When mandarin orange peel was used for a substrate of citric aCid fermentation by Aspergillus niger, principal enzyme activities were investigated. Not only the activity of polygalacturonase and pectin esterase being capable of digesting pectin and crude fiber of mandarin orange peel. but also that of carboxymethyl cellulase, xylanase and amylase was high. In carbohydrate metabolism, the activity of enzymes related in HMP pathway was higher than that in EMP pathway at the orange peel medium designed hereby rather than synthetic medium. Productivity of citric acid was significantly increased when the activity of citrate synthetase was high and 5imultaneously those of aconitase and NADP-dependent dehydrogenase were low.
Pectinesterase was isolated from the culture medium of Aspergillus japonicus and partially purified by DEAE-Sephadex batchwise, Sephadex G-75 gel filtration and ion exchange chromatography. The purified enzyme solution was completely free from polygalacturonase which depolymerizes pectin molecule. The ability of the pectinesterase to demethylate high methoxyl pectin was investigated. On 20 minute of incubation methoxyl content of low methoxyl pectin decreased from 88% to 6.93%. In general gel prepared with the pectin containing lower methoxyl content showed the lower value of percent sag, and showed the hieher Bel strength. Textural characteristics of pectin gel determined by Rheometer showed that as the methoxyl content was lowered, hardness and resilience of the gel were increased and cohesiveness was decreased. Apple juice containing HMP and organic acids can be converted into low methoxyl pectin apple jelly by the action of pectinesterase and addition of calcium ion. The strength of low methoxyl pectin apple jelly increased when it stored at room temperature.
The consequence analysis for the unconfined vapor cloud explosion(UVCE) accident by the continuous release of butane vapor was performed and effects of process parameters on consequences were analyzed in standard conditions. For the case of continuous release(87.8 kg/s) of butane vapor at 8 m elevated height in the debutanizing process of tile naphtha cracking plant operating at 877 kPa & 346.75 K, we found that combustion ranges of dispersed vapor estimated by HMP model were 11.2~120.2 m and overpressures estimated by TNT equivalency model at 200 m were about 37.35~55.1 kPa. Also, overpressures estimated by Model UVCE I based on advective travel time to $X_{LFL}$ were smaller than those estimated by Model UVCE IIbased on real travel time between $X_{UFL}$ and $X_{LFL}$. At the same time, damage intensities at 200 m and effect ranges by overpressure could be predicted. Furthermore, simulation results showed that effects of operating pressures on consequences were larger than those of operating temperatures and results of accidents were increased with increasing operating pressures. At this time, sensitivities of overpressures for UVCE accident by the continuous release were about 5 kPa/atm.
Very low methoxyl pectin (VLMP) has different physical and rheological properties compared to high and low methoxyl pectins (HMP and LMP). In this study, we produced LMP and VLMP by alkaline de-esterification, and investigated the structural and textural properties. Apple peel pectin was kept at pH 12 using 5.0 M NaOH solution for 3 and 24 h to produce LMP and VLMP, respectively. The molecular weight was decreased due to the removal of an esterified group in the pectin backbones by the alkali treatment, and the VLMP showed a higher calcium ion sensitivity which leads to the production of the gel with increased hardness. The result clearly showed that VLMP has the potential to improve the texture and stability in food products depending on their degree of esterification, and this result can be applied as a functional ingredient in food industrial area application to enhance the current commercial pectins.
Antimicrobial actions of reactive oxygen/nitrogen species (ROS/RNS) derived from products of NADPH oxidase and inducible nitric oxide (NO) synthase in host phagocytes inactivate various bacterial macromolecules. To cope with these cytotoxic radicals, pathogenic bacteria have evolved to conserve systems necessary for detoxifying ROS/RNS and repairing damages caused by their actions. In response to these stresses, bacteria also induce expression of molecular chaperones to aid in ameliorating protein misfolding. In this study, we explored the function of a newly identified chaperone Spy, that is localized exclusively in the periplasm when bacteria exposed to conditions causing spheroplast formation, in the resistance of Salmonella Typhimurium to ROS/RNS. A spy deletion mutant was constructed in S. Typhimurium by a PCR-mediated method of one-step gene inactivation with ${\lambda}$ Red recombinase, and subjected to ROS/RNS stresses. The spy mutant Salmonella showed a modest decrease in growth rate in NO-producing cultures, and no detectable difference of growth rate in $H_2O_2$ containing cultures, compared with that of wild type Salmonella. Quantitative RT-PCR analysis showed that spy mRNA levels were similar regardless of both stresses, but were increased considerably in Salmonella mutants lacking the flavohemoglobin Hmp, which are incapable of NO detoxification, and lacking an alternative sigma factor RpoS, conferring hypersusceptibility to $H_2O_2$. Results demonstrate that Spy expression can be induced under extreme conditions of both stresses, and suggest that the protein may have supportive roles in maintaining proteostasis in the periplasm where various chaperones may act in concert with Spy, thereby protecting bacteria against toxicities of ROS/RNS.
