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Fulltext The use of Caco-2 cells as an in vitro method to study bioavailability of iron

Maznah I

Malays J Nutr, 1999 Dec;5(1):31-45.
PMID: 22692356

Iron absorption is essential for the maintenance of iron levels in the body, since excretion is poorly regulated. Dietary factors can influence iron absorption including low molecular weight substances such as ascorbic acid which has been shown to enhance iron transport across mucosal cell monolayers. Both in vivo and in vitro work may be carried out to study iron absorption. Studies in vivo have the drawback of dealing with a complex system in which it is difficult to determine the relative importance of different factors. In vitro cell culture models could overcome this difficulty but attempts to establish differentiated enterocyte cell lines in culture have not been successful. However the Caco-2 line, derived from a colon carcinoma, is able to differentiate spontaneously when grown in standard culture conditions. The differentiated cells polarized, formed microvilli and T-junctions associated with the duodenal enterocytes brush border. This cell line thus represents an appropriate model for the study of transport mechanisms related to the intestinal barrier and can be used to study the absorption of nutrients especially iron in relation to dietary intake in particular pertaining to dietary factors that may affect absorption. In this work we have therefore used differentiated Caco-2 cells grown in bicameral chambers as a intestinal cell model to study the absorption of iron from different sources and compared it with INT 407 cells. Transfer of iron across the monolayers in the apical-to-basolateral direction has been found to be greater from feric lactoferrin than from iron citrate, while very little transport occurred from Fe-transferrin. It is concluded that in this in vitro study lactoferrin but not transferrin enhances mucosal iron transport. More importanty this study has also shown that Caco-2 can be used as an in vitro method to investigate not only iron bioavailability but can be applied to other minerals as well.
Fulltext Strobilanthes crispus leaves extracts (SCE) induced lipolysis and increased leptin level in diet-induced obese rats fed high-fat diet

Zawawi, N., Azizah, A. H., Maznah, I.

International Food Research Journal, 2016;23(3):1115-1122.
MyJurnal

The aim of the present study was to assess the anti-obesity and lipolysis effects of Strobilanthes
crispus leaves extract (SCE) in diet-induced obese (DIO) rats by administering 1% (w/w) of
SCE in drinking water. Methods: Normal Sprague-Dawley rats were induced obese using a
high-fat (HF) diet formulation for 14 weeks. DIO rats were subsequently treated with 1% (w/w)
SCE while the HF diet was switched to normal rat chow diet. Food intake, water intake and
bodyweight were measured weekly. Other parameters e.g. blood lipid profile were determined
in normal and DIO rats before and after treatment with SCE. Histopathological changes in the
liver were also observed after treatment. Results: Prior to treatment, DIO rats have significantly
higher (p
Fulltext Extraction and determination of oryzanol in rice bran of mixed herbarium UKMB; AZ 6807: MR 185, AZ 6808: MR 211, AZ6809: MR 29

Azrina, A., Maznah, I., Azizah, A.H.

International Food Research Journal, 2008;15(1):89-96.
MyJurnal

The level of total lipid and oryzanol content, an important antioxidant compound in locally produced bran was investigated. Total lipid in rice bran was extracted using 3:2 chloroform:methanol mixture yielding 16.4% fat. Oryzanol content was determined without saponification using a reverse-phase HPLC. Four fractions of oryzanol were successfully separated and quantitated. The 4 isomers were cycloartenyl ferulate, 24-methylene cycloartanyl ferulate, campestryl ferulate and mixtures of β–sitosteryl ferulate and cycloartanyl ferulate. The oryzanol content of local mixed varieties ranged from 23.7–43.0 mg g-1. The oryzanol concentration may depend on factors such as plant varieties, processing methods employed, extracting solvent used and ratio of extracting solvent to bran as well as extracting solvent temperatures. This study showed the potential of oryzanol extract from rice bran as a source of antioxidant.
Fulltext The effect of different milks and milk proteins on the growth of Bifidobacterium infantis ATCC 27920 in vitro

