• Title/Summary/Keyword: Metabolic enzymes

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Anesthetic management for dental surgery in a child with glycogen storage disease type IIIa: a case report

  • Bugra, Aykenar;Nedim, Cekmen
    • Journal of Dental Anesthesia and Pain Medicine
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    • v.22 no.6
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    • pp.451-455
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    • 2022
  • Glycogen storage disease (GSD) is a group of inherited disorders, which result in the deficiency of enzymes involved in glycogen metabolism, leading to an accumulation of glycogen in various organs. Deficiency of amylo-1-6-glicosidase (debranching enzyme) causes glycogen storage disease type III (GSD III). The main problems that anesthesiologists face in patients with GSD III include hypoglycemia, muscle weakness, delayed awakening due to abnormal liver function, possible difficulty in airway, and cardiomyopathy. In the face of these difficulties, airway preparation and appropriate glucose monitoring and support during the fasting period are important. The doses of the drugs to be used should be calculated considering the increased volume of distribution and decreased metabolic activity of the liver. We present the case of a child with GSD IIIa who underwent dental prosedation under general anesthesia. She was also being prepared for liver transplantation. This case was additionally complicated by the patient's serious allergic reaction to eggs and milk.

GC-MS Analysis of Ricinus communis, Pongamia pinnata, Datura metal, Azadirachta indica, Acalypha indica (leaf) Extract Using Methanol Extraction

  • J. Varshini premakumari;M. Job Gopinath
    • Mass Spectrometry Letters
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    • v.14 no.3
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    • pp.79-90
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    • 2023
  • Natural goods, especially therapeutic plants, are abundant in the World. Because they have the ability to provide all humanity with countless advantages as a source of medicines, medicinal plants are presently receiving more attention than ever. These plants' therapeutic efficacy is based on bioactive phytochemical components that have clear physiological effects on the human body. The drying process is crucial for the preparation of plant materials prior to extraction since freshly harvested plant materials include active enzymes that create active components, intermediates, and metabolic processes. Many of the phytoconstituents may be extracted using the semi-polar solvent methanol. The goal of the current work is to use the GC-MS gas chromatography- mass spectrometry technology to identify the phytochemicals and review their biological activity. In methanol leaf extract, 5 phytocompounds were found in Ricinus communis, 5 phytocompounds in Pongamia pinnata, 12 phytocompounds in Datura metal, 7 phytocompounds in Azadirachta indica, 11 phytocompounds in Acalypha indica.

Analysis of Flavonoid Components of Unripen Mandarin in Jeju Island and Change of Flavonoid Composition through Secondary Metabolism

  • Ho Bin Kim;Han Soo Kim;Moon Suk Choi;Jong Heon Kim;Min Sun Park;Mi Jung Kim
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2022.09a
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    • pp.105-105
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    • 2022
  • Unripen mandarin in Jeju Island is known to contain functional ingredients including various flavonoids. This Study was carried out to identify the components of Unripen mandarin extracts and Secondary metabolism by enzyme treatment on Unripen mandarin. We extracted Unripen mandarin using optimal extraction method and selected the most optimal enzyme among commercial enzymes for a Secondary metabolism. As a result, flavonoid components such as Hesperidine and Narirutin, which are known to be contained a lot in unripen mandarin, could be analyzed. However In this extraction method there were no other flavonoid components such as Nobiletin, Tangeretin known to contain in unripen mandarin. However as a result of secondary metabolism a new functional component called Prunin which was not known to be contained in unripen mandarin, was detected as a secondary metabolic product due to enzyme treatment. Through this, it can be confirmed that it would be possible to develop high-value-added products by enzyme treatment on unripen mandarin.

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Morphological Engineering of Filamentous Fungi: Research Progress and Perspectives

  • Zhengwu Lu;Zhiqun Chen;Yunguo Liu;Xuexue Hua;Cuijuan Gao;Jingjing Liu
    • Journal of Microbiology and Biotechnology
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    • v.34 no.6
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    • pp.1197-1205
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    • 2024
  • Filamentous fungi are important cell factories for the production of high-value enzymes and chemicals for the food, chemical, and pharmaceutical industries. Under submerged fermentation, filamentous fungi exhibit diverse fungal morphologies that are influenced by environmental factors, which in turn affect the rheological properties and mass transfer of the fermentation system, and ultimately the synthesis of products. In this review, we first summarize the mechanisms of mycelial morphogenesis and then provide an overview of current developments in methods and strategies for morphological regulation, including physicochemical and metabolic engineering approaches. We also anticipate that rapid developments in synthetic biology and genetic manipulation tools will accelerate morphological engineering in the future.

