Umah Rani A/P Kuppusamy

Abstract: Mushrooms have been used in folk medicine for thousands of years. In this study, the effect of β -glucan-rich extract of P. sajor-caju (GE) on lipid lowering and antioxidant potential was assessed in C57BL/6J mice fed on a high-fat diet. Obesity was induced in C57BL/6J mice by feeding a high-fat diet. The control groups in this study were ND (for normal diet) and HFD (for high-fat diet). The treated groups were ND240 (for normal diet) (240�mg/kg b.w) and HFD60, HFD120, and HFD240 (for high-fat diet), where the mice were administrated with three dosages of GE (60, 120, and 240�mg GE/kg b.w). Metformin (2�mg/kg b.w) served as positive control. GE-treated groups showed significantly reduced body weight, serum lipid, and liver enzymes levels. GE also attenuated protein carbonyl and lipid hydroperoxide levels by increasing the enzymic antioxidants (SOD, CAT, and GPx) activities in the mice. GE-treated groups induced the expression of hormone sensitive lipase (HSL) and adipose triglyceride lipase (ATGL) while downregulated the expression of peroxisome proliferator-activated receptor gamma (PPAR- γ ), sterol regulatory binding protein-1c (SREBP-1c), and lipoprotein lipase (LPL). Hence, GE prevented weight gain in the mice by inducing lipolysis and may be valuable in the formulation of adjuvant therapy for obesity.

Abstract: Receptor for advanced glycation end-product (RAGE) gene polymorphism 2245G/A is associated with diabetic retinopathy (DR). However, the mechanism on how it affects the disease development is still unclear.

Abstract: This study aimed to assess the circulating levels of activated nuclear factor kappa B p65 and monocyte chemotactic protein-1 in diabetic retinopathy patients who were taking antihyperglycemic and antihypertensive drugs.

Abstract: This study aims to investigate potential diabetic retinopathy (DR) risk factors by evaluating the circulating levels of pentosidine, soluble receptor for advanced glycation end-product (sRAGE), advanced oxidation protein product (AOPP) as well as glutathione peroxidase (GPx) and superoxide dismutase (SOD) activities in DR patients. A total of 235 healthy controls, 171 type 2 diabetic without retinopathy (DNR) and 200 diabetic retinopathy (DR) patients were recruited. Plasma was extracted for the estimation of pentosidine, sRAGE, AOPP levels and GPx activity whereas peripheral blood mononuclear cells were disrupted for SOD activity measurement. DNR and DR patients showed significantly higher levels of plasma pentosidine, sRAGE and AOPP but lower GPx and SOD activities when compared to healthy controls. The sRAGE/pentosidine ratio in DR patients was significantly lower than the ratio detected in DNR patients. Proliferative DR patients had significantly higher levels of plasma pentosidine, sRAGE, AOPP and sRAGE/pentosidine ratio than non-proliferative DR patients. High HbA1c level, long duration of diabetes and low sRAGE/pentosidine ratio were determined as the risk factors for DR. This study suggests that sRAGE/pentosidine ratio could serve as a risk factor determinant for type 2 DR as it has a positive correlation with the severity of DR.

Abstract: The pathogenesis of Blastocystis hominis in human hosts has always been a matter of debate as it is present in both symptomatic and asymptomatic individuals. A recent report showed that B. hominis isolated from an asymptomatic individual could facilitate the proliferation and growth of existing cancer cells while having the potential to downregulate the host immune response. The present study investigated the differences between the effects of symptomatic and asymptomatic derived solubilized antigen of B. hominis (Blasto-Ag) on the cell viability and proliferation of colorectal cancer cells. Besides that, the gene expression of cytokine and nuclear transcriptional factors in response to the symptomatic and asymptomatic B. hominis antigen in HCT116 was also compared. In the current study, an increase in cell proliferation was observed in HCT116 cells which led to the speculation that B. hominis infection could facilitate the growth of colorectal cancer cells. In addition, a more significant upregulation of Th2 cytokines observed in HCT116 may lead to the postulation that symptomatic Blasto-Ag may have the potential in weakening the cellular immune response, allowing the progression of existing tumor cells. The upregulation of nuclear factor kappa light chain enhancer of activated B cells (NF-κB) was observed in HCT116 exposed to symptomatic Blasto-Ag, while asymptomatic Blasto-Ag exhibited an insignificant effect on NF-κB gene expression in HCT116. HCT116 cells exposed to symptomatic and asymptomatic Blasto-Ag caused a significant upregulation of CTSB which lead to the postulation that the Blasto-Ag may enhance the invasive and metastasis properties of colorectal cancer. In conclusion, antigen isolated from a symptomatic individual is more pathogenic as compared to asymptomatic isolates as it caused a more extensive inflammatory reaction as well as more enhanced proliferation of cancer cells.

Abstract: Conflicting results have been reported in different populations on the association between two particular RAGE gene polymorphisms (-429T/C and -374T/A) and retinopathy in diabetic patients. Therefore this study was designed to assess the association between both gene polymorphisms with retinopathy in Malaysian diabetic patients. A total of 342 type 2 diabetic patients [171 without retinopathy (DNR) and 171 with retinopathy (DR)] and 235 healthy controls were included in this study. Genomic DNA was obtained from blood samples and the screening for the gene polymorphisms was done using polymerase chain reaction-restriction fragment length polymorphism approach. Overall, the genotype distribution for both polymorphisms was not statistically different (p>0.05) among the control, DNR and DR groups. The -429C minor allele frequency of DR group (12.0%) was not significantly different (p>0.05) when compared to DNR group (16.1%) and healthy controls (11.3%). The -374A allele frequency also did not differ significantly between the control and DNR (p>0.05), control and DR (p>0.05) as well as DNR and DR groups (p>0.05). This is the first study report on RAGE gene polymorphism in Malaysian DR patients. In conclusion, -429T/C and -374T/A polymorphisms in the promoter region of RAGE gene were not associated with Malaysian type 2 DR patients.

