Shuhaimi Mustafa

Gold nanoparticle sensor for the visual detection of pork adulteration in meatball formulation

We visually identify pork adulteration in beef and chicken meatball preparations using 20nm gold nanoparticles (GNPs) as colorimetric sensors. Meatball is a popular food in certain Asian and European countries. Verification of pork adulteration in meatball is necessary to meet the Halal and Kosher food standards. Twentynm GNPs change color from pinkish-red to gray-purple, and their absorption peak at 525nm is red-shifted by 30-50nm in 3mM phosphate buffer saline (PBS). Adsorption of single-stranded DNA protects the particles against salt-induced aggregation. Mixing and annealing of a 25-nucleotide (nt) single-stranded (ss) DNA probe with denatured DNA of different meatballs differentiated well between perfectly matched and mismatch hybridization at a critical annealing temperature. The probes become available in nonpork DNA containing vials due to mismatches and interact with GNPs to protect them from salt-induced aggregation. Whereas, all the pork containing vials, either in pure and mixed forms, consumed the probes totally by perfect hybridization and turned into grey, indicating aggregation. This is clearly reflected by a well-defined red-shift of the absorption peak and significantly increased absorbance in 550-800nm regimes. This label-free low-cost assay should find applications in food analysis, genetic screening, and homology studies.

Analysis of pork adulteration in commercial meatballs targeting porcine-specific mitochondrial cytochrome b gene by TaqMan probe real-time polymerase chain reaction

A test for assessing pork adulteration in meatballs, using TaqMan probe real-time polymerase chain reaction, was developed. The assay combined porcine-specific primers and TaqMan probe for the detection of a 109 bp fragment of porcine cytochrome b gene. Specificity test with 10 ng DNA of eleven different species yielded a threshold cycle (Ct) of 15.5 ± 0.20 for the pork and negative results for the others. Analysis of beef meatballs with spiked pork showed the assay can determine 100-0.01% contaminated pork with 102% PCR efficiency, high linear regression (r(2) = 0.994) and ≤ 6% relative errors. Residuals analysis revealed a high precision in all determinations. Random analysis of commercial meatballs from pork, beef, chicken, mutton and goat, yielded a Ct between 15.89 ± 0.16 and 16.37 ± 0.22 from pork meatballs and negative results from the others, showing the suitability of the assay to determine pork in commercial meatballs with a high accuracy and precision.

Corn Silk (Stigma Maydis) in Healthcare: A Phytochemical and Pharmacological Review

Corn silk (Stigma maydis) is an important herb used traditionally by the Chinese, and Native Americans to treat many diseases. It is also used as traditional medicine in many parts of the world such as Turkey, United States and France. Its potential antioxidant and healthcare applications as diuretic agent, in hyperglycemia reduction, as anti-depressant and anti-fatigue use have been claimed in several reports. Other uses of corn silk include teas and supplements to treat urinary related problems. The potential use is very much related to its properties and mechanism of action of its plant’s bioactive constituents such as flavonoids and terpenoids. As such, this review will cover the research findings on the potential applications of corn silk in healthcare which include its phytochemical and pharmacological activities. In addition, the botanical description and its toxicological studies are also included.

Nanoparticle sensor for label free detection of swine DNA in mixed biological samples

We used 40 ± 5 nm gold nanoparticles (GNPs) as colorimetric sensor to visually detect swine-specific conserved sequence and nucleotide mismatch in PCR-amplified and non-amplified mitochondrial DNA mixtures to authenticate species. Colloidal GNPs changed color from pinkish-red to gray-purple in 2 mM PBS. Visually observed results were clearly reflected by the dramatic reduction of surface plasmon resonance peak at 530 nm and the appearance of new features in the 620-800 nm regions in their absorption spectra. The particles were stabilized against salt-induced aggregation upon the adsorption of single-stranded DNA. The PCR products, without any additional processing, were hybridized with a 17-base probe prior to exposure to GNPs. At a critical annealing temperature (55 °C) that differentiated matched and mismatched base pairing, the probe was hybridized to pig PCR product and dehybridized from the deer product. The dehybridized probe stuck to GNPs to prevent them from salt-induced aggregation and retained their characteristic red color. Hybridization of a 27-nucleotide probe to swine mitochondrial DNA identified them in pork-venison, pork-shad and venison-shad binary admixtures, eliminating the need of PCR amplification. Thus the assay was applied to authenticate species both in PCR-amplified and non-amplified heterogeneous biological samples. The results were determined visually and validated by absorption spectroscopy. The entire assay (hybridization plus visual detection) was performed in less than 10 min. The LOD (for genomic DNA) of the assay was 6 µg ml(-1) swine DNA in mixed meat samples. We believe the assay can be applied for species assignment in food analysis, mismatch detection in genetic screening and homology studies between closely related species.

