Search results for: cholesterol oxidase
Commenced in January 2007
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Edition: International
Paper Count: 556

Search results for: cholesterol oxidase

166 Effect of Seasonal Variation on Two Introduced Columbiformes in Awba Dam Tourism Centre, University of Ibadan, Ibadan

Authors: Kolawole F. Farinloye, Samson O. Ojo

Abstract:

Two Columbiformes species were recently introduced to the newly established Awba Dam Tourism Centre [ADTC], hence there is need to investigate the effect of seasonal variation on these species with respect to hematological composition. Blood samples were obtained from superficial ulna vein of the 128 apparently healthy C. livia and C. guinea into tubes containing EDTA as anticoagulant. Thin blood smears (TBS) were prepared, stained and viewed under microscope. Values of Red Blood Cell (RBC) count, White Blood Cell (WBC) count, cholesterol (CH), Uric Acid (UA), Protein (PR), Mean Corpuscular Volume (MCV), Haemoglobin Content (HB), Blood Volume (BV), Plasma Glucose (PG) and Length/Width (L/W) ratio of red blood cells were assessed. The procedure was carried out on a seasonal basis (wet and dry seasons of 2013-2014). Data was analyzed using descriptive and inferential statistics. Lymphocyte count for C. livia was F3, 161 = 13.15, while for C. guinea was F3, 178 = 13.15. Heterophil, H/L ratio and Muscle score values for both species were (rs = -0.38, rs = -0.44), (rs = 0.51, rs = 0.31) (4, 3) respectively. Analyses also demonstrated a low WBC to RBC ratio (0.004: 25.3) in both species during the wet season compared to dry season, respectively. L/W varied significantly among sampling seasons i.e. wet (19.1% of BV, 12.6% of BV, 0.1% of BV) and dry (18.9% of BV, 12.7% of BV, 0.08% of BV). The level of HB in wet season (19.20±8.46108) is lower compared to dry season (19.70±8.48762). T-test also showed (wet=15.625, 0.111), (dry=12.125, 0.146) respectively, hence there is no association between species and haematological parameters. Species introduced were found to be haematologically stable. Although there were slight differences in seasonal composition, however this can be attributed to seasonal variation; suggesting little or no effect of seasons on their blood composition.

Keywords: seasonal variation, Columbiformes, Awba Dam tourism centre, University of Ibadan, Ibadan

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165 Spectrophotometric Detection of Histidine Using Enzyme Reaction and Examination of Reaction Conditions

Authors: Akimitsu Kugimiya, Kouhei Iwato, Toru Saito, Jiro Kohda, Yasuhisa Nakano, Yu Takano

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The measurement of amino acid content is reported to be useful for the diagnosis of several types of diseases, including lung cancer, gastric cancer, colorectal cancer, breast cancer, prostate cancer, and diabetes. The conventional detection methods for amino acid are high-performance liquid chromatography (HPLC) and liquid chromatography-mass spectrometry (LC-MS), but they have several drawbacks as the equipment is cumbersome and the techniques are costly in terms of time and costs. In contrast, biosensors and biosensing methods provide more rapid and facile detection strategies that use simple equipment. The authors have reported a novel approach for the detection of each amino acid that involved the use of aminoacyl-tRNA synthetase (aaRS) as a molecular recognition element because aaRS is expected to a selective binding ability for corresponding amino acid. The consecutive enzymatic reactions used in this study are as follows: aaRS binds to its cognate amino acid and releases inorganic pyrophosphate. Hydrogen peroxide (H₂O₂) was produced by the enzyme reactions of inorganic pyrophosphatase and pyruvate oxidase. The Trinder’s reagent was added into the reaction mixture, and the absorbance change at 556 nm was measured using a microplate reader. In this study, an amino acid-sensing method using histidyl-tRNA synthetase (HisRS; histidine-specific aaRS) as molecular recognition element in combination with the Trinder’s reagent spectrophotometric method was developed. The quantitative performance and selectivity of the method were evaluated, and the optimal enzyme reaction and detection conditions were determined. The authors developed a simple and rapid method for detecting histidine with a combination of enzymatic reaction and spectrophotometric detection. In this study, HisRS was used to detect histidine, and the reaction and detection conditions were optimized for quantitation of these amino acids in the ranges of 1–100 µM histidine. The detection limits are sufficient to analyze these amino acids in biological fluids. This work was partly supported by Hiroshima City University Grant for Special Academic Research (General Studies).

Keywords: amino acid, aminoacyl-tRNA synthetase, biosensing, enzyme reaction

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164 Renoprotective Effect of Alcoholic Extract of Bacopa monnieri via Inhibition of Advanced Glycation End Products and Oxidative Stress in Stz-Nicotinamide Induced Diabetic Nephropathy

Authors: Lalit Kishore, Randhir Singh

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Diabetic nephropathy (DN) is the major cause of morbidity among diabetic patients. In this study, the effect of Bacopa monnieri Linn. (Brahmi, BM), was studied in a Streptozotocin (STZ)-induced experimental rat model of DN. Diabetic nephropathy was induced in Male Wistar rats (body weight- 300± 10 gms) by single intra-peritoneal injection of STZ (45mg/kg, i.p.) after 15 min of Nicotinamide (230 mg/kg) administration. Different doses of alcoholic extract i.e. 100, 200 and 400 mg/kg was given for 45 days by oral gavage after induction of DN. Blood glucose level, serum insulin, glycosylated haemoglobin, renal parameters (serum urea, uric acid, creatinine and BUN) and lipid profile (total cholesterol, triglycerides, HDL, LDL and VLDL levels) were measured. Concentration of thiobarbituric acid reactive species (TBARS) and levels of antioxidant enzymes of reduced glutathione (GSH), superoxide dismutase (SOD), and catalase (CAT) were evaluated in the kidney, liver and pancreas. At the end of treatment period the alcoholic extract of BM reduced the elevated level of blood glucose, serum insulin, renal parameters, lipid levels, TBARS, AGE’s in kidney and significantly increased body weight, HDL and antioxidant enzymes in dose dependent manner as compared to diabetic control animals. These results suggested the BM possesses significant renoprotective activity.

Keywords: AGE's, lipid profile, oxidative stress, renal parameters

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163 GC-MS-Based Untargeted Metabolomics to Study the Metabolism of Pectobacterium Strains

Authors: Magdalena Smoktunowicz, Renata Wawrzyniak, Malgorzata Waleron, Krzysztof Waleron