Abstract: Chemotherapy can cause immunosuppression, which may trigger latent intestinal parasitic infections in stools to emerge. This study investigated whether intestinal parasites can emerge as opportunistic infections in breast and colorectal cancer patients (n=46 and n=15, respectively) undergoing chemotherapy treatment. Breast cancer patients were receiving a 5-fluorouracil/epirubicin/cyclophosphamide (FEC) regimen (6 chemotherapy cycles), and colorectal cancer patients were receiving either an oxaliplatin/5-fluorouracil/folinic acid (FOLFOX) regimen (12 cycles) or a 5-fluorouracil/folinic acid (Mayo) regimen (6 cycles). Patients had Blastocystis hominis and microsporidia infections that were only present during the intermediate chemotherapy cycles. Thus, cancer patients undergoing chemotherapy should be screened repeatedly for intestinal parasites, namely B. hominis and microsporidia, as they may reduce the efficacy of chemotherapy treatments.

Abstract: The receptor for advanced glycation end-products (RAGE) has been implicated in the pathogenesis of diabetic microvascular complications. The aim of this study was to investigate the association between 2245G/A gene polymorphism of the RAGE gene and retinopathy in Malaysian type 2 diabetic patients.

Abstract: Diabetic retinopathy is the most common diabetic eye disease, occurring in about 60% of type 2 diabetic patients. Other than known clinical risk factors, the influence of genes has been suggested as part of the development of diabetic retinopathy. We investigated the association of Gly82Ser, 1704G/T and 2184A/G polymorphisms in the RAGE gene with retinopathy in type 2 diabetic patients in Malaysia. Ninety-eight unrelated retinopathy patients and 185 unrelated healthy controls from all over Malaysia were recruited in this study. The allele and genotype frequencies of the three gene polymorphisms were investigated using PCR-RFLP. The allele frequency of the three polymorphisms did not differ significantly between the control and the retinopathy group (P > 0.05). Analysis of the frequency of GA+AA, GT+TT and AG+GG in the retinopathy group did not reveal significant differences (P > 0.05) compared to the control group. We conclude that RAGE gene Gly82Ser, 1704G/T and 2184A/G polymorphisms are not associated with retinopathy development in the Malaysian population.

Abstract: Pleurotus sajor-caju (P. sajor-caju) has been extremely useful in the prevention of diabetes mellitus due to its low fat and high soluble fiber content for thousands of years. Insulin resistance is a key component in the development of diabetes mellitus which is caused by inflammation. In this study, we aimed to investigate the in vivo efficacy of glucan-rich polysaccharide of P. sajor-caju (GE) against diabetes mellitus and inflammation in C57BL/6J mice fed a high-fat diet.

Abstract: In this study, we aimed to investigate the possible association between SLC2A1 26177A/G polymorphism and diabetic retinopathy (DR) in Malaysian patients with type 2 diabetes.

Abstract: Panus giganteus, a culinary and medicinal mushroom consumed by selected indigenous communities in Malaysia, is currently being considered for large scale cultivation. This study was undertaken to investigate the hepatoprotective effects of P. giganteus against thioacetamide- (TAA-) induced liver injury in Sprague-Dawley rats. The rats were injected intraperitoneally with TAA thrice weekly and were orally administered freeze-dried fruiting bodies of P. giganteus (0.5�or�1�g/kg) daily for two months, while control rats were given vehicle or P. giganteus only. After 60 days, rats administered with P. giganteus showed lower liver body weight ratio, restored levels of serum liver biomarkers and oxidative stress parameters comparable to treatment with the standard drug silymarin. Gross necropsy and histopathological examination further confirmed the hepatoprotective effects of P. giganteus. This is the first report on hepatoprotective effects of P. giganteus. The present study showed that P. giganteus was able to prevent or reduce the severity of TAA-induced liver injury.

Abstract: The chemical composition and in vitro antioxidant activity of aqueous butanol and ethyl acetate extracts of Pleurotus sajor-caju were investigated in this study. Twenty-two compounds comprising methyl esters, hydrocarbon fatty acids, ethyl esters, and sterols were identified in ethyl acetate extracts, while cinnamic acid, nicotinamide, benzeneacetamide, and 4-hydroxybenzaldyhde were identified in butanol extracts by gas chromatography-mass spectrometry and NMR analysis. The antioxidant activity was determined by a β-carotene bleaching method, ferric reducing antioxidant power, trolox equivalent antioxidant capacity, and lipid peroxidation assays, while the total phenolic content in P. sajor-caju was assessed by Folin-Ciocalteau's method. The aqueous and butanol extracts exhibited the highest antioxidant activity, corresponding to the total phenolic content. The subfractions from the ethyl acetate extract (EP1, EP2, EP3, and EP4), however, showed moderate antioxidant activity. The regular consumption of P. sajor-caju as a part of our diet may render nutritional and nutraceuticals benefits for good health.

Abstract: The present study compares water-soluble phenolic content (WPC) and antioxidant activities in Chinese long bean (Vigna unguiculata), bitter gourd (Momordica charantia), water convolvulus (Ipomoea aquatica) and broccoli (Brassica olearacea) prior to and after subjecting to boiling, microwaving and pressure cooking. The total antioxidant activity was increased in cooked water convolvulus, broccoli and bitter gourd, estimated based on the ferric reducing antioxidant power, the Trolox equivalent antioxidant capacity and 2,2-diphenyl-1-picryl-hydrazyl radical scavenging activity. Pressure cooking did not cause any significant decline in the antioxidant property. Boiling generally improved the overall antioxidant activity in all the vegetables. Correlation analysis suggests that WPC contributed to significant antioxidant activities in these vegetables. Thus, prudence in selecting an appropriate cooking method for different vegetables may improve or preserve their nutritional value.

Abstract: Water extract of Lentinus squarrosulus mycelia was analysed for nutritional content, antioxidant capacity, and antiulcer ability. The extract contains high protein (57.6�g/100�g) and low total fat (0.5�g/100�g) and is rich in magnesium (0.4�g/100�g), potassium (3.8�g/100�g), vitamins B(1) (1.42�mg/100�g), and B(3) (194.29�mg/100�g) with total phenolic content of 39.16�mg/100�g. The cupric reducing antioxidant capacity and 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity of the extract were A(450) of 0.20 ± 0.03 at 0.5�mg/ml and IC(50) of 14.29�mg/ml, respectively. Oral feeding of L. squarrosulus extract (250�mg/kg) offered significant gastric mucosal protection of Sprague-Dawley rats compared to cimetidine (50�mg/kg). The ulcer healing rate of ulcerated rats after 24, 48, and 72 hours of treatment was 82%, 90%, and 100%, respectively. The IL-1β level in the serum and the NF-κB level in the tissues indicate that the healing potential was associated with attenuation of proinflammatory cytokines.

