) exhibited a significant (p ≤ 0.05) suppression effect after inducing hypertension. To determine the optimum effective dose that will produce maximal reduction in blood pressure, ALP at three doses was fed to the rats after inducing hypertension. The results showed that the 800 mg/kg body weight dose significantly reduced blood pressure without noticeable negative physiological effect. In addition, there were no observable changes in the rats’ heart rate after oral administration of the ALP. It was concluded that Actinopyga lecanora proteolysate could potentially be used for the development of functional foods and nutraceuticals for prevention and treatment of hypertension.
PMCID: PMC5082324  PMID: 27706040
sea cucumber; ACE inhibition; normotensive rats; hypertension
2.  A higher sensitivity and efficiency of common primer multiplex PCR assay in identification of meat origin using NADH dehydrogenase subunit 4 gene 
A Common Primer Multiplex PCR (CP-M-PCR) was developed to detect meat origin of four groups of animal (pig, ruminant, avian and rabbit). This method demonstrated higher sensitivity and efficiency than the conventional multiplex PCR. In this approach, a common forward primer was designed in the 5′ end of a homologous region of mitochondrial NADH dehyrogenase subunit 4 (Nad 4) gene sequences of all the animal groups. Specific adapter reverse primers were designed by adding an adapter sequence at the 5′ end. The same adapter sequence was used as the common adapter reverse primer. The primers generated specific fragments of 267, 370, 504, and 548 bp lengths for pig, ruminant, avian and rabbit meats, respectively. The use of adapter sequence at the 5′ end of the common adapter reverse primers increased the efficiency of the amplification and the application of a common forward primer solved the complexity in multiplex PCR system. Bands of specific amplification can be detected in the PCR assays containing as low as 10−6 μM of adapter reverse primer. This result indicated that the sensitivity was tremendously increased as compared to the conventional multiplex PCR (10−3 μM). CP-M-PCR detection limit of the DNA samples was 0.1 ng for the four groups of meats. CP-M-PCR has greatly improved the sensitivity and efficiency of the PCR system for a more reliable and accurate outcome than conventional multiplex PCR system.
PMCID: PMC4486532  PMID: 26139881
Multiplex; Nad 4; Meat; Common primer
3.  Phytochemicals and Medicinal Properties of Indigenous Tropical Fruits with Potential for Commercial Development 
Hundreds of fruit-bearing trees are native to Southeast Asia, but many of them are considered as indigenous or underutilized. These species can be categorized as indigenous tropical fruits with potential for commercial development and those possible for commercial development. Many of these fruits are considered as underutilized unless the commercialization is being realized despite the fact that they have the developmental potential. This review discusses seven indigenous tropical fruits from 15 species that have been identified, in which their fruits are having potential for commercial development. As they are not as popular as the commercially available fruits, limited information is found. This paper is the first initiative to provide information on the phytochemicals and potential medicinal uses of these fruits. Phytochemicals detected in these fruits are mainly the phenolic compounds, carotenoids, and other terpenoids. Most of these phytochemicals are potent antioxidants and have corresponded to the free radical scavenging activities and other biological activities of the fruits. The scientific research that covered a broad range of in vitro to in vivo studies on the medicinal potentials of these fruits is also discussed in detail. The current review is an update for researchers to have a better understanding of the species, which simultaneously can provide awareness to enhance their commercial value and promote their utilization for better biodiversity conservation.
PMCID: PMC4906201  PMID: 27340420
4.  The inhibitory activity of cocoa phenolic extract against pro-inflammatory mediators secretion induced by lipopolysaccharide in RAW 264.7 cells 
SpringerPlus  2016;5:547.
Cocoa is a rich source of polyphenols that has been traditionally used as the treatment of several types of inflammation related disease. The response to inflammation comprises the consecutive release of mediators and the enlistment of circulating leukocytes, such as macrophages. Currently, Cocoa-derived polyphenolics have shown anti-inflammatory effects in vivo, but the therapeutic benefits in vitro remain unclear. Therefore, in this study, the effect of cocoa polyphenolic extract (CPE) on RAW 264.7 macrophage cells sensitized by lipopolysaccharide as in vitro inflammatory model was investigated. The anti-inflammatory activity of CPE was assessed by measuring its ability to inhibit the pro-inflammatory enzyme 5-lipoxygenase (5-LOX) and the pro-inflammatory mediators prostaglandin E2 (PGE2), reactive oxygen species (ROS), nitric oxide (NO) and tumor necrosis factor-alpha (TNF-α). The results show that CPE significantly inhibits 5-LOX activity (p < 0.01). In addition, CPE dose-dependently suppressed the production of PGE2, ROS, NO and TNF-α in RAW 264.7 cells. These data suggest that CPE may be used for the treatment of inflammation and it’s related-diseases.