Loh S, Maznah I

Malays J Nutr, 1999 Dec;5(1):61-70.
PMID: 22692359

Bifidobacteria is a well known bacteria that is found in abundance in the intestine of infants which provides several health and nutritional benefits. Realizing the many benefits of bifidobacteria to human, this study has been conducted with the objective to determine the growth promotional effect of different types of milk and milk proteins on Bifidobacterium species. One strains of Bifiodobacterium species that is B. infantis was used to study the growth promoting effect of human milk, cow's milk, goat's milk, milk based infant formula, soy-based infant formula, lactoferrin (1 mg/ml), lactoperoxidase (1p~g/ml), lysozyme (1 mg/ml) and the mixture of these three proteins. The growth promotion assay was done using the 96-well culture plates which consists of 200 (1 Trypticase-Peptone-Yeast extract (TPY) medium, 50 4 sample and 10 1il of bacteria inoculum. Control consists of PBS instead of the samples. The assay was incubated anaerobically at 370C for 18 hours before being spread on the agar plate containing TPY medium with agar. Comparison was made between the mean count (log cfu/ml) of different types of milks, between infant formula and between milk proteins. From the results, Oneway ANOVA test at P<0.05 showed that there was significant differences in the mean counts (log cfu/ml) between the milks (P = 0.0000). A similar trend was observed in the mean count (log cfu/mI) between the infant formulas (P = 0.0 124) and also between the milk proteins (P = 0.0005). Duncan Multiple Range tests showed that there was significant differences between all the milks and control and among the milks themselves. There was however, no significant difference among the two types of infant formulas. The milk proteins also showed significant differences between the proteins and control and among themselves except for lysozyme which showed no significant differences with lactoferrin. This study showed that the growth of B. infantis could be promoted by different kinds of milks and milk proteins in vitro. Comparing the differences in growth promoting effect between samples and control indicated that human milk has the highest growth promoting effect followed by cow's milk and the mixture of the three milk prtoeins. Lysozyme showed the lowest in term of differences in percentage of growth promoting effect among all these samples. In conclusion the findings of this study supported that human milk ios the best milk choice for infant in comparison to other types of milk in promoting the growth of bifidobacteria. In additon, this tudy also found that milk protein when used in combination may show better growth promoiotive effect than when used singly.
Fulltext Nutritional values and cooking quality of defatted Kenaf seeds yellow (DKSY) noodles

Zawawi, N., Gangadharan, P., Ahma Zaini, R., Samsudin, M.G., Karim, R., Maznah, I.

International Food Research Journal, 2014;21(2):603-608.
MyJurnal

Kenaf (Hibiscus cannabinus) seeds have always being wasted as agricultural waste. Recent studies revealed that the seeds contain high fiber. The purpose of this study is to develop defatted kenaf seeds yellow noodles (DKSY) and assess the nutritional and physicohemical properties of the noodles. Defatted kenaf flour at 25% and 75% were used to make DKSY noodles and compared to wheat yellow noodles (Control). Fresh DKSY noodles were analyzed for their nutritional and physiochemical properties. The ash and fiber contents increased in order of Control > 25% DKSY > 75% DKSY noodles. While total phenolic contents (TPC) was found to be higher in 75% DKSY noodles (138.30 ± 1.63 mg GAE/100 g) than Control noodles. Colour (L, b) and hardness decreased in order of Control > 25% DKSY > 75% DKSY indicating that DKSY noodles developed less quality than Control noodles. However, cooking loss values were found to be in the same order while cooking values exist in the opposite order indicating that. DKSY noodles have better noodle cooking quality. In conclusion, nutritional properties and noodle cooking quality of yellow noodles increased with higher concentration of defatted kenaf flour but the physicochemical properties were compromised. More research needs to be done in order to develop a formulation that can increase all of the attributes studied.
Fulltext Reliability and validity of the instrument used in the HELIC (healthy lifestyle in children) study of primary school children's nutrition knowledge, attitude and practice

Siti Sabariah, B., Zalilah, M.S., Norlijah, O., Normah, H., Maznah, I., Laily, P., et al.

Malays J Nutr, 2006;12(1):33-44.
MyJurnal

The objective of this paper is to report on the reliability and validity of a knowledge, attitude and practice instrument used among young primary school children. The instrument was developed as an evaluation tool in the HELIC study and consisted of 23 knowledge, 11 attitude and 10 practice items. A total of 335 Year 2 students from 4 randomly selected primary schools in Selangor and Wilayah Persekutuan participated in the HELIC study. Students were divided into small groups and an enumerator verbally administered the instrument to each group. Reliability for each construct (knowledge, attitude and practice) was estimated as item to total score correlation and internal consistency (Cronbach's alpha). Construct validity was determined through factor analysis and Pearson correlation. Results indicated that 3 attitude and 3 practice items did not correlate significantly to the total score (p>0.05). However, the deletion of these items did not significantly alter the Cronbach's alpha coefficients. Internal consistency was good for knowledge (a=0.68) but low for attitude (a=0.37) and practice (a=0.36) constructs. Based on factor analysis, 5 factor-solutions emerged for knowledge and 4 factor solutions for attitude and practice. Sufficient variance was obtained for the factors in knowledge (51.7%), attitude (51.2% and practice (51.0%). There were also significant positive correlations among the constructs ( ~ 4 . 0 1 ) . In conclusion, the instrument was valid and reliable, especially for the knowledge construct. Further improvements, particularly on the attitude and practice constructs, are needed in order for the instrument to be an effective assessment or evaluation tool in various settings.
Fulltext Bioavailability studies of nutraceuticals

Maznah I, Loh SP, Waffaa MH

Malaysian Journal of Medicine and Health Sciences, 2005;1(1):1-12.