Abstract: Nerve crush injury is a well-established axonotmetic model in experimental regeneration studies to investigate the impact of various pharmacological treatments. Hericium erinaceus is a temperate mushroom but is now being cultivated in tropical Malaysia. In this study, we investigated the activity of aqueous extract of H. erinaceus fresh fruiting bodies in promoting functional recovery following an axonotmetic peroneal nerve injury in adult female Sprague-Dawley rats by daily oral administration. The aim was to investigate the possible use of this mushroom in the treatment of injured nerve. Functional recovery was assessed in behavioral experiment by walking track analysis. Peroneal functional index (PFI) was determined before surgery and after surgery as rats showed signs of recovery. Histological examinations were performed on peroneal nerve by immunofluorescence staining and neuromuscular junction by combined silver-cholinesterase stain. Analysis of PFI indicated that return of hind limb function occurred earlier in rats of aqueous extract or mecobalamin (positive control) group compared to negative control group. Regeneration of axons and reinnervation of motor endplates in extensor digitorum longus muscle in rats of aqueous extract or mecobalamin group developed better than in negative control group. These data suggest that daily oral administration of aqueous extract of H. erinaceus fresh fruiting bodies could promote the regeneration of injured rat peroneal nerve in the early stage of recovery.

Abstract: This study was conducted to evaluate the effects of topical application of aqueous extract of Hericium erinaceus fruiting bodies (HEFB) on the rate of wound healing enclosure and histology of the healed wound. Five groups of male Sprague-Dawley rats were experimentally wounded in the posterior neck area. A uniform wound area of 2.00 cm in diameter, using a circular stamp, was excised from the nape of the dorsal neck of all rats with the aid of a round seal. The animal groups were topically treated, respectively, with 0.2 mL each of sterilized distilled water (sdH2O); Intrasite gel; and 20, 30, and 40 mg/mL HEFB. Macroscopically, those rats whose wounds were dressed with HEFB and those in the Intrasite gel-treated group healed earlier than those treated with sdH2O. Histological analysis of healed wounds dressed with HEFB showed less scar width at wound enclosure and the healed wound contained fewer macrophages and more collagen with angiogenesis, compared to wounds dressed with sdH2O. In conclusion, wounds dressed with HEFB significantly enhanced the acceleration of wound healing enclosure in rats.

Abstract: Beta-thalassaemia major causes severe anaemia and patients with it may be transfusion-dependent for life. Regular blood transfusions cause iron-overload that leads to oxidative damage which can hasten mortality. The objective of this research was to study the oxidant-antioxidant indices in beta-thalassaemia major patients at the University of Malaya Medical Centre (UMMC) who were on desferrioxamine-chelation or without chelation therapy. Blood was collected from 39 Chinese patients and 20 controls. Plasma and peripheral blood mononuclear cell lysates (PBMC) were extracted and biochemical tests to evaluate oxidative stress were performed. Oxidative stress was evident in these patients as advanced oxidized protein products (AOPP) and lipid hydroperoxides were elevated, whereas glutathione peroxidase activity and the ferric reducing antioxidant power (FRAP) were reduced. The catalase activity in the patients' PBMC was elevated, possibly as a compensatory mechanism for the reduced glutathione peroxidase activity in both red blood cells and PBMC. The lower FRAP and higher AOPP levels in the non-chelated patients compared with the chelated patients were indicative of a lower oxidative stress level in the chelated patients. The ferritin levels in the chelated and non-chelated patients were high and the mean levels of liver enzyme activities in the majority of patients were elevated regardless of chelation therapy. In conclusion, this study indicates that desferrioxamine chelation therapy does not normalize ferritin level but attenuates oxidative damage and improves total antioxidant level in Malaysian Chinese beta-thalassaemia major patients.

Abstract: The nutraceutical benefits of β-sitosterol (SIT) are well documented. The present study investigated the in vitro effects of SIT on adipogenesis, glucose transport, and lipid mobilization in rat adipocytes. Primary cultures of rat preadipocytes and differentiated adipocytes were used in this study. Glucose uptake was measured by the uptake of radio-labeled glucose. Adipogenesis and lipolysis were measured by oil-red-O and glycerol quantification methods, respectively. The expression of protein kinase B (Akt), glucose transporter 4 (GLUT4), hormone sensitive lipase (HSL), and phosphatidylinositol-3-kinase (PI3 K) genes in SIT-treated adipocytes were assessed by real-time reverse transcription polymerase chain reaction (RT-PCR). The data showed that SIT induced glucose uptake in adipocytes. It also stimulated adipogenesis in differentiating preadipocytes. Interestingly, although SIT displayed general insulin-mimetic activity by stimulating glucose uptake and adipogenesis, it also induced lipolysis in adipocytes. Furthermore, the SIT-induced lipolysis was not attenuated by insulin and co-incubation of SIT with epinephrine improved epinephrine-induced lipolysis. GLUT4 gene expression was highly down-regulated in SIT-treated adipocytes, compared to insulin-treated adipocytes, which was up-regulated. Insulin- and SIT-treated adipocytes showed similar levels of Akt, HSL, and PI3 K gene down-regulation. These observations suggest that the elevation of glucose uptake in SIT-treated adipocytes was unrelated to de novo synthesis of GLUT4 and the SIT-induced lipolysis is associated with the down-regulation of Akt and PI3K genes. The unique effects of SIT on the regulation of glucose uptake, adipogenesis, and lipolysis in adipocytes show that it has potential to be utilized in diabetes and weight management.