PMCID: PMC4850146  PMID: 27190746
Cocoa; Polyphenols treatment; Inflammation; 5-Lipoxygenase; Prostaglandin; RAW 264.7 macrophage cells
5.  Identification of phenolic compounds in polyphenols-rich extract of Malaysian cocoa powder using the HPLC-UV-ESI—MS/MS and probing their antioxidant properties 
The antioxidant components of cocoa powder, which is rich in polyphenols, were isolated using column chromatography and high performance liquid chromatography. Polyphenolic compounds were then characterized by high-performance liquid chromatography/Ultraviolet and electronspray ionization—tandem mass spectrometry (HPLC-UV-/ESI-MS-MS). As a result, five phenolic compounds were detected. In this study we also investigated scavenging or the total antioxidant capacity (%) of cocoa polyphenol (CP) fractionated from cocoa powder extract. 114.0 mg/g of gallic acid –equivalent phenolics and 94.3 mg/g catechin- equivalent flavonoids were quantified in this extract. Their free radical-scavenging activity was assessed by 1,1-diphenyl-2-picrylhydrazyl radical (DPPH) assay, β-carotene bleaching test, and xanthine oxidase inhibitory activity (OX). Total antioxidant capacity (TAC) was further assessed against the myoglobin-induced oxidation of 6-hydroxy-2, 5, 7, 8-tetramethylchroman-2-carboxylic acid (ABTS) and expressed as Trolox equivalent. A high correlation between TAC and phenolic contents indicated that phenolic compounds from cocoa were a major contributor of antioxidant activity (0.967 ≤ r ≤ 1.00). CP extract had significantly (P < 0.05) potential antioxidant activities with various concentrations. These results suggest that Polyphenols-rich cocoa extract possess prominent medical properties and can be exploited as natural drug to treat free radical associated diseases.
PMCID: PMC4375169  PMID: 25829590
Cocoa polyphenols; LC-UV; LC-MS/MS; Free radicals; Antioxidant activity
6.  Effect of Cosmos caudatus (Ulam raja) supplementation in patients with type 2 diabetes: Study protocol for a randomized controlled trial 
Type 2 diabetes mellitus is a major health threat worldwide. Cosmos caudatus is one of the medicinal plants used to treat type 2 diabetes. Therefore, this study aims to determine the effectiveness and safety of C. caudatus in patients with type 2 diabetes. Metabolomic approach will be carried out to compare the metabolite profiles between C. Caudatus treated diabetic patients and diabetic controls.
Methods and design
This is a single-center, randomized, controlled, two-arm parallel design clinical trial that will be carried out in a tertiary hospital in Malaysia. In this study, 100 patients diagnosed with type 2 diabetes will be enrolled. Diabetic patients who meet the eligibility criteria will be randomly allocated to two groups, which are diabetic C. caudatus treated(U) group and diabetic control (C) group. Primary and secondary outcomes will be measured at baseline, 4, 8, and 12 weeks. The serum and urine metabolome of both groups will be examined using proton NMR spectroscopy.
The study will be the first randomized controlled trial to assess whether C. caudatus can confer beneficial effect in patients with type 2 diabetes. The results of this trial will provide clinical evidence on the effectiveness and safety of C. caudatus in patients with type 2 diabetes.
Trial registration identifier: NCT02322268
PMCID: PMC4769500  PMID: 26920910
Cosmos caudatus; Type 2 diabetes mellitus; Glycemic status; Metabolomic; Herbal medicine; Functional food; Dietary Supplementation; Randomized controlled trial
7.  Protective effects of the extracts of Barringtonia racemosa shoots against oxidative damage in HepG2 cells 
PeerJ  2016;4:e1628.
Barringtonia racemosa is a tropical plant with medicinal values. In this study, the ability of the water extracts of the leaf (BLE) and stem (BSE) from the shoots to protect HepG2 cells against oxidative damage was studied. Five major polyphenolic compounds consisting of gallic acid, ellagic acid, protocatechuic acid, quercetin and kaempferol were identified using HPLC-DAD and ESI-MS. Cell viability assay revealed that BLE and BSE were non-cytotoxic (cell viabilities >80%) at concentration less than 250 µg/ml and 500 µg/ml, respectively. BLE and BSE improved cellular antioxidant status measured by FRAP assay and protected HepG2 cells against H2O2-induced cytotoxicity. The extracts also inhibited lipid peroxidation in HepG2 cells as well as the production of reactive oxygen species. BLE and BSE could also suppress the activities of superoxide dismutase and catalase during oxidative stress. The shoots of B. racemosa can be an alternative bioactive ingredient in the prevention of oxidative damage.