Limiting Concentrate during Growing Period Affect Performance and Gene Expression of Hepatic Gluconeogenic Enzymes and Visfatin in Korean Native Beef Calves

  • Chang, S.S.;Lohakare, J.D.;Singh, N.K.;Kwon, E.G.;Nejad, J.G.;Sung, K.I.;Hong, S.K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.2
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    • pp.202-210
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    • 2013
  • This study elucidated the effects of limited concentrate feeding on growth, plasma profile, and gene expression of gluconeogenic enzymes and visfatin in the liver of Hanwoo beef calves. The purpose of this study was to test that reducing the amount of concentrate would partially be compensated by increasing the intake of forage and by altering the metabolic status. The study utilized 20 Korean native beef calves (Hanwoo; 60 to 70 d of age) divided into two groups of 10 calves each for 158 d. Control group calves received the amount of concentrate as per the established Korean feeding standards for Hanwoo, whereas calves in the restricted group only received half the amount of concentrate as per standard requirements. Good quality forage (Timothy hay) was available for ad libitum consumption to both groups. Since calves were with their dam until 4 months of age in breeding pens before weaning, the intake of milk before weaning was not recorded, however, the concentrate and forage intakes were recorded daily. Body weights (BW) were recorded at start and on 10 d interval. Blood samples were collected at start and at 50 d interval. On the final day of the experiment, liver biopsies were collected from all animals in each group. The BW was not different between the groups at all times, but tended to be higher (p = 0.061) only at final BW in control than restricted group. Total BW gain in the control group was 116.2 kg as opposed to 84.1 kg in restricted group that led to average BW gain of 736 g/d and 532 g/d in respective groups, and the differences were significant (p<0.01). As planned, the calves in the control group had higher concentrate and lower forage intake than the restricted group. The plasma variables like total protein and urea were higher (p<0.05) in control than restricted group. The mRNA expressions for the gluconeogenic enzymes such as cytosolic phosphoenol pyruvate carboxykinase (EC 4.1.1.32) and pyruvate carboxylase (EC 6.4.1.1), and visfatin measured by quantitative real-time PCR in liver biopsies showed higher expression (p<0.05) in restricted group than control. Overall, restricting concentrate severely reduced the growth intensity and affected few plasma indices, and gene expression in liver was increased indicating that restricting concentrate in the feeding schemes during early growth for beef calves is not advocated.

Differential Expression of Xenobiotic-Matabolizing Enzymes by Benzylisothiazole in Association with Hepatotoxicity: Effects on Rat Hepatic Epoxide Hydrolase, Glutathione S-Transferases and Cytochrome P450s

  • Cho, Min- Kyung;Kim, Sang-Geon
    • Toxicological Research
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    • v.14 no.3
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    • pp.293-300
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    • 1998
  • Previous studies have shown that the heterocycles including thiazoles are efficacious in inducing phase phase II metabolizing enzyme as well as certain cytochrome P450s and that the inductin of these matabolizing enzymes by the heterocyclic agents is highly associated with their hepatotoxicity. In the present study, the effects of benzylisothiazole (BIT), which has a isothiazole moiety, on the expression of microsomal epoxide hydrolase (mEH), major glutathione S-transerases and cytochrome P450s were studied in the rat liver in association with its hepatotoxicity. Treatment of rats with BIT(1.17 mmol/kg, 1~3d) resulted in substantial increases in the mEH. rGSTA2, rGSTA2, rGSTM1 and rGSTM2 mRNA levels, whereas rGSTA3 and rGSTA5 mRNA levels were increased to much lesser extents. A time-course study showed that the mRNA levels of mEH and rGSTs were greater at 24hr after treatment than those after 3 days of consecutive treatment. Relative changes in mEH and rGST mRNA levels were consistent with those in the proteins, as assessed by Western immunoblot analysis. Hepatic cytochrom P450 levels were monitored after BIT treatment under the assumption that metabolic activation of BIT may affect expression of the enzymes in conjunction with hepatotoxicity. Immunoblot analysis revealed that cytochrome P450 2B1/2 were 3-to 4-fold induced in rats teatd with BIT(1.17 mmol/kg/day.3days), whereas P450 1A2, 2C11 and 3A1/2 levels were decreased to 20~30% of those in unteatd rats. P450 2E1 was only slightly decreased by BIT. Thus, the levels of several cytochrome P450s were suppressed by BIT treatment. Rats treated with BIT at the dose of 1.17mmol/kg for 3 days exhibited extensive multifocal nodular necrosis with moderate to extensive diffuse liver cell degeneration. No notable toxicity was observed in the kidney. These results showed that BIT induces mEH and rGSTs in the liver with increases in the mRNA levels, whereas the agent significantly decreased major cytochrome P450s. The changes in the detoxifying enzymes might be associated with the necrotic liver after consecutive treatment.