Abstract: Numerous studies have revealed the presence of oxidative stress in parasitic infections. However, such studies were lacking in the Malaysian population. Previously, we have provided evidence that oxidative stress is elevated in Malaysians infected with intestinal parasites. Stool examinations revealed that about 47.5% of them were infected with the polymorphic protozoa, Blastocystis hominis. However, they were found to have mixed infection with other intestinal parasites.

Abstract: Blastocystis hominis is one of the most common intestinal protozoan parasites in humans, and reports have shown that blastocystosis is coupled with intestinal disorders. In the past, researchers have developed an in vitro model using B. hominis culture filtrates to investigate its ability in triggering inflammatory cytokine responses and transcription factors in human colonic epithelial cells. Studies have also correlated the inflammation by parasitic infection with cancer. The present study provides evidence of the parasite facilitating cancer cell growth through observing the cytopathic effect, cellular immunomodulation, and apoptotic responses of B. hominis, especially in malignancy. Here we investigated the effect of solubilized antigen from B. hominis on cell viability, using peripheral blood mononuclear cells (PBMCs) and human colorectal carcinoma cells (HCT116). The gene expressions of cytokines namely interleukin 6 (IL-6), IL-8, tumor necrosis factor alpha, interferon gamma, nuclear factor kappa light-chain enhancer of activated B cells (a gene transcription factor), and proapoptotic genes namely protein 53 and cathepsin B were also studied. Results exhibited favor the fact that antigen from B. hominis, at a certain concentration, could facilitate the growth of HCT116 while having the ability to downregulate immune cell responses (PBMCs). Therefore, there is a vital need to screen colorectal cancer patients for B. hominis infection as it possesses the ability to enhance the tumor growth.

Abstract: The fact whether Blastocystis hominis can invade has always been in question. Apart from a few sporadic studies such as that done on gnotobiotic guinea pigs which showed surface invasion and mucosal inflammation of the host's intestine caused by B. hominis infection, no real documentation of invasion has been proven. Studies have shown that hyaluronidase is secreted during the penetration into the host's skin and gut by nematode parasites. Hyaluronidase activity in protozoa namely Entamoeba histolytica has also been described previously. This study attempts to determine hyaluronidase in urine samples of B. hominis-infected rats. The presence of hyaluronidase in urine provides an indirect evidence of invasion by B. hominis into colonic epithelium causing the degradation of extracellular matrix proteins namely hyaluronic acid (HA). HA is depolymerized by hyaluronidase which may be used by organisms to invade one another. In this study, the levels of urinary hyaluronidase of Sprague-Dawley rats infected with B. hominis were monitored for 30 days. Hyaluronidase levels in the infected rats were significantly higher on days 28 and 30 compared to the day before inoculation (P < 0.01 and P < 0.05, respectively). During this stage, parasitic burden in infected stools was also at a high level. Proinflammatory cytokines, interleukin-6 and interleukin-8, were also significantly higher (P < 0.05) in the serum of infected rats. The study demonstrates that since no other pathogen was present and that amoeboid forms of the parasites have been shown to exist previously, the elevated levels of hyaluronidase in this preliminary finding suggests that the organism is capable of having invasion or penetration activity in the hosts' intestine.

Abstract:

Abstract: Aqueous extracts from 20 Malaysian edible plants were screened for total phenolic content (TPC), antioxidant activity and genotoxic effects on freshly isolated human lymphocytes. Ferric reducing antioxidant power (FRAP) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging assays were performed to determine the antioxidant activity of the extracts. Single-cell gel electrophoresis (SCGE) or comet assay was carried out to determine the level of DNA damage in human lymphocytes. Of the 20 plant aqueous extracts tested, two exerted more than 50% DNA strand breaks (severe damage), nine exerted 25-50% strand breaks (moderate damage) and nine exerted <25% strand breaks (mild damage). Strong positive correlations between the extent of DNA damage and FRAP level (r = 0.816), DNA damage and TPC (r = 0.830) and DNA damage and DPPH radical scavenging activities (r = 0.859) were observed. It is evident from this study that plants rich in antioxidants have greater genotoxic effect. (C) 2010 Elsevier Ltd. All rights reserved.

Abstract: Hericium erinaceus, a temperate mushroom, is currently cultivated in Malaysia. As cultivation and processing conditions may affect the medicinal properties, antimicrobial and antioxidant properties of locally grown H. erinaceus have been investigated. The fruitbodies that were fresh, oven-dried or freeze-dried were extracted with methanol. Their properties were compared to those exhibited by mycelium extract of the same mushroom. Various extracts of H. erinaceus inhibited the growth of pathogenic bacteria but not of the tested fungus. Mycelium extract contained the highest total phenolic content and the highest ferric reducing antioxidant power (FRAP). The fresh fruitbody extract showed the most potent 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity. However, oven-dried fruitbody extract was excellent in reducing the extent of P-carotene bleaching. The total phenolic content and total antioxidant activity in the oven-dried fruitbody extract was high compared to the freeze-dried or fresh fruitbody extract. This may be due to generation and accumulation of Maillard�s reaction products (MRPs), which are known to have antioxidant properties. Thus, the consumption of H. erinaceus fruitbody grown in tropical conditions may have health promoting benefits. Furthermore, the production of H. erinaceus mycelium in submerged cultures may result in standardized antioxidant formulation for either human nutrition or therapy. Hence, it has been shown that the processing of fruitbody and not the cultivation conditions affects the selected bioactive properties of H. erinaceus.

Abstract: This study was aimed to evaluate the antioxidant properties of methanol extracts of fermented substrates optimised for antioxidant production by Marasmiellus sp. KUM 50061 mycelial biomass. Extract of fermented maize supplemented with (w/w) malt extract 4%, yeast extract 4% and rice bran 4% exhibited the highest 1,1diphenyl-2-picrylhydrazyl radical scavenging ability. The effective concentration of extract to scavenge 50% radicals was 1.875 mg mL-1. This formulation was chosen as the optimum substrate for antioxidant production by Marasmiellus sp. KUM 50061 mycelial biomass. The thiobarbituric acid reactive substance (TBARS) assay showed that the effective concentration to inhibit lipid peroxidation of buffered egg yolk by 50% was 6.00 mg mL-1. Total phenolics amounted to 31.41��1.56 mg GAE g-1 extract as measured by the Folin-Ciocalteau method.