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Pectobacterium spp. were previously classified into the Erwinia genus founded in 1917 to unite at that time all Gram-negative, fermentative, nonsporulating and peritrichous flagellated plant pathogenic bacteria. After work of Waldee (1945), on Approved Lists of Bacterial Names and bacteriology manuals in 1980, they were described either under the species named Erwinia or Pectobacterium. The Pectobacterium genus was formally described in 1998 of 265 Pectobacterium strains. Currently, there are 21 species of Pectobacterium bacteria, including Pectobacterium betavasculorum since 2003, which caused soft rot on sugar beet tubers. Based on the biochemical experiments carried out for this, it is known that these bacteria are gram-negative, catalase-positive, oxidase-negative, facultatively anaerobic, using gelatin and causing symptoms of soft rot on potato and sugar beet tubers. The mere fact of growing on sugar beet may indicate a metabolism characteristic only for this species. Metabolomics, broadly defined as the biology of the metabolic systems, which allows to make comprehensive measurements of metabolites. Metabolomics, in combination with genomics, are complementary tools for the identification of metabolites and their reactions, and thus for the reconstruction of metabolic networks. The aim of this study was to apply the GC-MS-based untargeted metabolomics to study the metabolism of P. betavasculorum in different growing conditions. The metabolomic profiles of biomass and biomass media were determined. For sample preparation the following protocol was used: extraction with 900 µl of methanol: chloroform: water mixture (10: 3: 1, v: v) were added to 900 µl of biomass from the bottom of the tube and up to 900 µl of nutrient medium from the bacterial biomass. After centrifugation (13,000 x g, 15 min, 4oC), 300µL of the obtained supernatants were concentrated by rotary vacuum and evaporated to dryness. Afterwards, two-step derivatization procedure was performed before GC-MS analyses. The obtained results were subjected to statistical calculations with the use of both uni- and multivariate tests. The obtained results were evaluated using KEGG database, to asses which metabolic pathways are activated and which genes are responsible for it, during the metabolism of given substrates contained in the growing environment. The observed metabolic changes, combined with biochemical and physiological tests, may enable pathway discovery, regulatory inference and understanding of the homeostatic abilities of P. betavasculorum.

Keywords: GC-MS chromatograpfy, metabolomics, metabolism, pectobacterium strains, pectobacterium betavasculorum

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162 Formulation and Anticancer Evaluation of Beta-Sitosterol in Henna Methanolic Extract Embedded in Controlled Release Nanocomposite

Authors: Sanjukta Badhai, Durga Barik, Bairagi C. Mallick

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In the present study, Beta-Sitosterol in Lawsonia methanolic leaf extract embedded in controlled release nanocomposite was prepared and evaluated for in vivo anticancer efficacy in dimethyl hydrazine (DMH) induced colon cancer. In the present study, colon cancer was induced by s.c injection of DMH (20 mg/kg b.wt) for 15 weeks. The animals were divided into five groups as follows control, DMH alone, DMH and Beta Sitosterol nanocomposite (50mg/kg), DMH and Beta Sitosterol nanocomposite (100 mg/kg) and DMH and Standard Silymarin (100mg/kg) and the treatment was carried out for 15 weeks. At the end of the study period, the blood was withdrawn, and serum was separated for haematological, biochemical analysis and tumor markers. Further, the colonic tissue was removed for the estimation of antioxidants and histopathological analysis. The results of the study displays that DMH intoxication elicits altered haematological parameters (RBC,WBC, and Hb), elevated lipid peroxidation and decreased antioxidants level (SOD, CAT, GPX, GST and GSH), elevated lipid profiles (cholesterol and triglycerides), tumor markers (CEA and AFP) and altered colonic tissue histology. Meanwhile, treatment with Beta Sitosterol nanocomposites significantly restored the altered biochemicals parameters in DMH induced colon cancer mediated by its anticancer efficacy. Further, Beta Sitosterol nanocomposite (100 mg/kg) showed marked efficacy.

Keywords: nanocomposites, herbal formulation, henna, beta sitosterol, colon cancer, dimethyl hydrazine, antioxidant, lipid peroxidation

Procedia PDF Downloads 164
161 The Expression of Lipoprotein Lipase Gene with Fat Accumulations and Serum Biochemical Levels in Betong (KU Line) and Broiler Chickens

Authors: W. Loongyai, N. Saengsawang, W. Danvilai, C. Kridtayopas, P. Sopannarath, C. Bunchasak

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Betong chicken is a slow growing and a lean strain of chicken, while the rapid growth of broiler is accompanied by increased fat. We investigated the growth performance, fat accumulations, lipid serum biochemical levels and lipoprotein lipase (LPL) gene expression of female Betong (KU line) at the age of 4 and 6 weeks. A total of 80 female Betong chickens (KU line) and 80 female broiler chickens were reared under open system (each group had 4 replicates of 20 chicks per pen). The results showed that feed intake and average daily gain (ADG) of broiler chicken were significantly higher than Betong (KU line) (P < 0.01), while feed conversion ratio (FCR) of Betong (KU line) at week 6 were significantly lower than broiler chicken (P < 0.01) at 6 weeks. At 4 and 6 weeks, two birds per replicate were randomly selected and slaughtered. Carcass weight did not significantly differ between treatments; the percentage of abdominal fat and subcutaneous fat yield was higher in the broiler (P < 0.01) at 4 and 6 week. Total cholesterol and LDL level of broiler were higher than Betong (KU line) at 4 and 6 weeks (P < 0.05). Abdominal fat samples were collected for total RNA extraction. The cDNA was amplified using primers specific for LPL gene expression and analysed using real-time PCR. The results showed that the expression of LPL gene was not different when compared between Betong (KU line) and broiler chickens at the age of 4 and 6 weeks (P > 0.05). Our results indicated that broiler chickens had high growth rate and fat accumulation when compared with Betong (KU line) chickens, whereas LPL gene expression did not differ between breeds.

Keywords: lipoprotein lipase gene, Betong (KU line), broiler, abdominal fat, gene expression

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160 Vitamin D Status in Tunisian Obese Patients

Authors: O. Berriche, R. Ben Othmen, H. Sfar, H. Abdesslam, S. Bou Meftah, S. Bhouri, F. Mahjoub, C. Amrouche, H. Jamoussi

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Introduction: Although current evidence emphasizes a high prevalence of vitamin D deficiency and an inverse association between serum 25-hydroxyvitamin D (25-OHD) concentration and obesity, no studies have been conducted in Tunisian obese. The objectives of our study were to estimate the vitamin D deficiency in obese, identify risk factors for vitamin D deficiency, demonstrating a possible association between vitamin D levels and metabolic parameters. Methods: This was a descriptive study of 100 obese 18-65 year-old. Anthropometric measurements were determined. Fasting blood samples were assessed for the following essays : serum calcium, 25 OH vitamin D, inorganic phosphorus, fasting glucose, HDL, LDL cholesterol and triglyceride. Insulin resistance was evaluated by fasting insulin, HOMA-IR and HOMA-ß. Consumption of foods riche in vitamin D, sunscreen use, wearing protective clothes and exposed surface were assessed through applied questionnaires. Results: The deficit of vitamin D (< 30 ng/ml) among obese was 98,8%. Half of them had a rate < 10ng/ml. Environmental factors involved in vitamin D deficiency are : the veil (p = 0,001), wearing protective clothes (p = 0,04) and the exposed surface (p = 0,011) and dietary factors are represented by the daily caloric intake (p = 0,0001). The percent of fat mass was negatively related to vitamin D levels (p = 0,01) but not with BMI (p = 0,11) or waist circumference (p = 0,88). Similarly, lipid and glucose profile had no link with vitamin D. We found no relationship between Insulin resistance and vitamin D levels. Conclusion: At the end of our study, we have identified a very important vitamin D deficiency among obese. Dosage and systematic supplementation should be applied and for that physician awareness is needed.