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Anaerobic Bacterial Degradation for the Effective Utilization of Biomass

  • Ohmiya, Kunio;Sakka, Kazuo;Kimura, Tetsuya
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.6
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    • pp.482-493
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    • 2005
  • Biomass is originally photosynthesized from inorgainic compounds such as $CO_2$, minerals, water and solar energy. Recent studies have shown that anaerobic bacteria have the ability to convert recalcitrant biomass such as cellullosic or chitinoic materials to useful compounds. The biomass containing agricultural waste, unutilized wood and other garbage is expected to utilize as feed, food and fuel by microbial degradation and other metabolic functions. In this study we isolated several anaerobic, cellulolytic and chitinolytic bacteria from rumen fluid, compost and soil to study their related enzymes and genes. The anaerobic and cellulolytic bacteria, Clostridium thermocellum, Clostridium stercorarium, and Clostridium josui, were isolated from compost and the chitinolytic Clostridium paraputrificum from beach soil and Ruminococcus albus was isolated from cow rumen. After isolation, novel cellulase and xylanase genes from these anaerobes were cloned and expressed in Escherichia coli. The properties of the cloned enzymes showed that some of them were the components of the enzyme (cellulase) complex, i.e., cellulosome, which is known to form complexes by binding cohesin domains on the cellulase integrating protein (Cip: or core protein) and dockerin domains on the enzymes. Several dockerin and cohesin polypeptides were independently produced by E. coli and their binding properties were specified with BIAcore by measuring surface plasmon resonance. Three pairs of cohesin-dockerin with differing binding specificities were selected. Two of their genes encoding their respective cohesin polypeptides were combined to one gene and expressed in E. coli as a chimeric core protein, on which two dockerin-dehydrogenase chimeras, the dockerin-formaldehyde dehydrogenase and the dockerin-NADH dehydrogenase are planning to bind for catalyzing $CO_2$ reduction to formic acid by feeding NADH. This reaction may represent a novel strategy for the reduction of the green house gases. Enzymes from the anaerobes were also expressed in tobacco and rice plants. The activity of a xylanase from C. stercorarium was detected in leaves, stems, and rice grain under the control of CaMV35S promoter. The digestibility of transgenic rice leaves in goat rumen was slightly accelerated. C. paraputrificum was found to solubilize shrimp shells and chitin to generate hydrogen gas. Hydrogen productivity (1.7 mol $H_2/mol$ glucos) of the organism was improved up to 1.8 times by additional expression of the own hydrogenase gene in C. paraputrficum using a modified vector of Clostridiu, perfringens. The hydrygen producing microflora from soil, garbage and dried pelletted garbage, known as refuse derived fuel(RDF), were also found to be effective in converting biomass waste to hydrogen gas.

