• Title/Summary/Keyword: feeding inhibition

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Optimization of Growth Medium and Poly-$\beta$-hydroxybutyric Acid Production from Methanol in Methylobacterium organophilum (메탄올로부터 Methylobacterium organophilum에 의한 Poly-$\beta$-hydroxybutyric Acid의 생산과 배지성분의 최적화)

  • Choi, Joon-H;Kim, Jung H.;M. Daniel;J.M. Lebeault
    • Microbiology and Biotechnology Letters
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    • v.17 no.4
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    • pp.392-396
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    • 1989
  • Methylobacterium organophilum, a facultative methylotroph was cultivated on a methanol as a sole carbon and energy source. The cell growth was affected by the various components of minimal synthetic medium and the medium composition was optimized with 0.5% (v/v) methanol at pH 6.8 and at 3$0^{\circ}C$. The maximum specific growth rate of M. organophilum was achieved to 0.26 hr$^{-1}$ in the optimized medium which has following composition: Methanol, 0.5% (v/v):(NH$_4$)$_2$SO$_4$, 1.0g/l:KH$_2$PO$_4$, 2.13g/l:KH$_2$PO$_4$, 1.305g/ι:MgSO$_4$.7$H_2O$. 45g/l and trace elements (CaCl$_2$.2$H_2O$, 3.3mg:FeSO$_4$.7$H_2O$, 1.3mg:MnSO$_4$.4$H_2O$, 130$\mu\textrm{g}$:ZnSO$_4$.5$H_2O$, 40$\mu\textrm{g}$:Na$_2$MoO$_4$.2$H_2O$, 40$\mu\textrm{g}$:CoCl$_2$.6$H_2O$, 40$\mu\textrm{g}$:H$_3$BO$_3$, 30$\mu\textrm{g}$ per liter). By the limitation of nitrogen and deficiency of Mn$^{+2}$ or Fe$^{+2}$, the cell growth was significantly repressed. Methanol greatly repressed the cell growth and the complete inhibition was observed at concentration above 4% (v/v). In order to overcome the methanol inhibition and to prevent the methanol limitation, intermittent feeding of methanol was conducted by a D.O.-stat technique. PHB production by M. organophilum was stimulated by deficiency of nutrients such as NH$_{4}^{+}$, SO$_{4}^{-2}$, $Mg^{+2}$, $K^{+}$, or PO$_{4}^{-3}$ in the medium. The maximum PHB content was obtained as 58% of dry cell weight under deficiency of potassium ion in the optimized synthetic medium.

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Purification and Characterization of Lactate Dehydrogenase Isozymes in Channa argus (가물치(Channa argus) 젖산탈수소효소 동위효소들의 정제 및 특성)

  • Park, Eun-Mi;Yum, Jung-Joo
    • Journal of Life Science
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    • v.20 no.2
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    • pp.260-268
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    • 2010
  • The lactate dehydrogenase (EC 1.1.1.27, LDH) isozymes in tissues from Channa argus were purified and characterized by biochemical, immunochemical and kinetic methods. The activity of LDH in skeletal muscle was the highest at 380.4 units and those in heart, eye and brain tissues were 13.4, 3,5 and 5.4 units, respectively. Citrate synthase (EC 4.1.3.7, CS) activity in heart tissue was the highest at 20.7 units. LDH/CS in skeletal muscle, heart, eye and brain tissues were 172.9, 0.6, 0.32 and 0.47. Protein concentration in skeletal muscle tissue was 14.7 mg/g and specific activities of LDH in skeletal muscle, heart, eye and brain tissues were 25.88, 0.79, 0.31 and 1.38 units/mg, respectively. Therefore, skeletal muscle tissue was anaerobic and heart tissue was aerobic. The LDH isozymes in tissues were identified by polyacrylamide gel electrophoresis, immunoprecipitation and Western blot with antiserum against $A_4$, $B_4$, and eye-specific $C_4$. LDH $A_4$, $A_3B$, $A_2B_2$. $AB_3$ and $B_4$ isozymes were detected in every tissue, $C_4$, $AC_3$, $A_2C_2$ and $A_3C$ were detected in eye tissue, and $A_3C$ was found in brain tissue. LDH $A_4$, $A_3B$, $A_2B_2$, $AB_3$, $B_4$, eye-specific $C_4$ isozymes were purified by affinity chromatography and Preparative PAGE Cells. The LDH $A_4$ isozyme was purified in the fraction from elution with $NAD^+$ containing buffer of affinity chromatography. Eye-specific $C_4$ isozyme was eluted right after $A_4$, after which $B_4$ isozyme was eluted with plain buffer. As a result, one part of molecular structures in $A_4$, $B_4$ and eye-specific $C_4$ were similar, but were different from each other in $B_4$ and $C_4$. Therefore the subunit A may be conservative in evolution, and the evolution of subunit B seems to be faster than that of subunit A. The activity of LDH $A_4$, $A_2B_2$, $B_4$, and eye-specific $C_4$ isozymes remained at 39.98, 21.28, 19.67 and 16.87% as a result of the inhibition by 10 mM of pyruvate, so the degree of inhibition was very high. The $Km^{PYR}$ values were 0.17, 0.27 and 0.133 mM in $A_4$, $B_4$ and eye-specific $C_4$ isozymes, respectively. The optimum pH of LDH $A_4$, $B_4$, eye-specific $C_4$, $A_2B_2$, $A_3B$, and $AB_3$ were pH 6.5, pH 8.5, pH 5.5, pH 6.0-6.5, pH 5.0 and pH 7.5. The $A_4$ and heterotetramer isozymes stabilized a broad range of pH. Especially, LDH activities in skeletal muscle tissue were high, resulting in a high degree of muscle activity.LDH metabolism in eye tissue seems to be converted faster from pyruvate to lactate by eye-specific $C_4$ isozyme as eye-specific $C_4$ have the highest affinity for pyruvate, and right after the conversion, oxidation of lactate was induced by $A_4$ isozyme. It was found that expression of Ldh-C, affinity to substrate and reaction time of $C_4$ isozyme were different according to the ecological environmental and feeding capturing patterns.