PMCID: PMC4734433  PMID: 26839752
Polyphenols; Barringtonia racemosa; HPLC-ESI-MS; Antioxidant enzymes; Lipid peroxidation; Oxidative stress
8.  An Investigation into the Antiobesity Effects of Morinda citrifolia L. Leaf Extract in High Fat Diet Induced Obese Rats Using a 1H NMR Metabolomics Approach 
Journal of Diabetes Research  2015;2016:2391592.
The prevalence of obesity is increasing worldwide, with high fat diet (HFD) as one of the main contributing factors. Obesity increases the predisposition to other diseases such as diabetes through various metabolic pathways. Limited availability of antiobesity drugs and the popularity of complementary medicine have encouraged research in finding phytochemical strategies to this multifaceted disease. HFD induced obese Sprague-Dawley rats were treated with an extract of Morinda citrifolia L. leaves (MLE 60). After 9 weeks of treatment, positive effects were observed on adiposity, fecal fat content, plasma lipids, and insulin and leptin levels. The inducement of obesity and treatment with MLE 60 on metabolic alterations were then further elucidated using a 1H NMR based metabolomics approach. Discriminating metabolites involved were products of various metabolic pathways, including glucose metabolism and TCA cycle (lactate, 2-oxoglutarate, citrate, succinate, pyruvate, and acetate), amino acid metabolism (alanine, 2-hydroxybutyrate), choline metabolism (betaine), creatinine metabolism (creatinine), and gut microbiome metabolism (hippurate, phenylacetylglycine, dimethylamine, and trigonelline). Treatment with MLE 60 resulted in significant improvement in the metabolic perturbations caused obesity as demonstrated by the proximity of the treated group to the normal group in the OPLS-DA score plot and the change in trajectory movement of the diseased group towards the healthy group upon treatment.
PMCID: PMC4698747  PMID: 26798649
9.  Effect of microbial cell preparation on renal profile and liver function among type 2 diabetics: a randomized controlled trial 
The beneficial effect of probiotics on renal profile and liver function has been reported among patients with chronic kidney disease and fatty liver respectively. However, its effect on renal profile and liver function among type 2 diabetic individuals has not been fully understood. To investigate the effect of microbial cell preparation on renal profile and liver function tests among type 2 diabetic individuals.
A randomized, double-blind, parallel-group, controlled clinical trial was conducted on a total of 136 type 2 diabetics age 30-70 years old in a teaching hospital in Kuala Lumpur, Malaysia. Subjects were randomly assigned to receive microbial cell preparation (N = 68) or a placebo (N = 68) for 12 weeks. The outcomes measured at baseline, week 6, and week 12 and included changes in renal profile (Sodium, Potassium, Urea, Creatinine, Glomerular Filtration Rate), and liver function tests (Albumin, Total Protein, Alkaline Phosphatase, Alanine Aminotransferase, Aspartate Aminotransferase). Intention to treat (ITT) analysis was performed on all the recruited subjects, while per protocol (PP) analysis was conducted on those who completed the trial with good compliance.
The urea levels significantly declined in the probiotic group. Serum urea levels reduced from 4.26 mmol/L to 4.04 mmol/L in Probiotic Group while it increased in Placebo Group from 4.03 mmol/L to 4.24 mmol/L. These changes were significant between groups in ITT analysis (p = 0.018). Other parameters did not change significantly between groups.
12 weeks supplementation with daily dosage of 6 × 1010 Colony Forming Units of multi-strain microbial cell preparation significantly improved urea levels.
Trial registration
(Clinical trials: #NCT01752803)
Electronic supplementary material
The online version of this article (doi:10.1186/s12906-015-0952-5) contains supplementary material, which is available to authorized users.
PMCID: PMC4676823  PMID: 26654906
Microbial cell preparation; Probiotics; Urea; Renal profile; Liver function tests; Type 2 diabetes; Lactobacillus; Bifidobacterium
10.  Eight Weeks of Cosmos caudatus (Ulam Raja) Supplementation Improves Glycemic Status in Patients with Type 2 Diabetes: A Randomized Controlled Trial 
Objectives. Optimizing glycemic control is crucial to prevent type 2 diabetes related complications. Cosmos caudatus is reported to have promising effect in improving plasma blood glucose in an animal model. However, its impact on human remains ambiguous. This study was carried out to evaluate the effectiveness of C. caudatus on glycemic status in patients with type 2 diabetes. Materials and Methods. In this randomized controlled trial with two-arm parallel-group design, a total of 101 subjects with type 2 diabetes were randomly allocated to diabetic-ulam or diabetic controls for eight weeks. Subjects in diabetic-ulam group consumed 15 g of C. caudatus daily for eight weeks while diabetic controls abstained from taking C. caudatus. Both groups received the standard lifestyle advice. Results. After 8 weeks of supplementation, C. caudatus significantly reduced serum insulin (−1.16 versus +3.91), reduced HOMA-IR (−1.09 versus +1.34), and increased QUICKI (+0.05 versus −0.03) in diabetic-ulam group compared with the diabetic controls. HbA1C level was improved although it is not statistically significant (−0.76% versus −0.37%). C. caudatus was safe to consume. Conclusions. C. caudatus supplementation significantly improves insulin resistance and insulin sensitivity in patients with type 2 diabetes.
