Malaysian Cocoapsasir.upm.edu.my/id/eprint/57608/1/0001.pdf · bentuk atau sebarang cara elektronik, mekanik, penggambaran semula, rakaman dan sebagainya tanpa terIebih dahulu mendapat

Malaysian Cocoaor ChocolatesA story of antioxidants and more ...

Profesor Dr. Amin Ismail3ac. Food Science &'Technology (UPM), PhD (Food Chemistry & Biochemistry) (UPM), FNSM

14 AUGUST 2015

Dewan Kuliah UtamaFakulti Perubatan dan Sains Kesihatan

Universiti Putra Malaysia

Universiti Putra Malaysia PressSerdang • 2015

http://www.penerbit.upm.edu.my

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Contents

Abstract 1

Introduction 3

Polyphenols in Cocoa and Cocoa-derived Products 7

Antioxidant Capacity and Health Benefits of CocoaPolyphenols 14

• Cocoa polyphenols and antioxidant capacity 16• Cocoa polyphenols and diabetes mellitus 21• Cocoa polyphenols and inflammatory mediators 25• Cocoa polyphenols and cancer 27• Cocoa polyphenols and osteoporosis 29• Cocoa polyphenols and cardiovascular 31• Cocoa polyphenols and obesity 35

Molecular Mechanisms Underlying the Health Benefitsof Cocoa Polyphenols 44

Conclusion 47

References 49

Biography 57

Acknowledgments 61

List of Inaugural Lectures 63

ABSTRACTFor centuries, cocoa has been recognized as a rich source ofdietary antioxidants, especially polyphenols. It is known not onlyfor its good flavour but also for its health benefits. Cocoa hasdrawn increased attention because of its antioxidant propertiesand marked effects in the prevention of various oxidative stress-associated diseases. In the last few years, research and developmenton polyphenols and extracts from cocoa and cocoa-derivedproducts, such as cocoa powder, cocoa liquor and chocolates,has become a major area of health and medical related research.The recommended human diet contains a significant amount ofpolyphenols, as they are assumed to be 'antioxidants' that scavengeexcessive harmful free radicals arising from normal metabolicprocesses. In vitro as well as in vivo data indicate that polyphenolspresent in Malaysian cocoa may have antioxidant capacity, anti-diabetes and anti-inflammatory properties and also promote ananti-obesity phenotype. Over the last 2-3 years, there have beenexciting new developments which have shone more light on the invivo mechanisms behind the health benefits of Malaysian cocoa.Our studies showed that cocoa polyphenols not only function asantioxidants but also as non-antioxidants. Several molecular targets(e.g., nuclear factor Kappa B (NF-kB), activated protein-I (AP-I)peroxisome proliferator-activated receptors (PPARs) and liver Xreceptors (LXRs)) have been recently identified and may partlyexplain the potential benefits of cocoa in relation to combatingobesity-associated diseases. Cocoa polyphenols have been reportedto regulate lipid metabolism by inducing metabolic gene expressionand activating transcription factors that regulate the expression ofnumerous genes, many of which play an important role in energymetabolism. Further studies have been performed to investigate theprotective effects of cocoa polyphenols against metabolic diseases

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Malaysian Cocoa of Chocolates: A Story of Antioxidants and More ...

which include acting as an antioxidant or suppressing transcriptionfactors that antagonize lipid accumulation. Recent evidencesuggests that polyphenols also have indirect antioxidant effectsthrough the induction of endogenous protective enzymes. Evidenceof potential benefits through polyphenolic-mediated regulation ofcellular processes such as inflammation is also increasing, and thesesignalling effects may occur at concentrations which are muchlower than those required for effective radical scavenging. Thus,polyphenol-rich cocoa products may potentially diminish obesity-mediated metabolic diseases by multiple mechanisms, therebyattenuating chronic inflammation. Our findings from in-vitro andin-vivo studies on the health benefits of Malaysian cocoa-derivedproducts, suggest that the intake of cocoa polyphenols could lead toreduced disease risk. Moreover, the consumption of a balanced dietthat includes a variety of polyphenol-rich food sources is importantfor the promotion of health.

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INTRODUCTIONThe chronology of the history of cocoa begins in 2,000 B.C. Thisdate is attributed by historians to the oldest drinking cups and platesever discovered in Latin America in a small village in the Uhiavalley in Honduras, where cocoa played a central role. In 200 -900AD, cocoa was one of the main products in Mayan agriculture andreligion (Barry Callebaut, 200S). The word cacao is derived fromthe Olmec and the subsequent Mayan languages (Kakaw), and thename chocolate is derived from Olmec/Mayan etymo~ogy. In 1773,the cocoa tree was named Theobroma cacao, which can be attributedto the mythical background ofthe tree, and literally means "Food ofGods" (Barry Callebaut, 200S). The medicinal uses of cocoa havebeen traced to Mexican sources, and more than 150 uses of cocoain medicines have been documented (Dillinger et aI., 2000). Later,in 1630, in Europe, cocoa was used as a medicine rather than as adelicious foodstuff. The use of chocolate to stimulate the normalfunctions of the spleen and other digestive system functions hadbeen recognized. Additionally, during the ISth century, chocolatewas regularly mixed into medications for many diseases, includingthe common cold, cough and high blood pressure (Barry Callebaut,200S). The first evidence of the health benefits of cocoa and itsproducts comes from the Kuna Indians, a native population livingon the coast of Panama. The Kunas consumed adequate amountsof cocoa daily, usually evenly mixed with salt. Physical and clinicalinvestigations found that the Kunas indeed had lower blood pressureand no age-dependent decreases in kidney functions. Furthermore,mortality due to cardiovascular diseases was markedly lower in thisnative population, as compared to other citizens.

Cocoa is a food with many uses. Cocoa generally refers tococoa powder, which is made from roasted cocoa beans. This is notto be confused with Theobroma cacao, which literally refers to the

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cocoa tree. Good quality cocoa beans are produced through two vitalsteps, namely primary and secondary processing. The primary stepsare fermentation and drying, whilst secondary processing involvescleaning and cracking, roasting, grinding, pressing (optional) andtempering.

There are different types of cocoa and cocoa-derived productsavailable in the market. Cocoa powder and cocoa liquor are used asingredients in chocolate manufacturing, along with cocoa butter andmilk. Dark and milk chocolates are made from a mixture of cocoa

.r-liquor and sl._lgar,but the latter involves the addition of cocoa butter.White chocolate, however, is not 'real' chocolate because it is madefrom a mixture of cocoa butter, milk and sugar. Dark chocolate isbitter and less sweet compared to milk and white chocolate. Darkchocolate is considered one of the major contributors of antioxidantsto the American diet, after fruits and vegetables. Cocoa beansand their derived-products (Figurel) have been used as the majoringredient in chocolate and cocoa powder for a long period of time.

Malaysia was ranked the world's fifth largest producerof cocoa beans in 2012. Today, Malaysia is one of the mainproducers of cocoa-based products in the world and the biggestin Asia. The utilization and exploitation of cocoa extract and/orpolyphenol substances (fine chemicals) from Malaysian beanscan contribute to the nation's economic development. It willbenefit the Malaysian cocoa industry to exploit and diversify theuse of cocoa extract, which is a rich source of polyphenols. Thestandardized polyphenol-rich extract and its fine chemicals can beexploited for the development of functional foods, nutraceuticalsand pharmaceuticals. In Malaysia, the development of the cocoaindustry has begun and a variety of cocoa products are nowavailable in the local market. With the growing interest in healthfoods, products that incorporate cocoa polyphenol extract have an

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added value due to the potential health benefits. Our local cocoaindustry, especially the grinding sector, has grown steadily over theyears and should continue at grow at a faster rate to advance thisagro-industry sector. Due to a shortage in the production of cocoabeans in Malaysia, our country imports approximately 44,000 tonsannually from Indonesia (most of the beans are under-fermentedand inexpensive). The manufacturers usually blend these beanswith those from West Africa and Ghana (expensive well-fermentedbeans) to meet their requirements. Through partial polyphenolextraction, which can reduce excessive astringent and improve thebitter taste, valuable polyphenol substances can be produced asby-products which can be utilized for human benefit. Our studycan help improve the competitiveness of cocoa beans from Asianregions, especially Malaysian beans, in global markets.

In the year 2000, our research group reported that Malaysiancocoa beans and their derived products had significant antioxidantcapacity compared to other cocoa beans (Azizah et al., 2007). Thusthe Malaysian cocoa beans and their derived products could helpdecrease chronic disease risk factors (Abbe et al., 2009; Abbe et al.,2008a; Azizah et al. 2007; Ruzaidi et al. 2005; Amin et al. 2004b;Amin et al. 2004c).

Cocoa beans are reported to have a high content of antioxidantpolyphenols, which comprise 12-18% of the dry weight of wholebeans, compared to other foods. Besides polyphenols, proteins playan important role in its flavour quality (Amin et al., 2002a, b).

Polyphenols or phenolics are one of the non-nutrient groupsfound in plants, and they are characterized by the presence ofone or more hydroxyl groups on aromatic rings. They are foundwidely in the plant kingdom and are the secondary metabolites ofplants. They are involved in the defence against DV and pathogens.More than 8,000 phenolic compounds have been reported as being

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antioxidants (Bravo, 2000). Nutritionally, polyphenol-rich foodscan strongly inhibit iron absorption. Due to the chelating propertiesof these compounds, they can bind more or less strongly to ironin the intestinal lumen and influence iron absorption. Polyphenolcompounds were not considered necessary from a nutritional pointof view during the early years due to their mutagenic and genotoxicactivities. However, data from epidemiological studies suggest thathigh dietary intake of polyphenols is associated with decreased riskof a range of diseases, including cardiovascular diseases, certaincancers and neurodegenerative diseases. Epidemiological studiesalso demonstrate that regular dietary intake of cocoa polyphenolsreduces the risk of coronary heart disease and stroke and is inverselyassociated with the risk of cardiovascular diseases (Hooper et aI.,2012).

Fruits of Theobroma cacao Dried cocoa beans (raw cocoa)

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Cocoa powder Cocoa liquor/mass

Chocolates

Figure 1 Fruits of the Theobroma cocoa tree and their derived products.

Polyphenols in Cocoa and Cocoa-derived Products

Food generally contains complex mixtures of polyphenols andnumerous factors may affect the content of polyphenols in plantsand plant products. Our studies have reported that Malaysian fruits,

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vegetables, legumes and their by-products are rich in polyphenols(Chew et al., 2011; Emmy Hainida et al., 2008 & 2009; Khoo& Amin, 2008; Prasad et al., 2010; Marina et al., 2009; Sun etal., 2013; Tan et al., 2010a, b; Zabidah et al., 2014; Azrina et al.,2010; Fouad et al., 2011; Sadeq et al., 2012; Kong et al., 2010a,b& 2011). Flavonoids are one of the polyphenol groups in foodswhich have attracted great interest since the 1990s. Cocoa is a richsource of dietary flavanol (a sub-class of flavonoids). Epicatechinand catechin are the major dietary flavanols in cocoa beans (Aliet al., 2014a; Abbe &Amin, 2010; Azizah et al., 2007) (Figure 2).Some chocolates (especially dark chocolates) remain good'sourcesof polyp henoIs, especially flavonol (Figure 3). Researchers are verymuch interested in cocoa polyphenols due to their potent antioxidantproperties as well as their credible effects in the prevention ofvarious free radicals associated diseases.

erOH

HO~~ 0 •••••

.& OH.. H .

erOH.Hl(CX~0 ......

.& OH

H

Figure 2(a) Catechin Figure 2(b) Epichatechin

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.JX

Figure 2(c) Procyanidins dimer and trimer in cocoa

Figure 2 Major polyphenol compounds (catechin, epicatechin andprocyanidins) in cocoa. (Source: Ali et al., 2014a; Abbe & Amin, 2010;

Azizah et al., 2007)

1200951

1000GI.!:!III

~8OO.~•~ 800'0cGIs::~ 400S

200

odark

chocolate(4Og)

milkchocolate

(4Og)

chocolate homemademilk (240 mil chocolate

milk (240 ml)

homemadehot cocoa

(180ml)

hot cocoamixes(180 mil

Figure 3: Polyphenols content in chocolate products(Source: Visioli et al., 2009)

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The total intake of polyphenol-rich food in humans is aboutIg/day with some large inter individual variability (Nobutomo etal. 2010). A recent study by Buijsse et al. (2010) among Germanadults suggested that 7.5 g/day intake of chocolate (24% ofintake from dark chocolate) is associated with lower systolic anddiastolic blood pressure and a 10% lower 8 years risk of stroke. Inthe Dutch population cocoa products contribute up to 20% of thetotal flavonoids intake in adults, while in children the percentageis even higher. However, the mean daily intake of polyphenols in

!

the Spanish diet was estimated at between 2590 and 3016 mg/person/day. It is important to realize that the biological actionsand health benefits of cocoa polyphenols depend mainly on theirbioavailability, metabolism and amount consumed. However,previous studies of various classes of polyp heno Is have shown thatbioavailability varies widely from one polyphenol to another, so themost abundant polyphenols in our diet are not necessarily thoseleading to the highest levels of active metabolites in target tissues.The impacts of food compositions on the bioavailability ofpolyphenols have not been clearly investigated. The bioavailabilityof all phenolics is sti1llargely unexplored, which demands furtherinvestigation, especially with regards to its functions. For example,independent of the doses of chocolate and cocoa ingested, only0.5% of catechin was recovered in free unbound form from humanplasma and urine. However, scientists hypothesize that someindividuals could have better absorption and tissue distributionthan others, possibly because of particular polymorphisms. Thiscould be the mechanism that explains the high variability in thelevels of flavonoid absorption that have been published (Borcherset al., 2000). Currently there is experimental evidence that cocoapolyphenols can act as dietary signals for direct interaction withDNA and gene expression (Manach et al.'::2005).Th'ere has,

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however, been relatively little study of the molecular mechanismsunderlying the protective effects of cocoa polyphenols on energymetabolism and relevant gene expression profiles. Thus, the possiblemechanisms involved in the cellular uptake of cocoa polyphenols,as well as, its cellular concentrations and distribution, have to beextensively clarified.