Keywords: insulinresistance, risk factors, obesity, vitamin D

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159 Beneficial Effect of Chromium Supplementation on Glucose, HbA1C and Lipid Variables in Individuals with Newly Onset Type-2 Diabetes

Authors: Baljinder Singh, Navneet Sharma

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Chromium is an essential nutrient involved in normal carbohydrate and lipid metabolism. It influences glucose metabolism by potentiating the action as taking part in insulin signal amplification mechanism. A placebo-controlled single blind, prospective study was carried out to investigate the effect of chromium supplementation on blood glucose, HbA1C and lipid profile in newly onset patients with type-2 diabetes. Total 40 newly onset type-2 diabetics were selected and after one month stabilization further randomly divided into two groups viz. study group and placebo group. The study group received 9 gm brewer’s yeast (42 μ Cr) daily and the other placebo group received yeast devoid of chromium for 3 months. Subjects were instructed not to change their normal eating and living habits. Fasting blood glucose, HbA1C and lipid profile were analyzed at beginning and completion of the study. Results revealed that fasting blood glucose level significantly reduced in the subjects consuming yeast supplemented with chromium (197.65±6.68 to 103.68±6.64 mg/dl; p<0.001). HbA1C values improved significantly from 9.51±0.26% to 6.86±0.28%; p<0.001 indicating better glycaemic control. In experimental group total cholesterol, TG and LDL levels were also significantly reduced from 199.66±3.11 to 189.26±3.01 mg/dl; p<0.02, 144.94±8.31 to 126.01±8.26; p<0.05 and 119.19±1.71 to 99.58±1.10; p<0.001 respectively. These data demonstrate beneficial effect of chromium supplementation on glycaemic control and lipid variables in subjects with newly onset type-2 diabetes.

Keywords: type-2 diabetes, chromium, glucose, HbA1C

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158 Evaluation of Insulin Sensitizing Effects of Different Fractions from Total Alcoholic Extract of Moringa oleifera Lam. Bark in Dexamethasone-Induced Insulin Resistant Rats

Authors: Hasanpasha N. Sholapur, Basanagouda M.Patil

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Alcoholic extract of the bark of Moringa oleifera Lam. (MO), (Moringaceae), has been evaluated experimentally in the past for its insulin sensitizing potentials. In order to explore the possibility of the class of phytochemical(s) responsible for this experimental claim, the alcoholic extract was fractionated into non-polar [petroleum ether (PEF)], moderately non-polar [ethyl acetate (EAF)] and polar [aqueous (AQF)] fractions. All the fractions and pioglitazone (PIO) as standard (10mg/kg were p.o., once daily for 11 d) were investigated for their chronic effect on fasting plasma glucose, triglycerides, total cholesterol, insulin, oral glucose tolerance and acute effect on oral glucose tolerance in dexamethasone-induced (1 mg/kg s.c., once daily for 11 d) chronic model and acute model (1 mg/kg i.p., for 4 h) respectively for insulin resistance (IR) in rats. Among all the fractions tested, chronic treatment with EAF (140 mg/kg) and PIO (10 mg/kg) prevented dexamethasone-induced IR, indicated by prevention of hypertriglyceridemia, hyperinsulinemia and oral glucose intolerance, whereas treatment with AQF (95 mg/kg) prevented hepatic IR but not peripheral IR. In acute study single dose treatment with EAF (140 mg/kg) and PIO (10 mg/kg) prevented dexamethasone-induced oral glucose intolerance, fraction PEF did not show any effect on these parameters in both the models. The present study indicates that the triterpenoidal and the phenolic class of phytochemicals detected in EAF of alcoholic extract of MO bark may be responsible for the prevention of dexamethasone-induced insulin resistance in rats.

Keywords: Moringa oleifera, insulin resistance, dexamethasone, serum triglyceride, insulin, oral glucose tolerance test

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157 Effects of Boron Compounds in Rabbits Fed High Protein and Energy Diet: A Metabolomic and Transcriptomic Approach

Authors: Nuri Başpınar, Abdullah Başoğlu, Özgür Özdemir, Çağlayan Özel, FundaTerzi, Özgür Yaman

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Current research is targeting new molecular mechanisms that underlie non-alcoholic fatty liver disease (NAFLD) and associated metabolic disorders like nonalcoholic steatohepatitis (NASH). Forty New Zealand White rabbits have been used and fed a high protein (HP) and energy diet based on grains and containing 11.76 MJ/kg. Boron added to 3 experimental groups’ drinking waters (30 mg boron/L) as boron compounds. Biochemical analysis including boron levels, and nuclear magnetic resonance (NMR) based metabolomics evaluation, and mRNA expression of peroxisome proliferator-activated receptor (PPAR) family were performed. LDL-cholesterol concentrations alone were decreased in all the experimental groups. Boron levels in serum and feces were increased. Content of acetate was in about 2x higher for anhydrous borax group, at least 3x higher for boric acid group. PPARα mRNA expression was significantly decreased in boric acid group. Anhydrous borax attenuated mRNA levels of PPARα, which was further suppressed by boric acid. Boron supplementation decreased the degenerative alterations in hepatocytes. Except borax group other boron groups did not have a pronounced change in tubular epithels of kidney. In conclusion, high protein and energy diet leads hepatocytes’ degenerative changes which can be prevented by boron supplementation. Boric acid seems to precede in this effectiveness.

Keywords: high protein and energy diet, boron, metabolomics, transcriptomic

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156 Eugenol Effects on Metabolic Syndrome Induced Liver Damages

Authors: Fatemeh Kourkinejad Gharaei, Tahereh Safari, Zahra Saebinasab

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Metabolic syndrome (MetS) is a set of risk factors associated with cardiovascular diseases, atherosclerosis, and type 2 diabetes. Nonalcoholic fatty liver disease (NAFLD) is the most important liver disorder in metabolic syndrome. High fructose consumption increases the risk of NAFLD. Eugenol shows anti-thrombotic, insulin-sensitive, fat-reducing effects. This study was designed to investigate the protective role of eugenol in NAFLD caused by metabolic syndrome. Methods: Thirty male Wistar rats were randomly divided into five groups; group 1, drinking water intake animals; group 2, fructose, group 3, fructose+eugenol solvent; group 4, fructose+ eugenol 50mg/kg and group 5, fructose+ eugenol 100mg/kg. At the end of the experiment, after 12 hours of fasting and under anesthesia, blood samples were taken for measurement of fast blood glucose (FBS), SGOT, AGPT, LDL, HDL, cholesterol, triglyceride. Results: FBG significantly increased in group 2 compared to group 1 (p < 0.001); however, it significantly decreased in groups 4 and 5 compared to group 2 (p < 0.05). SGOT and SGPT levels significantly increased in group 2 compared to drinking water alone (p < 0.001). However, SGOT and SGPT levels significantly decreased in groups 4 and 5. MDA and LTDS significantly increased in group 2 compared with drinking water alone (p < 0.01), while MDA and LTDS decreased in 4 and 5 groups compared to group 2 (p < 0.05), which confirms the pathology results related to the liver damage. Conclusion: Eugenol has protective effects on the liver and fat accumulation in liver cells.