PMCID: PMC4680046  PMID: 26713097
11.  Angiotensin-I Converting Enzyme (ACE) Inhibitory and Anti-Oxidant Activities of Sea Cucumber (Actinopyga lecanora) Hydrolysates 
In recent years, food protein-derived hydrolysates have received considerable attention because of their numerous health benefits. Amongst the hydrolysates, those with anti-hypertensive and anti-oxidative activities are receiving special attention as both activities can play significant roles in preventing cardiovascular diseases. The present study investigated the angiotensin-I converting enzyme (ACE) inhibitory and anti-oxidative activities of Actinopyga lecanora (A. lecanora) hydrolysates, which had been prepared by alcalase, papain, bromelain, flavourzyme, pepsin, and trypsin under their optimum conditions. The alcalase hydrolysate showed the highest ACE inhibitory activity (69.8%) after 8 h of hydrolysis while the highest anti-oxidative activities measured by 2,2-diphenyl 1-1-picrylhydrazyl radical scavenging (DPPH) (56.00%) and ferrous ion-chelating (FIC) (59.00%) methods were exhibited after 24 h and 8 h of hydrolysis, respectively. The ACE-inhibitory and anti-oxidative activities displayed dose-dependent trends, and increased with increasing protein hydrolysate concentrations. Moreover, strong positive correlations between angiotensin-I converting enzyme (ACE) inhibitory and anti-oxidative activities were also observed. This study indicates that A. lecanora hydrolysate can be exploited as a source of functional food owing to its anti-oxidant as well as anti-hypertension functions.
PMCID: PMC4691087  PMID: 26690117
Actinopyga lecanora; anti-oxidative; ACE inhibitory
12.  Potential medicinal benefits of Cosmos caudatus (Ulam Raja): A scoping review 
Cosmos caudatus is widely used as a traditional medicine in Southeast Asia. C. caudatus has been reported as a rich source of bioactive compounds such as ascorbic acid, quercetin, and chlorogenic acid. Studies have shown that C. caudatus exhibits high anti-oxidant capacity and various medicinal properties, including anti-diabetic activity, anti-hypertensive properties, anti-inflammatory responses, bone-protective effect, and anti-microbial activity. This review aims to present the potential medicinal benefits of C. caudatus from the available scientific literature. We searched PubMed and ScienceDirect database for articles published from 1995 to January 2015. Overall, 15 articles related to C. caudatus and its medicinal benefits are reviewed. All these studies demonstrated that C. caudatus is effective, having demonstrated its anti-diabetic, anti-hypertensive, anti-inflammatory, bone-protective, anti-microbial, and anti-fungal activity in both in vitro and animal studies. None of the studies showed any negative effect of C. caudatus related to medicinal use. Currently available evidence suggests that C. caudatus has beneficial effects such as reducing blood glucose, reducing blood pressure, promoting healthy bone formation, and demonstrating anti-inflammatory and anti-microbial properties. However, human clinical trial is warranted.
PMCID: PMC4746859  PMID: 26929767
Anti-diabetic; anti-hypertensive; anti-inflammatory; anti-microbial; anti-oxidant; bone-protective; Cosmos caudatus; medicinal effect
13.  Phenolic composition, antioxidant, anti-wrinkles and tyrosinase inhibitory activities of cocoa pod extract 
Cocoa pod is an outer part of cocoa fruits being discarded during cocoa bean processing. Authors found out that data on its usage in literature as cosmetic materials was not recorded in vast. In this study, cocoa pod extract was investigated for its potential as a cosmetic ingredient.
Cocoa pod extract (CPE) composition was accomplished using UHPLC. The antioxidant capacity were measured using scavenging assay of 1,2-diphenyl-2-picrylhydrazyl (DPPH), β-carotene bleaching assay (BCB) and ferric reducing antioxidant power (FRAP). Inhibiting effect on skin degradation enzymes was carried out using elastase and collagenase assays. The skin whitening effect of CPE was determined based on mushroom tyrosinase assay and sun screening effect (UV-absorbance at 200-400 nm wavelength).
