HbE dan diabetis melitus lapuran akhir projek jangka

Glycated hemoglobins (HbAiiHbAic) serves as an indicator of diabetic control and hence the effectiveness of treatment. Studies on the methods available have shown that factors such as hemoglobin concentration and the presence of high lipid in the blood may affect the validity of the results. Atypical hemoglobins such as HbS and HbC have also been shown to affect the determination of glycated hemoglobins. However no study has been done on the effect of HbE , the hemoglobin variant that is present in the South East Asian population. Thus this study was unde~ to determine the effect of HbE on fu~ - determination ofHbAI/HbAlc by 4 commercial kits and also to determine its prevalence among diabetic subjects in Kelantan. Blood was taken from 58 normal subjects (23 males and 35 females) and 63 HbE (31 males and 32 females) heterozygous subjects. All subjects ,bad no history of diabetes mellitus. They had random blood glucose values < 7.8 mmolll and hemoglobin (hb) values between 7 to 24 g/dl. The EDTA plasma collected were tested for HbAI and HbAlc using 4 commercial kits i.e. Eagles Diagnostics, Boehringer Mannheim (BM), Diastat and Ames DCA 2000. The tests were done within one week and samples were stored at 4 oc before analysis. Results showed that the mean± s.d HbAI levels in normal vs HbE heterozygous subjects using Eagles Diagnostics, Boehringer Mannheim and Diastat kits were 6.9 ± 1.1% vs 9.5± 3.9%; 7.1 ± 1.2% vs 14.8 ± 9.3% and 8.3 ± 3.8% vs 13.5 ± 10.8% respectively. All were significantly different (p<O.OOI). The mean± s.d HbAlc levels for normal vs HbE heterozygous subjects determined using Ames DCA 2000 and Diastat kits were 5.1 ± 0.5% vs 5.3 ± 2.7% and 6.1 ±.).9% vs 10.7 ± 8.7% respectively. The results measured using the Diastat kit were significantly different (p<0.001) while those measured using the Ames DCA 2000 kit were not significantly different (p = 0.5). Therefore the cation-exchange chromatography based kits (Eagles Diagnostic, 7 . Boehringer Mannheim and Diastat) were affected by the presence of HbE. This - interference was minimised by dilution. The use of~pecific antibody (Ames) did not have any influence on HbA 1 c determination. The prevalence of HbE among diabetics were also studied. Blood was taken from 202 diabetic patients (121 females and 81 males; aged 46.2 ± 15.2 years) who were attending the outpatient diabetes clinic, Hospital USM. 28 patients (13.9%) were confirmed to carry HbE by hemoglobin electrophoresis. This consists of 4 males and 24 females. The finding is consistent with the prevalence ofHbE heterozygotes in the general population. In conclusio~ this study showed that HbE affect the determination ofHbAl/HbAlc by kits using cation exchange chromatography and that the interference may be diluted out The prevalence of HbE among diabetes mellitus patients is similar to that of the general population

γ-Tocotrienol prevents oxidative stress-induced telomere shortening in human fibroblasts derived from different aged individuals

The effects of palm γ-tocotrienol (GGT) on oxidative stress-induced cellular ageing was investigated in normal human skin fibroblast cell lines derived from different age groups; young (21-year-old, YF), middle (40-year-old, MF) and old (68-year-old, OF). Fibroblast cells were treated with γ-tocotrienol for 24 hours before or after incubation with IC50 dose of H2O2 for 2 hours. Changes in cell viability, telomere length and telomerase activity were assessed using the MTS assay (Promega, USA), Southern blot analysis and telomere repeat amplification protocol respectively. Results showed that treatment with different concentrations of γ-tocotrienol increased fibroblasts viability with optimum dose of 80 µM for YF and 40 µM for both MF and OF. At higher concentrations, γ-tocotrienol treatment caused marked decrease in cell viability with IC50 value of 200 µM (YF), 300 µM (MF) and 100 µM (OF). Exposure to H2O2 decreased cell viability in dose dependent manner, shortened telomere length and reduced telomerase activity in all age groups. The IC50 of H2O2 was found to be; YF (700 µM), MF (400 µM) and OF (100 µM). Results showed that viability increased significantly (p < 0.05) when cells were treated with 80 µM and 40 µM γ-tocotrienol prior or after H2O2-induced oxidative stress in all age groups. In YF and OF, pretreatment with γ-tocotrienol prevented shortening of telomere length and reduction in telomerase activity. In MF, telomerase activity increased while no changes in telomere length was observed. However, post-treatment of γ-tocotrienol did not exert any significant effects on telomere length and telomerase activity. Thus, these data suggest that γ-tocotrienol protects against oxidative stress-induced cellular ageing by modulating the telomere length possibly via telomerase