The identification of the polyphenols in cocoa products (cocoabeans, cocoa powder, chocolate and cocoa liquor) is the first stepin the utilization of these compounds in the preparation ofnutraceuticals and functional foods. In our studies (Aliet al., 2014a;Abbe &Amin, 2010), polyphenols from cocoa were extracted fromdefatted ~ocoa products (Figure 4).'

Column chromatography

Fractions (Fl-F2-F3)

HPLC+LC-MS

Figure 4 Isolation steps of crude polyphenols from cocoa products

(Source: Abbe & Amin, 2010)

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Polyphenol compounds in each fraction were detected byusing high-performance liquid chromatography (HPLC) (Agilent1100, Palo Alto, USA) equipped with a quaternary pump, autoinjector, degasser and DAD. A reversed-phase C 18 column (Alltech,Licosphere, United States) (250 mm x 4 mm, 5 urn I.D) was usedfor the separation of bioactive compounds and gradient elution of(A) water - trifluoroacetic acid (99.9: 0.1, v/v) and (B) acetonitrile- trifluoroacetic acid (99.9: 0.1, v/v) (Nobutomo et aI., 2010). Alinear tendency elution ofO - 10% (A) for 5 min, 10 - 25% (A) for25 min and 25 - 100% (A) for 5 min, with a flow rate or 0.8 mLl~~ - .-.min, was applied to the analysis along with UV spectra recordedfrom 280 - 360 run. The amount of catechin, epicatechin, gallicacid, protocatechuic acid and chlorogenic acid (mg/g fraction) werequantitatively assessed based on external standards (100 - 1000ug/ml). The eluent was well analyzed by ESI - MS - MS usingan electron spray ionization tandem mass spectrometer (FinniganLCQ Advantage MAX ion-trap mass spectrometer) operating in anegative mode. Cocoa contains five compounds, namely, catechin,epicatechin, gallic acid, protocatechuic acid and chIorogenic acid(Table 1).

.. Table 1 Flavonoids and phenolic acids in cocoa powder

Peaks Con>pound,. RT(min) Concentrations mlll!l CP

Gallic acid 5.9 0.84"'0.4$2 Prulocatcchuic acid 10.1 18.8~2.403 Chloro¥enic acid 12.8 1.18~0.334 Epicatcchin 13.$ 1.39",0.14$ Catechin 11.03 1.32,*,0.47

Phenol ic acid amount 114 mg/IIFlavonoid amount 94.9$ lng/IIYield extraCt" 23.7$ gllOO II

The d..ta ...... !liven _ mean z SO (n -3)·Yleld (percent) [solvent extracts \Oft (g)lsample \Oft (g)) >C 100

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The cocoa polyphenols were then analyzed by HPLC/uV-ESI-MS/MS through molecular ion and mass (Figure 5). At the sametime, the HPLC-MS/MS results were further confirmed usingour standard library information. Through our standard libraryinformation search (e.g. Peak retention times, UV spectrum, [M-H] (rn'), molecular mass and ESI-MS/MS data), the five phenoliccompounds that had been investigated using the HPLC-UV werefurther confirmed in HPLC-MS-MS.

RT: 0.00·65.32

1 109.05

245.1

179.07

244.9

319.29

100 200 300 400mJz

500 600 700

Figure 5 Polyphenol compounds in cocoa by LC-MSIMS with ESI.

(Source: Ali et aI., 2014a)

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Antioxidant Capacity and Health Benefits of CocoaPolyphenolsIt has been reported that the health properties of cocoa productscan be attributed to their antioxidant capacity, and the antioxidantcapacity itself is related to the presence of cocoa polyphenols. Overthe last decade numerous studies have shown the health effectsand phytotherapy of polyphenols from cocoa in vitro and in vivo(Abbe & Amin, 2013; Abbe & Amin, 2008b; Ali et aI., 2014b).Most of the experimental and epidemiological outcomes led to thehypothesis that such health benefits might be linked, at least inpart, to flavonoids, a large subgroup of the heterogeneous groupof polyp heno Is. Moreover, most of the outcomes of the studies oncocoa polyphenols were related to the health effects of polyphenolicstowards the antioxidant status, endothelial function, blood pressure,insulin resistance, inflammatory process and platelet function(Figure 6). Strictly speaking, the previous studies emphasized on thepreventive role ofpolyphenolics, which means that it is the effectsbefore the development of disease or dysfunction. However, somestudies have also investigated the therapeutic role of polyphenolics,meaning its effects after the onset of disorders. In our works, bothprotection and pharmacological activities were assessed, including,cell culture, experimental animals and human studies. Recently,we have shifted from classical research onto molecular, cellularand functional genomics analysis using sophisticated techniquesto unveil the molecular mechanism by which polyphenols preventor ameliorate metabolic diseases with special emphasis on obesityand related diseases, such as, diabetes, osteoporosis, vascular andinflammation disorders.

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ImprovedVascularFunction

Other: Anti-Inflammatory

Figure 6 Potential Health benefits of cocoa polyphenols (Source: Ali etal., 2014b; Abbe &Amin, 2008b; Abbe & Amin, 2013)

The. medicinal properties of polyphenols against many healthproblems have been widely studied. Existing information on thebenefits of polyphenols is mainly focussed on its preventive benefits,but how about its therapeutic benefits if abnormalities have alreadyoccurred in the cardiovascular system? Currently, there is limiteddata on the therapeutic benefits of cocoa supplement ingestion toknow how well it benefits those with existing diseases. Our studieswere mainly focused on the beneficial effects of polyphenol-richcocoa extracts on a number of chronic non-communicable diseases,in vitro and in vivo, including one human intervention trial, asfollows:-

• Cocoa polyphenols and antioxidant capacityCocoa polyphenols and diabetes mellitus.Cocoa polyphenols and inflammatory mediators.

••

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• Cocoa polyphenols and cancer• Cocoa polyphenols and osteoporosis• Cocoa polyphenols and cardiovascular disorders• Cocoa polyphenols and obesity

The interrelationship between the above mentioned diseasesand the health effects of cocoa polyphenols are discussed furtherin the following sections.

Cocoa Polyphenols and Antioxidant CapacityAntioxidants can be defined as any substances which when presentat low concentrations compared with those of an oxidisable substratesignificantly delays or prevents the oxidation of that substrate. Thecommon dietary antioxidants include ~-carotene, vitamin E, vitaminC and selenium. Apart from these components, polyphenols arealso reported to have antioxidant properties, which can be foundnaturally in fruits, vegetables (Amin &Tan, 2002; Amin & Cheah,2003; Amin et aI., 2004a; Amin & Lee, 2005; Amin et aI., 2006;Amin &Mukrizah, 2006; Lee at aI., 2007; Tiong et aI., 2010; Khooet aI., 2011), legumes (Hasnah et aI., 2009) and beverages such astea, cocoa and dark chocolates (Zabidah et aI., 2011; Ruzaidi et aI.,2008a, b; Azizah et aI., 2007; Abbe & Amin, 2007).

A study was conducted to investigate the antioxidant capacityand total phenolic and (-) epicatechin contents of cocoa beansfrom different countries, namely, Malaysia, Ghana, Cote d'Ivoireand Sulawesi. Several methods are available to assess antioxidantcapacity/activity, based on chemical and biological principles (Aminet aI., 2013). In our efforts antioxidant capacity was assayed usingfour different assays, namely, ~-carotene bleaching, scavengingactivity, ferric reducing/antioxidant potential (FRAP) and trolox

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equivalent antioxidant capacity (TEAC) methods. To estimate thecontent of total phenolics, an assay using Folin-Ciocalteu reagentwas used. Further, high-performance liquid chromatography(HPLC) was used to determine the (-) epicatechin content. Theantioxidantcapacity/activity of the cocoa bean based on ~-carotenebleaching method followed the order of Cote d'Ivoire > Malaysia> Ghana> Sulawesi. The Ghanaian beans exhibited the highestscavenging activity, followed by Cote d'Ivoirian, Malaysian andSulawesian. Malaysian beans showed significant highest value (p< 0.05) in phenolic content followed by the Sulawesian, Ghanaianand Cote d'Ivoirian. Sulawesian beans exhibited significant highest(p < 0.05) amount of epicatechin content among the studied beans(Figure 7). The results indicated that different assays revealeddifferent antioxidant values. Moreover, the cocoa beans extractsfrom the four different countries of origin showed differentantioxidant capacities. A positive and high correlation was foundbetween total phenolics and antioxidant potential (FRAP) (Azizah etaI., 2007). Antioxidant capacity of cocoa beans could be contributedby phenolic substances, through its reducing potential. Moreover,(-) epicatechin content showed a positive and high correlation withantioxidant capacity, thus, indicating that, (-) epicatechin could beone of the phenolic substances that contributes towards antioxidantcapacity. The results indicated that the antioxidant capacity andtotal phenolic and (-) epicatechin content of Malaysian beans werecomparable to that of the Ghanaian and Cote d'Ivoirian beans(Azizah et aI., 2007).

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1400

...e 1200cooXco0 1000u0uell

800C>C>...;;..§. 600e:cli: 400•.!:!Cl. 200wZ

" 0Malaysia (M) Cote d'lvolre Sulawesi (S)

(CD)

Figure 7 (-) Epicatechin content of cocoa beans extract.

Ghana (G)

(Source: Azizah et al., 2007)

Furthermore, Cheng et al. (2009) showed that Malaysian darkchocolates exhibited the highest phenolic and flavonoid contents,followed by, milk and white chocolates. Flavonols were the majorflavonoids detected in dark chocolates. Theobromine was detectedin dark and milk chocolates but not in white chocolates. A very highcorrelation (r= 0.93) between total phenolic and flavonoid contentsindicates that the major phenolic compound in dark chocolates isfrom the flavonoid classes. As far as nutrition and health promotionare concerned, dark chocolates would be recommended over milkand white chocolates owing to their higher amount of antioxidantphenolic compounds. Abbe and Amin (2010) found that thepresence of methylxanthines could reduce antioxidant capacity offlavonoids in cocoa powder.

Although cocoa polyphenols do not appear to have anysignificant in vivo antioxidant capacity, some of our datademonstrated that they can protect cells from oxidative stressin vitro, at physiologically relevant concentrations. Our studies

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indicate that polyphenols extracted from cocoa powder may inducethe activity of endogenous antioxidant enzymes in vivo and thuspossess an indirect antioxidant effect.

Cocoa, a naturally occurring plant containing variousfunctional compounds, has been used to determine the cytotoxicityand antioxidant efficacy in 3T3fibroblast cells (Ranneh et al.,2014). In our study, ABTS and ORAC assays were deployed as acomprehensive analysis for evaluating the antioxidant capacity of thecocoa polyphenol extracts (CP) in vitro (Figs. 8& 9). Pretreatmentof cells with 250, 500, 1000 mg/ml of CPE completely preventedany toxicity of3T3 cells and enhanced antioxidant capacity. The CPhad significant (P < 0.05) potential antioxidant activities comparedwith the Trolox equivalent. The high correlation between antioxidantcapacity and phenolic contents indicated that phenolic compoundsfrom cocoa were a major contributor of antioxidant activity (0.967s r s 1.00). These results show that treatment of3T3 cells in culturewith CPE confers the cells significant protection against oxidation.

2S

20

.,. lS

.~

.1:::9i 10

5

00 250 soo 1,000

CPE I'&Iml +mcells

Figure 8 Antioxidant capacity of3T3 cells treated with CPE (250,500 and 1000 ug/ml) compared with Trolox using ABTS. The positivecontrol contained 3T3 cells with Trolox while the negative control

contained 3T3 cells with ABTS alone.(Source: Ranneh et al., 2014)

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7000

J!! 6000

B 5000'"0 4000...i: 3000.!!

i 2000

u 1000~ 00

Trelox +MPH t 1000cells

sao 250 AAPH + cells

(PE tmCellsIlIIml

Figure 9 Total Antioxidant ability of3T3 cells treated with CPE (250,500 and 1000 ug/ml) compared with Trolox

(Source: Ranneh et al., 2014)

The positive control contained 3T3 cells with Trolox while thenegative control contained 3T3 cells withAAPH alone. Results areexpressed as means ±SD (n=3). Differences between means weresignificantly different at p < 0.05. Tukey's test: *P<0.05 ,**P< 0.01versus AAPH. TE: Trolox equivalent per gram of extract.

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Cocoa Polyphenols and Diabetes MellitusFindings from a systematic review and meta-analysis of randomizedtrials show the potential health benefits of cocoa and chocolateon flow-mediated dilation (FMD), insulin and insulin resistance(Hooper et al., 2012). An observational study by Jacques et al.(2013), among members of the Framingham Heart Study Offspringcohort, reported that higher dietary flavonol intake is associated withlower incidence of Type 2 diabetes. Although promising effects ofcocoa polyphenols on biomarkers of non-corrimunicable diseaserisk have emerged, limited studies have been done on the possiblemechanisms behind the actions of cocoa polyphenols (Hollman etal.,2011)

Cocoa polyphenols induce the activity of endogenousantioxidant enzymes and thereby exert an indirect antioxidanteffect. Our work investigated the role of polyphenols in activatingthe antioxidant enzymes and suppression of lipid peroxidationmarkers in rats (Abbe et al., 2008a). The effects of cocoa extractcontaining polyphenols, prepared from cocoa powder, on the liverenzymes' antioxidant parameters of obese-diabetic (Ob-db) ratswere assessed. The Ob-db rats were developed using a high-fatdiet (49% fat, 32% carbohydrates and 19% protein from totalenergy, kcal) for 3 months, followed by a low dose (35 mg/kg bodyweight) streptozotocin (STZ) injection. The rats were given cocoapolyphenols-rich extract (600 mglkg body weight/day) for4 weeks.The oxidative stress biomarkers (8-isoprostane) were significantly (p< 0.05) reduced after cocoa supplementation (Figure 10). Further,superoxide dismutase activity was enhanced in the Ob-db ratscompared to that in the non-supplemented rats (Figure 11). Theresults showed that cocoa supplementation had an effect on theprevention of lipid peroxidation and in enhancing the antioxidantdefense system.

