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Fulltext Brewers’ rice, a by-product from rice processing, has antip

Tan, B. L., Norhaizan, M. E., Hazilawati, H., Roselina, K.

International Food Research Journal, 2016;23(3):1182-1189.
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Colon carcinogenesis is a malignant tumor, and is well-known as the third leading cancer,
which contributes to high mortality and morbidity worldwide. Brewers’ rice, known locally as
temukut, consists of a mixture of broken kernels with rice bran and rice germ which is a byproduct
produced in the rice industry. Although extensive studies on the anti-cancer properties
of rice bran, published data on the cytotoxicity of brewers’ rice are very limited. The present
study was conducted to evaluate the apoptosis induction capability of the water extract of
brewers’ rice (WBR) on human colorectal cancer (HT-29) cell line. The HT-29 cells were
treated with various concentrations (16, 32, and 64 μg/mL) of WBR for 24 and 48 hours. The
morphological analysis of apoptotic cells was evaluated using inverted light microscope and
fluorescence microscope. The apoptotic HT-29 cells was evaluated using Annexin V-FITC and
propidium iodide (PI) staining apoptosis test and cell cycle analyses. The data obtained were
evaluated using a one-way analysis of variance (ANOVA) and P < 0.05 was considered statistical
significant. Overall analyses indicated that WBR induced typical characteristics of apoptosis in
HT-29 cells, including nuclear fragmentation (NF), nuclear compaction (NC), apoptotic bodies
(AB), cellular shrinkage (CS), and chromatin condensation (CC), as visualized under inverted
light microscope and fluorescence microscope. Cell cycle analyses and Annexin V-FITC and
propidium iodide (PI) staining apoptosis test using flow cytometry revealed that WBR induced
apoptotic population in HT-29 cells. In this study, our findings provide clear evidence that
WBR inhibits the growth of HT-29 cells via induction of apoptosis. Taken together, we suggest
that WBR may be a potential candidate for the prevention and treatment of colorectal cancer.
Effects of Ficus deltoidea aqueous extract on hematological and biochemical parameters in rats

Fazliana MS, Muhajir H, Hazilawati H, Shafii K, Mazleha M

Med J Malaysia, 2008 Jul;63 Suppl A:103-4.
PMID: 19025006

Aqueous extract of Ficus deltoidea var. agustifolia was examined for the subchronic toxicity effects in rats. Groups of 10 rats were given the extract daily by oral gavage for 90 days at 0 (control), 100 and 300mg/kg/body weight, respectively. Blood samples were collected upon sacrificed and analysed for haemogram and biochemistry. The results showed there were no significant changes of the blood parameters in all treated groups compared to the control.
Fulltext Improving the methods for isolation of monocyte and establishing macrophage cell culture in caprine model

Shaqinah NN, Mazlina M, Zamri-Saad M, Hazilawati H, Jasni S

Cytotechnology, 2016 Aug;68(4):1655-9.
PMID: 25511802 DOI: 10.1007/s10616-014-9836-7

Monocytes are widely used for immunological research, especially in the study of innate immune system. Although methods for isolation of human monocytes have been established, the procedure for non-human monocyte has not been well developed. This paper describes an improved method for isolation of monocyte and the subsequent macrophage cultivation from caprine blood. Monocytes were isolated from 16 ml of heparinized caprine blood using double density methods; the Ficoll and Percoll. The number of monocytes obtained was 5.12 ± 0.89 × 10(7) cells/ml at 70 % purity. The isolated monocytes were maintained in 10 % fetal bovine serum-enriched Dulbecco's Modified Eagle Medium for maturation to form macrophage cell culture. At the end of the experiment, the harvested macrophage was 2.48 ± 0.33 × 10(6) cells/ml.
Fulltext Brewers' Rice: A By-Product from Rice Processing Provides Natural Hepatorenal Protection in Azoxymethane-Induced Oxidative Stress in Rats

Tan BL, Norhaizan ME, Hairuszah I, Hazilawati H, Roselina K

Oxid Med Cell Longev, 2015;2015:539798.
PMID: 26257841 DOI: 10.1155/2015/539798

Brewers' rice, which is known locally as temukut, is a mixture of broken rice, rice bran, and rice germ. Our present study was designed to identify the effect of brewers' rice on the attenuation of liver and kidney damage induced by azoxymethane (AOM). Alanine transaminase (ALT), alkaline phosphatase (ALP), aspartate transaminase (AST), creatinine, and urea were evaluated to understand potential hepatoprotective effects and the ability of brewers' rice to attenuate kidney pathology induced by AOM treatment. Liver and kidney tissues were evaluated by hematoxylin and eosin (H&E) staining. Overall analyses revealed that brewers' rice improved the levels of serum markers in a manner associated with better histopathological outcomes, which indicated that brewers' rice could enhance recovery from hepatocyte and kidney damage. Taken together, these results suggest that brewers' rice could be used in future applications to combat liver and kidney disease.
DNA damage and adduct formation in immune organs of developing chicks by polycyclic aromatic hydrocarbons