LC-MS/MS data showed the presence of carboxylic acid, phenolic acid, fatty acid, flavonoids (flavonol and flavones), stilbenoids and terpenoids in CPE. Results for antioxidant activity exhibited that CPE possessed good antioxidant activity, based on the mechanism of the assays compared with ascorbic acid (AA) and standardized pine bark extract (PBE); DPPH: AA > CPE > PBE; FRAP: PBE > CPE > AA; and BCB: BHT > CPE > PBE. Cocoa pod extract showed better action against elastase and collagenase enzymes in comparison with PBE and AA. Higher inhibition towards tyrosinase enzyme was exhibited by CPE than kojic acid and AA, although lower than PBE. CPE induced proliferation when tested on human fibroblast cell at low concentration. CPE also exhibited a potential as UVB sunscreen despite its low performance as a UVA sunscreen agent.
Therefore, the CPE has high potential as a cosmetic ingredient due to its anti-wrinkle, skin whitening, and sunscreen effects.
PMCID: PMC4195981  PMID: 25292439
Cocoa pod extract; Antioxidant; Anti-wrinkles; Skin whitening; Sunscreen agent; Cosmetic ingredient
14.  The Effectiveness of Rambutan (Nephelium lappaceum L.) Extract in Stabilization of Sunflower Oil under Accelerated Conditions 
Antioxidants  2014;3(2):371-386.
The oxidative properties of sunflower oil supplemented with rambutan extract, (crude extract and its fractionated fraction, SF II) in comparison with synthetic antioxidant were investigated. The supplemented sunflower oils were stored under accelerated conditions for 24 days at 60 °C. For every 6-day interval, the oxidative properties of the supplemented sunflower oil were evaluated based on the following tests, namely peroxide value, p-anisidine value, Thiobarbituric Acid Reactive Substances (TBARS) assay, iodine value and free fatty acids. The total oxidation (TOTOX) values were also calculated based on the peroxide values and p-anisidine values. Rambutan extract is a potential source of antioxidant. The oxidative activities of the extracts at all concentrations were significantly (p < 0.05) higher than the control. Generally, the partially fractionated fraction was more effective than the crude extract. With a 2-year storage period at ambient temperature, the fractionated fraction of the extract, SF II at 300 ppm, was observed to work more effectively than the synthetic antioxidant, t-Tocopherol, and it possessed a protective effect comparable with butylatedhydrioxynanisole (BHA). Therefore, rambutan extract could be used as a potential alternative source of antioxidant in the oil industry or other fat-based products to delay lipid oxidation.
PMCID: PMC4665487  PMID: 26784877
rambutan; oxidative stability; peroxide value; p-anisidine value; iodine value; TOTOX; free fatty acid
15.  Inhibition of Oxidative Stress and Lipid Peroxidation by Anthocyanins from Defatted Canarium odontophyllum Pericarp and Peel Using In Vitro Bioassays 
PLoS ONE  2014;9(1):e81447.
Canarium odontophyllum, also known as CO, is a highly nutritious fruit. Defatted parts of CO fruit are potent sources of nutraceutical. This study aimed to determine oxidative stress and lipid peroxidation effects of defatted CO pericarp and peel extracts using in vitro bioassays. Cell cytotoxic effect of the CO pericarp and peel extracts were also evaluated using HUVEC and Chang liver cell lines. The crude extracts of defatted CO peel and pericarp showed cytoprotective effects in t-BHP and 40% methanol-induced cell death. The crude extracts also showed no toxic effect to Chang liver cell line. Using CD36 ELISA, NAD+ and LDL inhibition assays, inhibition of oxidative stress were found higher in the crude extract of defatted CO peel compared to the pericarp extract. Hemoglobin and LDL oxidation assays revealed both crude extracts had significantly reduced lipid peroxidation as compared to control. TBARS values among defatted CO pericarp, peel, and cyanidin-3-glucoside showed no significant differences for hemoglobin and LDL oxidation assays. The protective effects of defatted CO parts, especially its peel is related to the presence of high anthocyanin that potentially offers as a pharmaceutical ingredient for cardioprotection.