Keywords: eugenol, fructose, metabolic syndrome, nonalcoholic fatty liver disease

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155 Antihyperglycemic Potential of Chrysin and Diosmin alone or in Combination against Streptozotocin-Induced Hyperglycemia in Rats: Anti-Inflammatory and Antioxidant Mechanisms

Authors: Sally A. El Awdan, Gehad A. Abdel Jaleel, Dalia O Saleh, Manal Badawi

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Background: Diabetes is a metabolic disease that affects a wide range of people worldwide and results in serious complications. Streptozotocin (STZ) causes selective cytotoxicity in the pancreatic β-cell, and it has been extensively used to induce diabetes mellitus in rats. The present study investigated the effects of diosmin and chrysin alone or in combination with each other on glucose level and on liver in STZ diabetic rats. Methods: In this study, rats were divided into six experimental groups (normal, untreated STZ-diabetic (60 mg/kg B.W., IP), treated STZ-diabetic with glycazide (10 mg/kg B.W, oral), treated STZ-diabetic with diosmin (100 mg/kg B. W., oral), treated STZ-diabetic with chrysin (80 mg/kg B.W., oral), treated STZ-diabetic with diosmin (50 mg/kg B.W, oral) + chrysin (40 mg/kg B.W., oral). After 2 weeks blood samples were withdrawn and glucose was measured. Animals were anaesthetized with an intraperitoneal injection of sodium pentobarbital (60 mg/kg), and sacrificed for dissecting liver. Results: Throughout the experimental period, all treatments significantly (P<0.05) lowered serum glucose, AST, ALT, triglyceride, cholesterol, IL-6, TNF-α and IL-1β. Moreover, the treated diabetic rats showed higher levels of reduced glutathione (P<0.05) in the liver compared to the diabetic control rats and inhibited diabetes-induced elevation in the levels of malondialdehyde in liver. The results of this study clearly demonstrated that diosmin and chrysin possess several treatment-oriented properties, including the control of hyperglycemia, antioxidant effects and anti-inflammatory effects. Conclusion: Considering these observations, it appears that diosmin and chrysin may be a useful supplement to delay the developmentof diabetes and its complications.

Keywords: diabetes, streptozocin, chrysin, rat, diosmin, cytokines

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154 Prevalence of Cyp2d6 and Its Implications for Personalized Medicine in Saudi Arabs

Authors: Hamsa T. Tayeb, Mohammad A. Arafah, Dana M. Bakheet, Duaa M. Khalaf, Agnieszka Tarnoska, Nduna Dzimiri

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Background: CYP2D6 is a member of the cytochrome P450 mixed-function oxidase system. The enzyme is responsible for the metabolism and elimination of approximately 25% of clinically used drugs, especially in breast cancer and psychiatric therapy. Different phenotypes have been described displaying alleles that lead to a complete loss of enzyme activity, reduced function (poor metabolizers – PM), hyperfunctionality (ultrarapid metabolizers–UM) and therefore drug intoxication or loss of drug effect. The prevalence of these variants may vary among different ethnic groups. Furthermore, the xTAG system has been developed to categorized all patients into different groups based on their CYP2D6 substrate metabolization. Aim of the study: To determine the prevalence of the different CYP2D6 variants in our population, and to evaluate their clinical relevance in personalized medicine. Methodology: We used the Luminex xMAP genotyping system to sequence 305 Saudi individuals visiting the Blood Bank of our Institution and determine which polymorphisms of CYP2D6 gene are prevalent in our region. Results: xTAG genotyping showed that 36.72% (112 out of 305 individuals) carried the CYP2D6_*2. Out of the 112 individuals with the *2 SNP, 6.23% had multiple copies of *2 SNP (19 individuals out of 305 individuals), resulting in an UM phenotype. About 33.44% carried the CYP2D6_*41, which leads to decreased activity of the CYP2D6 enzyme. 19.67% had the wild-type alleles and thus had normal enzyme function. Furthermore, 15.74% carried the CYP2D6_*4, which is the most common nonfunctional form of the CYP2D6 enzyme worldwide. 6.56% carried the CYP2D6_*17, resulting in decreased enzyme activity. Approximately 5.73% carried the CYP2D6_*10, consequently decreasing the enzyme activity, resulting in a PM phenotype. 2.30% carried the CYP2D6_*29, leading to decreased metabolic activity of the enzyme, and 2.30% carried the CYP2D6_*35, resulting in an UM phenotype, 1.64% had a whole-gene deletion CYP2D6_*5, thus resulting in the loss of CYP2D6 enzyme production, 0.66% carried the CYP2D6_*6 variant. One individual carried the CYP2D6_*3(B), producing an inactive form of the enzyme, which leads to decrease of enzyme activity, resulting in a PM phenotype. Finally, one individual carried the CYP2D6_*9, which decreases the enzyme activity. Conclusions: Our study demonstrates that different CYP2D6 variants are highly prevalent in ethnic Saudi Arabs. This finding sets a basis for informed genotyping for these variants in personalized medicine. The study also suggests that xTAG is an appropriate procedure for genotyping the CYP2D6 variants in personalized medicine.

Keywords: CYP2D6, hormonal breast cancer, pharmacogenetics, polymorphism, psychiatric treatment, Saudi population

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153 Effect of Zingerone on High-Fructose Diet-Indeuced Metabolic Derangements in Growing Sprague-Dawley Rats

Authors: Nondumiso Lushozi, Busisani Lembede, Eliton Chivandi

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Consumption of fructose increases the risk of obesity, nonalcoholic fatty liver disease (NAFLD) and metabolic syndrome in children. Zingerone which is found in ginger has antidiabetic and antiobesogenic properties. Therefore, the aim of the study was to investigate the potential of orally administered zingerone to protect growing Sprague-Dawley rats (mimicking growing children) against high-fructose diet-induced metabolic derangements. Forty, 21-day old female Sprague-Dawley rats were randomly allocated and administered the following four treatments for 12 weeks: group I: standard rat chow (SR) + plain water (PW) + plain gelatine cube (PC). group II: SR + 20% (w/v) fructose solution (FS) + PC. group III: SR + FS + 100 mg/kg/day of fenofibrate in gelatine cube. group IV: SR+ FS + 20 mg/kg/day of zingerone in gelatine cube. The rats’ triglyceride, cholesterol, insulin & adiponectin concentration, visceral fat liver lipid content, homoeostasis model assessment of insulin resistance (HOMA-IR) and ability to handle glucose were determined. Oral administration of zingerone significantly increased (P<0.001) visceral fat and liver lipid content (P<0.001), respectively. Results from the study revealed that administration of 20% fructose solution did not induce metabolic dysfunction, however the zingerone treatment increased visceral fat and liver lipid content, all these lipid abnormalities are typical features of the metabolic syndrome, therefore the current study suggests that zingerone has no effect on metabolic dysfunction in adolescent females.

Keywords: antidiabetic, metabolic syndrome, zingerone, antiobesogenic

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152 Japanese Quail Breeding: The Second in Poultry Industry

Authors: A. Smaï, H. Idouhar-Saadi, S. Zenia, F. Haddadj, A. Aboun, S.Doumandji

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The quail is the smallest member of the order fowl. His captive breeding has been practiced for centuries by the Japanese. Knowing that in the literature, it is mentioned that the end of lay is noted for the age of 6 months, our work has revealed a good egg production by females aged up to 35 weeks of age. In the same vein, our study focused on various parameters such as weight, diet and the number of eggs laid and this in order to better know the potential production and reproduction of domestic quail. Egg production has started from the 8th week of age of breeding, crop them and their counts are conducted daily basis until the age of 35 weeks. Indeed, biometric parameters are studied such as weight, length, and the largest diameter, the shape index, the index of shell, in order to analyze the physical condition of eggs by females of age. Until the age of 22 weeks, the eggs have maintained good biometric features. Japanese quail are best producing eggs. Hatchability is also considered. They are excellent poultry yields, since they begin laying eggs in two months and can provide abundant nesting with females over 8 months in our study. Other farms results reveal conclusions. Indeed, one aspect remains to be developed; it is the analysis of nutritional and therapeutic values of eggs over the age of females. The latter, given their wealth is a dietary supplement of animal origin with dietary value (it contains 0 cholesterol) that characterizes the quail eggs. Raising quail among other reproduction requires minimal when compared to other domestic birds space, this is the second breeding, in terms of importance after the chicken. Therefore, in the case of a farm that works exclusively in the production of eggs, requires minimal work and free space, as well as reduced costs.