Nisha AR, Hazilawati H, Mohd Azmi ML, Noordin MM

Toxicol. Mech. Methods, 2017 Mar;27(3):215-222.
PMID: 28030985 DOI: 10.1080/15376516.2016.1273432

Polycyclic aromatic hydrocarbons (PAHs) are persistent pollutants and chemically a class of structurally similar chemical compounds characterized by the presence of fused aromatic rings. This research was undertaken to find out immunotoxic effects produced by pyrene, phenanthrene and fluoranthene. These chemicals were injected into developing chicks at three dose levels (0.2, 2 and 20 mg per kg) through allantioc route to rule out possible mechanisms involved in immunotoxicity. DNA adduct produced by PAHs in immune organs were analyzed by DNA adduct enzyme-linked immunosorbent assay (ELISA) kit and DNA damage was assessed by comet assay. A significant increase in the DNA adduct levels was found in thymus and bursa in 2 mg and 20 mg dose levels of pyrene, fluoranthene and phenanthrene treated groups, whereas those in spleen simulated the value of controls. Comet assay indicated that PAHs especially pyrene, fluoranthene and phenanthrene were capable of inducing increased level of comet parameters in thymus at all the dose levels. Bursa of Fabricius and spleen also showed a gradual rise in comet parameters corresponding to all dose levels, but the increase was more marked as in thymus. Thus, it can be concluded that DNA adducts produced by PAHs lead to single-strand breaks and reduced DNA repair, which ultimately begin a carcinogenic process. Hence, this experiment can be considered as a strong evidence of genotoxic potential of PAHs like pyrene, phenanthrene and fluoranthene in developing chicks.
The induction of aryl hydrocarbon receptor (AHR) in immune organs of developing chicks by polycyclic aromatic hydrocarbons

Nisha AR, Hazilawati H, Mohd Azmi ML, Noordin MM

Toxicol. Mech. Methods, 2018 Jul;28(6):461-466.
PMID: 29606035 DOI: 10.1080/15376516.2018.1459992

Polycyclic aromatic hydrocarbons are pollutants which are persistent in nature. The aryl hydrocarbon receptor is a ligand-activated cytosolic transcription factor activated by xenobiotics. The objective was to isolate and identify AHR mRNA transcript in immune organs of developing chicks and to interpret the correlation between AHR induction and dose of PAHs. Specific pathogen free embryonated eggs on day nine were inoculated with solutions of pyrene, phenanthrene, and fluoranthene dissolved in tricaprylin (vehicle) through the allantoic route at three dose levels: 0.2 mg/kg, 2 mg/kg, and 20 mg/kg. A 650 base pair product was observed by RNA extraction and reverse transcription PCR from thymus, bursa of Fabricius and spleen on 21st day. When AHR concentration was analyzed by ELISA in these organs, pyrene showed maximum potency in inducing AHR in thymus. Fluoranthene made highest concentration of AHR in bursa of Fabricius. None of these chemicals caused an increase in AHR concentration in spleen.
Fulltext Pathological changes and bacteriological assessments in the urinary tract of pregnant goats experimentally infected with Brucella melitensis

Mazlina M, Khairani-Bejo S, Hazilawati H, Tiagarahan T, Shaqinah NN, Zamri-Saad M

BMC Vet Res, 2018 Jun 25;14(1):203.
PMID: 29940976 DOI: 10.1186/s12917-018-1533-x

BACKGROUND: This study was conducted to investigate the pathological changes and distribution of B. melitensis in the urinary tract of pregnant goats following acute experimental infection. Six Jamnapari crossbred does in their third trimester of pregnancy were randomly assigned into two groups; Group 1 was uninfected control and Group 2 was inoculated conjunctival with 0.1 mL of the inoculums containing 109 cfu/mL of live B. melitensis. All does were sacrificed 30 days post-inoculation before the kidney, ureter, urinary bladder, urethra and vaginal swab were collected for isolation of B. melitensis. The same tissue samples were fixed in 10% neutral buffered formalin for hematoxylin and eosin, and immunoperoxidase staining.
RESULTS: None of the goats showed clinical signs or gross lesions. The most consistent histopathology finding was the infiltration of mononuclear cells, chiefly the macrophages with few lymphocytes and occasionally neutrophils in all organs along the urinary tract of the infected goats of Group 2. Other histopathology findings included mild necrosis of the epithelial cells of the renal tubules, congestion and occasional haemorrhages in the various tissues. Kidneys showed the most severe lesions. Immunoperoxidase staining revealed the presence of B. melitensis within the infiltrating macrophages and the epithelium of renal tubules, ureter, urethra and urinary bladder. Most extensive distribution was observed in the urinary bladder. Brucella melitensis was successfully isolated at low concentration (3.4 × 103 cfu/g) in the various organs of the urinary tract and at high concentration (2.4 × 108 cfu/mL) in the vaginal swabs of all infected goats. Although B. melitensis was successfully isolated from the various organs of the urinary tract, it was not isolated from the urine samples that were collected from the urinary bladder at necropsy.
CONCLUSION: This study demonstrates the presence of low concentrations of B. melitensis in the organs of urinary tract of pregnant does, resulting in mild histopathology lesions. However, B. melitensis was not isolated from the urine that was collected from the urinary bladder.
Fulltext Antihypercholesterolemic and antioxidant efficacies of zerumbone on the formation, development, and establishment of atherosclerosis in cholesterol-fed rabbits