PMCID: PMC3886967  PMID: 24416130
16.  Antioxidative and Cardioprotective Properties of Anthocyanins from Defatted Dabai Extracts 
This study aimed to determine anthocyanins and their antioxidative and cardioprotective properties in defatted dabai parts. Anthocyanins in crude extracts and extract fractions of defatted dabai peel and pericarp were quantified using UHPLC, while their antioxidant capacity and oxidative stress inhibition ability were evaluated by using DPPH and CUPRAC assays as well as linoleic acid oxidation system, hemoglobin oxidation, and PARP-1 inhibition ELISA. Cardioprotective effect of the defatted dabai peel extract was evaluated using hypercholesterolemic-induced New Zealand white rabbits. Six anthocyanins were detected in the defatted dabai peel, with the highest antioxidant capacities and oxidative stress inhibition effect compared to the other part. The defatted dabai peel extract has also inhibited lipid peroxidation (plasma MDA) and elevated cellular antioxidant enzymes (SOD and GPx) in the tested animal model. Major anthocyanin (cyanidin-3-glucoside) and other anthocyanins (pelargonidin-3-glucoside, malvidin-3-glucoside, cyanidin-3-galactoside, cyanidin-3-arabinoside, and peonidin-3-glucoside) detected in the defatted dabai peel are potential future nutraceuticals with promising medicinal properties.
PMCID: PMC3867864  PMID: 24368926
17.  Impaired of a non-DNA dependent methylation status decides the fat decision of bone marrow-derived C3H10T1/2 stem cell 
SpringerPlus  2013;2:590.
A decrease in the lineage commitment of multipotent Mesenchymal stem cells (MSC) to the bone forming osteoblast lineage and an increase in the commitment to the fat forming adipocyte lineage is more common in bone marrow of elderly persons. A link between methylation status and MSC differentiation remains unclear. Therefore, we hypothesize that hypomethylation may decide the fate decisions of MSC. In the current study, murine bone marrow derived-C3H10T1/2 stem cell was used to examine the role of methylation mechanism on the differentiation potential of stem cells into osteoblasts or adipocytes. C3H10T1/2 cells were treated with Periodate oxidized adenosine (Adox), an inhibitor of S-adenosylhomocysteine-dependent hydrolase (SAHH), which in turn block the non-DNA methylation pathway. The effect of hypomethylation on C3H10T1/2 stem cell differentiation was determined by measuring the alkaline phosphates activity and the degree of mineralization as well as Oil-red O staining and lipid content. The ratio of S-adenosylmethionine (SAM) and S-adenosylhomocysteine (SAH) was determined as a metabolic indicator of cellular methylation potential. It was clearly observed that hypomethylation significantly (P < 0.05) reduces SAM: SAH ratio, alkaline phosphates activity, calcification and thereby, osteoblast differentiation. Conversely, adipocyte differentiation was stimulated by hypomethylation. Altogether, our data suggest that non-DNA hypomethylation changes the differentiation potential of C3H10T1/2 stem cells for less osteogenic and more adipogenic.
PMCID: PMC3833906  PMID: 24294542
Methylation mechanism; C3H10T1/2 stem cell; S-adenosylmethionine; S-adenosylhomo-cysteine; Osteablast; Adipocyte
18.  Phytochemicals and Antioxidant Capacity from Nypa fruticans Wurmb. Fruit 
Nypa fruticans Wurmb. is one of the important underutilized fruit of Malaysia, which lacks scientific attention. Total phenolics, flavonoid content, and antioxidant capacities from endosperm extracts of Nypa fruticans (unripe and ripe fruits) were evaluated. Endosperm extract of unripe fruits (EEU) exhibited the highest phenolics (135.6 ± 4.5 mg GAE/g), flavonoid content (68.6 ± 3.1 RE/g), and antioxidant capacity. Free radical scavenging capacity of EEU as assessed by 2-2′-azino-bis (3-ethylbenz-thiazoline-6-sulfonic acid (ABTS) and 1,1-diphenyl-2-picryl hydrazyl (DPPH) radicals showed inhibitory activity of 78 ± 1.2% and 85 ± 2.6%, respectively. Beta carotene bleaching coefficient of EEU was higher (2550 ± 123), when compared to endosperm extract of ripe fruits (1729 ± 172). Additionally, EEU exhibited high antioxidant capacity by phosphomolybdenum method and ferric reducing antioxidant power values. Eight phenolic compounds from Nypa fruticans endosperm extracts were identified and quantified by ultra-high-performance liquid chromatography. Chlorogenic acid, protocatechuic acid, and kaempferol were the major phenolic compounds. Thus this fruit could be used as a potential source of natural antioxidant.
PMCID: PMC3654328  PMID: 23710209
19.  Quantitative Determination of Fatty Acids in Marine Fish and Shellfish from Warm Water of Straits of Malacca for Nutraceutical Purposes 
BioMed Research International  2012;2013:284329.