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I c_.... .FlnaI

350b

Figure 10 Plasma 8- isoprostane of experimental rats

Baseline, week-13; after high-fat diet for 12 weeks followedby low-dose of STZ injection; Final, week-I7. ND,normal diet;Diabetic-NO, normal diet + low-dose STZ injection; Ob, obese;Ob-db, obese-diabetic; Ob-db + cocoa, Ob-db + 600 mg cocoalkgbody weight.

(a) 30 •

125..f 20

1'5t 10

Ii 5

0lID O~ Ob

Figure 11 Superoxide dismutase activity of experimental rats.(Source: Abbe et al., 2008a)

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More than 80% of diabetic patients are overweight or obese. Thisgroup of obesity-related metabolic diseases is also known as themetabolic syndrome. Insulin resistance is a pre-diabetic statecharacterized by diminished tissue sensitivity to endogenous insulin,leading to chronic elevation of blood glucose levels. There are in-vitro and in-vivo methods to evaluate the antidiabetic properties ofcocoa. The effects of cocoa extract containing polyphenols, preparedfrom cocoa powder, on the oral glucose tolerance test (OGTT),blood glucose and insulin levels ofOb-db rats were assessed. Cocoaextract (600 mg/kg body weight/day) was supplemented to the ratsfor 4 weeks. The results indicated that there were no significantdifferences in fasting plasma glucose and insulin levels after 4weeks of cocoa extract administration (Table 2). OGTT revealedthat cocoa supplementation in Ob-db rats significantly (p < 0.05)reduced plasma glucose at 60 and 90 min compared to that in thenon-supplemented Ob-db rats (Figure 12). The results showed thatcocoa supplementation had an effect on postprandial glucose controlbut not in the long term. Improved insulin sensitivity in vitro studyusing BRIN-BDI Icell lines has also been reported (Ruzaidi et al.,2005). Our studies also found that other useful substances in cocoainclude polyphenols, cocoa peptides and amino acids, which haveanti diabetic properties (Sarmadi et al., 2012; Sarmadi et al., 2011;Sarmadi & Amin, 2010).

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35

;;:30oE 25.s)20"15

1.. 10


•..

o

6

O+-----~~----~------~------~------~----~o 30 60

,",-(min)

15 90i'-"

120

Figure 12 Oral glucose tolerance test (OGTT) results of the rats.(Source: Abbe et al., 2008a)

ND, normal diet; Diabetic-ND, normal diet + low-dose STZinjection; Ob, obese; Ob-db, obese-diabetic; Ob-db + cocoa, Ob-db+ 600 mgcocoa/kg body weight.

Table 2 Glucometabolism Parameters of the Experimental Rats(Source: Abbe et al., 2008a)

. rats

glucose level (mmol/l) insulin level (pmol/l)

baseline- final" baseline final7.06± 0.78a 7.68± O.SOa 209.01± 60.3a 242.65± 42.56a14.93± 3.76b 20.33± 3.39b 281.87± 49.53b 179.18± 60.17C

NODiabetic-NO

Ob 8.16± O.83a 7.08± 0.49a 375.46± 6O.65c,d316.12± 70.12dOb-db 18.41±l.63c 15.72±3.07c 129.45±29.24e 112.18±4O.82eOb-db+ 18.78± 1.19c 16.21±4.18c 132.95± SO.60e 123.29± 57.198cocoa

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Cocoa Polyphenols and Inflammatory MediatorsReactive oxygen species (ROS) are produced as a normal productof cellular metabolism of oxygen in all aerobic organisms. Manystudies have indicated the deleterious effect ofROS in deterioratinghealth. In addition, a wide range of diseases associated withinflammation are correlated with a high production of ROS. Forcenturies, cocoa has been a rich source of dietary polyphenols. Ithasbeen known not only for its good taste but also for its health effects.The consumption of polyp heno Is-rich foods likecocoa or its derivedproducts has traditionally been used to reduce inflammation-relateddiseases. The effect of cocoa polyphenols extract (CP) on RAW264.7 macrophage cells sensitized by lipopolysaccharide (LPS)as in vitro inflammatory model was tested in our study (Rannehet aI., 2014). The treatment of LPS-stimulated cells with CP atconcentrations of up to 1000 ug/ml did not affect the viability ofthe cells compared with the untreated LPS-stimulated cells. Theanti-inflammatory activity of CP was assessed by measuring itsability to inhibit the pro-inflammatory enzyme 5-lipoxygenase (5-LOX) and the pro-inflammatory mediators prostaglandin E2(PGEi),reactive oxygen species (ROS), nitric oxide (NO) and tumornecrosisfactor-alpha (TNF-a). The results show that CP significantlyinhibits 5-LOX activity (p<0.01) (Figurel3). Additionaly, CPdose-dependently suppressed the production of PGE2, ROS, NOand TNF-a in the RAW 264.7 cells (Figure 14). Collectively, ourresults provide interesting insights on the beneficial effects of CPin the prevention and maintenance of the inflammation mediatedreactive oxygen species in vitro.

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1.2g

'"~ 081; .

e~ 0.6o11 0.4«

0.2

oo 15.625 31.5 62.5 125 250 500 1000

CPE concentration II&Iml

Figure 13 Inhibition ofS-LOX by cocoa polyphenols extract(Source: Ranneh et al., 2014)

----_._. ._------- ---------_.-3 e

e 2.5c0

2In..1;~ 1.Sc.....s 1

;i 0.5

00

e

15.63 31.25 62.5 125 250 SOD 1000

CPE concentration II&Iml

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Figure 14 Effects ofCPE on the LPS-stimulated production ofinflammatory mediators in RAW 264.7 cells

(Source: Ranneh et al., 2014)

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Cocoa Polyphenols and CancerThe effect of cocoa polyphenols extract (CP) on tumour markerenzymes, which are, alkaline phosphatase (ALP), gamma-glutamyl transpeptidase (GGT), glutathione-S-transferase (GST)and glutathione reductase (GR) activities, in the plasma and!or liver of hepatocarcinogenesisrats, which were induced withdiethylnitrosamine (DEN) and 2-acetylaminofluorene (AAF), wasdetermined. The findings showed that CP could lower the activityof tumour marker enzymes in rats during hepatocarcinogenesis(Amin et al., 2004b). Based on the results obtained, polyphenolcompounds present in the cocoa liquor, extracted using ethanol,showedP?tential in decreasing the severityofhepatocarcinogenesis.

Colorectal cancer (CRC) is the third most common malignancyin males and the second most common cancer occurringworldwide. Chronic colonic inflammation is a known risk factorfor CRC. Cocoa contains many polyphenolic compounds thathave beneficial effects in humans. Pandurangan et al. (2015)assessed the antioxidant properties of cocoa on CRC, the mousemodel of azoxymethane (AOM)/dextran sulfate sodium (DSS)-induced colitis-associated cancer, focusing on the activation ofNrf2 signaling. Mice were treated withAOMIDSS and randomizedto receive either a control diet or a 5 and 10% cocoa diet duringthe study period. On day 62 of the experiment, the entire colonwas processed for biochemical and histopathological examination(Figure 15). Increased levels of malondialdehyde (MDA) wereobserved in the AOMlDSS-induced mice. However, subsequentadministration of cocoa decreased the MDA. Enzymatic andnon-enzymatic antioxidants, such as, superoxide dismutase,catalase, glutathione peroxidase and glutathione reductase, weredecreased in the AOMlDSS mice. Cocoa treatment increases theactivities/levels of enzymatic and non-enzymatic antioxidants.Inflammatory mediators, such as, inducible nitric oxide synthase

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(iNOS) and cyclooxygenase (COX)-2, were elevated during AOM!DSS-induction, and treatment with 5 and 10% cocoa effectivelydecreased the expression of iNOS and COX-2. The NF-E2-relatedfactor 2 and its downstream targets, such as NQOl and UDP-GT,were increased by the cocoa treatment. The results of our studysuggest that cocoa may merit further clinical investigation as achemo preventive agent that helps prevent colorectal cancer.

Figure 15 Histopathology of colonic lesions(Source: Pandurangan et aI., 2015)

(Control): normal colonic specimen depicting intact crypt architecture.(AOM/DSS) group of mice with tubular growth pattern and severecellular polymorphism resembling human tubular adenoma with highgrade intra-epithelial neoplasia. (AOMlDSS 1 5% cocoa) group showedan almost complex reduction in tumor size. (AOMIDSS 1 10% cocoa)the hyperplastic colonic mucosa exhibits reduced branching and buddingin crypts. (10% cocoa) showed similar pattern in{he control.

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Cocoa Polyphenols and OsteoporosisOsteoporosis is a skeletal disease characterized by low bone massand microarchitectural deterioration of bone tissue, resultingin increased fragility and subsequently, enhanced fracture risk.Postmenopausal osteoporosis is the most widespread cause ofbone loss relating to age, in which the ovarian hormone deficiencyis a major risk factor. To delay or avoid functional impairments, itis crucial to focus on early prevention of osteoporosis. Hormonetherapy is the most common approach in the management ofosteoporosis. However, estrogen is reported to be related tooccurrences of breast cancer and vascular thrombosis. Fruits andvegetables can have significant long-term health benefits sincethey contain of a variety of phytochemicals, or naturally occurringbioactive non-nutrients, besides vitamins and minerals. Certainphenolic compounds can also be applied as anti-inflammatory andbone-sparing agents. In line with this, various studies have beenconducted in the search for new anti osteoporotic agents from naturalsources, such as, dried plums, blueberries, oranges, green tea andonions. Our studies reported that tempeh may also provide readilyavailable polyphenols and calcium for the studied population ofwomen at risk of low bone mass (Hasnah et aI., 20 I0).

Among the various sources of polyphenols, cocoa has beenthe focus of research attention, mostly due to its high polyphenolcontent. A vast number of studies have indicated its health benefits.However, no research was found that studied the effect of cocoa onbone. In our study, it was hypothesized that cocoa polyphenols maybe effectively involved in bone metabolism and prevention of boneloss. Meanwhile, cocoa is also rich in minerals like magnesium,zinc, potassium and copper and therefore, it was assumed thatcocoa minerals may also positively contribute to bone health. Inour study female Sprague-Dawley rats were fed with 6% (C6%)

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or 12% (C12%) cocoa diet (w/w) after ovariectomy for 95 days,and compared with ovariectomized and sham-operated controls.Based on our results, bone mineral density (BMD) and bone mineralcontent (BMC) of the femur of the C12% rats were significantly(p<0.05) lower than that of all the other groups (Table 3). Cocoaat lower concentrations did not exert any adverse effects on BMDwhile cocoa diet was seen to have significantly elevated vertebraBMD and BMC. Based on these results it is presumed that theeffects of cocoa on BMD are moderate and site-specific (Sarmadi,2015)., It is evident that certain inflammatory cytokines like IL-1,TNF and IL-6. play key roles in pathogenesis of osteoporosis.Elevated levels of serum inflammatory cytokines (TNF -a, IL-1 p,and IL-6) were reported in postmenopausal women. Likewise, inour study, ovariectomy significantly enhanced serum levels of IL-1,TNF and IL-6. However, cocoa dose-dependently attenuated thelevels of these inflammatory biomarkers. In addition, mRNA levelsof IL-6, IL-1 and TNF were higher in the OVX group as comparedwith the sham and cocoa groups. Expression ofIL-1, TNF and IL-6was down-regulated by the cocoa diet. Cocoa can attenuate suchinflammatory conditions by down-regulating IL-1, IL-6 and TNF-agenes as well as by decreasing the serum levels of these markers(Sarmadi, 2015). Cocoa had no effect on whole body BMD and itincreased bone density in the 4th lumbar vertebrae. Additionally,cocoa had no effects at low dose levels and adverse effects at highdose levels on bone density of the femur in the ovariectomized rats.Both concentrations of cocoa reduced inflammatory biomarkers. Wethus assume that cocoa has potential to modulate bone metabolismthrough its anti-inflammatory properties.

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Table 3 Effects of cocoa on the whole body, femur, vertebraBMDandBMC

(Source: Sarmadi, 2015)

Sham OVX C6% C12%

Whole body

BMD (g/cm') o.rzao.oos- O.IS4±O.OOSb O.147±O.OOSb O.IS2±O.OOSb

BMC(g) II.S±O.387 IO.6±O.387 IO.43±O.387 IO.49±O.387

Femur

BMD (g/cm/) O.278±O.OSc O.2SS±O.03a O.26±O.04a O.24±O.Olb

BMC (g), O.6±O.OI6c O.S±O.OI6a O.S03±O.O 17a 0.44±O.O 16b

L4

BMD (g/cm') O.39±O.04c O.19±O.08b O.23±O.O7l" O.23±O.082a

BMC(g) O.2±O.088c O.IO±O.09b O.IS3±O.08a O.lS6±O.O 12a

Values are mean±se (n=4-S). OVX: Ovariecomized group, C6%: OVX fed 6% of cocoa(w/w), CI2%: OVX fed 6% of cocoa (w/w). Within a row values with different letters aresignificantly different (p<O.OS)

Cocoa Polyphenols and CardiovascularPopulation studies have shown that plant polyphenol is inverselycorrelated with mortality from cardiovascular disease andnumerous dietary flavonoids have been shown to beneficiallyimpact atherosclerosis, including lipoprotein oxidation, bloodplatelet aggregation and vascular reactivity. Based on our reviewarticles, there are many studies that reported that cocoa flavonoidshad shown a similar degree in being protective against CVD dueto antioxidant, anti-platelet and anti-inflammatory effects, as wellas increasing HDL-c, lowering blood pressure and improvingendothelial functions (Abbe &Amin, 2008b; Abbe &Amin, 2013;Kurlandsky et aI., 2006; Visioli et aI., 2011).