Kang Donghee;Khil Lee-Yong;park Kwangsik;Lee Byung-Hoon;Moon Chang-Kiu
Environmental Analysis Health and Toxicology
/
v.20
no.1
/
pp.75-85
/
2005
This study was aimed to know the effect of cadmium chloride (CdCl₂) on glucose transport in L6 myotube and its action mechanism. CdCl₂ increased the 2-deoxy- (l-3H)-D-glucose (2-DOG) uptake 1.9 and 2.4 fold at 10 and 25 μM respectively. To investigate the stimulating-mechanism of glucose transport induced by CdCl₂, the wortmannin and PD98059 were used as PI3K (phosphatidylinositol 3-kinase) inhibitor and MAPK inhibitor respectively, which did not affect 2-DOG uptake. This fact suggests that CdCl₂ induced 2-DOG uptake may not be concerned to the insulin signalling pathway. Whereas nifedipine, a calcium channel blocker, and trifluoperazine, a calmodulin inhibitor, were found to inhibit the 2-DOG uptake stimulted by CdCl₂. In addition, we also measured the ROS (reactive oxygen species) production and GSH level in L6 myotube to investigate the correlation between the glucose uptake and ROS. CdCl₂(25 μM) increased ROS generation approximately 1.5 fold and changed the cellular GSH level, but GSSG/GSH ratio remained unchanged. CdCl₂ stimulated 2-DOG uptake and ROS generation were inhibited by N-acetylcystein. And BSO pretreatment, a potent inhibitor of γ-GCS, resulted in the dramatic decrease of 2-DOG uptake and also the increase of the sensitivity to cadmium cytotoxicity. The obtained results suggest that CdCl₂-stimulated glucose uptake might be based on the activation of HMP shunt as an antioxidant defense mechanism of the cells.
To elucidate the relationship between chemical structure and biological activity of alantolactone, and also to investigate the relationship between the growth of cells and the respiration of Chlorella pyrenoidosa affected by alantolactone, alantolactone and isoalantolactone were isolated from Inula helenium L., and di-, and tetrahydroalantolactones were prepared by the hydrogenation. At a concentration of 5$\times$10-5M alantolactone, the growth rate of Chlorella was greatly reduced. The viability of cells was also reduced over 50% within 2 hr at a concentration of 2.5$\times$10-4M alantolactone. However, oxygen uptake was increased by 20% over 3 hr. And 14CO2 production from glucose-1-14C, glucose-6-14C and 14C-acetate-U.L. was also increased by alantolactone. Biological activityof alantolactone was significantly reduced by cysteine, reduced glutathione or cystine but not by tryptophan or histidine. It was detected by spectrophotometrically and by TLC that alantolactone was also reacted with thiols except cystine. The solution of alantolactone reached with thiol gave the UV absorption spectrum of $\alpha$-saturated ${\gamma}$-lactone, and most of SH groups were disappeared by the addition reaction. From the reaction mixture of alantolactone and cysteine, a lactone adduct was isolated and purified. Isoalantolactone had shown similar activity as alantolactone, however, it was appeared that di-, and tetrahydroalantolactones were not only inactive biologically but also in vitro. It was concluded that there was no correlationship between increased respiration rate and mortality of Chlorella. During the respiration TCA cycle was activated, however it was uncertain that the activation of EMP or HMP was also appeared. Alantolactone and isoalantolactone were biologically active compounds but others were inactive. The reactivity of $\alpha$-methylene ${\gamma}$-lactone moiety toward SH group was principally responsible for its biological activity in sesquiterpene lactones.
The metabolic patterns of C-1 and C-6-carbon atoms of glucose were observed in the tissue homogenates of the Ehrlich ascites tumor tissue which was incubated for 3 hours in the Dubnuff metabolic shaking incubator. $C^{14}-1-and\;C^{14}-6-glucose$ were used as tracers. The glucose media in which tissue homogenate was incubated was kept at a concentration of 200mg% glucose of carrier and appropriate amount of $C^{14}-1-or\;C^{14}-6-tracer$. At the end of 3 hour incubation, respiratory $CO_2$ samples trapped by alkaline which is placed in the tenter well of incubation flask were analyzed for the total $CO_2$ production rates and their radioactivities. The tissue homogenate samples after incubation were analyzed for their concentrations of glucose, lactate, pyruvate and glycogen and calculations were made on the glucose consumption rate, pyruvate and lactate accumulation rates. The following results were obtained. Data obtained in each group are as follows: 1. In the tissue homogenate, which was incubated with $C^{14}-1-glucose as a substrate, total $CO_2$ production rate averaged $19.0{\pm}5.0{\mu}M/hr/gm$ and the mean specific activity of respiratory $CO_2$ was $840{\pm}296\;cpm/mgC.$ Relative specific activity (RSA) which means the fraction of $CO_2$ derived from medium $C^{14}-1-glucose$ to total $CO_2$ production rate was calculated by ratio of SA of respiratory $CO_2$ and medium $C^{14}-1-glucose.$ RSA was $14.3{\pm}5.0%,$ Accordingly actual $CO_2$ production rate from medium $C^{14}-1-glucose$ showed a mean value of $2.79{\pm}1.35\;{\mu}m$ of which amount was equivalent to the mean value of total glucose consumption rate $(RGDco_2)$, namely, $5.1{\pm}1.3%.$ Lactate and pyruvate appearance rates averaged $7.13{\pm}1.26\;and\;0.21{\pm}0.02{\mu}M/hr/gm,$ respectively. Assuming that these 3 carbon compounds appeared in the medium were derived from glucose, calculations were made that relative glucose disappearance rate into lactate $(RGD_L)$ was $38.0{\pm}5.4%\;and\;RGD_P$ was $1.23{\pm}0.03%.