METABOLITES PROFILE OF COLORECTAL CANCER CELLS AT DIFFERENT STAGES

Objective: The aim of this study is to characterize the metabolite profiles of colorectal cancer (CRC) cells of different stages of the disease to understandthe pathophysiological changes that may help to identify prevention strategies as well as the sites for potential therapeutic drug actions.Methods: Six CRC cell lines of different stages (classified using the Dukes classification) were used, and they are SW 1116 (stage A), HT 29 and SW480 (stage B), HCT 15 and DLD-1 (stage C), and HCT 116 (stage D). Metabolites were extracted using methanol and water, and metabolic profiling wasperformed using liquid chromatography-mass spectrometry. Mass profiler professional software was used for statistical analysis.Results: There were 111,096 compounds detected across the samples, and 24 metabolites were identified to be significantly different betweenthe CRC stages. Most notably, there were eight metabolites that were significantly upregulated in the more advanced stages (B, C, and D) comparedwith Stage A. These metabolites include flavin mononucleotide, l-methionine, muricatacin, amillaripin, 2-methylbutyroylcarnitine, lumichrome,hexadeconoic acid, and lysoPE (0:0/16:0).Conclusion: This study showed that the expressions of metabolites at different stages of CRC were different, which represent the metabolic changesoccurring as CRC advances. The knowledge may help identify biomarkers for the staging of CRC, which could improve its prognosis as well as providea basis for the development of therapeutic interventions

Thymoquinone prevents β-amyloid neurotoxicity in primary cultured cerebellar granule neurons

Thymoquinone (TQ), a bioactive constituent of Nigella sativa Linn (N. sativa) has demonstrated several neuropharmacological attributes. In the present study, the neuroprotective properties of TQ were investigated by studying its anti-apoptotic potential to diminish β-amyloid peptide 1–40 sequence (Aβ1–40)-induced neuronal cell death in primary cultured cerebellar granule neurons (CGNs). The effects of TQ against Aβ1–40-induced neurotoxicity, morphological damages, DNA condensation, the generation of reactive oxygen species, and caspase-3, -8, and -9 activation were investigated. Pretreatment of CGNs with TQ (0.1 and 1 μM) and subsequent exposure to 10 μM Aβ1–40 protected the CGNs against the neurotoxic effects of the latter. In addition, the CGNs were better preserved with intact cell bodies, extensive neurite networks, a loss of condensed chromatin and less free radical generation than those exposed to Aβ1–40 alone. TQ pretreatment inhibited Aβ1–40-induced apoptosis of CGNs via both extrinsic and intrinsic caspase pathways. Thus, the findings of this study suggest that TQ may prevent neurotoxicity and Aβ1–40-induced apoptosis. TQ is, therefore, worth studying further for its potential to reduce the risks of developing Alzheimer’s disease

Expression profile of late responsive genes induced by spatial learning task: involvement of pathways in locomotion and memory

The altered molecular mechanisms by experimental therapies for neurodegenerative diseases itself could be overlapped with genes induced by the behavioral task. We employed the microarray platform to identify the differentially expressed late responsive genes in medial temporal lobes of four Morris Water Maze (MWM) trained and untrained BALB/c mice. After MWM training, the mice were sacrificed to obtain their brains’ medial temporal lobes. The total RNA was extracted from the tissues and global mRNA gene expression analysis was performed using Affymetrix GeneChip ®Mouse Gene 1.0 ST Array. There were 3635 (62.2%) up-regulated genes and 2206 (37.8%) down-regulated genes at p-values of < 0.05. These genes were operationally defined as late memory-related genes and behavior-related genes indicating that behavioral learning has a significant impact on the gene expression of the medial temporal lobes. From the pathway analysis, the network of memory and locomotion genes, and the guanylate cyclase pathway were identified as one of the most interesting pathways. The qPCR validation showed that the genes NMDA receptor 2a (Nmda2a) and cAMP dependent protein kinase type I beta regulatory subunit (Prkar1b) were up-regulated while adenylate cyclase 5 (Adcy5) was down-regulated. We proposed that the involvement of guanylate cyclase pathway in the long-term potentiation lasted at least up to three days after the MWM test. Present study suggested that the molecular mechanisms followed by spatial learning task could be altered up to three days or even longer, it could be overlapped with genes induced by further invented experimental therapy