Abstract: Oxidative stress has been implicated as an important pathogenic factor in the pathophysiology Of various life-threatening diseases such as cancer, cardiovascular diseases and diabetes. It occurs when the production of free radicals (generated during aerobic metabolism, inflammation, and infections) overcome the antioxidant defences in the body. Although previous Studies have implied that oxidative stress is present in serum of patients with parasitic infection there have been no Studies confirming oxidative stress levels in the Malaysian population infected with intestinal parasites. Three biochemical assays namely hydrogen peroxide (H(2)O(2)), lipid peroxidation (LP) and advanced oxidative protein product (AOPP) assays Were carried out to measure oxidative stress levels in the urine of human subjects whose stools,were infected with parasites Such as Blastocystis hominis, Ascaris, Trichuris, hookworm and microsporidia. The levels of H(2)O(2), AOPP and LP were Significantly higher (P<0.001, P<0.05 and P<0.05 respectively) in the parasite-infected subjects (n = 75) compared to the controls (n = 95). In conclusion, the study provides evidence that oxidative stress is elevated in humans infected by intestinal parasites. This study may influence future researchers to consider free radical-related pathways to be a target in the interventions of new drugs against parasitic infection and related diseases.

Abstract: Peripheral nerve injury represents a huge burden to society. Following peripheral nerve injury, improved behavioral outcome may be the most important evidence of functionality of axonal regeneration after any repair strategy. Nerve-crush injury is a well-established axonotmetic model in experimental regeneration studies to investigate the impact of various pharmacological treatments. Hericium erinaceus is a temperate mushroom but is now being cultivated in tropical Malaysia. In this study, we investigated the activity of aqueous extract of H. erinaceus fresh fruitbodies in promoting functional recovery following an axonotmetic peroneal nerve injury in adult female Sprague-Dawley rats with a long-term view toward the possible use of this mushroom in the treatment of nerve injury. Functional recovery was assessed in the behavioral experiment by walking-track analysis and toe-spreading reflex. The peroneal functional index (PFI) was determined before surgery and after surgery, as the rats showed signs of recovery. Analysis of the PFI indicated that the return of hind-limb function occurred by day 10 to 14 and by day 14 to 17 in the treated and control (nontreated) groups, respectively. Normal toe-spreading in the crushed limb was achieved by day 7 to 10 and day 12 to 17 in the treated and control group, respectively. These results suggest that daily administration of aqueous extract of H. erinaceus fresh firuitbodies has a beneficial effect on the recovery of injured rat peroneal nerve in the early stages of regeneration. The PFI and toe-spreading reflex improved faster in the treated group than in the nontreated group.

Abstract: Auricularia auricula-judae is currently grown in Malaysia. In the present study, the methanolic extracts from fruit bodies (fresh, oven-dried, and freeze-dried) and mycelium of A. auricula-judae were evaluated for their antioxidant capacities based on 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity and ferric reducing antioxidant power (FRAP) assay. The total phenolic content in the extracts were also measured. The extract of freeze-dried fruit bodies of A. auricula-judae had potent DPPH free radical scavenging activity with a 50% effective concentration of 2.87 mg/mL, whereas the FRAP value of A. auricula-judae mycelium was 5.22 micromol of FeSO(4).7H(2)O equivalents/g of mycelium sample. Further, a positive correlation (R(2) = 0.7668) between FRAP level of A. auricula-judae extracts and the total phenolic contents was observed. Thus the method of processing of fresh fruit bodies had an effect on the antioxidant potential of A. auricula-judae.

Abstract: This study assessed several common oxidative indices in subjects infected with intestinal parasites, as well as in colorectal cancer (CRC) patients both with and without intestinal parasites.

Abstract: Oxidative stress has been implicated as an important pathogenic factor in the pathophysiology of various life-threatening diseases such as cancer, cardiovascular diseases and diabetes. It occurs when the production of free radicals (generated during aerobic metabolism, inflammation, and infections) overcome the antioxidant defences in the body. Although previous studies have implied that oxidative stress is present in serum of patients with parasitic infection there have been no studies confirming oxidative stress levels in the Malaysian population infected with intestinal parasites. Three biochemical assays namely hydrogen peroxide (H2O2), lipid peroxidation (LP) and advanced oxidative protein product (AOPP) assays were carried out to measure oxidative stress levels in the urine of human subjects whose stools were infected with parasites such as Blastocystis hominis, Ascaris, Trichuris, hookworm and microsporidia. The levels of H2O2, AOPP and LP were significantly higher (P<0.001, P<0.05 and P<0.05 respectively) in the parasite-infected subjects (n=75) compared to the controls (n=95). In conclusion, the study provides evidence that oxidative stress is elevated in humans infected by intestinal parasites. This study may influence future researchers to consider free radical-related pathways to be a target in the interventions of new drugs against parasitic infection and related diseases.

Abstract: This study aimed to use non-invasive methods to assess and compare the levels of oxidative indices and non-enzymatic antioxidants in breast and colorectal cancer (CRC) patients. Various studies have reported on lipid peroxidation, hydrogen peroxide (H(2)O(2)) and ferric-reducing antioxidant power (FRAP) levels in the serum of cancer patients but this is the first report that highlights the significance of urinary-advanced oxidative protein product (AOPP) in cancer patients.

Abstract: Lentinula edodes (Berk) Pegler, commonly known as Shiitake mushroom has been used as medicinal food in Asian countries, especially in China and Japan and is believed to possess strong immunomodulatory property. In the present study, the methanolic extract of the fruit bodies of L. edodes was investigated for cytoprotective effect against H2O2-induced cytotoxicity in human peripheral blood mononuclear cells (PBMCs) by measuring the activities of xanthine oxidase (XO) and glutathione peroxidase (GPx) . H2O2 at a concentration of 5 microM caused 50% inhibition of PBMCs viability. The extract improved the PBMC viability and exerted a dose-dependent protection against H2O2-induced cytotoxicity. At 100 microg/ml of extract concentration, the cell viability increased by 60% compared with the PBMCs incubated with H2O2 alone. The extract also inhibited XO activity in PBMC, while showing moderate stimulatory effect on GPx. However, in the presence of H2O2 alone, both the enzyme activities were increased significantly. The GPx activity increased, possibly in response to the increased availability of H2O2 in the cell. When the cells were pretreated with the extract and washed (to remove the extract) prior to the addition of H2O2, the GPx and XO activities as well as the cell viability were comparable to those when incubated with the extract alone. Thus, it is suggested that one of the possible mechanisms via which L. edodes methanolic extract confers protection against H2O2-induced oxidative stress in PBMC is by inhibiting the superoxide-producing XO and increasing GPx activity which could rapidly inactivate H2O2.