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Male New Zealand White rabbits were fed with a specially designedcholesterol-rich diet for 12 weeks to study the effects of CP onplasma lipids concentrations, atherosclerotic plaque formationand other biomarkers and parameters. Rats were not used as theanimal model because of their resistance or lower sensitivity tothe atherogenic effects of a cholesterol-rich diet. The phenoliccontent of CP was about 70 mg of ECE/g extract as assayed byFolin-Ciocalteu method. Based on our previous findings, CP isrich in polyphenol content (Ruzaidi et al. 2005; Amin et al. 2004a;Amin et ~l. 2004b). The dosage used in this study was. based onan aforementioned study which stated that about 2610 mg of totalpolyphenol showed significant increase in the lag phase of LDLto be oxidized ex vivo (Baba et al. 2000). Our findings showedthat the administration of CP did not only stabilize the plaque,but also reduced the area of aorta occupied with atheroscleroticlesions (Figure 16- 20). Furthermore, the present study showedthat, administration ofCP to hypercholesterolemic rabbits reducedapproximately 50% of total area of atherosclerotic lesions comparedto that in the untreated hypercholesterolemic rabbits (CP group).The development of atherosclerotic lesion is dependent on theaccumulation of fatty streaks. Fatty streaks are developed by the.~.. .

scavenging process of oxidized lipid molecules (e.g. LDL-c) bymacrophages (monocytes). The results indicated that polyphenol-rich extract prepared from Malaysian cocoa has potential to preventthe risk of atherosclerosis (Faizul, 2007).

We also conducted a study to determine the effects of Malaysiancocoa, in beverage form, on biochemical parameters among healthysubjects in UPM. A randomized and cross-over design studyinvolving 37 living and healthy subjects was carried out over 9weeks. Our results showed that plasma total cholesterol of subjectsin the treatment group decreased (p<0.05) whereas their antioxidant

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enzymes increased significantly (p<0.05) at the end of the study(Suryati, 2010). The consumption of 18 g of cocoa beverage dailyfor 9 weeks had improved several biomarkers associated withcardiovascular diseases. Thus, Malaysian cocoa may possibly helpin preventing the promotion of oxidative stress.

Figure 16 Intimal Surfaces of the Aorta from Control Rabbit (CNgroup) showing Sudan IV Stainable Lipid Deposits. There was no

Atherosclerotic Lesions as shown by these staining.(Source: Faizul, 2007)

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Figure 17 Intimal Surfaces of the Aorta from HypercholesterolemicRabbit (CP group) showing Lipid Deposit Stain with Sudan IV

Atherosclerotic Lesions were indicated by Concentrated Brick RedColors.

(Source: Faizul, 2007)


Figure 18 Intimal Surfaces of the Aorta from HypercholesterolemicRabbit Treated with Cocoa Extract (300 mg CE group) showing LipidDeposit Stained with Sudan IV Atherosclerotic Lesions were indicated

by Concentrated Brick Red Colors.

Figure 19 Intimal Surfaces of the Aorta from HypercholesterolemicRabbit Treated with Cocoa Extract (600 mg CE group) showing LipidDeposit Stain with Sudan IV Atherosclerotic Lesions were indicated by

Concentrated Brick Red Colors.(Source: Faizul, 2007)

Figure 20 Intimal Surfaces of the Aorta from HypercholesterolemicRabbit Treated with Cocoa Extract (800 mg CE group) showing LipidDeposit Stain with Sudan IV Atherosclerotic Lesions were indicated by

Concentrated Brick Red Colors.

(Source: Faizul, 2007)

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Cocoa polyphenols and ObesityCocoa, the fruit of the Theobroma cacao plant, is traditionally used infolk medicine as a pharmaceutical for blood pressure reduction andcardiovascular disease prevention. The nuclear receptor peroxisomeproliferator-activated receptor gamma (PPAR-y) is widely known toimprove insulin sensitivity and is therefore being used as a majordrug target for the treatment of type 2 diabetes mellitus. A study wasdone to investigate the anti-diabetic/anti-obesity effects of cocoapolyphenol-rich extract (CP) (Farhana et al., 2015). The rats receivedeither normal diet, high-fat diet or high-fat diet with additionalcocoa polyphenols for 8 weeks. At the end of the treatment, bodyweight, plasma glucose and insulin were measured. Additionally,mRNA and protein levels of PPARy in skeletal muscle and whiteadipose tissue were also measured. Compared to the high-fat dietgroup, increases in body weight, plasma glucose and insulin weresignificantly suppressed for the CP-treated groups. Compared tothe high-fat diet group, the PPARy mRNA level was significantlyhigher in both the skeletal muscle and the white adipose tissue forthe CP groups (Figure 21 & 22). Protein expression ofPPARy in theCP groups was also significantly higher as compared to the high-fat diet group. The anti-diabetic mechanism of CP along with themetformin hypoglycemic drug partially attributed to the increasedexpression of PPARy in the skeletal muscle and adipose tissues.These results suggest that CP could be a useful phytomedicine agentfor alleviating insulin resistance.

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PPARy

•1.2

1

~ 0.8Cl ..ul!.. E.~ i!cO. ~0.6-;.8'00co: 0.,c-

8:: 0.4,

0.2

oN Ob Ob-db Ob-db + cocoa Ob-db +

metforminGroup orralS

Figure 21 mRNA levels PPAR-y in the adipose tissue of differentgroups of rats

(Source: Farhana et al., 2015)

The mean values of cDNA copies were corrected with respect to thereference gene (~-actin). The presence of a single RT-PCR product wasverified by 2% gel electrophoresis for its specificity. * P< 0.05 vs normal(N).

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PPAR-y

Beta actin

180

160

2"140'8~'C 120.2..~ 100

=e 80i...~ 60jOJ

40II:

20

0control(N)

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kDa

kDa

Ob Ob-db + cocoa Ob-db+metformin

Ob-db

Group orrall

Figure 22. Expression ofPPAR-y protein in rat adipose tissue

(Source: Farhana et al., 2015)

The protein extracts were subjected to immunoblotting by the PPAR-yantibody, and levels were normalized y-actin. Levels in the control werearbitrarily assigned a value of 100.0. All values are significantly differentat the level of P< 0.05. Protein contents were determined using thebicinchoninic acid method with BSA as a standard. * P< 0.05 vs normal(N).

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Development of obesity diseases due to dietary factors, particularlya high fat intake, is of increasing global concern. This study wasconducted to investigate the pharmacological activities of cocoapolyphenols (CP) on visceral obesity in diet-induced obese ratsand the possible mechanisms involved. Sprague-Dawley (SD)rats were fed a low-fat (LF) or high-fat (HF) diet for 10 weeks.Thereafter, the HFD rats (n=lO/group) were treated with a dose of600 mg/kg b.wlday CP (HFD+CPs) for 4 weeks. Lipid contents andmorphological changes in the liver and mesenteric white adipose

l~

tissue ~ere analyzed. The gene encoding AMPK-activated proteinkinase and protein expression of phosphorylated (AMPK aJ~) weremeasured using real time-PCR and western blotting. The mRNAexpression level oflipogenic key enzymes (Acaca, Fasn, Meat andScd-1) and lipolysis key enzymes (CPT!, Acox1) were investigated.In addition, upstream transcription factors (PPARu, PPARy, ClEBPu and SREBP-1c) were also examined.

Our findings showed that CP treatment improved visceraladiposity, adipocytes hypertrophy and liver steatosis inducedby chronic HFD feeding in rats (Ali et al., 2015). AMPKaJ~phosphorylation in the liver and adipose tissue ofHFD+CP-treatedr~ts was activated compared with the HFD-fed rats. The expressionof lipogenesis related-genes were found to have decreased whileexpression level of oxidation -related genes was increased comparedwith that in the HFD-fed rats (Figures 23- 25). The results partiallyunraveled the ameliorative effect of CP treatment on obesitybiomarkers by inhibiting lipogenesis and promoting lipolysisthrough activation of the AMPK pathway (Ali et al., 2015).

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700 --LFD

600. ..!::! : ---HFD

S 500l:of 400>....0

1%1 300

200

100

02 3 4 5 6 7 a 9 10

FHding peroids (W«ks)

700

~6005:. SOO10~;400f"111300

200

100

0LFD

••

HFD HFD+CPs

Figure 23 Effects ofCP treatment on body weight ofHFD-inducedobese rats

(Source: Ali et al., accepted for publication)

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LFD HFD HFD+CPs


A) Visceral adipose tissue

B) Liver

LFD HFD HFD+CPs

35

30

3 25

.§, 20Ji;1:

15

10

5

0LFD HFD HFD+CPs

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C) Epididymal adipose tissue

LFD HFD HFD+CPs

20

18

15

14

0lil2

.£,w..,:< 8

5

4

0LFD HFD HFD+CPs

Figure 24 Effect of Cl' treatment on adipose tissues and liver mass inHFD-induced obese rats

(Source: Ali et aI., accepted for publication)

CA)Whole visceral adipose tissue; CB)Liver; and Cc) Epididymal adiposetissue were obtained from rats after fasting overnight at the end of thestudy and weighed and photographed. L, liver; MES-WAT, mesentericwhite adipose tissue.

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(A)

3.5

g.;;

3~Il§.. 2.5.~10~

2

1.5

(B)

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1

0.5

oScdl Acoxl PPARCl Fasn CPTI Prkaal

LFD HFD HFD+CPs

p-AMPK-al

p-AMPK-a2

GAPDH protein

CGAPOH

·LFD

·HFO+CPs

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(C)

2.5p-AMPK-a2 protein

0.5

olFD HFD HFD+CPs

p-AMPK-fli

p-AMPK-fl2

Figure 25 Effect of CP on mRNA expression of lipid metabolism-related genes in liver and AMPK protein expression

(Source: Ali et al., accepted for publication)

(A) The values of gene levels were normalized to the value of GAD PH,which was set to 1. (B) Liver tissue was homogenized and the lysates weresubjected to western blot analysis for AMPK-~1/2. (C) The intensity ofeach band was quantified using western bolt densitometry measurement.

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Molecular Mechanisms Underlying the HealthBenefits of Cocoa PolyphenolsCocoa polyphenols (CP) have been shown to exhibit hypolipidemicactions, suggesting that CP holds great promise for correctionof lipid abnormalities. Indeed, recent research demonstrates thebeneficial effect of CP on obesity and its mediation of bloodpressure, insulin resistance and vascular and inflammation relateddiseases. Although still debatable, there is a range of potentialmechanisms through which CP might exert its benefits on obesityhealth. This work investigated the hepatic genetic expressionpatterns in high-fat diet (HFD)-induced obese rats using themicroarray system. Rats were fed either a low fat or HFD for 12weeks. After diet intervention, HFD rats (n= 10/group) were treatedwith 600 mglkg bw/day CP (HFD+CP) for 4 weeks. As comparedto the HFD group those receiving CP treatment showed significantdecrease in lipids in the liver and in body weight as well as visceralfat accumulation. DNA microarray analysis resulted in a differentialexpression of 862 genes out of the 12,282 genes expressed in theliver. The differential expression patterns of selected genes wereconfirmed with real-time-PCR. Metabolic pathway analysis viabioinformatics tools showed that in the CP group the genes inlipid lipolysis, primarily in fatty acid oxidation, were up-regulatedwhereas the genes in lipid synthesis pathways were down-regulated(Figure 26). Our data provides new insights into the possiblemechanisms behind the actions of CP on the management of obesitymarkers in rats fed a HFD (Figure 27).

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,___._ ••••••• ._.._'08!958L-..j_ ••••••• ~"_P111159i'd$;_-.J

••••••• A_6._POl"'9~

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r-r-:__ ••••••••• _6._'Ot0989(C!P.alt.GC!l3SI)

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••••••• A_"_''"''''••••••• ~.3_PIIl1'(PIl1g")

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••••••• A_"_Pl.>155.......~.._'''' ..,(_••••••• ~ .. _PS5)tl.L-..j_ ••••••• ~6._po;"I1~~

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~ c <> c c

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~ ~ . .StltdtltC'ff'Itlftt. _~W{2l(upancl~~Gfll,"inallof2condLC""'" CE" _

GtntUlt ""'_""tlX ....__ gtft ..... ctl,.,.._o~

Figure 26 Hierarchical clustering of HFD and HFD + CP treated ratgroups based on micro array gene expression profiling

(Source: Ali et al., 2015)

Heat maps were obtained using the Cluster-Tree view program withinthe GeneSpring GX (Agilent) platform. Red = upregulation, green =downregulation and black = unchanged. The 862 differentially expressedgenes according to their relative expression levels and ontology.