Effect of pre-germination time of brown rice on serum cholesterol levels of hypercholesterolaemic rats.

BACKGROUND Brown rice is unpolished rice with immeasurable benefits for human health. Brown rice (BR) and pre-germinated brown rice (PGBR) are known to contain various functional compounds such as gamma-oryzanol, dietary fibre and gamma-aminobutyric acid (GABA). In the present study, the experimental diets containing BR and PGBR (24, 48 h pre-germination) were used to investigate the influence of pre-germination time of brown rice on blood cholesterol in Sprague-Dawley male rats. RESULTS Hypercholesterolaemia and elevation of LDL-cholesterol were successfully ameliorated by the experimental diets containing BR and PGBR (24 and 48 h pre-germination). As compared to the control sample, the pre-germination time had a significant (P < 0.05) effect on blood cholesterol of Sprague-Dawley male rats. It was also found that the significantly (P < 0.05) better effect on lipid profile of hypercholesterolaemic rats was observed by prolonging the pre-germination time. As compared to non-germinated brown rice, the germinated brown rice showed the higher cardio-protective effect on hypercholesterolaemic Sprague-Dawley male rats. CONCLUSION The present study suggests that the changes of blood cholesterol can be mainly modulated by using the PGBR rather than BR. The prolonging of pre-germination time led to an increase in the bioactive components, thereby providing a more efficient functional diet affecting the high blood cholesterol. This study suggests that PGBR can be used instead of BR and polished rice in the human diet.

Baseline distributions and sources of Polycyclic Aromatic Hydrocarbons (PAHs) in the surface sediments from the Prai and Malacca Rivers, Peninsular Malaysia

In this study, the surface sediments of the Malacca and Prai Rivers were analyzed to identify the distributions, and sources of Polycyclic Aromatic Hydrocarbons (PAHs). The total PAH concentrations varied from 716 to 1210 and 1102 to 7938 ng g(-1)dw in the sediments of the Malacca and Prai Rivers, respectively. The PAH concentrations can be classified as moderate and high level of pollution in the sediments of the Malacca and Prai Rivers, respectively. The comparison of PAHs with the Sediment Quality Guidelines (SQGs) indicates that the PAHs in the sediments of the Malacca and Prai Rivers may have the potential to cause adverse toxicity effects on the sampled ecosystems. The diagnostic ratios of individual PAHs indicate both petrogenic- and pyrogenic-origin PAHs with dominance of pyrogenic source in both rivers. These findings demonstrate that the environmental regulations in Malaysia have effectively reduced the input of petrogenic petroleum hydrocarbons into rivers.

Nanobiosensor for detection and quantification of DNA sequences in degraded mixed meats

A novel class of nanobiosensor was developed by integrating a 27-nucleotide AluI fragment of swine cytochrome b (cytb) gene to a 3-nm diameter citrate-tannate coated gold nanoparticle (GNP). The biosensor detected 0.5% and 1% pork in raw and 2.5- h autoclaved pork-beef binary admixtures in a single step without any separation or washing. The hybridization kinetics of the hybrid sensor was studied with synthetic and AluI digested real pork targets from moderate to extreme target concentrations and a sigmoidal relationship was found. Using the kinetic curve, a convenient method for quantifying and counting target DNA copy number was developed. The accuracy of the method was over 90% and 80% for raw and autoclaved pork-beef binary admixtures in the range of 5-100% pork adulteration. The biosensor probe identified a target DNA sequence that was several-folds shorter than a typical PCR-template. This offered the detection and quantitation of potential targets in highly processed or degraded samples where PCR amplification was not possible due to template crisis. The assay was a viable alternative approach of qPCR for detecting, quantifying and counting copy number of shorter size DNA sequences to address a wide ranging biological problem in food industry, diagnostic laboratories and forensic medicine.