Keywords: Japanese quail, reproduction, eggs, biometrics, reproductive age

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151 A Case Study: Effect of Low Carbs High Fats Diet (Also Known as LCHF Diet) Combined with Fried Foods in Extra Virgin Olive Oil in Patient with Type 2 Diabetes and Central Obesity

Authors: Cristian Baldini

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‘Diabesity’ is a term for diabetes occurring in the context of obesity. The positive effect of LCHF diets (low-carb, high-fat diets) is well documented: LCHF diets are at least as effective as other dietary strategies for reducing body weight, improving glycaemic control, and reducing both hyperinsulinaemia and blood glucose (reduction of HbA1c) in type 2 diabetes and have unique positive effects on blood lipid concentrations and cardiovascular risk factors. Also, in obese insulin-resistant women, food fried in extra-virgin olive oil significantly reduced both insulin and C-peptide responses after a meal. This case study shows that if combined, both dietary strategies produce a strong effect on blood glucose, resulting in a “forced” reduction of exogenous insulin injection to avoid the problem of hypoglycaemia. Blood tests after three months of this dietary treatment show how HbA1c, triglycerides, and blood lipid profile (LDL, HDL, Total Cholesterol) are improved despite the reduction of exogenous insulin injection of 80% with a parallel body weight decrease of 15%. For continuous glucose monitoring (CGM), the patient used FreeStyle Libre before and after the dietary treatment. In order to check general body functions and glycosuria, the patient used the urine test Multistix 10 SG Siemens.

Keywords: diabetes, obesity, diabesity, fat, fried foods

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150 Gonadal Maturation in Pen Shells Pinna Rudis and Pinna Nobilis Stimulated by Reproductive Neuropeptides

Authors: Ntalamagka N., Sanchis-Benlloch P. J., Mayoral-Serrano R., Tena-Medialdea J., García-March J. R.

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The pen shell Pinna nobilis population has declined dramatically since 2016 due to die-off events observed in the whole extent of the Mediterranean Sea associated with the protozoan Haplosporidium pinnae. As of 2019, it is considered a critically endangered species. Due to its ecological importance and its endangered status, several initiatives have been developed for its salvation and recovery. This research is an effort to understand and control its reproduction under captivity. As a limited number of Pinna nobilis individuals could be used for experimentation, the possibility of using the Pinna rudis as a model animal was explored. The molecular mechanism that regulates the reproduction of both species is unknown; consequently, transcriptomic analysis was performed to identify neuropeptides that are expressed in the key regulatory tissues of the visceral ganglia and gonads of both species. Neuropeptides form an important group of signaling peptides that regulate reproductive, behavioral and physiological functions in molluscs. In total, 17 neuropeptide precursors were identified in P. nobilis and 14 in P. rudis transcriptomes; 14 of them were identical in both species. This affinity verified the genetic similarity of these species at the reproduction level. APGWamide, buccalin, ELH and GnRH were tested in P. rudis and demonstrated their capacity to advance gonadal maturation and trigger spawning while spawning was recorded in P. nobilis after the usage of APGWamide and buccalin. The neuropeptides were administered using intramuscular injection and cholesterol implants following relative literature as well as a new method was developed for external administration without the use of anesthesia using a mathematical model. The know-how of this research will not only lead to the survival of the species but also will narrow the horizons of broodstock conditioning of other similar species.

Keywords: neuropeptides, Pinna nobilis, reproduction, transcriptomics

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149 Dietary Habit and Anthropometric Status in Hypertensive Patients Compared to Normotensive Participants in the North of Iran

Authors: Marjan Mahdavi-Roshan, Arsalan Salari, Mahbobeh Gholipour

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Hypertension is one of the important reasons of morbidity and mortality in countries, including Iran. It has been shown that hypertension is a consequence of the interaction of genetics and environment. Nutrients have important roles in the controlling of blood pressure. We assessed dietary habit and anthropometric status in patients with hypertension in the north of Iran, and that have special dietary habit and according to their culture. This study was conducted on 127 patients with newly recognized hypertension and the 120 normotensive participants. Anthropometric status was measured and demographic characteristics, and medical condition were collected by valid questionnaires and dietary habit assessment was assessed with 3-day food recall (two weekdays and one weekend). The mean age of participants was 58 ± 6.7 years. The mean level of energy intake, saturated fat, vitamin D, potassium, zinc, dietary fiber, vitamin C, calcium, phosphorus, copper and magnesium was significantly lower in the hypertensive group compared to the control (p < 0.05). After adjusting for energy intake, positive association was observe between hypertension and some dietary nutrients including; Cholesterol [OR: 1.1, P: 0.001, B: 0.06], fiber [OR: 1.6, P: 0.001, B: 1.8], vitamin D [OR: 2.6, P: 0.006, B: 0.9] and zinc [OR: 1.4, P: 0.006, B: 0.3] intake. Logistic regression analysis showed that there was not significant association between hypertension, weight and waist circumference. In our study, the mean intake of some nutrients was lower in the hypertensive individuals compared to the normotensive individual. Health training about suitable dietary habits and easier access to vitamin D supplementation in patients with hypertension are cost-effective tools to improve outcomes in Iran.

Keywords: hypertension, north of Iran, dietary intake, weight

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148 Effects of the Supplementation of Potassium Humate at Different Levels to the Dairy Cows' Concentrated Mix during Dry Period on Early Lactation Yield Parameters and Dam/Calf Immunity

Authors: Cangir Uyarlar, E. Eren Gultepe, I. Sadi Cetingul, Ismail Bayram

Abstract:

This study was conducted to investigate the effect of humic acid (Potassium Humate) at different levels on rations on the effects of both maternal and offspring health, metabolic parameters and immunity levels in transition dairy cows. For this purpose, 50 Holstein dairy cows divided 5 trial groups. Experimental groups were designed as follows: A) Control (0% Humas); B) 0.5 Humas (0,5% in concentrated diet); C) 1 Humas (1% in concentrated diet), D) 1,5 Humas (1,5% in concentrated diet), E) 2 Humas (2% in concentrated diet), respectively. The study lasted from the first day of the dry period to postpartum 30th day. Diets were prepared as isocaloric and isonitrogenic. In the experiment, the day on which the animals gave birth was accepted as ‘0 (zero)’ and blood was taken from tail vein (v. coccygea) at -60, -53, -46, -39, -32, -25, -18, -11, -4, 0, ; Colostrum samples were taken on days 0, 1 and 2; Blood samples were taken on days 0, 1, 2, 15 and 30 from the juguler vein (v. jugularis) of the new born calves. Total blood leukocyte, Lymphocyte, Monocyte, granulocytes, Hemoglobin, Hematocrit, MCV, MCH, MWC, RDW, PLT, MPV, PDW, PCT, NEFA, BHBA, Glucose, Total Cholesterol , Triglyceride, LDL, HDL, VLDL, ALT, AST, ALP, GGT and Total IgG levels and colostrum IgG levels were determined in this experiment. The results suggest that although the supplementation of humic acid at 2% level adversely affected to production parameters, the addition of humic acid (potassium humate) to the concentrate mix during the dry period (particularly 0.5 and 1% levels) may provide an increasing on mother and the offspring immunity, some improving on serum metabolism parameters and enhancing the milk production.