This study was conducted to quantitatively determine the fatty acid contents of 20 species of marine fish and four species of shellfish from Straits of Malacca. Most samples contained fairly high amounts of polyunsaturated fatty acids (PUFAs), especially alpha-linolenic acid (ALA, C18:3 n3), eicosapentaenoic acid (EPA, C20:5 n3), and docosahexaenoic acid (DHA, C22:6 n3). Longtail shad, yellowstripe scad, and moonfish contained significantly higher (P < 0.05) amounts of eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and alpha-linolenic acid (ALA), respectively. Meanwhile, fringescale sardinella, malabar red snapper, black pomfret, Japanese threadfin bream, giant seaperch, and sixbar grouper showed considerably high content (537.2–944.1 mg/100g wet sample) of desirable omega-3 fatty acids. The polyunsaturated-fatty-acids/saturated-fatty-acids (P/S) ratios for most samples were higher than that of Menhaden oil (P/S = 0.58), a recommended PUFA supplement which may help to lower blood pressure. Yellowstripe scad (highest DHA, ω − 3/ω − 6 = 6.4, P/S = 1.7), moonfish (highest ALA, ω − 3/ω − 6 = 1.9, P/S = 1.0), and longtail shad (highest EPA, ω − 3/ω − 6 = 0.8, P/S = 0.4) were the samples with an outstandingly desirable overall composition of fatty acids. Overall, the marine fish and shellfish from the area contained good composition of fatty acids which offer health benefits and may be used for nutraceutical purposes in the future.
PMCID: PMC3591149  PMID: 23509703
20.  Early G1 cyclin-dependent kinases as prognostic markers and potential therapeutic targets in esophageal adenocarcinoma 
Clinical Cancer Research  2011;17(13):4513-4522.
Chromosomal gain at 7q21 is a frequent event in esophageal adenocarcinoma (EAC). However, this event has not been mapped with fine resolution in a large EAC cohort and its association with clinical endpoints and functional relevance are unclear.
Experimental design
We used a cohort of 116 patients to fine map the 7q21 amplification using SNP microarrays. Prognostic significance and functional role of 7q21 amplification and its gene expression were explored.
Amplification of the 7q21 region was observed in 35% of tumors with a focal, minimal amplicon containing 6 genes. 7q21 amplification was associated with poor survival and analysis of gene expression identified CDK6 as the only gene in the minimal amplicon whose expression was also associated with poor survival. A low level amplification (10%) was observed at the 12q13 region containing the CDK6 homolog, CDK4. Both amplification and expression of CDK4 correlated with poor survival. A combined model of both CDK6 and CDK4 expression is a superior predictor of survival than either alone. Specific knockdown of CDK4 and/or CDK6 by siRNAs shows that they are required for proliferation of EAC cells and that their function is additive. PD-0332991 targets the kinase activity of both molecules and suppresses proliferation and anchorage-independence of EAC cells through activation of the pRB pathway.
We suggest that CDK6 is the driver of 7q21 amplification and that both CDK4 and CDK6 are prognostic markers and bona fide oncogenes in EAC. Targeting these molecules may constitute a viable new therapy for this disease.
PMCID: PMC3390776  PMID: 21593195
Esophageal adenocarcinoma; CDK6; CDK4; PD-0332991
21.  Antiatherosclerotic Effect of Canarium odontophyllum Miq. Fruit Parts in Rabbits Fed High Cholesterol Diet 
The effect of C. odontophyllum (CO) fruit parts was investigated in hypercholesterolemic rabbits. Forty-nine rabbits, which were randomly divided into seven groups of seven animals (n = 7), received a diet containing different parts of CO fruit parts for 8 weeks. The groups were as follows: (1) normal diet: NC group and (2) hypercholesterolemic diet: PC, HS (10 mg/kg/day simvastatin), HPO (20 g kg−1 oil extracted from the pulp of CO), HKO (20 g kg−1 oil extracted from the kernel of CO), HF (50 g kg−1 fullfat pulp of CO), and HD (50 g kg−1 defatted pulp of CO). Among these groups, rabbits receiving defatted pulp of CO showed the greatest cholesterol lowering effect as it had reduced plasma LDL-C, TC, and thiobarbiturate reactive substance (TBARS) levels as well as atherosclerotic plaques. The presence of high dietary fiber and antioxidants activity are potential factors contributing to the cholesterol lowering effect. Consequently, these results indicate the potential use of CO defatted pulp as a cholesterol lowering and antioxidant agent.