Effects of Phytase and Carbohydrases Supplementation to Diet with a Partial Replacement of Soybean Meal with Rapeseed Meal and Cottonseed Meal on Growth Performance and Nutrient Digestibility of Growing Pigs

  • Shim, Y.H.;Chae, B.J.;Lee, J.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.9
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    • pp.1339-1347
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    • 2003
  • An experiment was conducted to investigate the effects of microbial phytase ($Natuphos^{(R)}$) supplementation in combination with carbohydrases (composed of enzymes targeted to soybean meal (SBM) dietary components such as $\alpha$-galactosides and galactomannans; $Endo-Power^{(R)}$) to corn-soybean meal based diet (CSD) and complex diet (CD) with a partial replacement of SBM with rape seed meal (RSM) and cotton seed meal (CSM) on growth performance and nutrient digestibility of growing pigs. A total of 168 growing pigs averaging $13.18{\pm}1.77kg$ of initial body weight was arranged as a $2{\times}2$ factorial design with main effects of diet types (corn-SBM based diet (CSD) and complex diets (CD; 5% of SBM was replaced with 2.5% of RSM and 2.5% of CSM in diet for phase I (0 to 3 weeks) and 6% of SBM was replaced with 3% of RSM and 3% of CSM in diet for phase II (4 to 7 weeks))) and enzyme supplementation (none and 0.1% of phytase (500 FTU/kg diet) and 0.1% of carbohydrases). The diet with enzyme application were formulated to have a 0.18% unit lower aP than diets without enzyme application. Each treatment had three replicates with 14 pigs per replicate. To determine supplementation effect of phytase and carbohydrases on ileal amino acid digestibility of SBM, RSM and CSM, a total of 18 T-cannulated pigs (initial body weight; $13.52{\pm}1.24kg$) were assigned to six dietary treatments in the present study. Dietary treatments in metabolic trial included 1) SBM diet, 2) SBM diet+with enzymes (phytase (500 FTU/kg) and carbohydrases at 0.1%, respectively), 3) CSM diet, 4) CSM diet+enzymes, 5) RSM diet and 6) RSM diet+enzymes. During whole experimental period (0 to 7 wks), there was no difference in growth performance between diets (CSD and CD). However, dietary phytase and carbohydrases supplementation significantly improved gain/feed ratio (G:F) of growing pigs. During the phase II (4-7 weeks), dietary phytase and carbohydrases supplementation significantly improved all fecal nutrient digestibilities (Dry matter (DM), gross energy (GE), crude protein (CP), crude fat (CF), calcium (Ca) and phosphorus (P)). Dietary phytase and carbohydrases supplementation improved significantly overall ileal amino acid digestibilities of SBM, RSM and CSM based diets (p<0.05). The simultaneous inclusion of phytase and carbohydrases in both of CSD and CD reduced feed cost per kg body weight gain (FCG). Also, results suggest that 2.5 to 3% of RSM and CSM, respectively, might be used as a protein source in growing pig diets without having an adverse effect on the growth performance and nutrient digestibility and simultaneous phytase and carbohydrases addition improves nutritional value of SBM, RSM and CSM by improving ileal amino acid digestibilities.

Effects of Selenium, Copper and Magnesium on Antioxidant Enzymes and Lipid Peroxidation in Bovine Fluorosis

  • Han, Bo;Yoon, Soonseek;Su, Jingliang;Han, H.R.;Wang, Mei;Qu, Weijie;Zhong, Daibin
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.12
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    • pp.1695-1699
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    • 2004
  • The antioxidant enzymes, lipid peroxidation and free radicals assessment were made of the effects of selenium, copper and magnesium on bovine endemic fluorosis under high fluoride, low selenium and low copper productive conditions. Thirty-two beef cattle were selected from high fluoride area, and randomly divided into four groups with eight cattle each as follows: (1) high fluoride control group (HFC); (2) supplemented group with 0.25 mg/kg selenium (HFSe); (3) supplemented group with 15 mg/kg copper (HFCu) and (4) supplemented group with 0.25 mg/kg selenium+15 mg/kg copper+1 mg/kg magnesium (HFSeCuMg) per day for 83 days. Moreover, eight beef cattle were selected from non-high fluoride area as normal control group. Blood samples were collected from cattle on 0 d, 30 d and 83 d respectively, to analyze the enzyme activities and concentration of GSH-px, CAT, SOD, MDA and free radicals. The results showed that the contents of free radicals and MDA in HFC group were significantly higher, and the whole blood GSH-px, CAT, erythrocyte SOD activities were lower than the normal control group. Free radicals, metabolic imbalance and antioxidant disorder therefore, play an important role in fluorosis. However, GSH-px, CAT and SOD activities in HFSe group and HFSeCuMg group at 30 d and 83 d were markedly higher than the same groups at the 0 d and the HFC group at the same time. Likewise, there was a corresponding reduction in the contents of free radicals and MDA. These findings indicated that supplementation with selenium, copper and magnesium elevated high fluoride bovine antioxidant enzymes, and decreased MDA and free radicals contents. But, the activities of supplementation selenium group did not increase until day 83. These results demonstrated that fluorosis was associated with lower serum Se and Cu levels than in the control, and it was therefore concluded that fluorosis is associated with decreased serum levels of these minerals. Long-term high fluoride intake under productive condition enhances oxidative stress in the blood, thereby disturbing the antioxidant defense of cattle. Increased oxidative stress could be one of the mediating factors in the pathogenesis of toxic manifestations of fluoride. It is benefical for high fluoride cattle supplemented with proper selenium, copper and magnesium to increase fluoride excretion and obtain the protective impact of the activity of oxidative enzymes, and to decrease lipid peroxidation and free radicals contents.