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Lipogenic genes +FA synthesis

1/TG accumulation '"

Figure 27 Possible molecular mechanisms of the obesity attenuatingeffect of cocoa polyphenols (Cl') in the liver

PPARs, peroxisome proliferator-activated receptors alpha; LXRs,liver X receptors alpha; PPRE, peroxisome proliferator response

element; LXRE, liver X receptor response element; FA, fatty acid; TG,triglyceride. The symbols -Lt .. represent inhibition, up-regulation and

down-regulation, respectively,

(Source: Ali et al., 2015)

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CONCLUSIONSOur scientific studies, as well as those of others, have clearly shownthat the consumption of cocoa and its derived products whichcontain antioxidant polyphenols could provide protection againstdiet-related diseases. Malaysian cocoa polyphenols have antioxidantand non-antioxidant properties that endow them with various healthbenefits against several pathological disorders. In addition, cocoapolyphenol extracts may possess potential hypoglycaemic andhypocholesterolaemic properties and contribute to prevention ofthe risk of atherosclerosis, enhancement of antioxidant enzymesand modulation of bone metabolism through its anti-inflammatoryproperties and exert anti-inflammation effects towards oxidativestress.

In our current research, we have used comprehensivetranscriptomic analysis to understand the molecular mechanismsbehind the ameliorative effects of cocoa polyphenols. Cocoapolyphenols may hold great promise for therapeutic applicationssuch as in the treatment oflipid abnormalities and as lipid-loweringagents to attenuate obesity-associated diseases inhumans. However,the research needs to go further in establishing the effects ofpolyphenols from cocoa on carbohydrate and fat metabolism.

Collectively, in an animal model of diet-induced obesity, cocoapolyphenol administration improved some metabolic parametersand decreased both adiposity and body weight gain. Throughmicro array and qPCR analysis, genome-wide profiling of mRNAexpression in the liver and visceral adipose tissue showed that cocoapolyphenol treatment increased the expression of genes involved infatty acid oxidation while decreasing the expression of lipogenicgenes. However, further research is needed on the regulation ofsuch nuclear receptors by cocoa polyphenols to determine whichspecific polyphenols (flavonols or phenolic acids) are involved.

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Consumption of cocoa and cocoa-derived products in the formof chocolate (milk or dark) or as an ingredient in foods maysignificantly contribute to human intake of dietary polyphenolsand health. It must however be noted that high sugar content andthe presence of other compounds may hinder the healthful effectsof cocoa. Moderate intake of cocoa products (dark chocolate) ishighly recommended owing to its high antioxidant polyphenolsand low sugar content. Furthermore, our studies did not showconclusive proof of the most effective dose of cocoa consumptionthat ~:omotes beneficial effects towards our health;

It should be noted that limitations in nutrition research areunavoidable due to the challenges of time, confounding factors andthe high cost of arrays, which can prevent large-scale studies. In theresearch described in this lecture series, an important mechanismthat is still not fully understood is whether cocoa polyphenolsonly have a local effect in the tissues which are tested or whetherthey also affect other tissues. Another key question is whether theimpacts of the cocoa polyphenol treatment on lipid metabolismare particularly mediated solely by the activation of transcriptionregulatory factors or also include other biological factors. In~ddition to. these challenges, determining the definite mechanismin living organisms is highly complicated. The complex process invivo involves many variations due to different genetic backgroundsbetween animal models and/or hormonal feedback due to dietsupplementation. These physiological factors could undeniablyinfluence the genome-wide expression of metabolic genes involvedin lipid metabolism. Therefore, an analysis of a wide array of genesin the genome under different conditions and a single polyphenolhas to be conducted in the future.

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REFERENCESAbbe Maleyki, M. J., Azrina, A., Amin, I. (2007). Assessment of

antioxidant capacity and phenolic content of selected commercialbeverages. Malaysian Journal of Nutrition, 13(2), 149-159.

Abbe Maleyki, M. J., Amin, I., Chong, P. P., Muhajir, H., Syed Hasbullah,S. K. (2008a). Effects of cocoa extract on glucometabolism, oxidativestress, and antioxidant enzymes in obese-diabetic (Ob-db) rats. Journalof Agricultural and Food Chemistry, 56, 7877-7884.

Abbe Maleyki, M. J., Amin, I. (2008b). Polyphenols in cocoa and cocoaproducts: Is there a link between antioxidant properties and health? -Review. Molecules, 13(9),2190-2219.

Abbe Maleyki, M. J., Amin, I., Chong, P. P., Muhajir, H., Syed Hasbullah,S. K.' (2009). Effects of cocoa extract containing polyphenols andmethylxanthines on biochemical parameters of obese-diabetic rats.Journal of the Science of Food and Agriculture, 88, 130-137.

Abbe Maleyki, M. J., Amin I. (2010). Antioxidant properties of cocoapowder. Journal of Food Biochemistry, 34, 111-128.

Abbe Maleyki, MJ., Amin, I. (2013). Cocoa and Cocoa-derived Products:Dietary Polyphenols and Beyond. Polyphenols: Chemistry, DietarySources and Health Benefits. Nova Science Publishers, New York,USA. ISBN: 978-1-62081-809-1.

Ali, E, Ranneh, Y., Amin, I., Norhaizan, M. E. (2014a). Identificationof phenolic compounds in polyphenols-rich extract of Malaysiancocoa powder using the HPLC-UV-ESI-MSIMS and probing theirantioxidant properties. Journal of Food Science Technology; DOl10.1007/s13197-013-1187-4.

Ali, E,Amin, I., Kersten, S. (2014b). Molecular mechanisms underlyingthe potential antiobesity-related diseases effect of cocoa polyphenols- Review. Molecular Nutrition and Food Research, 58(1), 33-48.

Ali, E, Amin, I., Norhaizan, M. E., Chong P. P. (2015). Transcriptomicsexpression analysis to unveil the molecular mechanisms underlyingthe cocoa polyphenol treatment in diet-induced obesity rats. Genomics,105(1),23-30.

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Amin, I., Tan, S.H. (2002). Antioxidant activity of selected commercialseaweeds. Malaysian Journal of Nutrition, 8, 167-177.

Amin, I., Jinap, S., Jamilah, B., Harikrishna, K., Biehl, B. (2002a). Analysisofvicilin (7S)-class globulin in coco cotyledons from various genetic.origins. Journal of the Science of Food and Agriculture, 82,728-732.

Amin, I., Jinap, S., Jamilah, B., Harikrishna, K. and Biehl, B. (2002b).Oligopeptide patterns produced from Theobroma cacao L of variousgenetic origins. Journal of the Science of Food and Agriculture, 82,733-737.

Amin, I., Cheah, S.E (2003). Determination of vitamin C, ~-caroteneand riboflavin contents in five green vegetables 'organically andconventionally grown. Malaysian Journal of Nutrition, 9,31- 39.

Amin, I., Zamaliah, M.M., Chin, W.E (2004a). Total antioxidant activityand phenolic content in selected vegetables. Food Chemistry, 87,581-586.

Amin, I., Koh, B.K., Asmah, R. (2004b). Effect of cacaa liquor extract ontumor marker enzymes during chemical hepatocarcinogenesis in rats.Journal of Medicinal Food, 7(1),7 - 12.

Amin, I., Faizul, H.A., Azli R. (2004c). Effect of cocoa powder extracton plasma glucose levels in hyperglycaemic rats. Nutrition and FoodScience, 34(3), 116 - 121.

Amin, I., Lee, w.Y. (2005). Effect of different blanching times on.. antioxidant properties in selected cruciferous vegetables. Journal of

the Science of Food and Agriculture, 85, 2314-2320.

Amin, I., Norazaidah, Y., Emmy Hainida, K. I. (2006). Antioxidantactivity and phenolic content of spinach species (Amaranthus sp.).Food Chemistry, 94,47-52.

Amin I., Mukrizah, O. (2006). Antioxidant capacity of methanolic andwater extracts prepared from food processing by-products. Journal ofthe Science of Food and Agriculture, 86, 778-784.

Amin, I., Azrina, A., Khoo, H. E., Prasad, N., Kong K. W. (2013).AntioxidantAssays: Principle, Theory &Techniques.l!niversity PutraMalaysia Pr~~s. ' ..

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Andres-Lacueva, C., Monagas, M., Khan, N., Izquierdo-Pulido, M., Urpi-Sarda, M., Permanyer, J., Lamuela-Raventos, R.M. (2008). Flavanoland flavonol contents of cocoa powder products: Influence of themanufacturing process. Journal of Agricultural and Food Chemistry,56, 3111~31l7.

Azizah, 0., Amin, I., Nawalyah, A. G., Ilham, A. (2007). Antioxidantcapacity. and phenolic content of cocoa beans. Food Chemistry, 100,1523-1530.

Azrina, A., Nurul Nadiah, M. N., Amin, I. (2010) Antioxidant propertiesof methanolic extract of Canarium odontophyllum fruit.1nternationalFood Research Journal, 17, 319-326.

Baba, S., Osakabe, N., Natsume, M., Yasuda.A; Takizawa, T., Nakamura,T., Terao, J. (2000). Cocoa powder enhances the level of antioxidativeactivity in rat plasma. British Journal of Nutrition, 84, 673-680.

Barry Callebaut. History of chocolates. 2008; http://www.barry-callebaut.cornl1589.

Borchers, A. A.T., Keen, C.L., Hannum, S.M., Gershwin, M.E. (2000).Cocoa and chocolate composition, bioavailability, and healthimplications. Journal of medicinal Food, 3, 77-105.

Boyer, J.,Brown, D., Liu, R. H. (2004). Uptake of quercetin and quercetin3-glucoside from whole onion and apple peel extracts by Caco-2 cellmonolayers. Journal ofAgricultural andFood Chemistry, 52, 7172-79.

Bravo, L. (2000). Polyphenols: chemistry, dietary sources, metabolism,and nutritional significant. Nutrition Review, 56, 317-333.

Buijsee, B., Weikert, C., Drogan, D., Bergmann, M., Boieng, H. (2010).Chocolate consumption in relation to blood pressure and risk ofcardiovascular disease in German adults. European Heart Journal,31,1616- 23

Cheng,C. M., Abbe Maleyki, M. J., Amin I. (2009). Phenolic andtheobromine contents of commercial dark, milk and white chocolateson the Malaysian markets. Molecules, 14,200-209.

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Chew, L.Y., Khoo, H.E., Amin, I., A Azrina, A, C.Y. Lau (2011). Analysisof phenolic compounds of dabai (Canarium odontophyllum Miq.)fruits by high performance liquid chromatography. Food AnalyticalMethod, 5(1), 126- 137.

Dillinger, T.L., Barriga, P., Escarcega, S., Jimenez, M., Lowe, D.S.,Grivetti, L.E. (2000). Food of the Gods: Cure for humanity? Acultural history of the medicinal and ritual use of chocolate. Journalof Nutrition, 130(8), 2057S-20nS.

Emmy Hainida, K. I., Amin, I., Normah, H., Norhaizan, M. E., Ainul, Z.(2008). Effects of defatted dried roselle (Hibiscus sabdariffa L.) seedpowder on lipid profiles of hypercholesterolemia rats. Journal of theScience of Food and Agriculture, 88,1043-1050.

Emmy Hainida, K. I., Khoo, H. E., Abbe Maleyki, M. 1.,Amin, I., Salma, I.,Azrina, A., Halimatul Saadiah, M. N., Norzatol Akmar, M. D., RuzaidiAzli, M. M. (2009). Antioxidant capacity and total phenolic contentof Malaysian underutilized fruits. Journal of Food Composition andAnalysis, 22, 388-393.

Faizul, H.A (2007). Effect of polyphenol extract prepared from Malaysiancocoa powder on atherosclerosis. Master Thesis, UPM.

Farhana, A., Ali, E, Amin, I., Chong, P. P., Muhajir, H. (2015). Cocoapolyphenol- rich extract enhances the expression level ofPPAR-y inskeletal muscle and adipose tissue of obese-diabetic rats fed a high- fatdiet. International Journal of Pharmacology, 10, 3923.

Fouad, A H., Amin, I., Azizah, A. H., Azrina, A (2011). Identificationand quantification of phenolic compounds in bambangan (Mangiferapajang Kort.) peels and their free radical scavenging activity. Journalof Agricultural and Food Chemistry, 59, 9102-9111.

Hasnah, H., Amin, I., Suzana, S., Azrina, A., Loh, S. P. (2010). Apparentbioavailability ofisoflavones in urinary excretions of postmenopausalMalay women consuming tempeh compared with milk. InternationalJournal of Food Science and Nutrition, 62(60), 642-650.

Hasnah, H., Amin, I., Azrina, A., Suzana, S., Loh, S. P. (2009). Daidzeinand gene stein contents in tempeh and selected soy products. FoodChemistry,115, 1350-1356. .

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Hollman, P. C. H., Cassidy A., Comte, B., Heinone, M., Richelle, M.,Richling, E., Seferani, M., Scalbert, A., Sies, H., Vidry, S. (2011).The biological revalance of direct antioxidant effect of polyphenolsfor cardiovascular health in human is not established. The Journal ofNutrition, 141(5), 989S-1009S.

Hooper, L., Kay, C., Abdelhamid, A., Kroon, P. A., Cohn, J. S., Rimm, E.B., Cassidy, A. (2012). Effects of chocolate, cocoa, and flavan-J-olson cardiovascular health: a systematic review and meta-analysis ofrandomized trials. The American Journal of Clinical Nutrition, 95,740- 51.

Jacques, P. E, Cassidy, A., Rogers, G., Peterson, J. J., Meigs, J. B., Dwyer,J, T. (2013). Higher dietary flavonol intake is associated with lowerincidence of type 2 diabetes. The Journal of Nutrition, 143(9), 1474-1480.

Khoo, H. E., Amin, I. (2008). Determination of daidzein and genisteinin Mangifera fruit. Malaysian Journal of Nutrition, 14(2), 189-198.

Khoo, H. E., Prasad, K. N., Kong, K. W.,Amin I. (2011) Carotenoids andits isomers in fruits and vegetables. Molecules. 16, 1710-1738.