Keywords: humic acid, dairy cow, calf, immunity

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147 Subclinical Renal Damage Induced by High-Fat Diet in Young Rats

Authors: Larissa M. Vargas, Julia M. Sacchi, Renata O. Pereira, Lucas S. Asano, Iara C. Araújo, Patricia Fiorino, Vera Farah

Abstract:

The aim of this study was to evaluate the occurrence of subclinical organ injuries induced by high-fat diet. Male wistar rats (n=5/group) were divided in control diet group (CD), commercial rat chow, and hyperlipidic diet (30% lipids) group (HD) administrated during 8 weeks, starting after weaning. All the procedures followed the rules of the Committee of Research and Ethics of the Mackenzie University (CEUA Nº 077/03/2011). At the end of protocol the animals were euthanized by anesthesia overload and the left kidney was removed. Intrarenal lipid deposition was evaluated by histological analyses with oilred. Kidney slices were stained with picrosirius red to evaluate the area of the Bowman's capsule (AB) and space (SB), and glomerular tuft area (GT). The renal expression of sterol regulatory element–binding protein (SREBP-2) was performed by Western Blotting. Creatinine concentration (serum and urine) and lipid profile were determined by colorimetric kit (Labtest). At the end of the protocol there was no differences in body weight between the groups, however the HD showed a marked increase in lipid deposits, glomeruli and tubules, and biochemical analysis for cholesterol and triglycerides. Moreover, in the kidney, the high-fat diet induced a reduction in the AB (13%), GT (18%) and SB (17%) associated with a reduction in glomerular filtration rate (creatinine clearance). The renal SRBP2 expression was increased in HD group. These data suggests that consumption of high-fat diet starting in childhood is associated with subclinical renal damage and function.

Keywords: high-fat diet, kidney, intrarenal lipid deposition, SRBP2

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146 Impact of Environmental Changes on Blood Parameters in the Pelophylax ridibundus

Authors: Murat Tosunoglu, Cigdem Gul, Nurcihan Hacioglu, Nurdan Tepeova

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Amphibian and Reptilian species are influenced by pollution and habitat destruction. Blood parameters of Amphibia species were particularly affected by the negative environmental conditions. Studied frog samples 36 clinically normal Pelophylax ridibundus individuals were captured along the Biga Stream between April–June 2014. When comparing our findings with the Turkish legislation (Water pollution control regulation), the 1. Locality of the Biga stream in terms of total coliform classified as "high quality water" (Coliform: 866.66 MPN/100 mL), while the 2. locality was a "contaminated water" (Coliform: 53266.66 MPN/100 mL). Blood samples of the live specimens were obtained in the laboratory within one day of their capture. The blood samples were taken from the etherized frogs by means of ventriculus punctures, via heparinized hematocrit capillaries. Hematological and biochemical analyses based on high quality water and contaminated water, respectively, are as follows: Red blood cell count (444210.52-426846.15 per cubic millimeter of blood), white blood cell count (4215.78-4684.61 per cubic millimeter of blood), hematocrit value (29.25-29.43 %), hemoglobin concentration (7.76-7.22 g/dl), mean corpuscular volume (637.64-719.99 fl), mean corpuscular hemoglobin (184.78-174.75 pg), mean corpuscular hemoglobin concentration (29.44-24.82 %), glucose (103.74-124.13 mg/dl), urea (87.68-81.72 mg/L), cholesterol (148.20-197.39 mg/dl), creatinine (0.29-0.28 mg/dl), uric acid (10.26-7.55 mg/L), albumin (1.13-1.39 g/dl), calcium (11.45-9.70 mg/dl), triglyceride (135.23-155.85 mg/dl), total protein (4.26-3.73 g/dl), phosphorus (6.83-17.86 mg/dl), and magnesium (0.95-1.06 mg/dl). The some hematological parameters in P. ridibundus specimens are given for the first time in this study. No water quality dependent variation was observed in clinic hematology parameters measured.

Keywords: Pelophylax ridibundus, hematological parameters, biochemistry, freshwater quality

Procedia PDF Downloads 370
145 Relationship Between Muscle Mass and Insulin Resistance in Cirrhotic Patients with Hepatitis B

Authors: Eyüp S. Akbas, Betul Ayaz, Beyza S. Haksever, Sema Basat

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We aimed to evaluate the relationship between insulin resistance, muscle mass and muscle strength in patients with Hepatitis B virus-related cirrhosis. In our study, there were 65 patients with hepatitis B virus-related cirrhosis in Child A and B group and 65 healthy control individual. Control group was chosen between patients who admitted to the internal medicine clinic and had no pathological values in a routine examination. Muscle mass index was calculated with bioimpedance analysis for both groups to determine muscle strength and muscle mass. Handgrip strength, arm, and calf circumference were measured. In both groups, HOMA-IR was calculated to determine insulin resistance. Homeostatic Model Assessment of Insulin Resistance (HOMA-IR) value was detected 3,47±3,80 in the study group and 1,83±1,20 in control group. There were significant differences between the two groups in arm circumference, fasting insulin, fasting glucose, HOMA-IR, High-density lipoprotein (HDL) and total cholesterol parameters. The correlation coefficient between muscle mass and insulin resistance was statistically insignificant, especially in the study group. In healthy individuals group and all the groups, there wasn’t a correlation between muscle mass and insulin resistance. The upper limit for HOMA-IR was determined as 3,2. In control group, %78,9 of individuals were in HOMA-IR ( < 3.2) group and %21,1 of them were in ( ≥ 3,2) group. In study group, %68,3 of individuals were in HOMA-IR ( < 3,2) group and %31.7 were in HOMA-IR ( ≥ 3,2) group. In our study, we did not find a relationship between muscle mass and insulin resistance in patients with liver cirrhosis. In the study group, we detected a positive relationship between muscle mass, handgrip strength, and calf circumference. We did not find a relationship between insulin resistance and handgrip strength in our study.

Keywords: cirrhosis, hepatitis B, Insulin resistance, muscle mass

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144 Luteolin Exhibits Anti-Diabetic Effects by Increasing Oxidative Capacity and Regulating Anti-Oxidant Metabolism

Authors: Eun-Young Kwon, Myung-Sook Choi, Su-Jung Cho, Ji-Young Choi, So Young Kim, Youngji Han

Abstract:

Overweight and obesity have been linked to a low-grade chronic inflammatory response and an increased risk of developing metabolic syndrome including insulin resistance, type 2 diabetes mellitus and certain types of cancers. Luteolin is a dietary flavonoid with anti-inflammatory, anti-oxidant, anti-cancer and anti-diabetic properties. However, little is known about the detailed mechanism associated with the effect of luteolin on inflammation-related obesity and its complications. The aim of the present study was to reveal the anti-diabetic effect of luteolin in diet-induced obesity mice using “transcriptomics” tool. Thirty-nine male C57BL/6J mice (4-week-old) were randomly divided into 3 groups and were fed normal diet, high-fat diet (HFD, 20% fat) and HFD+0.005% (w/w) luteolin for 16 weeks. Luteolin improved insulin resistance, as measured by HOMA-IR and glucose tolerance, along with preservation action of pancreatic β-cells, compared to the HFD group. Luteoiln was significantly decreased the levels of leptin and ghrelin that play a pivotal role in energy balance, and the macrophage low-grade inflammation marker sCD163 (soluble Cd antigen 163) in plasma. Activities of hepatic anti-oxidant enzymes (catalase and glutathione peroxidase) were increased, while the levels of plasma transaminase (GOT and GPT) and oxidative damage markers (hepatic mitochondria H2O2 and TBARS) were markedly decreased by luteolin supplementation. In addition, luteolin increased oxidative capacity and fatty acid utilization by presenting decrease in enzyme activities of citrate synthase, cytochrome C oxidase and β-hydroxyacyl CoA dehydrogenase and UCP3 gene expression compared to high-fat diet. Moreover, our microarray results of muscle also revealed down-regulated gene expressions associated with TCA cycle by HFD were reversed to normal level by luteolin treatment. Taken together, our results indicate that luteolin is one of bioactive components for improving insulin resistance by increasing oxidative capacity, modulating anti-oxidant metabolism and suppressing inflammatory signaling cascades in diet-induced obese mice. These results provide possible therapeutic targets for prevention and treatment of diet-induced obesity and its complications.