Hemn HO, Noordin MM, Rahman HS, Hazilawati H, Zuki A, Chartrand MS

Drug Des Devel Ther, 2015;9:4173-208.
PMID: 26347047 DOI: 10.2147/DDDT.S76225

Owing to the high incidence of cholesterol-induced cardiovascular disease, particularly atherosclerosis, the current study was designed to investigate the preventive and therapeutic efficacies of dietary zerumbone (ZER) supplementation on the formation and development of atherosclerosis in rabbits fed with a high cholesterol diet. A total of 72 New Zealand white rabbits were divided randomly on two experimental studies carried out 8 weeks apart. The first experiment was designed to investigate the prophylactic efficacy of ZER in preventing early developed atheromatous lesion. The second experimental trial was aimed at investigating the therapeutic effect of ZER in reducing the atherosclerotic lesion progression and establishment. Sudanophilia, histopathological, and ultrastructural changes showed pronounced reduction in the plaque size in ZER-medicated aortas. On the other hand, dietary supplementation of ZER for almost 10 weeks as a prophylactic measure indicated substantially decreasing lipid profile values, and similarly, plaque size in comparison with high-cholesterol non-supplemented rabbits. Furthermore, the results of oxidative stress and antioxidant biomarker evaluation indicated that ZER is a potent antioxidant in suppressing the generation of free radicals in terms of atherosclerosis prevention and treatment. ZER significantly reduced the value of malondialdehyde and augmented the value of superoxide dismutase. In conclusion, our data indicated that dietary supplementation of ZER at doses of 8, 16, and 20 mg/kg alone as a prophylactic measure, and as a supplementary treatment with simvastatin, significantly reduced early plague formation, development, and establishment via significant reduction in serum lipid profile, together with suppression of oxidative damage, and therefore alleviated atherosclerosis lesions.
Brewers' rice attenuated aberrant crypt foci developing in colon of azoxymethane-treated rats

Tan BL, Norhaizan ME, Pandurangan AK, Hazilawati H, Roselina K

Pak J Pharm Sci, 2016 Jan;29(1):205-12.
PMID: 26826813

Brewers' rice is one of abundant agricultural waste products in the rice industry. The present study is designed to investigate the potential of brewers' rice to inhibit the development of aberrant crypt foci (ACF) in colon of azoxymethane (AOM)-treated rats. The effects on the attenuation of hepatic toxicity and kidney function enzymes were also evaluated. Male Sprague-Dawley rats were randomly divided into five groups: (G1) normal; (G2) AOM alone; and (G3), (G4), and (G5), which were AOM fed with 10%, 20%, and 40% (w/w) of brewers' rice, respectively. The rats in group 2-5 were injected intraperitoneally with AOM (15 mg/kg body weight) once weekly for two weeks. After 8 weeks of treatment,the total number of ACF/colon and the number of ACF in the distal and middle colon were significantly reduced in all treatment groups compared to G2 (p<0.05). Brewers' rice decreased the number of ACF with dysplastic morphology in a dose-dependent manner. Alkaline phosphatase (ALP) level in G5 was significantly lower compared to the G2 (p<0.05). In conclusion, this study found the potential value of brewers' rice in reducing the risk of cancer susceptibility in colon.
Fulltext Water extract of brewers' rice induces apoptosis in human colorectal cancer cells via activation of caspase-3 and caspase-8 and downregulates the Wnt/β-catenin downstream signaling pathway in brewers' rice-treated rats with azoxymethane-induced colon carcinogenesis

Tan BL, Norhaizan ME, Huynh K, Heshu SR, Yeap SK, Hazilawati H, et al.