Kong, K.W., Hock, E. K., Prasad K. N., Amin I., Chin, P. T., Nor ER.(2010a). Revealing the power of the natural red pigment Iycopene.Molecules. 15,959-987. .

Kong, K.W., Abdu,1 R. 1.1, Seok, T., Prasad K. N., Amin I. (2010b).Response surface optimization of extraction for phenolics andflavonoids from pink guava puree industrial refinery by-product.InternationalJournal of Food Science and Technology, 45, 1739-1745.

Kong, K.W., Chew, L.Y., Lau, C.Y., Prasad, K.N., Sun, J., Amin, I.,Hosseinpoursarinadi, B. (2011). Nutritional constituents andantioxidant properties of Malaysian indigenous kembayau fruits(Dacryodes rostrata) in Sarawak. Food Research International, 44,2332-2338.

Kurlandsky, S. B., Stote, K. S. (2006). Cardioprotective effects of chocolateand almond consumption in healthy women. Nutrition Research,26(10),509-516.

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Lee, W. Y., Emmy Hainida, K. I., Abbe Maleyki, M. 1.,Amin, I. (2007).Antioxidant capacity and phenolic content of selected commerciallyavailable cruciferous vegetables. Malaysian Journal of Nutrition,13(I),71-S0.

Manach, C., Williamson, G., Morand, C., Scalbert,A, Remesy, C. (2005).Bioavailability and bioefficacy of poly-phenols in humans. Review of97 bioavailability studies. The American Journal of Clinical Nutrition,81(1),230S-242S.

MarinaAB., Yaakub B.C.M., Nazimah S.AH,Amin I. (2009). Antioxidantcapacity and phenolic acids of virgin coconut oil. International Journalof Food Science and Nutrition, 60(1),114-123. ...

Nobutomo, I., Takahiro, T., Takehiro, 0., Kiyomi, I., Wataru, O. Kiyoshi,S. (2010). Anti-obesity and anti-diabetic effects of acacia polyphenolin obese diabetic KKAy mice fed high-fat diet Evidence-BasedComplementary and Alternative Medicine, 241,1-10.

Pandurangan,A. K., Saadatdoust, Z., Harnzah, H., Amin, I. (2015). Dietarycocoa protects against colitis-associated cancer by activating the Nrf2/Keapl pathway. BioFactors, 41(1), 1-14.

Prasad, K.N., Chew, L.Y., Kong, K.W., Khoo, H.E., Amin, I., Azlan, A.(2010). Antioxidant capacities of peel, pulp and seed fractions ofCanarium odontopyllum Miq. Fruit. Journal of Biomedicine andBiotechnology, JBB/S71379: I-S .

. Ranneh, Y., Ali, E, Fadel, A (2014). The antioxidant activity of cocoapolyphenolic extract- treated 3T3 fibroblast cells. InternationalJournal of Toxicological and Pharmacoligical Research, 6(1),14- 19.

Ruzaidi, A., Amin, I., Nawalyah, A G., Hamid, M., & Faizul, H. A (2005).The effect of Malaysian cocoa extract on glucose levels and lipidprofiles in diabetic rats. Journal ofEthnopharmacology, 98(1), 55-60.

Ruzaidi Azli, M. M., Abbe Maleyki, M. 1., Amin, I., Nawalyah, A. G.,Muhajir, H. (200Sa). Protective effect of polyphenol-rich extractprepared from Malaysian cocoa (Theobroma cacao) on glucose levelsand lipid profiles in streptozotocin-induced diabetic rats. Journal ofthe Science of Food and Agriculture, 88, .1442 -1447.

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Ruzaidi Azli, M. M., Abbe Maleyki, M. J., Amin, I., Nawalyah, A.G.,Muhajir, H., Pauliena, M.B.S.M.J., Muskinah, M.S. (2008b)Hypoglycaemic properties of Malaysian cocoa (Theobroma cacao)polyphenols-rich extract. International Food Research Journal, 15(3),305-312.

Sadeq, H. A., Amin, I.,Yazid, M. M., Shuhaimi, M., Rokiah, M. Y., Fouad,A. H. (2012). Purification, characterization and antioxidant activity ofpolysaccharides extracted from the fibrous pulp of Mangifera pajangfruits. LWT - Food Science and Technology, 48(2), 291-296.

Sarmadi, B.H., Amin, I. (2010). Antioxidative peptides from food proteins- Review. Peptides, 31, 1949-1956 ..

Sarmadi, B.H., Amin, I., Muhajir, H. (2011). Antioxidant and ACEinhibitory activities of cocoa autolysates. Food Research International,44, 290-296.

Sarmadi, B.H., Farhana, A., Muhajir, H., Nazamid, S., Azizah, A. H.,Amin I. (2012). Hypoglycemic effects of cocoa (Theobroma cacaoL.) autolysates. Food Chemistry, 134,905-911.

Sarmadi B.H. (2015). Effects of cocoa-rich diet on prevention of bone lossin ovariectomized rat model of osteoporosis. In PhD. Thesis, UPM

Sun, J., Li, C., Li, Z., Liao, E, Prasad, K. N., Amin, I. (2013). Polyphenolsin major tropical fruits of Sapindaceae family. Polyphenols: Chemistry,Dietary Sources and Health Benefits. Nova Science Publishers, NY,USA. ISBN: 978-1-62081-809-1.

Suryati, M. A. (2010). Effect of Malaysian cocoa powder beverageconsumption on biochemical parameters in healthy subjects. InMasterScience Thesis, UPM.

Tan, S. T., Mohd, H. J., Amin I. (201 Oa).Antioxidant properties of fresh,powder and fiber products of Mangiferafoetida fruits. InternationalJournal of Food Properties. 13,682-691.

Tan, S. T., Prasad K. N., Amin, I. (201 Ob)Antioxidant capacity, phenolicsand isoflavones in soybean by-products. Food Chemistry, 123, 583-589

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Tiong, N. w., Prasad, K. N., Bao, Y., Amin I. (2010). Bioactivitysubstance contents and antioxidant capacity of raw and blanchedvegetables. Innovative Food Science and Emerging Technologies.11, 464-469.

Visioli, E, Berneart, H., Corti, R., Ferri, C., Heptinstall, S., Molinari, E.,Poli, A., Seferani, M., Smith, H. J., Vinson, J. A., Violi, E, Paoletti,R. (2009). Chocolate, lifestyle, and health. Critical Reviews in FoodScience Nutrition, 49, 299-312.

Visioli, E, Lastra, C. A. D. L., Andres- Lacueva, C., Aviram, M., Calhau,C., Cassano, A., Archivo, M. D., et. al. (2011). Polyphenols and Humanhealth: A prospectus, Critical Reviews in Food Science and Nutrition,51(6),524-546.

Zabidah, A. A., Kong, K.w., Amin, I. (2011). Antioxidant properties oftropical juices and their effects on in vitro hemoglobin and low densitylipoprotein (LDL) oxidations. International Food Research Journal,18, 545-552.

Zabidah, A. A., Fouad, A. H., Amin, I., Barakatun Nisak, M. Y., Muhajir,H. (2014). Chemical compositions, antioxidative and antidiabeticproperties of underutilized vegetable palm hearts from Plectocomiopsisgeminiflora and Eugeissona insignis. Journal of Agricultural FoodChemistry, 62(9),2077-2084.

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BIOGRAPHYProf. Dr. Amin bin Ismail was born in Kubang Pasu, Kedah on 11April 1969. Dr. Amin started his career as a tutor in July 2000 inthe Department of Nutrition and Dietetics (formerly known as theDepartment of Nutrition and Health Sciences), Faculty of Medicineand Health Sciences, Universiti Putra Malaysia. Subsequently, inDecember 2000, he was appointed as a lecturer in the department.Dr. Amin was promoted to senior lecturer in 2004 and AssociateProfessor in May 2006, in the same department. In December2010, he was appointed to the rank of full professor in the field of"Food Chemistry and Biochemistry". From2005 - 2006, he servedas the head of the Food Quality Laboratory, Centre of StrategicStudies and Food Innovation in UPM. He was then selected tolead the Laboratory of Halal Science Research (formerly knownas the Laboratory of Analysis and Authentication), Halal ResearchProducts Institute, Universiti Putra Malaysia, and was also theProgramme Coordinator for the Bachelor of Science (Nutrition andCoriununity Health) programme in the Department of Nutritionand Dietetics from 2009 - 2012. He is currently the Deputy Dean(Graduate Student Affairs & Industry and Community Relations)at the Faculty of Medicine and Health Sciences and has held thisposition since 2012.

Dr. Amin has 15 years of teaching experience and more than 19years of research experience. His abilities and full commitment to .administrative duties are well known within his department, faculty,UPM, Ministry of Health Malaysia (MOH) and other professionalbodies. At the university, national and international levels, hehas been appointed as chairman, coordinator, adviser, secretary,facilitator and member of several committees, student programs,research mentoring programmes and conferences. He has beenappointed to the Programme Advisory Panel for the Master's in

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Nutrition programme at UKM and is actively involved as a memberofthe MQA Panel for assessing undergraduate programmes relatedto nutrition and food science at various universities in Malaysia.

Dr. Amin is currently a Fellow of the Nutrition Society ofMalaysia and was an Assistant Honorary Secretary ofthe NutritionSociety of Malaysia (2001 - 2006). He is also a member of theInternational Society for Nutraceuticals and Functional Foods.

Dr. Amin's research focus areas include the health benefitsof antioxidant polyphenols from Malaysian cocoa products, the

I

utilization of food processing by-products and antioxidant andnutritional analysis. In the past, he has received several researchgrants (more than RM 1 Million) from different ministries andUPM for projects related to antioxidants and polyphenols derivedfrom fruits and cocoa, as well as the utilization of food processingby-products for functional foods. He has also collaborated withseveral research institutes and industries - the Malaysia CocoaBoard (MCB), the Forestry Research Institute of Malaysia (FRIM),the Malaysian Agricultural Research Development Institute(MARDI), Golden Hope Fruits and Beverages Industry Sdn. Bhd,Kotra Pharma (M) Sdn. Bhd, and the International Atomic Energy

..Agency (IAEA). Dr. Amin has received several research awards atthe faculty, UPM, national and international levels. He was alsoawarded the Young Scholar Award in 2005 and received a Fellowshipaward from the United Nations University in 2001.

Dr. Amin has graduated 10 PhD and 15 MSc students. Themajority ofthem have published in Q 1& Q2 journals and have wonvarious national and international awards. To date, Dr. Amin hasmore than 150 articles in peer-reviewed journals, book chapters,proceedings and research/consultancy reports. He has had twopatents issued and several pending patents. Since 2000, he haspresented more than 100 papers at national and international

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conferences. Dr. Amin's publications have been cited more than2,331 times, and as of May 2015 his h-index is 24 (based onSCOPUS).

Dr. Amin was appointed as an Editorial Board member for"Food Chemistry" (a Q1journal, published by Elsevier) from 2007till20 13. Currently, he is an Associate Editor for "Food Chemistry."Known for his expertise in polyphenols and antioxidant research,Dr. Amin has reviewed more than 200 manuscripts being consideredfor publication in peer-reviewed journals (more than 60 manuscriptsfor lSI journals) from 2004 - 2014. He has been appointed as anexternal examiner by both local and foreign universities (Universityof New South Wales, University of Newcastle, University of My soreand University of Madras) for MS and PhD thesis adjudicationand academic promotion. Since 2012, he has been appointed asa Visiting Researcher at the Guangxi Academy of AgriculturalSciences, Nanning, China.

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Amin Ismail

ACKNO\VLEDGEMENTSFirst and foremost, thank you to Allah AI-Mighty for the blessings

on all my professional work as an academician.

The realization of this inaugural lecture would not have beenpossible without the support and contribution of many individualswho provided their kind help in one way or the other.

My appreciation to Prof. Dato' Dr. Mohd Fauzi Hj. Ramlan,the Vice Chancellor; Prof. Dr. Abdul Jalil Nordin, the Dean; andAssociate Prof. Dr Norhaizan Mohd Esa, the Department Head, fortheir support.

My heartfelt gratitude and appreciation goes to all my formergraduate students (Dr. Faisal Ali, Dr. Bahareh, Dr. Aida Hamimi,Dr. Chang Sui Kiat, Dr. Fouad Hassan, Dr. Sadeq Hassan, Dr.Tan Seok Tyug, Dr. Hasnah Haron, Dr. Muhammad Ibrahim, Dr.Nurulhidayah, Faizul Helmi, Emmy Hainida, Ruzaidi, Azizah,Kong Kin Weng, Khoo Hock Eng, Chew Lye Yee, Abbe Maleyki,Suryati, Naeem, Hanaffi, Alia, Aina, Nur Azira, Nadia and Zabidah)and my current students for their contributions to my academic andresearch activities.

I express my deep gratitude and heartfelt thanks to my Ph.D.supervisor, Prof. Dr. Jinap Selamat, for her incessant guidance andcoaching in research when I was her student. I am thankful to myinternational colleagues, Prof. Dr. Feereidon Shahidi from Canada,Prof. Dr. Chin Kun Wang from Taiwan and Prof. Dr. Manohar Gargfrom Australia, for their continuous support since our first meetingin 2000.

My special gratitude goes to Prof. Dr. Norlijah Othman(former Dean), all colleagues, academic and non-academic staff,at the Department of Nutrition and Dietetics, for their support andcooperation. I bow and doff my cap to all of you. Special thanks

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goes to Prof. Dr. Maznah Ismail, the former department head,for her continuous support and motivation when I first joined thedepartment in 2000. I would not have been a member of UPM ifit had not been for her decision.