Functional properties and characterization of dietary fiber from mangifera pajang kort. Fruit pulp

A dried high fiber product from bambangan (Mangifera pajang Kort.) fruit pulp was prepared and evaluated for proximate composition, functional properties, and soluble and insoluble dietary fiber composition. Mangifera pajang fibrous (MPF) consisted of 4.7% moisture, 0.8% fat, 4% protein, and 30 mg total polyphenol per g of dry sample, and 9, 79 and 88% soluble, insoluble and total dietary fiber, respectively. Water holding capacity, oil holding capacity, swelling, and solubility were found to be 9 g/g dry sample, 4 g/g dry sample, 16 mL/g dry sample, and 11%, respectively. The glucose dialysis retardation index of MPF was approximately double that of cellulose fiber. Soluble dietary fiber contained mannose, arabinose, glucose, rhamnose, erythrose, galactose, xylose, and fucose at 1.51, 0.72, 0.39, 0.16, 0.14, 0.05, 0.04, and 0.01%, respectively, with 5.8% uronic acid, while insoluble dietary fiber was composed of arabinose (18.47%), glucose (4.46%), mannose (3.15%), rhamnose (1.65%), galactose (1.20%), xylose (0.99%), and fucose (0.26%) with 15.5% uronic acid and 33.1% klason lignin. These characteristics indicate that MPF is a rich source of dietary fiber and has physicochemical properties which make it suitable as an added ingredient in various food products and/or dietetic, low-calorie high-fiber foods to enhance their nutraceutical properties.

Prebiotic Activity of Polysaccharides Extracted from Gigantochloa Levis (Buluh beting) Shoots

Bamboo shoot crude polysaccharides (BSCP) extracted from the shoots of Gigantochloa levis gave about 3.27 ± 0.18% on dry basis and a very minute percentage of protein (0.02 ± 0.01%). The molecular weight of BSCP estimated by gel chromatography was found to be around 7.49 × 103 Da, while the molecular weights of purified fractions (F1 to F5) were around 1550.96, 1471.63, 1685.78, 1691.61 and 1551.67 Da, respectively. The FTIR spectrum of BSCP revealed the possibility that the extract contains β-glucan, which can be considered a valuable compound for the medical and food industries. These relate to the resistance of BSCP towards artificial human gastric juice which is more than 99%. Prebiotic activity tested using BSCP as a carbon source showed significant increase in the growth of B. animalis ATCC 1053, B. longum BB 536 and L. acidophilus ATCC 4356 as compared to the use of FOS. Survivality of S. choleraesuis JCM 6977 was found to be slower in both BSCP and FOS. Study conducted reflects a good sign for the BSCP to be exploited as a promising prebiotic.

Hypocholesterolaemic effect of yoghurt containing Bifidobacterium pseudocatenulatum G4 or Bifidobacterium longum BB536

The effect of a yoghurt supplement containing Bifidobacterium pseudocatenulatum G4 or Bifidobacterium longum BB536 on plasma lipids, lipid peroxidation and the faecal excretion of bile acids was examined in rats fed a cholesterol-enriched diet. After 8 weeks, the rats in the positive control (PC) group who were fed the cholesterol-enriched diet showed significant increases in plasma total cholesterol (TC), low-density lipoprotein (LDL) cholesterol, and malondialdehyde (MDA). However, groups fed a cholesterol-enriched diet supplemented with yoghurt containing B. pseudocatenulatum G4 or B. longum BB536 had significantly lower plasma TC, LDL-C, very-low-density lipoprotein (VLDL) cholesterol, and MDA than had the PC group after 8 weeks of treatment. In addition, faecal excretion of bile acids was markedly increased in the rats fed the yoghurt containing B. pseudocatenulatum G4 or B. longum BB536 as compared to the PC and NC groups.