Keywords: anti-oxidant metabolism, diabetes, luteolin, oxidative capacity

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143 Exploring the Safety of Sodium Glucose Co-Transporter-2 Inhibitors at the Imperial College London Diabetes Centre, UAE

Authors: Raad Nari, Maura Moriaty, Maha T. Barakat

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Introduction: Sodium-glucose co-transporter-2 (SGLT2) inhibitors are a new class of oral anti-diabetic drugs with a unique mechanism of action. They are used to improve glycaemic control in adults with type 2 diabetes by enhancing urinary glucose excretion. In the UAE, there has been certainly an increased use of these medications. As with any new medication, there are safety considerations related to their use in patients with type two diabetes. A retrospective study was conducted at the three main centres of the Imperial College London Diabetes Centre. Methodology: All patients in electronic database (Diamond) from October 2014 to October 2017 were included with a minimum of six months usage of sodium glucose co-transporter inhibitors that comprise canagliflozin, dapagliflozin and empagliflozin. There were 15 paired sample biochemical and clinical correlations. The analysis was done at the start of the study, three months and six months apart. SPSS version 24 was used for this study. Conclusion: This study of sodium glucose co-transporter-2 inhibitors used showed significant reductions in weight, glycated haemoglobin A1C, systolic and diastolic blood pressures. As the case with systematic reviews, there were similar changes in liver enzymes, raised total cholesterol, low density lipopoptein and high density lipoprotein. There was slight improvement in estimated glomerular filtration rate too. Our analysis also showed that they increased in the incidence of urinary tract symptoms and incidence of urinary tract infections.

Keywords: SGLT2 inhibitors dapagliflozin empagliflozin canagliflozin, adverse effects, amputation diabetic ketoacidosis DKA, urinary tract infection

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142 Efficacy of Ethanolic Extract of Aerva javanica Aerial Parts in the Amelioration of CCl4-Induced Hepatotoxicity and Oxidative Damage in Rats

Authors: Mohammad K. Parvez, Ahmed H. Arbab, Mohammed S. Al-Dosari, Adnan J. Al-Rehaily

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We investigated ex vivo and in vivo antioxidative and hepatoprotective effect of Aerva javanica. Total ethanol extract of A. javanica aerial parts was prepared, and tested on DCFH-toxicated HepG2 cell in CCl4-injured Wistar rats. MTT-assay was used to determine cell viability, and serum biochemical markers of liver injury as well as histopathology were performed. In vitro DPPH and β-carotene free-radical scavenging assay and phytochemical screening of the extract was done. Furthermore, A. javanica total extract was standardized and validated by HPTLC method. While DCFH-injured cells were recovered to about 56.7% by 100 microg/ml of the extract, a 200 microg/ml dose resulted in hepatocytes recovery by about 90.2%. Oral administration of the extract (100 and 200 mg/kg.bw/day) significantly normalized the serum SGOT, SGPT, GGT, ALP, bilirubin, cholesterol, HDL, LDL, VLDL, TG and MDA levels, including tissue NP-SH and TP in CCl4-injured rats. In addition, the histopathology of dissected liver also revealed that A. javanica cured the tissue lesion compared to reference drug, Silymarin. In vitro assays revealed strong free-radical scavenging ability of the extract and presence of alkaloids, flavonoids, tannins, sterols and saponins where Rutin, a well-known antioxidant flavonoid was identified. Our finding therefore, suggests the therapeutic potential of A. javanica in various liver diseases. However, isolation of the active principles, their mechanism of action and other therapeutic contribution remain to be addressed.

Keywords: Aerva javanica, antioxidant, hepatoprotection, rutin

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141 Obesity, Metabolic Syndrome and Related Risk Behaviors Among Thai Medical Students of Thammasat University

Authors: Patcharapa Thaweekul, Paskorn Sritipsukho

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Background: During the training period of the 6-year medical curriculum, medical students seem to have many risk behaviors of developing obesity. This study aims to demonstrate the prevalence and risk behavior of obesity and related metabolic disorders among the final-year medical students of Thammasat University as well as the change in nutritional status during studying program. Methods: 123 participants were asked to complete the self-report questionnaires. Weight, height, waist circumference and blood pressure were obtained. Blood samples were drawn for total cholesterol, high-density lipoprotein, low-density lipoprotein, triglycerides and plasma glucose. Body weight and height of the medical students in the first year were obtained from the medical report at the entry. Results: The prevalence of overweight and obesity at the entry to medical school was 22.0% and increased to 30.1% in the final year. Two obese students (5.4%) was diagnosed as metabolic syndrome. During 6-year curriculum, the BMI gained in male medical students were more significant as compared to female students (1.76±1.74 and 0.43±1.82 kg/m2, respectively; p <.001). The current BMI is significantly correlated with the BMI at entry. Serum LDL-C in the overweight/obese students was significantly higher as compared to the normal weight and underweight group. Sleep deprivation was a significantly frequent behavior in the overweight/obese students. Conclusion: Medical students, as having high-risk behaviors, should be assessed for the nutritional status and metabolic parameters. Medical schools should promote the healthy behaviors to increase the healthy eating and exercise habits and reduced the risk behaviors among them.

Keywords: medical students, metabolic syndrome, obesity, risk behaviors

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140 Rapid Plasmonic Colorimetric Glucose Biosensor via Biocatalytic Enlargement of Gold Nanostars

Authors: Masauso Moses Phiri

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Frequent glucose monitoring is essential to the management of diabetes. Plasmonic enzyme-based glucose biosensors have the advantages of greater specificity, simplicity and rapidity. The aim of this study was to develop a rapid plasmonic colorimetric glucose biosensor based on biocatalytic enlargement of AuNS guided by GOx. Gold nanoparticles of 18 nm in diameter were synthesized using the citrate method. Using these as seeds, a modified seeded method for the synthesis of monodispersed gold nanostars was followed. Both the spherical and star-shaped nanoparticles were characterized using ultra-violet visible spectroscopy, agarose gel electrophoresis, dynamic light scattering, high-resolution transmission electron microscopy and energy-dispersive X-ray spectroscopy. The feasibility of a plasmonic colorimetric assay through growth of AuNS by silver coating in the presence of hydrogen peroxide was investigated by several control and optimization experiments. Conditions for excellent sensing such as the concentration of the detection solution in the presence of 20 µL AuNS, 10 mM of 2-(N-morpholino) ethanesulfonic acid (MES), ammonia and hydrogen peroxide were optimized. Using the optimized conditions, the glucose assay was developed by adding 5mM of GOx to the solution and varying concentrations of glucose to it. Kinetic readings, as well as color changes, were observed. The results showed that the absorbance values of the AuNS were blue shifting and increasing as the concentration of glucose was elevated. Control experiments indicated no growth of AuNS in the absence of GOx, glucose or molecular O₂. Increased glucose concentration led to an enhanced growth of AuNS. The detection of glucose was also done by naked-eye. The color development was near complete in ± 10 minutes. The kinetic readings which were monitored at 450 and 560 nm showed that the assay could discriminate between different concentrations of glucose by ± 50 seconds and near complete at ± 120 seconds. A calibration curve for the qualitative measurement of glucose was derived. The magnitude of wavelength shifts and absorbance values increased concomitantly with glucose concentrations until 90 µg/mL. Beyond that, it leveled off. The lowest amount of glucose that could produce a blue shift in the localized surface plasmon resonance (LSPR) absorption maxima was found to be 10 – 90 µg/mL. The limit of detection was 0.12 µg/mL. This enabled the construction of a direct sensitivity plasmonic colorimetric detection of glucose using AuNS that was rapid, sensitive and cost-effective with naked-eye detection. It has great potential for transfer of technology for point-of-care devices.