BMC Complement Altern Med, 2015;15:205.
PMID: 26122204 DOI: 10.1186/s12906-015-0730-4

Brewers' rice, is locally known as temukut, is a mixture of broken rice, rice bran, and rice germ. The current study is an extension of our previous work, which demonstrated that water extract of brewers' rice (WBR) induced apoptosis in human colorectal cancer (HT-29) cells. We also identified that brewers' rice was effective in reducing the tumor incidence and multiplicity in azoxymethane (AOM)-injected colon cancer rats. Our present study was designed to identify whether WBR confers an inhibitory effect via the regulation of upstream components in the Wnt signaling pathway in HT-29 cells. To further determine whether the in vitro mechanisms of action observed in the HT-29 cells inhibit the downstream signaling target of the Wnt/β-catenin pathway, we evaluated the mechanistic action of brewers' rice in regulating the expressions and key protein markers during colon carcinogenesis in male Sprague-Dawley rats.
Fulltext Brewers' rice modulates oxidative stress in azoxymethane-mediated colon carcinogenesis in rats

Tan BL, Norhaizan ME, Huynh K, Yeap SK, Hazilawati H, Roselina K

World J Gastroenterol, 2015 Aug 7;21(29):8826-35.
PMID: 26269672 DOI: 10.3748/wjg.v21.i29.8826

To investigate the mechanistic action of brewers' rice in regulating the Wnt/nuclear factor-kappa B (NF-κB)/Nrf2-signaling pathways during colon carcinogenesis in male Sprague-Dawley rats.
Antigenic distribution, pathological changes, antibody response and serological detection in non-pregnant goats following experimental infection by Brucella melitensis

Mazlina M, Khairani-Bejo S, Hazilawati H, Shaqinah NN, Zamri-Saad M

Transbound Emerg Dis, 2021 Jul;68(4):2028-2038.
PMID: 32979887 DOI: 10.1111/tbed.13850

This study describes the pathological changes, antibody response, isolation and distribution patterns following exposure of non-pregnant goats to live Brucella melitensis. Eighteen healthy adult female goats were divided into two equal groups. Group 1 was infected via conjunctival sac with 109 cfu/ml of B. melitensis, while Group 2 was similarly exposed to sterile PBS. Serum and swabs from the eyes and vagina were collected at 5-day intervals. On days 15, 30 and 75 post-infection, 3 goats from each group were killed before the conjunctiva, ovary, oviduct, uterine horn, uterine body and vagina, the submandibular, prescapular and supramammary lymph nodes, the mammary gland, liver, spleen, urinary bladder and synovial membranes were collected for bacterial isolation and pathological study. Exposure of non-pregnant goats to B. melitensis did not produce clinical signs and gross lesions but produced mild necrosis and inflammation in the lymph nodes, the organs of reproductive tract, the mammary gland and urinary bladder. In general, microscopic lesions were most severe in the D75 goats, followed by D30 and D15 goats. Brucella melitensis was most frequent and significantly (p
Fulltext Subacute Oral Toxicity Assesment of Ethanol Extract of Mariposa christia vespertilionis Leaves in Male Sprague Dawley Rats

Nurul SAS, Hazilawati H, Mohd RS, Mohd FHR, Noordin MM, Norhaizan ME

Toxicol Res, 2018 Apr;34(2):85-95.
PMID: 29686770 DOI: 10.5487/TR.2018.34.2.085

The term Butterfly tea refers to decoction of Mariposa christia vespertilionis leaves which is widely consumed by cancer patients throughout Malaysia and has gained a huge popularity among Malaysians, not only cancer patients but also researchers to discover the real potential of this plant. Herein, the study is aimed at evaluating the possible toxicity in 28-day subacute oral toxicity of ethanolic extract M. christia vespertilionis in male Sprague Dawley rats. The 28-day subacute toxicity study was conducted to detect the no-observed adverse effect level (NOAEL). In this study, a total of 30 rats were divided into the control, 5% DMSO (vehicle), low dose (75 mg/kg), medium dose (125 mg/kg) and high dose (250 mg/kg) groups. The extract was administered daily from day 1 until day 28. At the end of the study, the animals were humanely sacrificed and assessed for the effect extract of Mariposa christia vespertilionis leaves on body weight and relative organ weights and haematological, biochemical and histopathological parameters. The haematological and serum biochemical parameters for the assessment of kidney and liver injuries were carried out. Results of haematological and serum biochemistry results showed no changes in the control and treated groups. In the histopathology, evaluation of kidney tissues in all treated groups showed no significant (p > 0.05) lesions. In contrast to kidney, liver tissues showed significant differences (p < 0.05) in lesions observed in low dose (430 mg), medium dose (700 mg) and high dose (1480 mg) groups with very mild, mild and mild to moderate lesion of hepatic necrosis, in the respective groups, and very mild hepatic degeneration and hepatitis were scored in all three groups.

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