Despite increasing interest in in nutraceuticals and their potential health benefits, not much is known about bioavailability of most of these nutraceutical compounds. Although they are considered dietary supplements and are subjected to a limited form of regulation, there is, however, a need to improve the efficacy and safety of these nutraceuticals. Additional research which defines the pharmacology, stability and bioavailability of these products is expected to gain strength and may offer a better understanding of their applicability in the prevention of disease conditions. This article reviews some aspects of nutraceutical bioavailability with examples from our work on the absorption studies of minerals from spirulina (Arthrospira plantensis) and gamma-oryzanol from rice bran (Oryza sativa) extract which employed human colon carcinoma (Caco-2) cell line and in vivo bioassays using animal models. Bioavailability of iron from spirulina was compared with its common source FeS04. Using the in vitro digestion protocol in combination with Caco-2 cell culture system, spirulina showed a high iron bioavailability compared to FeS04. The presence of other dietary factors (calcium, ascorbic acid, zinc, tannin and caffeine) was found to be not as significant as ferrous sulphate in affecting the iron uptake from spirulina. In vivo study showed that the efficacy of iron repletion in anaemic rats was enhanced in groups fed either commercial or cultured spirulina with improved haematological parameters of iron status. Further work on the behaviour and distribution of radiolabelled iron from spirulina has shown that iron-59 retained in the GIT of mice was lower in spirulina group compared to FeS04. Bioavailability study of gamme oryzanol was similarly conducted using Caco-2 cell as in vitro system and rabbit as in vivo model with the application of different formulations of gamma oryzanol in comparison with the natural form. Both systems showed that gamma oryzanol in its natural oil was poorly absorbed. However, when converted to other formulations, gamma oryzanol bioavailability was greatly increased by as much as 200 and 33 times more from the emulsion and microspheres respectively. These findings suggest that the efficacy of nutraceuticals in particular plant derived products which contain many phytochemicals should be assessed in terms of not only their potential health benefits such as antioxidant action but also their bioavailability in order to provide a more wholesome picture of their potential.
Nutritional composition and total collagen content of three commercially important edible jellyfish

Khong NM, Yusoff FM, Jamilah B, Basri M, Maznah I, Chan KW, et al.

Food Chem, 2016 Apr 1;196:953-60.
PMID: 26593577 DOI: 10.1016/j.foodchem.2015.09.094

The study aimed to evaluate nutraceutical potential of three commercially significant edible jellyfish species (Acromitus hardenbergi, Rhopilema hispidum and Rhopilema esculentum). The bell and oral arms of these jellyfishes were analyzed for their proximate composition, calorific value, collagen content, amino acid profile, chemical score and elemental constituent. In general, all jellyfish possessed low calorific values (1.0-4.9 kcal/g D.W.) and negligible fat contents (0.4-1.8 g/100 g D.W.), while protein (20.0-53.9 g/100 g D.W.) and minerals (15.9-57.2g/100g D.W.) were found to be the richest components. Total collagen content of edible jellyfish varied from 122.64 to 693.92 mg/g D.W., accounting for approximately half its total protein content. The dominant amino acids in both bell and oral arms of all jellyfish studied includes glycine, glutamate, threonine, proline, aspartate and arginine, while the major elements were sodium, potassium, chlorine, magnesium, sulfur, zinc and silicon. Among the jellyfish, A. hardenbergi exhibited significantly higher total amino acids, chemical scores and collagen content (p<0.05) compared to R. hispidum and R. esculentum. Having good protein quality and low calories, edible jellyfish is an appealing source of nutritive ingredients for the development of oral formulations, nutricosmetics and functional food.
Fulltext Characterization of rice bran wax policosanol and its nanoemulsion formulation