$ Therefore, about 43.3% of the total glucose consumed were accounted for by conversion into the respiratory $CO_2$, lactate and pyruvate. 2. In the second group, which was incubated with $C^{14}-1-glucose$ as a substrate, glucose consumption rate, lactate and pyruvate appearance rates showed almost the same order as the values of the $C^{14}-1-glucose$ substrate group. However, RSA was remarkably decreased showing a mean value of $1.02{\pm}0.13%.$ This fact means that the C-6 carbon of glucose take the minor part in the oxidative metabolism of glucose. The glycogen level in both substrate tissue homogenate showed less than 0.3% of tissue weight. These low value suggested that there was an inhibition of carbohydrate synthesis in the Ehrlich ascites tumor tissue. 3. The catabolic pathway of glucose in the tumor tissue were analyzed on the basis of Bloom's principle from the values of RSA. It was found that in the tumor tissue more than 90% of $CO_2$ derived from glucose were oxidized via the alternate pathway other than principal EMP-TCA cycle such as hexose monophosphate pathway (HMP). From the data described above, it was assumed that in the Ehrlich tumor tissue anaerobic glycolysis proceeds normally although, the oxidation of products of anaerobic glycolysis via the TCA cycle is inhibited resulting in the accumulation of lactate and almost all of oxidative energy from glucose is released by oxidative pathway such as HMP.
Background: The safety of herbal products is an important issue in Korea. It is more complicated because of the concomitant use of conventional western drugs and herbal medicine, including prescribed medicine, folk herbs and food supplements. Although both western and traditional Korean medical doctors have studied on the safety of HMP, their results did not show relevant or consistent conclusions because of the poor quality of studies, including bias. Objectives: The aim of this study was to review the studies in Korea related to drug induced liver injury (DILI) systematically. We tried to estimate the proportion of causative materials of DILI and evaluate the clinical difference of DILIs from different materials. Methods: Systematic searches were performed on MedRIC and MEDLIS in Korea. The extraction of data as well as selective screening of the studies was carried out independently by two of the authors. There were no restrictions on the types of publication, including grey literature. Results: Twenty-one articles were included. Of them only 7 adapted prospective design and only 6 were published in peer-reviewed journals. The proportion of conventional drugs associated with hepatotoxic injuries in all DILI ranged from 15.8% to 83.3% and that of herbal medicine was from 28.9% to 44.7%. However, the criteria for herbal medicine were not clearly defined and concomitant medications were not fully investigated in most studies. There were limited objective data in the clinical differences of liver injuries from different materials and their results were conflicting. Conclusions: The causality assessments regarding DILI of herbal medicine were not performed properly and causative materials were misclassified in most of the studies published in Korea. These make the safety issue still ambiguous because of the limitations and lack of objectivity of the studies. More rigorous studies are required for clearly addressing these conflicting issues with cooperative investigation between traditional Korean and western medicine.
Metabolic responses to the protein-free, high-carbohydrate diet and subsequent food restriction on the same diet at the level of 50% and 75% has been studied on the adult albino rats. The energy source was either corn starch or sugar. In experiment I, adult male rats weighing $509{\pm}8g$ were divided into two groups 10 rats each. Rats fed on the stock diet served as a control. Rats of restriction group received a protein free diet until they reduced their weight down to 400g and continue on a protein-calorie restriction diet until they reduced their weight down to 300g. In experiment II, 28 adult male rats and the same numbers of female rats weighing $329{\pm}5g$ and $223{\pm}4g$ respectively were divided into four groups, 7 males and females in each. Rats fed on a stock diet were sacrificed at the point when others started a protein free diet. These were served as the control. The protein free group received a protein free diet ad libitum for 4 weeks. The 50% restriction group and 75% restriction group were fed on a protein free diet coupled with food restriction at levels of 50% and 75% respectively for 3 weeks. In the result of this study: 1. The rate of body weight changes was similar between the males and the females. Feeding protein free diet ad lib. initiated a rapid weight lost of approximately 25% and protein free diet coupled with food restriction showed 37-43% reduction of their initial weight. 2. There was no significant differences in the value of the N concentration in liver, spleen, brain and muscle between controls and experimental groups. 3. Rats fed on protein free diet showed 1/10 value of the control in the nitrogen excretion in urine. However female showed less N excretion than male. 4. Observing blood picture, the effects of protein depletion and calorie restriction were not appeared any remarkable changes. 5. There was no sign of fatty liver which might result from protein depletion and calorie restriction. 6. Following semi-starvation, FAO and HMP-DH total enzyme activity was reduced, but activity per unit weight was relatively stable.
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