Keywords: colorimetric, gold nanostars, glucose, glucose oxidase, plasmonic

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139 Linking Metabolism, Pluripotency and Epigenetic Changes during Early Differentiation of Embryonic Stem Cells

Authors: Arieh Moussaieff, Bénédicte Elena-Herrmann, Yaakov Nahmias, Daniel Aberdam

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Differentiation of pluripotent stem cells is a slow process, marked by the gradual loss of pluripotency factors over days in culture. While the first few days of differentiation show minor changes in the cellular transcriptome, intracellular signaling pathways remain largely unknown. Recently, several groups demonstrated that the metabolism of pluripotent mouse and human cells is different from that of somatic cells, showing a marked increase in glycolysis previously identified in cancer as the Warburg effect. Here, we sought to identify the earliest metabolic changes induced at the first hours of differentiation. High-resolution NMR analysis identified 35 metabolites and a distinct, gradual transition in metabolism during early differentiation. Metabolic and transcriptional analyses showed the induction of glycolysis toward acetate and acetyl-coA in pluripotent cells, and an increase in cholesterol biosynthesis during early differentiation. Importantly, this metabolic pathway regulated differentiation of human and mouse embryonic stem cells. Acetate delayed differentiation preventing differentiation-induced histone de-acetylation in a dose-dependent manner. Glycolytic inhibitors upstream of acetate caused differentiation of pluripotent cells, while those downstream delayed differentiation. Our data suggests that a rapid loss of glycolysis in early differentiation down-regulates acetate and acetyl-coA production, causing a loss of histone acetylation and concomitant loss of pluripotency. It demonstrate that pluripotent stem cells utilize a novel metabolism pathway to maintain pluripotency through acetate/acetyl-coA and highlights the important role metabolism plays in pluripotency and early differentiation of stem cells.

Keywords: pluripotency, metabolomics, epigenetics, acetyl-coA

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138 Effects of in Ovo Injection of Royal Jelly on Hatchability, One-Day Old Chickens Quality, Total Antioxidant Status and Blood Lipoproteins

Authors: Amin Adeli, Maryam Zarei

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Background and purpose: Royal jelly (RJ) is a natural product with anti-hyperlipidemic and antioxidant properties. In ovo administration of RJ may improve lipid profile and antioxidant properties. This study was conducted to evaluate, for first time, the effects of in ovo injection of the RJ on hatchability, one-day old chick quality, total antioxidant status and blood lipoproteins. Methods: 400 incubating eggs produced by Ross 308 strain (52 weeks of age in first stage of production) were prepared and assigned into 4 groups (n=100) and 4 replications per group (n=25). These 4 groups were injected by the following pattern: 1) 0.1 ml normal saline (control), 2) 0.1 mg RJ+0.1 ml normal saline, 3) 0.2 mg RJ+0.1 ml normal saline, and 4) 0.3 mg RJ+0.1 ml normal saline. Injections were performed using a laminar flow system Lipid profile, antioxidant properties, hatchability, and one-day old chicken quality were assessed. Results: The administration of RJ at concentration of 0.1increased the percentage of hatchability compared to concentration of 0.2 and control, significant differences have not been observed among groups for quality scores (P>0.05). The results showed that in ovo injection of the RJ did not have any significant effects on lipid profile; but administration of the RJ only decreased High-density lipoprotein (HDL cholesterol, HDL-C) (P<0.05). The results showed that injection of the RJ at concentration of 0.3 increased total antioxidant capacity (TAC) compared to control group (p<0.05). Injection of the RJ progressively increased gluthation peroxidase (GPx) activity (p<0.05). The results showed that injection of the RJ decreased superoxide dismutase (SOD) compared to control group (p<0.05). Conclusion: In ovo injection of the RJ at the highest concentration increased TAC and GPx, but it did not have significant effects on lipid profile. Future studies are needed to investigate the effects of the RJ on the above-mentioned mechanisms.

Keywords: antioxidant enzymes, chicken quality, hatchability, royal jelly

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137 Effect of 8 Weeks of Intervention on Physical Fitness, Hepatokines, and Insulin Resistance in Obese Subjects

Authors: Adela Penesova, Zofia Radikova, Boris Bajer, Andrea Havranova, Miroslav Vlcek

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Background: The aim of our study was to compare the effect of intensified lifestyle intervention on insulin resistance (HOMA-IR), alanine aminotransferase (ALT), aspartate aminotransferase (AST), and Fibroblast growth factor (FGF) 21 after 8 weeks of lifestyle intervention. Methods: A group of 43 obese patients (13M/30F; 43.0±12.4 years; BMI (body mass index) 31.2±6.3 kg/m2 participated in a weight loss interventional program (NCT02325804) following an 8-week hypocaloric diet (-30% energy expenditure) and physical activity 150 minutes/week. Insulin sensitivity was evaluated according to the homeostasis model assessment of insulin resistance (HOMA-IR) and insulin sensitivity indices according to Matsuda and Cederholm were calculated (ISImat and ISIced). Plasma ALT, AST, Fetuin-A, FGF 21, and physical fitness were measured. Results: The average reduction of body weight was 6.8±4.9 kg (0-15 kg; p=0.0006), accompanied with a significant reduction of body fat amount of fat mass (p=0.03), and waist circumference (p=0.02). Insulin sensitivity has been improved (IR HOMA 2.71±3.90 vs 1.24±0.83; p=0.01; ISIMat 6.64±4.38 vs 8.93±5.36 p ≤ 0.001). Total, LDL cholesterol, and triglycerides decreased (p=0.05, p=0.04, p=0.04, respectively). Physical fitness significantly improved after intervention (as measure VO2 max (maximal oxygen uptake) (p ≤ 0.001). ALT decreased significantly (0.44±0.26 vs post 0.33±0.18 ukat/l, p=0.004); however, AST not (pre 0.40±0.15 vs 0.35±0.09 ukat/l, p=0.07). Hepatokine Fetuin-A significantly decreased after intervention (43.1±10.8 vs 32.6±8.6 ng/ml, p < 0.001); however, FGF 21 levels tended to decrease (146±152 vs 132±164 pg/ml, p=0.07). Conclusion: 8-weeks of diet and physical activity intervention program in obese otherwise healthy subjects led to an improvement of insulin resistance parameters and liver marker profiles, as well as increased physical fitness. This study was supported by grants APVV 15-0228; VEGA 2/0161/16.

Keywords: obesity, diet, exercice, insulin sensitivity

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