Tocotrienol rich fraction supplementation improved lipid profile and oxidative status in healthy older adults: A randomized controlled study

<p>Abstract</p> <p>Background</p> <p>Vitamin E supplements containing tocotrienols are now being recommended for optimum health but its effects are scarcely known. The objective was to determine the effects of Tocotrienol Rich Fraction (TRF) supplementation on lipid profile and oxidative status in healthy older individuals at a dose of 160 mg/day for 6 months.</p> <p>Methods</p> <p>Sixty-two subjects were recruited from two age groups: 35-49 years (n = 31) and above 50 years (n = 31), and randomly assigned to receive either TRF or placebo capsules for six months. Blood samples were obtained at 0, 3^rdand 6^thmonths.</p> <p>Results</p> <p>HDL-cholesterol in the TRF-supplemented group was elevated after 6 months (p < 0.01). Protein carbonyl contents were markedly decreased (p < 0.001), whereas AGE levels were lowered in the > 50 year-old group (p < 0.05). Plasma levels of total vitamin E particularly tocopherols were significantly increased in the TRF-supplemented group after 3 months (p < 0.01). Plasma total tocotrienols were only increased in the > 50 year-old group after receiving 6 months of TRF supplementation. Changes in enzyme activities were only observed in the > 50 year-old group. SOD activity was decreased after 3 (p < 0.05) and 6 (p < 0.05) months of TRF supplementation whereas CAT activity was decreased after 3 (p < 0.01) and 6 (p < 0.05) months in the placebo group. GPx activity was increased at 6 months for both treatment and placebo groups (p < 0.05).</p> <p>Conclusion</p> <p>The observed improvement of plasma cholesterol, AGE and antioxidant vitamin levels as well as the reduced protein damage may indicate a restoration of redox balance after TRF supplementation, particularly in individuals over 50 years of age.</p

Metabolomic characterization of colorectal cancer cell lines highlighting stage-specific alterations during cancer progression

Introduction: Metabolomic studies on various colorectal cancer (CRC) cell lines have improved our understanding of the biochemical events underlying the disease. However, the metabolic profile dynamics associated with different stages of CRC progression is still lacking. Such information can provide further insights into the pathophysiology and progression of the disease that will prove useful in identifying specific targets for drug designing and therapeutics. Thus, our study aims to characterize the metabolite profiles in the established cell lines corresponding to different stages of CRC. Methods: Metabolite profiling of normal colon cell lines (CCD 841 CoN) and CRC cell lines corresponding to different stages, i.e., SW 1116 (stage A), HT 29 and SW 480 (stage B), HCT 15 and DLD-1 (stage C), and HCT 116 (stage D), was carried out using liquid chromatography-mass spectrometry (LC-MS). Mass Profiler Professional and Metaboanalyst 4.0 software were used for statistical and pathway analysis. METLIN database was used for the identification of metabolites. Results: We identified 72 differential metabolites compared between CRC cell lines of all the stages and normal colon cells. Principle component analysis and partial least squares discriminant analysis score plot were used to segregate normal and CRC cells, as well as CRC cells in different stages of the disease. Variable importance in projection score identified unique differential metabolites in CRC cells of the different stages. We identified 7 differential metabolites unique to stage A, 3 in stage B, 5 in stage C, and 5 in stage D. Conclusion: This study highlights the differential metabolite profiling in CRC cell lines corresponding to different stages. The identification of the differential metabolites in CRC cells at individual stages will lead to a better understanding of the pathophysiology of CRC development and progression and, hence, its application in treatment strategies

Gamma-tocotrienol acts as a BH3 mimetic to induce apoptosis in neuroblastoma SH-SY5Y cells