My heartfelt appreciation for the love, continuous support,encouragement and great patience of my wife Fatimah Omar andthe love and endurance of my children Amir Farhan, Amir Safuan,Alya Izzati and Amir Haziq. I am also grateful to my mother, CheRokiah Md Isa, my late father, Ismail Dee, my sisters, brothers and

I

in-laws for their love, unconditional support and doa.I also take this opportunity to thank all my colleagues/

collaborators at the)Faculty of Medicine and Health Sciencesand other facult~nd institutes for their continuous support andknowledge shanng. Special thanks goes to my former lecturers atthe Faculty of Food Science and Technology (Prof. Dr. Jinap, Prof.Dr. Hasanah, Prof. Dr. Jamilah, Prof. Azizah, Prof Nazamid, Assoc.Prof Dr. Kharidah, Prof. Russly, Prof. Dato' Mohd. Yazid) for theirguidance and encouragement when I was enrolled with the facultyin 1989 as an undergraduate student.

I would also like to thank the CoSComm, UPM Press and the.. special committee headed by Dr. Nurul Husna Shafie who workedvery hard to ensure the success of this inaugural event - Thank you

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LIST OF INAUGURAL LECTURES

1. Prof. Dr. Sulaiman M. Yassin 9. Prof. Ir. Dr. Mohd. Zohadie BardaieThe Challenge to Communication Engineering TechnologicalResearch in.Extension Developments Propelling Agriculture22 July 1989 into the 21st Century

28 May 19942. Prof. Ir. Abang Abdullah Abang Ali

Indigenous Materials and Technology 10. Prof. Dr. Shamsuddin Jusopfor Low Cost Housing Rock, Mineral and Soil30 August 1990 18 June 1994

3. Prof. Dr. Abdul Rahman Abdul Razak II. Prof. Dr. Abdul Salam AbdullahPlant Parasitic Nematodes, Lesser Natural Toxicants Affecting AnimalKnown Pests of Agricultural Crops Health and Production30 January 1993 29 JUlie 1994

4. Prof. Dr. Mohamed Suleiman 12. Prof. Dr. Mohd. YusofHusseinNumerical Solution of Ordinary Pest Control: A Challenge in AppliedDi.I.forential Equations: A Historical EcologyPerspective 9July 1994I I December 1993

13. Prof. Dr. Kapt. Mohd. Ibrahim Haji5. Prof. Dr. Mohd. Ariff Hussein Mohamed

Changing Roles of Agricultural Managing Challenges in FisheriesEconomics Development through Science and5 March 1994 Technology

23 July 19946. Prof. Dr. Mohd. Ismail Ahmad

Marketing Management: Prospects 14. Prof. Dr. Hj. Amat Juhari Moainand Challenges for Agriculture Sejarah Keagungan Bahasa Melayu6April1994 6 August 1994

7. Prof. Dr. Mohamed Mahyuddin Mohd. IS. Prof. Dr. Law Ah TheemDahan Oil Pollution in the Malaysian SeasThe Changing Demandfor Livestock 24 September 1994Products20 April 1994 16. Prof. Dr. Md. Nordin Hj. Lajis

Fine Chemicals from Biological8. Prof. Dr. Ruth Kiew Resources: The Wealth from Nature

Plant Taxonomy, Biodiversity and 21 January 1995ConservationII May 1994 17. Prof. Dr. Sheikh Omar Abdul Rahman

Health, Disease and Death inCreatures Great and Small25 February 1995

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18. Prof. Dr. Mohamed Shariff MohamedDinFish Health: An Odyssey through theAsia - Pacific Region25 March 1995

19. Prof. Dr. Tengku Azmi Tengku IbrahimChromosome Distribution andProduction Performance of WaterBuffaloes6 May 1995

20. Prof. Dr. Abdul Hamid MahmoodBahasa Melayu sebagai Bahasa Ilmu-Cabaran dan Harapan10 June 1995

21. Prof. Dr. Rahim Md. sail)Extension Educationfor; .Industrialising Malaysia: Trends.Priorities and Emefging Issues22 July 1995

22. Prof. Dr. Nik Muhammad NikAbd.MajidThe Diminishing Tropical Rain Forest:Causes. Symptoms and Cure19August 1995

23. Prof. Dr. Ang Kok JeeThe Evolution of an EnvironmentallyFriendly Hatchery Technology forUdang Galah, the King of FreshwaterPrawns and a Glimpse into the Futureof Aquaculture in the 21st Century14 October 1995

24. Prof. Dr. Sharifuddin Haji AbdulHamidManagement of Highly Weathered AcidSoils for Sustainable Crop Production28 October 1995

25. Prof. Dr. Yu Swee YeanFish Processing and Preservation:Recent Advances and FutureDirections •.9 December 1995

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26. Prof. Dr. Rosli MohamadPesticide Usage: Concern and Options10 February 1996

27. Prof. Dr. Mohamed Ismail AbdulKarimMicrobial Fermentation andUtilization of AgriculturalBioresources and Wastes in Malaysia2 March 1996

28. Prof. Dr. Wan Sulaiman Wan HarunSoil Physics: From Glass Beads toPrecision Agriculture16 March 1996 .,

29. Prof. Dr. Abdul Aziz Abdul RahmanSustained Growth and SustainableDevelopment: Is there a Trade-Off 1orMalaysia13 April 1996

30. Prof. Dr. Chew Tek AnnSharecropping in PerfectlyCompetitive Markets: A Contradictionin Terms27 April 1996

31. Prof. Dr. Mohd. Yusuf Sulaiman. Back to the Future with the Sun18 May 1996

32. Prof. Dr. Abu Bakar SallehEnzyme Technology: The Basis forBiotechnological Development8 June 1996

33. Prof. Dr. Kamel Ariffin Mohd. AlanThe Fascinating Numbers29June 1996

34. Prof. Dr. Ho Yin WanFungi: Friends or Foes

~ 27 July 1996

35~0f. Dr. Tan Soon GuanGenetic Diversity of Some SoutheastAsianAnimals: Of Buffaloes andGoats and FIShes Too10August 1996

36. Prof. Dr. Nazaruddin Mohd. JaJiWill Rural Sociology Remain Relevantin the 21st Century?21 September 1996

37. Prof. Dr. Abdul Rani BahamanLeptospirosis-A ModelforEpidemiology, Diagnosis and Controlof Infectious Diseases16 November 1996

38. Prof. Dr. Marziah MahmoodPlant Biotechnology - Strategies forCommercialization21 December 1996

39. Prof. Dr. Ishak Hj. OmarMarket Relationships in the MalaysianFish Trade: Theory and Application22 March 1997

40. Prof. Dr. Suhaila MohamadFood and Its Healing Power12 April 1997

41. Prof. Dr. Malay Raj MukerjeeA Distributed CollaborativeEnvironment for Distance LearningApplications17 June 1998

42. Prof. Dr. Wong Kai ChooAdvancing the Fruit Industry inMalaysia: A Need to Shift ResearchEmphasisIS May 1999

43. Prof. Dr. Aini IderisAvian Respiratory andImmunosuppressive Diseases-A FatalAttraction10July 1999

44. Prof. Dr. Sariah MeonBiological Control of Plant Pathogens:Harnessing the Richness of MicrobialDiversity14August 1999

AminIsmail

45. Prof. Dr. Azizah HashimThe Endomycorrhiza: A FutileInvestment?23 October 1999

46. Prof. Dr. Noraini Abdul SamadMolecular Plant Virology: The WayForward2 February 2000

47. Prof. Dr. MuhamadAwangDo We Have Enough Clean Air toBreathe?7 April 2000 .

48. Prof. Dr. Lee Chnoong KhengGreen Environment, Clean Power24 June 2000

49. Prof. Dr. Mohd. Ghazali MohayidinManaging Change in the AgricultureSector: The Needfor InnovativeEducational Initiatives12 January 2002

50. Prof. Dr. Fatimah Mohd. ArshadAnalisis Pemasaran Pertaniandi Malaysia: Keperluan AgendaPembaharuan26 January 2002

51. Prof. Dr. Nik Mustapha R. AbdullahFisheries Co-Management: AnInstitutional Innovation TowardsSustainable Fisheries Industry28 February 2002

52. Prof. Dr. Gulam Rusul Rahmat AliFood Safety: Perspectives andChallenges23 March 2002

53. Prof. Dr. Zaharah A. RahmanNutrient Management Strategies forSustainable Crop Production in AcidSoils: The Role of Research UsingIsotopes13 Apri12002

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54. Prof. Dr. Maisom AbdullahProductivity Driven Growth: Problems& Possibilities27 April 2002

55. Prof. Dr. Wan Omar AbdullahImmunodiagnosis and Vaccination forBrugian Filariasis: Direct Rewardsfrom Research Investments6 June 2002

56. Prof. Dr. Syed Tajuddin Syed HassanAgro-ento Bioinformation: Towardsthe Edge of Reality22 June 2002

57. Prof. Dr. Dahlan Ismail ~Sustainability of Tropical 'Animal-Agricultural Production Systems:Integrotion of Dynamic-ComplexSystems27 June 2002

58. Prof. Dr. Ahmad ZubaidiBaharumshahThe Economics of Exchange Rates inthe East Asian Countries26 October 2002

59. Prof. Dr. Shaik Md. Noor Alam S.M.HussainContractual Justice in Asean: AComparative View of Coercion31 October 2002

60. Prof. Dr. Wan Md. Zin Wan YunusChemical Modification of Polymers:Current and Future Routes forSynthesizing New PolymericCompounds9 November 2002

61. Prof. Dr. Annuar Md. NassirIs the KLSE Efficient? Efficient MarketHypothesis vs Behavioural Finance23 November 2002

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62. Prof. Ir. Dr. Radin Umar Radin SohadiRoad Safety Interventions in Malaysia:How Effective Are They?21 February 2003

63. Prof. Dr. Shamsher MohamadThe New Shares Market: RegulatoryIntervention. Forecast Errors andChallenges26 April 2003

64. Prof. Dr. Han Chun KwongBlueprintfor Transformation orBusiness as Usual? A StructurationalPerspective of the Knowledge-BasedEconomy in Malaysia31 May2003

65. Prof. Dr. Mawardi RahmaniChemical Diversity of MalaysianFlora: Potential Source of RichTherapeutic Chemicals26July 2003

66. Prof. Dr. Fatimah Md. YusotfAn Ecological Approach: A ViableOption for Aquaculture Industry inMalaysia9 August 2003

67. Prof. Dr. Mohamed Ali RajionThe Essential Fatty Acids-Revisited23 August 2003

68. Prof. Dr. Azhar Md. ZainPsychotheraphy for Rural Malays -Does it Work?13 September 2003

69. Prof. Dr. Mohd. Zamri SaadRespiratory Tract Injection:Establishment and Contral27 September 2003

70. Prof. Dr. Jinap Selamat"Cocoa-Wonders for Chocolate LoversIF~bruary 2004.

71. Prof. Dr. Abdul Halim ShaariHigh Temperature Superconductivity:Puzzle & Promises13 March 2004

72. Prof. Dr. Yaakob Che ManOils and Fats Analysis - RecentAdvances and Future Prospects27 March 2004

73. Prof. Dr. Kaida KhalidMicrowave Aquametry: A GrowingTechnology24 April 2004

74. Prof. Dr. Hasanah Mohd. GhazaliTapping the Power of Enzymes-Greening the Food IndustryII May2004

75. Prof. Dr. YusoflbrahimThe Spider Mite Saga: Quest forBiorational Management Strategies22 May2004

76. Prof. Datin Dr. Sharifah Md. NorThe Education of At-Risk Children:The Challenges Ahead26 June 2004

77. Prof. Dr. Ir. Wan Ishak Wan IsmailAgricultural Robot: A New TechnologyDevelopmentfor Agro-Based Industryl4August 2004

78. Prof. Dr. Ahmad Said SajapInsect Diseases: ResourcesforBiopesticide Development28 August 2004

79. Prof. Dr. Aminah AhmadThe Interface of Work and FamilyRoles: A Questfor Balanced LivesI I March 2005

80. Prof. Dr. Abdul Razak AlimonChallenges in Feeding Livestock:From Wastes to Feed23 April 2005

AminIsmail

81. Prof. Dr. HajiAzimi Hj. HamzahHelping Malaysian Youth MoveForward: Unleashing the PrimeEnablers29 April 2005

82. Prof. Dr. Rasedee AbdullahIn Search of An Early Indicator ofKidney Disease27 May2005

83. Prof. Dr. Zulkifli Hj. ShamsuddinSmart Partnership: Plant-Rhizobacteria Associations17 June 2005

84. Prof. Dr. Mohd KhanifYusopFrom the Soil to the TableI July 2005

85. Prof. Dr. Annuar KassimMaterials Science and Technology:Past. Present and the Future8 July 2005

86. Prof. Dr. Othman MohamedEnhancing Career DevelopmentCounselling and the Beauty of CareerGames12 August 2005

87. Prof. Ir. Dr. MohdAmin Mohd SoomEngineering Agricultural WaterManagement Towards PrecisionFraming26 August 2005

88. Prof. Dr. Mohd Arif SyedBioremediation-A Hope Yetfor theEnvironment?9 September 2005

89. Prof. Dr. Abdul Hamid Abdul RashidThe Wonder of Our NeuromotorSystem and the TechnologicalChallenges They Pose23 December 2005

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90. Prof. Dr. Norhani AbdullahRumen Microbes and Some of TheirBiotechnological Applications27 January 2006

91. Prof. Dr. Abdul Aziz SahareeHaemorrhagic Septicaemia in Cattleand Buffaloes: Are We Ready forFreedom?24 February 2006

92. Prof. Dr. Kamariah Abu BakarActivating Teachers' Knowledge andLifelong Journey in Their ProfessionalDevelopment3 March2006