Ishaka A, Umar Imam M, Mahamud R, Zuki AB, Maznah I

Int J Nanomedicine, 2014;9:2261-9.
PMID: 24872689 DOI: 10.2147/IJN.S56999

Policosanol, a mixture of long-chain alcohols found in animal and plant waxes, has several biological effects; however, it has a bioavailability of less than 10%. Therefore, there is a need to improve its bioavailability, and one of the ways of doing this is by nanoemulsion formulation. Different droplet size distributions are usually achieved when emulsions are formed, which solely depends on the preparation method used. Mostly, emulsions are intended for better delivery with maintenance of the characteristics and properties of the leading components. In this study, policosanol was extracted from rice bran wax, its composition was determined by gas chromatography mass spectrophotometry, nanoemulsion was made, and the physical stability characteristics were determined. The results showed that policosanol nanoemulsion has a nanosize particle distribution below 100 nm (92.56-94.52 nm), with optimum charge distribution (-55.8 to -45.12 mV), pH (6.79-6.92) and refractive index (1.50); these were monitored and found to be stable for 8 weeks. The stability of policosanol nanoemulsion confers the potential to withstand long storage times.
Fulltext Estrogen receptor modulatory effects of germinated brown rice bioactives in the uterus of rats through the regulation of estrogen-induced genes

Muhammad SI, Maznah I, Mahmud RB, Saeed MI, Imam MU, Ishaka A

Drug Des Devel Ther, 2013;7:1409-20.
PMID: 24324328 DOI: 10.2147/DDDT.S50861

The expression of genes regulated by estrogen in the uterus was studied in ovariectomized (OVX) rats treated with germinated brown rice (GBR) bioactives, and compared to Remifemin or estrogen at different doses to identify the regulation of these genes in the uterus and their molecular mechanisms.
Improved collagen extraction from jellyfish (Acromitus hardenbergi) with increased physical-induced solubilization processes

Khong NMH, Yusoff FM, Jamilah B, Basri M, Maznah I, Chan KW, et al.

Food Chem, 2018 Jun 15;251:41-50.
PMID: 29426422 DOI: 10.1016/j.foodchem.2017.12.083

Efficiency and effectiveness of collagen extraction process contribute to huge impacts to the quality, supply and cost of the collagen produced. Jellyfish is a potential sustainable source of collagen where their applications are not limited by religious constraints and threats of transmittable diseases. The present study compared the extraction yield, physico-chemical properties and toxicology in vitro of collagens obtained by the conventional acid-assisted and pepsin-assisted extraction to an improved physical-aided extraction process. By increasing physical intervention, the production yield increased significantly compared to the conventional extraction processes (p  .05) while retaining high molecular weight distributions and polypeptide profiles similar to those extracted using only acid. Moreover, they exhibited better appearance, instrumental colour and were found to be non-toxic in vitro and free of heavy metal contamination.
Edible Bird's Nest Attenuates Menopause-Related Bone Degeneration in Rats via Increaing Bone Estrogen-Receptor Expression

Hou ZP, Tang SY, Ji HR, He PY, Li YH, Dong XL, et al.

Chin J Integr Med, 2021 Apr;27(4):280-285.
PMID: 31872369 DOI: 10.1007/s11655-019-3209-1

OBJECTIVE: To investigate the mechanistic basis for the attenuation of bone degeneration by edible bird's nest (EBN) in ovariectomized rats.
METHODS: Forty-two female Sprage-Dawley rats were randomized into 7 groups (6 in each group). The ovariectomized (OVX) and OVX + 6%, 3%, and 1.5% EBN and OVX +estrogen groups were given standard rat chow alone, standard rat chow +6%, 3%, and 1.5% EBN, or standard rat chow +estrogen therapy (0.2mg/kg per day), respectively. The sham-operation group was surgically opened without removing the ovaries. The control group did not have any surgical intervention. After 12 weeks of intervention, blood samples were taken for serum estrogen, osteocalcin, and osteoprotegerin, as well as the measurement of magnesium, calcium abd zinc concentrations. While femurs were removed from the surrounding muscles to measure bone mass density using the X-ray edge detection technique, then collected for histology and estrogen receptor (ER) immunohistochemistry.
RESULTS: Ovariectomy altered serum estrogen levels resulting in increased food intake and weight gain, while estrogen and EBN supplementation attenuated these changes. Ovariectomy also reduced bone ER expression and density, and the production of osteopcalcin and osteorotegerin, which are important pro-osteoplastic hormones that promote bone mineraliztion and density. Conversely, estrogen and EBN increased serum estrogen levels leading to increased bone ER expression, pro-osteoplastic hormone production and bone density (all P<0.05).
CONCLUSION: EBN could be used as a safe alternative to hormone replacement therapys for managing menopausal complications like bone degeneration.
Fulltext Germinated brown rice (GBR) reduces the incidence of aberrant crypt foci with the involvement of beta-catenin and COX-2 in azoxymethane-induced colon cancer in rats

Latifah SY, Armania N, Tze TH, Azhar Y, Nordiana AH, Norazalina S, et al.

Nutr J, 2010 Mar 26;9:16.
PMID: 20346115 DOI: 10.1186/1475-2891-9-16

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

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