Metabolic engineering of Vit C: Biofortification of potato

  • Upadhyaya, Chandrama P.;Park, Se-Won
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2010.10a
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    • pp.14-14
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    • 2010
  • Vitamin C (ascorbic acid) is an essential component for collagen biosynthesis and also for the proper functioning of the cardiovascular system in humans. Unlike most of the animals, humans lack the ability to synthesize ascorbic acid on their own due to a mutation in the gene encoding the last enzyme of ascorbate biosynthesis. As a result, vitamin C must be obtained from dietary sources like plants. In this study, we have developed two different kinds of transgenic potato plants (Solanumtuberosum L. cv. Taedong Valley) overexpressing strawberry GalUR and mouse GLoase gene under the control of CaMV 35S promoter with increased ascorbic acid levels. Integration of the these genes in the plant genome was confirmed by PCR and Southern blotting. Ascorbic acid(AsA) levels in transgenic tubers were determined by high-performance liquid chromatography(HPLC). The over-expression of these genes resulted in 2-4 folds increase in AsA intransgenic potato and the levels of AsA were positively correlated with increased geneactivity. The transgenic lines with enhanced vitamin C content showed enhanced tolerance to abiotic stresses induced by methyl viologen(MV), NaCl or mannitol as compared to untransformed control plants. The leaf disc senescence assay showed better tolerance in transgenic lines by retaining higher chlorophyll as compared to the untransformed control plants. Present study demonstrated that the over-expression of these gene enhanced the level of AsA in potato tubers and these transgenics performed better under different abiotic stresses as compared to untransformed control. We have also investigated the mechanism of the abiotic stress tolerance upon enhancing the level of the ascorbate in transgenic potato. The transgenic potato plants overexpressing GalUR gene with enhanced accumulation of ascorbate were investigated to analyze the antioxidants activity of enzymes involved in the ascorbate-glutathione cycle and their tolerance mechanism against different abiotic stresses under invitro conditions. Transformed potato tubers subjected to various abiotic stresses induced by methyl viologen, sodium chloride and zinc chloride showed significant increase in the activities of superoxide dismutase(SOD, EC 1.15.1.1), catalase, enzymes of ascorbate-glutathione cycle enzymes such as ascorbate peroxidase(APX, EC 1.11.1.11), dehydroascorbate reductase(DHAR, EC 1.8.5.1), and glutathione reductase(GR, EC 1.8.1.7) as well as the levels of ascorbate, GSH and proline when compared to the untransformed tubers. The increased enzyme activities correlated with their mRNA transcript accumulation in the stressed transgenic tubers. Pronounced differences in redox status were also observed in stressed transgenic potato tubers that showed more tolerance to abiotic stresses when compared to untransformed tubers. From the present study, it is evident that improved to lerance against abiotic stresses in transgenic tubers is due to the increased activity of enzymes involved in the antioxidant system together with enhanced ascorbate accumulated in transformed tubers when compared to untransformed tubers. At moment we also investigating the role of enhanced reduced glutathione level for the maintenance of the methylglyoxal level as it is evident that methylglyoxal is a potent cytotoxic compound produced under the abiotic stress and the maintenance of the methylglyoxal level is important to survive the plant under stress conditions.

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