Abstract: Health scenarios are constantly evolving, particularly in developing countries but little is known regarding the health status of indigenous groups in Malaysia. This study aims to elucidate the current health status in four indigenous populations in the country, who by and large been left out of mainstream healthcare developments.

Abstract: The effects of Carica papaya leaf (CPL) aqueous extract on alcohol induced acute gastric damage and the immediate blood oxidative stress level were studied in rats. The results showed that gastric ulcer index was significantly reduced in rats pretreated with CPL extract as compared with alcohol treated controls. The in vitro studies using 2,2-Diphenyl-1-Picryl-Hydrazyl (DPPH) assay showed strong antioxidant nature of CPL extract. Biochemical analysis indicated that the acute alcohol induced damage is reflected in the alterations of blood oxidative indices and CPL extract offered some protection with reduction in plasma lipid peroxidation level and increased erythrocyte glutathione peroxidase activity. Carica papaya leaf may potentially serve as a good therapeutic agent for protection against gastric ulcer and oxidative stress.

Abstract: Crude and hexane leaf extracts of Apama tomentosa (Aristolochiaceae) showed inhibitory effect on ovarian adenocarcinoma (CaOV-3) cells cultured in vitro with IC50 of 46.13��1.09 �ľg/ml and 18.50��0.50 �ľg/ml respectively. The crude extract exerted a mild inhibitory effect on peripheral blood mononuclear cells (PBMCs). Hexane extract and palmitone did not exert any significant effect on the PBMC proliferation. Subsequent isolation of the leaf extract led to the isolation and identification of palmitone (16-hentriacontanone). This compound exhibited inhibitory effect (IC50=17.70��1.86 �ľg/ml) on CaOV-3 but was ineffective on PBMCs. This plant compound can potentially help to treat ovarian cancer.

Abstract: Traditionally, the fruits and seeds of Syzygium jambolanum (Lam.) DC [Family: Myrtaceae] have been used to treat diabetes mellitus. Insulin is known to inhibit lipolysis induced by epinephrine as well as promote lipogenesis and glucose uptake adipocytes. The present study evaluates the insulin-like properties of S. jambolanum leaf methanolic extract through lipogenic, anti-lipolytic and glucose uptake activities. Methanolic leaf extract of S. jambolanum induced significant lipogenesis in the dose range of 0.1-10 �ľg mL 1 with the highest activity (114%) at 10 �ľg mL 1. This extract (1 �ľg mL 1 inhibited 29% of epinephrine (1 �ľM) induced lipolysis and enhanced the inhibitory effect of insulin against epinephrine induced lipolysis. This extract also inhibited (64% inhibition) of isoproterenol (1 �ľM) ( -adrenergic agonist) induced lipolysis. However, the extract did not enhance the antilipolytic action of propanolol ( -adrenergic antagonist). Thus, it is possible to speculate that the mode of action of this extract may involve competitive binding at -adrenergic receptor binding sites. S. jambolanum (0.1-10 �ľg mL 1) exerted a dose-dependant glucose uptake activity and further enhanced insulin mediated glucose uptake in primary rat adipocytes. These results confirm that S. jambolanum methanolic leaf extract has insulin-like properties and may be useful as potential therapeutic agent in the management of hyperglycemia.

Abstract: The flavonoid naringenin is commonly found in citrus fruits and tomato. The main aim of this study was to assess the in vitro insulin-like effects of naringenin using rat adipocyte primary culture. The adipogenic, lipolytic and glucose uptake activities of naringenin in primary rat adipocytes were assessed using Oil Red O, glycerol, and 2-deoxy-D-[2,6-3H]glucose uptake quantitative assays respectively. Naringenin exhibited significant increase of lipogenesis in the presence and absence of insulin in primary rat preadipocytes. At the concentration range of 0.01 â�� 100 �ľM, naringenin inhibited 50 % of epinephrine induced lipolysis in rat adipocytes and enhanced insulin�s antilipolytic activity. Naringenin (100 �ľM) stimulated 163 % glucose uptake in rat adipocytes (compared to untreated cells) and this was significantly higher than the insulin mediated glucose uptake at similar concentration. Thus, naringenin may play an important role as an adjuvant and/or alternative to insulin therapy for the management of diabetes mellitus. [ABSTRACT FROM AUTHOR] Copyright of Internet Journal of Alternative Medicine is the property of Internet Scientific Publications LLC and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder�s express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Abstract: Mycelia of Marasmiellus sp. (KUM 50061) were grown on maize for antioxidant production. This formed the mycelial biomass which was then extracted with methanol. Antioxidant activity of methanolic extract was analysed by the TBARS assay, using egg yolk or palm cooking oil as a source of lipid, rather than the conventional rat liver microsomes or linoleic acid. Results showed that at low concentrations of extracts, inhibition of lipid peroxidation in buffered egg yolk was marginal, but significant inhibitory response was evident as the concentrations was increased. The concentration of extract of fermented maize that caused 50% inhibition of lipid peroxidation of buffered egg yolk was 6 mg/ml. Results also indicated a decrease in peroxidation in heated cooking oil supplemented with dried extract compared to unsupplemented cooking oil. The concentration range of dried extract supplementation was 0.2-5 mg/ml. Increasing the extract concentration did not significantly alter the inhibition of peroxidation. The inhibition effect was still evident even at the lowest concentration tested, and was found to be better than catechin and BHA. This pattern of observation was consistent over the 12-day period of observation. Therefore, the possibility of substituting synthetic antioxidants such as BHA and BHT, which are known to be carcinogenic, with antioxidants of natural origin is suggested. (c) 2007 Elsevier Ltd. All rights reserved.