Bcl-2 family proteins are crucial regulators of apoptosis. Both pro- and antiapoptotic members exist, and overexpression of the latter facilitates evasion of apoptosis in many cancer types. Bcl-2 homology domain 3 (BH3) mimetics are small molecule inhibitors of antiapoptotic Bcl-2 family members, and these inhibitors are promising anticancer agents. In this study, we report that gamma-tocotrienol (γT3), an isomer of vitamin E, can inhibit Bcl-2 to induce apoptosis. We demonstrate that γT3 induces cell death in human neuroblastoma SH-SY5Y cells by depolarising the mitochondrial membrane potential, enabling release of cytochrome c to the cytosol and increasing the activities of caspases-9 and -3. Treatment of cells with inhibitors of Bax or caspase-9 attenuated the cell death induced by γT3. Simulated docking analysis suggested that γT3 binds at the hydrophobic groove of Bcl-2, while a binding assay showed that γT3 competed with a fluorescent probe to bind at the hydrophobic groove. Our data suggest that γT3 mimics the action of BH3-only protein by binding to the hydrophobic groove of Bcl-2 and inducing apoptosis via the intrinsic pathway in a Bax- and caspase-9-dependent manner

Comparison of non-alcoholic fatty liver disease (NAFLD) model using diet-induced NAFLD mice with genetically modified mice

Prevalence of non-alcoholic fatty liver disease (NAFLD) is increasing steadily every year affecting all population both Western and Asian countries. The current treatments available for NAFLD are non-conclusive warranting newer effective pharmacological agents. Newly formulated agents require prior testing using animal models. However, in developing countries, these models are often costly. The possibility of using more affordable animal model in local settings should be investigated. In this study, ten Institute of Cancer Research (ICR) and seven B6.Cg-LepOb/J leptin-knockout (JAX) male mice were recruited. Five ICR and all JAX mice were subjected to high-fat diet (60% kcal fat) and remaining ICR mice were given standard diet (SD) for six weeks. Body weight and food intake were measured weekly while abdominal circumference, random blood glucose and liver span were measured at the end of the HFD study. Livers collected were subjected to histology assessment. Compared to ICR group, JAX group presented with significantly higher body weight (58 ± 0.72, p<0.05), larger body weight changes (16.57 ± 0.81, p<0.05), more HFD intake (197.14 ± 0.812, p<0.05) and larger abdominal circumference (11.79 ± 0.34: p<0.05). Liver from JAX group appeared with general steatosis and presentation of high-grade panacinar steatosis, low number of lobular inflammations and minimal fibrosis. Liver of ICR mice showed Zone 3 steatosis with high number of lobular inflammations without fibrosis. The NAFLD characteristics presented in JAX group suggested that B6.Cg-LepOb/J mice developed characteristics of NAFLD resembling human while ICR is suitable NAFLD model resembling human population resilient towards NAFLD

Effect of vitamin E (Tri E®) on antioxidant enzymes and DNA damage in rats following eight weeks exercise

<p>Abstract</p> <p>Background</p> <p>Exercise is beneficial to health, but during exercise the body generates reactive oxygen species (ROS) which are known to result in oxidative stress. The present study analysed the effects of vitamin E (Tri E^®) on antioxidant enzymes; superoxide dismutase (SOD), glutathione peroxidase (GPx), catalase (Cat) activity and DNA damage in rats undergoing eight weeks exercise.</p> <p>Methods</p> <p>Twenty four <it>Sprague-Dawley </it>rats (weighing 320-370 gm) were divided into four groups; a control group of sedentary rats which were given a normal diet, second group of sedentary rats with oral supplementation of 30 mg/kg/d of Tri E^®, third group comprised of exercised rats on a normal diet, and the fourth group of exercised rats with oral supplementation of 30 mg/kg/d of Tri E^®. The exercising rats were trained on a treadmill for 30 minutes per day for 8 weeks. Blood samples were taken before and after 8 weeks of the study to determine SOD, GPx, Cat activities and DNA damage.</p> <p>Results</p> <p>SOD activity decreased significantly in all the groups compared to baseline, however both exercised groups showed significant reduction in SOD activity as compared to the sedentary groups. Sedentary control groups showed significantly higher GPx and Cat activity compared to baseline and exercised groups. The supplemented groups, both exercised and non exercised groups, showed significant decrease in Cat activity as compared to their control groups with normal diet. DNA damage was significantly higher in exercising rats as compared to sedentary control. However in exercising groups, the DNA damage in supplemented group is significantly lower as compared to the non-supplemented group.</p> <p>Conclusions</p> <p>In conclusion, antioxidant enzymes activity were generally reduced in rats supplemented with Tri E^®probably due to its synergistic anti-oxidative defence, as evidenced by the decrease in DNA damage in Tri E^®supplemented exercise group.</p