PMCID: PMC3395265  PMID: 22811751
22.  Ficus deltoidea: A Potential Alternative Medicine for Diabetes Mellitus 
Ficus deltoidea from the Moraceae family has been scientifically proven to reduce hyperglycemia at different prandial states. In this study, we evaluate the mechanisms that underlie antihyperglycemic action of Ficus deltoidea. The results had shown that hot aqueous extract of Ficus deltoidea stimulated insulin secretion significantly with the highest magnitude of stimulation was 7.31-fold (P < 0.001). The insulin secretory actions of the hot aqueous extract involved K+ ATP channel-dependent and K+ ATP-channel-independent pathway. The extract also has the ability to induce the usage of intracellular Ca2+ to trigger insulin release. The ethanolic and methanolic extracts enhanced basal and insulin-mediated glucose uptake into adipocytes cells. The extracts possess either insulin-mimetic or insulin-sensitizing property or combination of both properties during enhancing glucose uptake into such cells. Meanwhile, the hot aqueous and methanolic extracts augmented basal and insulin-stimulated adiponectin secretion from adipocytes cells. From this study, it is suggested that Ficus deltoidea has the potential to be developed as future oral antidiabetic agent.
PMCID: PMC3372277  PMID: 22701507
23.  Protective Effect of Pulp Oil Extracted from Canarium odontophyllum Miq. Fruit on Blood Lipids, Lipid Peroxidation, and Antioxidant Status in Healthy Rabbits 
The aim of this paper was to compare the effects of pulp and kernel oils of Canarium odontophyllum Miq. (CO) on lipid profile, lipid peroxidation, and oxidative stress of healthy rabbits. The oils are rich in SFAs and MUFAs (mainly palmitic and oleic acids). The pulp oil is rich in polyphenols. Male New Zealand white (NZW) rabbits were fed for 4 weeks on a normal diet containing pulp (NP) or kernel oil (NK) of CO while corn oil was used as control (NC). Total cholesterol (TC), HDL-C, LDL-c and triglycerides (TG) levels were measured in this paper. Antioxidant enzymes (superoxide dismutase and glutathione peroxidise), thiobarbiturate reactive substances (TBARSs), and plasma total antioxidant status (TAS) were also evaluated. Supplementation of CO pulp oil resulted in favorable changes in blood lipid and lipid peroxidation (increased HDL-C, reduced LDL-C, TG, TBARS levels) with enhancement of SOD, GPx, and plasma TAS levels. Meanwhile, supplementation of kernel oil caused lowering of plasma TC and LDL-C as well as enhancement of SOD and TAS levels. These changes showed that oils of CO could be beneficial in improving lipid profile and antioxidant status as when using part of normal diet. The oils can be used as alternative to present vegetable oil.
PMCID: PMC3366250  PMID: 22685623
24.  Plants' Metabolites as Potential Antiobesity Agents 
The Scientific World Journal  2012;2012:436039.
Obesity and obesity-related complications are on the increase both in the developed and developing world. Since existing pharmaceuticals fail to come up with long-term solutions to address this issue, there is an ever-pressing need to find and develop new drugs and alternatives. Natural products, particularly medicinal plants, are believed to harbor potential antiobesity agents that can act through various mechanisms either by preventing weight gain or promoting weight loss amongst others. The inhibition of key lipid and carbohydrate hydrolyzing and metabolizing enzymes, disruption of adipogenesis, and modulation of its factors or appetite suppression are some of the plethora of targeted approaches to probe the antiobesity potential of medicinal plants. A new technology such as metabolomics, which deals with the study of the whole metabolome, has been identified to be a promising technique to probe the progression of diseases, elucidate their pathologies, and assess the effects of natural health products on certain pathological conditions. This has been applied to drug research, bone health, and to a limited extent to obesity research. This paper thus endeavors to give an overview of those plants, which have been reported to have antiobesity effects and highlight the potential and relevance of metabolomics in obesity research.
PMCID: PMC3362029  PMID: 22666121
25.  Evaluation of Minerals Content of Drinking Water in Malaysia 
The Scientific World Journal  2012;2012:403574.
The drinking and mineral water samples obtained from different geographical locations had concentrations of the selected minerals lower than the standard limits, except for manganese, arsenic, and fluoride. The concentrations of manganese and arsenic in two mineral water samples were slightly higher than the standard international recommended limits. One mineral water sample had a fluoride concentration higher than the standard limits, whereas manganese was not detected in nine drinking and mineral water samples. Most of the selected minerals found in the tap water samples were below the international standard limits, except for iron and manganese. The concentrations of iron and manganese in the tap water samples were higher than the standard limits, which were obtained from one and three of the studied locations, respectively. The potable water obtained from various manufacturers and locations in Peninsular Malaysia is safe for consumption, as the minerals concentrations were below the standard limits prescribed by the Malaysian Food Regulations of 1985. The data obtained may also provide important information related to daily intake of these minerals from drinking water.
PMCID: PMC3353278  PMID: 22649292

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