Abstract: Cyrtandra cupulata Ridl. (Gesneriaceae) is local medicinal plant, also known as �Bebangun� or �Kabut� by the East Malaysian Ibans. It is indicated for post partum malaise and known to be an abortifacient, among other uses. ��-Sitosterol is a known phytoestrogen with many reported bioactivities including anticancer. This study examined the effect of ��-sitosterol isolated from C. cupulata through bioactivity-guided purification monitored by the growth inhibitory effect on MCF-7, an estrogen receptor-positive breast cancer cell line. Our results demonstrated that ��-sitosterol dose-dependently inhibited the growth of MCF-7 cells. Treated cells showed a 1.53-fold increase of DEVDase activity, indicating elevated caspase activity. Thus, the growth inhibitory effect of ��-sitosterol isolated from C. cupulata appeared to be mediated by caspase-induced apoptosis.

Abstract: Medicinal plants have widely been used to treat different kinds of ailments including cancer. Syzygium jambolanum, Linn (Myrtaceae) is a tropical plant used traditionally in the treatment of diabetes. However the effect of this plant on cancer has not been reported. Breast cancer is the most prevalent type of cancer among women and the incidence of this cancer has increased significantly around the world. In the present study, aqueous extract of S. jambolanum leaves and methanolic extract of its fruits were tested on breast cancer (MCF-7 and MDA-MB-231) cell lines for anti-proliferative effect. The MTT (3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay was used to quantitate the viable cells. The aqueous extract of S. jambolanum leaves and the methanolic extract of the fruits exerted dose-dependent inhibition (dose range of 100-500 �źg/ml) on both cell lines. The leaf extract exerted more potent inhibitory effect on the MDA-MB-231 (estrogen negative) cell line as well as the MCF-7 (estrogen positive) compared with the methanolic extract of the fruit. The concentration of leaf extract that caused 50% inhibition (IC50) of MDA-MB-231 was 270 �źg/ml. The IC50 for the rest of the incubations could not be determined.

Abstract: The excess accumulation of advanced glycation end products (AGEs) contributes to the chronic complications of type 2 diabetes mellitus (DM) and renal failure. Biopsy specimens (n = 184) of arterial (n = 92) and venous (n = 92) tissues were obtained (radial artery and cephalic vein) from end-stage renal disease (ESRD) patients with or without DM and normal healthy subjects (n = 12) requiring surgery (trauma patients). Immunohistochemical assessment of the blood vessels revealed the presence of pentosidine (AGE marker) in both veins and arteries in 72% of the ESRD patients. The percentage of arteries and veins that showed positive pentosidine staining in ESRD patients with type 2 DM alone was 100% and 92% respectively, in the non-diabetic ESRD patients it was < 70% (for arteries and veins), and in the ESRD patients with hypertension as an additional co-morbidity to type 2 DM it was 70% and 82%, respectively. The veins of ESRD patients with DM showed a strong (+++) positive staining and very strong (++++) positive staining was observed in the patients with DM and hypertension. Only mild (+) or moderate (++) pentosidine staining intensity was observed in the arteries of ESRD patients without or with comorbidities, respectively. The accumulation of AGE in the vein rather than the artery may be a better reflection of the extent of complications of ESRD.

Abstract: The trace elements copper, zinc, and selenium are important immune modulators and essential cofactors of the antioxidant enzymes. In the present study, the proliferative effect of human peripheral mononuclear cells (PBMCs) that have been exposed to copper, zinc, and selenium and the corresponding activities of antioxidant enzymes, namely superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase, were determined. Zinc and copper stimulated the PBMC proliferation in a dose-dependent manner within the dose range 25-200 micromol/L. SOD and GPx activities in PBMCs exposed to zinc were inhibited, whereas catalase activity was unaffected. All the three antioxidant enzymes in the cells exposed to copper were inhibited. Selenium exerted more potent inhibition of the cell proliferation while causing stimulation of the antioxidant enzymes at the lowest dose (25 micromol/L) than at the highest dose (200 micromol/L) tested. A significant negative correlation was observed between proliferation and antioxidant enzyme (SOD and GPx) activities in trace-element-exposed PBMC. The present findings substantiate the importance of trace elements as immune modulators and the involvement of enzymatic antioxidant system in the immune cell regulation.

Abstract: Increased oxidative stress and oxidative damage are present in Type 2 diabetes mellitus (DM). The aim of this study was to assess the oxidative stress levels in the three major ethnic groups in Malaysia and to study the association between glycaemic control and oxidant-antioxidant levels in these patients.

Abstract: Comparisons of oxidative indices and total antioxidant status between end-stage renal disease (ESRD) patients with or without diabetes is scant, especially in the Asian population.

Abstract: The present study was designed to explore the relationship between lipid peroxidation and antioxidant enzymes in young Malaysian insulin dependant diabetes mellitus (IDDM) patients. Indicative parameters of lipid peroxidation, activities of antioxidant enzymes and diabetes parameters were evaluated in single blood samples from 30 young type 1 diabetic patients and 30 healthy control subjects. Antioxidant enzymes namely superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) were significantly decreased while plasma malondialdehyde (MDA), an indicator for lipid peroxidation was significantly increased in IDDM patients compared to control subjects. Positive correlations between HbA1c and MDA; fasting blood glucose (FBG) and MDA and negative correlations between HbA1c and SOD; MDA and SOD were observed in these patients. No significant correlation existed between HbA1c and fasting blood glucose, GPx or CAT in the diabetic patients. The strong correlations found between lipid peroxidation, antioxidant enzymes and diabetes parameters confirms the existence of oxidative stress in our IDDM patients.