93. Prof. Dr. Borhanuddin JOhd. AliInternet Unwired24 March 2006

94. Prof. Dr. Sundararajan ThilagarDevelopment and Innovation in theFracture Management of Animals31 March 2006

95. Prof. Dr. Zainal Aznam Md. JelanStrategic Feedingfor a SustainableRuminant Farming19May2006

96. Prof. Dr. Mahiran BasriGreen Organic Chemistry: Enzyme atWork14 July2006

97. Prof. Dr. Malik Hj. Abu HassanTowards Large Scale UnconstrainedOptimization20 April 2007

98. Prof. Dr. Khalid Abdul RahimTrade and Sustainable Development:Lessons from Malaysia s Experience22 June2007

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99. Prof. Dr. Mad Nasir ShamsudinEconometric ModellingforAgricultural Policy Analysis andForecasting: Between Theory andReality13 Juiy2007

100. Prof. Dr. Zainal Abidin MohamedManaging Change - The Fadsand The Realities: A Look atProcess Reengineering. KnowledgeManagement and Blue OceanStrategy9 November 2007

101. Prof. Ir. Dr. Mohamed DaudExpert Systems for EnvironmentalImpacts and Ecotourism Assessments23 November 2007

102. Prof. Dr. Saleha Abdul AzizPathogens and Residues; How Safeis Our Meat?30 November 2007

103. Prof. Dr. Jayum A. JawanHubungan Sesama Manusia7 December 2007

104. Prof. Dr. Zakariah Abdul RashidPlanningfor Equal IncomeDistribution in Malaysia: A GeneralEquilibrium Approach28 December 2007

105. Prof. Datin Paduka Dr. KhatijahYusoffNewcastle Disease virus: A Journeyfrom Poultry to CancerII January 2008

106. Prof. Dr. Dzulkefly Kuang AbdullahPalm Oil: Still the Best ChoiceI February 2008

107. Prof. Dr. Elias SaionProbing the Microscopic Worlds byIOnizing Radiation22 February 2008

108. Prof. Dr. MohdAli HassanWaste-to-Wealth ThroughBiotechnology: For Profit, Peopleand Planet28 March 2008

109. Prof. Dr. Mohd MaarofH. A. MoksinMetrology at Nanoscale: ThermalWave Probe Made It Simple11 April 2008

110. Prof. Dr. Dzolkhifli OmarThe Future of Pesticides Technologyin Agriculture: Maximum Target Killwith Minimum Collateral Damage25 April 2008

111. Prof. Or. Mohd. Yazid Abd. ManapProbiotics: Your Friendly GutBacteria9 May 2008

112. Prof. Dr. Hamami SahriSustainable Supply of Wood andFibre: Does Malaysia have Enough?23 May2008

113.Prof. Dato' Dr. Makhdzir MardanConnecting the Bee Dots20 June 2008

114. Prof. Dr. Maimunah IsmailGender & Career: Realities andChallenges25 Ju1y2008

115. Prof. Dr. Nor Aripin ShamaanBiochemistry of Xenobiotics:Towards a Healthy Lifestyle and SafeEnvironment1 August 2008

116. Prof. Dr. Mohd Yunus AbdullahPenjagaan Kesihatan Primer diMalaysia: Cabaran Prospek danImplikasi dalam Latihan danPenyelidikan Perubatan sertaSains Kesihatan di Universiti PutraMalaysia8 August 2008

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117. Prof. Dr. Musa Abu HassanMemanfaatkan Teknologi Maklumat& Komunikasi ICT untuk Semua15 August 2008

118. Prof. Dr. Md. Salleh Hj. HassanRole of Media in Development:Strategies, Issues & Challenges22 August 2008

119. Prof. Dr. Jariah MasudGender in Everyday Life10 October 2008

120 Prof. Dr. Mohd Shahwahid HajiOthmanMainstreaming Environment:Incorporating Economic Valuationand Market-Based Instruments inDecision Making24 October 2008

121. Prof. Dr. Son RaduBig Questions Small Worlds:Following Diverse Vistas31 October 2008

122. Prof. Dr. Russly Abdul RahmanResponding to Changing Lifestyles:Engineering the Convenience Foods28 November 2008

123. Prof. Dr. Mustafa Kamal MohdShariffAesthetics in the Environment anExploration of Environmental:Perception Through LandscapePreference9 January 2009

124. Prof. Dr. Abu Daud SilongLeadership Theories, Research& Practices: Farming FutureLeadership Thinking16 January 2009

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125. Prof. Dr. Azni IdrisWaste Management, What is theChoice: Land Disposal or Biofuel?23 January 2009

126. Prof. Dr. Jamilah BakarFreshwater Fish: The OverlookedAlternative30 January 2009

127. Prof. Dr. Mohd. Zobir Hussein. The Chemistry of Nanomaterial andNanobiomaterial6 February 2009

Prof. Ir. Dr. Lee Teang ShuiEngineering Agricultural: WaterResources )0 )

20 February 20/ .

129. Prof. Dr. Ghizan Saleh

128.

Crop Breeding: Exploiting Genes forFood and Feed6 March2009

130. Prof. Dr. Muzafar Shah HabibullahMoney Demand27 March 2009

131. Prof. Dr. Karen Anne CrouseIn Search of Small Active Molecules3April2009

132. Prof. Dr. Turiman SuandiVolunteerism: Expanding theFrontiers of Youth Development17 April 2009

133. Prof. Dr. Arbakariya AriffIndustrializing Biotechnology: Rolesof Fermentation and BioprocessTechnology8 May 2009

134. Prof. Ir. Dr. Desa AhmadMechanics of Tzllage Implements12 June 20Q9

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135. Prof. Dr. W. Mahmood MatYunusPhotothermal and Photoacoustic:From Basic Research to IndustrialApplications10 July 2009

136. Prof. Dr. Taufiq Yap Yun HinCatalysis for a Sustainable World7 August 2009

137 Prof. Dr. Raja Noor Zaliha RajaAbd. RahmanMicrobial Enzymes: From Earth toSpace9 October 2Q09

138 Prof. Ir. Dr. Barkawi SahariMaterials, Energy and CNGDIVehicle Engineering6 November 2009

139. Prof. Dr. Zulkifli IdrusPoultry Welfare in ModernAgriculture: Opportunity or Threat?13 November 2009

140. Prof. Dr. Mohamed Hanafi MusaManaging Phosphorus: Under AcidSoils Environment8 January 2010

141. Prof. Dr. Abdul Manan Mat JaisHaruan Channa striatus a DrugDiscovery in an Agro-IndustrySetting12 March 2010

142. Prof. Dr. Bujang bin Kim HuatProblematic Soils: In Search forSolution19 March 2010

143. Prof. Dr. Samsinar Md SidinFamily Purchase Decision Making:Current Issues & Future Challenges16April2010

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144. Prof. Dr. Mohd Adzir Mahdi 154. Prof. Dr. Seow Heng FongLightspeed: Catch Me If You Can Are there "Magic Bullets "for4 June 2010 Cancer Therapy?

II February 20 II145. Prof. Dr. Raha Hj. Abdul Rahim

Designer Genes: Fashioning Mission ISS. Prof. Dr. Mohd Azmi Mohd LilaPurposed Microbes Biopharmaceuticals: Protection,IS June 2010 Cure and the Real Winner

IS February 20 II146. Prof. Dr. Hj. Hamidon Hj. Basri

A Stroke of Hope, A New Beginning 156. Prof. Dr. Siti Shapor Siraj2 July 2010 Genetic Manipulation in Farmed

Fish: Enhancing Aquaculture147. Prof. Dr. Hj. Kamaruzaman Jusoff Production

Going Hyperspectral: The "Unseen" 25 March 20 IICaptured?16 July 2010 157. Prof. Dr. Ahmad Ismail

Coastal Biodiversity and Pollution:14S. Prof. Dr. Mohd Sapuan Sal it A Continuous Conflict

Concurrent Engineeringfor 22 April 20 I IComposites30 july 2010 ISS. Prof. Ir. Dr. Norman Mariun

Energy Crisis 2050? Global149. Prof. Dr. Shattri Mansor Scenario and Way Forwardfor

Google the Earth: What's Next? MalaysiaIS October 2010 10 June 2011

ISO. Prof. Dr. Mohd Basyaruddin Abdul 159. Prof. Dr. Mohd Razi IsmailRahman Managing Plant Under Stress: AHaute Couture: Molecules & Challenge for Food SecurityBiocatalysts IS July 201129 October 20 I0

160. Prof. Dr. Patimah IsmaillSI. Prof. Dr. Mohd. Hair Bejo Does Genetic Polymorphisms Affect

Poultry Vaccines: An Innovation for Health?Food Safety and Security 23 September 20 II12 November 2010

161. Prof. Dr. Sidek Ab. Aziz152. Prof. Dr. Umi Kalsom Yusuf Wonders of Glass: Synthesis,

Fern of Malaysian Rain Forest Elasticity and Application3 December 20 I0 7 October 20 I I

153. Prof. Dr. Ab. Rahim Bakar 162. Prof. Dr. Azizah OsmanPreparing Malaysian Youths for The Fruits: Nutritious, Colourful, YetWorld of Work: Roles of Technical Fragile Gifts of Natureand Vocational Education and 14 October 20 IITraining (rVET)14 January 2011

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163. Prof. Dr. Mohd. Fauzi RamlanClimate Change: Crop Performanceand PotentialII November 20 II

164. Prof. Dr. Adem KilicrnanMathematical Modeling withGeneralized Function25 November 2011

165. Prof. Dr. Fauziah OthmanMy Small World: In BiomedicalResearch23 December 20 II

166. Prof. Dr. Japar Sidik BujangThe Marine A .: , s, Seagrass23 March 2012

167. Prof. Dr. Zailin~s imAir Quality and Children'sEnvironmental Health: Is OurFuture Generation at Risk?30 March 2012

168. Prof. Dr. ZainalAbidin MohamedWhere is the Beef? liantage Pointform the Livestock Supply Chain27 April 2012

169. Prof. Dr. Jothi Malar PanandamGenetic Characterisation of AnimalGenetic Resourcesfor SustaninableUtilisation and Development30 November 2012

170. Prof. Dr. Fatimah Abu BakarThe Good The Bad & Ugly of FoodSafety: From Molecules to Microbes7December2012

171. Prof. Dr. Abdul Jalil NordinMy Colour fit! Sketches from Scratch:Molecular Imaging5 April2013

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172. Prof. Dr. Norlijah OthmanLower Respiratory Infections inChildren: New Pathogens, OldPathogens and The Way Forward19 April2013

173. Prof. Dr. Jayakaran MukundanSteroid-like Prescriptions EnglishLanguage Teaching Can Ill-afford26 April 2013

174. Prof. Dr. Azmi ZakariaPhotothermals Affect Our Lives7 June 20l}

175. Prof. Dr. Rahinah IbrahimDesign Informatics21 June 2013

176. Prof. Dr. Gwendoline Ee ChengNatural Products from MalaysianRainforestsI November 2013

177. Prof. Dr. Noor Akma IbrahimThe Many Facets of StatisticalModeling22 November 2013

178. Prof. Dr. Paridah Md. TahirBonding with Natural Fibres6 December 2013

179. Prof. Dr. Abd. Wahid HaronLivestock Breeding: The Past, ThePresent and The Future9 December 2013

180. Prof. Dr. Aziz ArshadExploring Biodiversity & FisheriesBiology: A Fundamental Knowledgefor Sustainabale Fish Production24 January 2014

181. Prof. Dr. Mohd Mansor IsmailCompetitiveness of BeekeepingIndustry in Malaysia21. March 2014

182. Prof. Dato' Dr. Tai Shzee YewFood and Wealth from the Seas:Health Checkfor the MarineFisheries of Malaysia25 April 2014

183. Prof. Datin Dr. Rosenani Abu BakarWaste to Health: Organic WasteManagementfor Sustainable SoilManagement and Crop Production9 May 2014

184. Prof. Dr. Abdul Rahman OmarPoultry Viruses: From Threat toTherapy23 May 2014

185. Prof. Or. Mohamad Pauzi ZakariaTracing the Untraceable:Fingefprinting Pollutants throughEnvironmental Forensics13 June 2014

186. Prof. Dr. -Ing. Ir. RenuganthVaratharajooSpace System Trade-offs: TowardsSpacecraft Synergisms15 August 2014

187. Prof. Dr. Latiffah A. LatiffIranformas! Kesihatan Wanita keArah Kesejahteraan Komuniti7 November 2014

188. Prof. Dr. Tan Chin PingFat and Oils for a Healthier Future:Makro, Micro and Nanoscales21 November2014

189. Prof. Dr. Suraini Abd. AzizLignocellulosic Biofuel: A WayForward28 November2014

190. Prof. Dr. Robiah YunusBiobased Lubricants: Harnessingthe Richness of AgricultureResources30 January 2015

Amin Ismail

190. Prof. Dr. Khozirah ShaariDiscovering Future Cures fromPhytochemistry to Metabolomics13 February 2015

191. Prof. Dr. Tengku Aizan Tengku AbdulHamidPopulation Ageing in Malaysia: AMosaic of Issues, Challenges andProspects13 March 2015

192. Prof. Datin Dr. Faridah HanumIbrahimForest Biodiversity: Importance ofSpecies Composition Studies27 March 2015

192. Prof. Dr. Mohd Salleh KamarudinFeeding & Nutritional Requirementsof Young FishIOApril2015

193. Prof. Dato' Dr. Mohammad ShatarSabranMoney Boy: Masalah Sosial EraGenerasi Y8 Mei 2015

194. Prof. Dr. Aida Suraya Md. YunusDeveloping Students' MathematicalThinking: How Far Have We Come?5 June 2015

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