Abstract: Quercetin and fisetin, two naturally occurring bioflavonoids mobilized lipids and enzymes in the absence or presence of epinephrine in intact rat adipocytes. Dose-(0-250 microM) and time-(0-2 hr) course studies, showed that they stimulated phosphodiesterase (PDE) activity and simultaneously exert cyclic AMP accumulation. These bioflavonoids when present either singly or together with epinephrine stimulated the membrane-bound PDE but not the cytosolic PDE. The stimulation may act as a feedback mechanism to terminate the cyclic AMP effects. The action of theophylline, a known lipolytic agent (exerting its effects through antagonism of adenosine A1 receptor as well as PDE inhibition) was not potentiated by either fisetin or quercetin. However, the flavonoids potentiated epinephrine or isoproterenol- (a specific beta-adrenoreceptor agonist) induced lipolysis. Their effects were inhibited by propranolol (a beta-receptor antagonist). These results suggest that the flavonoids act synergistically with epinephrine on beta-adrenergic receptor and not through phosphodiesterase inhibition to stimulate adipocyte lipolysis. Increase in membrane phospholipid methylation occurred as a consequence of the epinephrine and/or quercetin/fisetin actions, and it correlated with the cellular accumulation of cyclic AMP.

Abstract: The plant natural products namely tannic acid and the flavonoids luteolin, kaempferol and apigenin exerted potent inhibition on the hyaluronidase enzyme. They were able to neutralize the haemorrhage induced by Crotalus adamenteus venom in mice dose-dependently from 0.12-2.4 mg/kg subcutaneously. In addition, they were able to antagonize the lethal activity of the venom when injected subcutaneously into mice and the order of potency was: tannic acid > luteolin = kaempferol > apigenin. Tannic acid (24 mg/kg, subcutaneously) was able to reduce significantly the venom-induced elevation of blood creatine kinase activity. It could also prolong the survival time(s) of mice, when injected immediately after the administration of venom. Experimental evidence is presented for the first time that malondialdehyde in liver and kidney is significantly elevated as a result of rattlesnake venom poisoning and that this effect can be controlled by tannic acid.

Abstract: Hesperetin was able to inhibit both epinephrine- and theophylline-induced lipolysis in rat adipocytes. At concentrations of 125 microM and 250 microM, it could augment the inhibitory effect of low insulin concentration (17 microU/2ml) on epinephrine-induced lipolysis. The basal cyclic AMP levels and phosphodiesterase activities in intact adipocytes were unaltered by hesperetin. The antilipolytic action of hesperetin in adenosine deaminase pretreated adipocytes was abolished. Hesperetin could only prevent lipolysis but not reverse the initial lipolytic activity stimulated through the adenosine deaminase pretreatment of the adipocytes. In pertussis or cholera toxin (PT or CT) pretreated adipocytes, the antilipolytic action of hesperetin was unaffected. From the data obtained it is possible to conclude that the antilipolytic action of hesperetin is not mediated through the inhibition of cyclic AMP level nor through CT- or PT-sensitive G proteins. However, the involvement of hesperetin in preventing the accessibility of cellular receptors to the actions of lipolytic agents may be implicated.

Abstract: Thirty-one flavonoids were tested for their effects on low Km phosphodiesterase with cyclic AMP as the substrate. Quercetin, luteolin, scutellarein, phloretin and genistein showed inhibitory potencies comparable to or greater than 3-isobutyl-2-methylxanthine (EC50 30-50 microM). Only four compounds namely, catechin, epicatechin, taxifolin and fustin stimulated the enzyme activity (stimulatory EC50 130-240 microM). The most potent phosphodiesterase (PDE) inhibitors were aglycones that had a C2.3 double bond, a keto group at C4 and hydroxyls at C3' and/or C4'. However, when the C-ring is opened then the requirement for the C2.3 double bond is eliminated. The same series of flavonoids were also tested for their lipolytic activity. The structural features required for effective synergistic lipolysis (with epinephrine) were generally similar to that required for potent PDE inhibition except that, for lipolytic activity, an intact C-ring was necessary. Fisetin and quercetin having the above-mentioned structure showed a dose- and time-dependent increase in lipolysis which was synergistic with epinephrine. Only butein and hesperetin showed inhibition of epinephrine-induced lipolysis, and their effect was dose-dependent. A time-course study indicated that hesperetin was able to delay the lipolytic action of epinephrine. It is most likely that the lipolytic effects of these compounds were not a result of PDE inhibition, as the orders of potency for the two activities had poor correlation. Apparently, the effective lipolytic flavonoids were also potent PDE inhibitors but not all the PDE inhibitors were able to induce lipolysis.

Abstract: In vitro studies showed that the flavonoid aglycones apigenin, luteolin and kaempferol inhibited the hyaluronidase activity of five different venoms dose-dependently. They were also able to delay the venom action when injected into mice. Naringenin, catechin and flavonoid glycosides had no effect. The flavonoids with unsubstituted hydroxyl groups at C-positions 5, 7 and 4', a double bond between carbons 2 and 3, as well as a ketone group at position 4, exhibited potent inhibitory actions on the venom hyaluronidases.

Abstract: The order of decreasing potency for five most potent flavonoids as inhibitors of hyaluronidase was found to be: condensed tannin less than luteolin less than apigenin less than kaempferol less than silybin. Kinetic studies of these inhibitors showed that their mode of inhibition was competitive. Aglycones were stronger inhibitors than their corresponding glycosides. The following flavonoid structure conferred potent inhibitory effect: a double bond between carbons 2 and 3; unsubstituted hydroxyl groups at positions 5, 7 and 4' and a ketone group at position 4.

Abstract: Cyclic AMP phosphodiesterase (PDE) activity was assayed in the plasma membrane, mitochondrial and microsomal fractions of rat brain. The specific activity of the enzyme was highest in the plasma membrane fraction followed by mitochondrial and then the microsomal fraction. Phosphodiesterase activity of all three fractions was reduced after pretreatment with lecithinase C (PCase) from Clostridium perfringens but less markedly affected by the pretreatment with sphingomyelinase (SMase) from human placenta. The PDE activity of the plasma membrane fraction was more sensitive to PCase treatment compared with the other two particulate fractions, which showed only a slight loss of activity. Temperature seemed to affect PDE activity of the plasma membrane. The enzyme was quite stable at 30 degrees C but its activity dropped by approximately 46% at 37 degrees C after 90 min of incubation. Pretreatment of the plasma membrane at 30 degrees C with PCase at a concentration of more than 5 U caused a marked loss of PDE activity and the decrease in activity reached a plateau at concentrations above 10 U.