Tre Effect of UW Solution for Protection of Ischemic Injury in Free Myocutaneous Flaps of the Rabbit (가토 근피판에서 허혈성 손상 방지를 위한 UW 관류제 사용의 효과)

  • Suh Woo-Suk;Kwun Woo-Heung;Kim Sang-Woon;Lee Su-Jung;Kwun Koing-Bo
    • Korean Journal of Head & Neck Oncology
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    • v.9 no.1
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    • pp.3-9
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    • 1993
  • The benficial effects for perfusion in the preservation of free flaps have been controversial in the clinical and experimental field until now. This study was undertaken to observe the effect of UW solution. a recently developed. high molecular weight. organ perfusion solution. for protection of ischemic injury in normothermic free myocutaneous flaps. Forty rabbits were used in this sutdy. A 1x2x1cm sized gastrocnemius myocutaneous flap based on the feeding vessel from common femoral artery was made. The author set up the ischemic time for 12 hours in these flaps. The flap was washed out with normal saline(control grop, n=10), urokinase(comparative group I, n=10), UW solution before ischemic time(comparative group II, n=10) and UW solution before ischemic time and pentoxifylline before reperfusion(comparative group III, n=10). Afterthen, reperfusion was made for 12 hours. After this procedure, we checked the degree of ischemia and necrosis of myocutaneous flap by gross finding, electrical stimulation test of muscle, triphenyltetrazolium chloride staining and wet/dry weight ratio. The degree of necrosis of comparative group II and III were lesser than control and urokinase group in gross finding(p<0.05). In the electrical stimulation test of muscle, there was no statistical difference between control($1.76{\pm}1.01$) and urokinase($2.36{\pm}\1.02$) group however the muscular power of comparative group II($3.54{\pm}0.93$) and III($3.49{\pm}1.37gm/mm^2$) demonstrated significantly higher than control group(p<0.05). The ischemic findings were found in seven cases of control group and three cases of urokinase group but there were no ischemic findings in comparative group II and III in TIC stain(p<0.05). In the wet/dry weight ratio of flaps in order to evaluate the tissue edema. there was no statistical difference between control($4.55{\pm}0.29$) and III($3.75{\pm}0.48$) were scored significantly lesser than control and urokinase group (p<0.05). These results suggest that perfusion washout with UW solution improves the viability of normothermic free myocutaneous flap by inhibition of cellular swelling.

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Effect of Oral Administration of Egg Yolk on Cholesterol Metabolism in Rats (난황 경구투여가 랫드의 콜레스테롤 대사에 미치는 영향)

  • Bang, Han-Tae;Hwangbo, Jong;Park, Sang-Oh;Park, Byung-Sung
    • Journal of the Korean Applied Science and Technology
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    • v.31 no.2
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    • pp.255-264
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    • 2014
  • This study was performed to elucidate the biochemical mechanism of metabolism on reducing blood lipid, by oral administration of egg yolk in rats. A total of 36 Sprague Dawley male rats were randomized into four treatment groups, according to a randomized block design. Each group was further divided into three repeat cages, with each repeat cage comprising of 3 rats. The animals were orally administered with egg yolk once a day, while feeding the same purified pellet diet for 6 weeks. The four treatment groups were: C(control, saline 1.0 g), T1(pork belly oil 1.0 g), T2(egg yolk 1.0 g), T3(pork belly oil 1.0 g and egg yolk 1.0 g alternating every week). The measured parameters in each group are listed as follows in the order of highest to the lowest: daily average gain of body weight(T1>T3>T2>C); blood triglyceride and total cholesterol(T1>C>T3>T2) HDL-C (T2>C>T3>T1); and LDL-C (T1>T3>C>T2). AST and ALT, which are the index of liver function, were the highest in T1 but was lowest in T2. The weights of the liver, spleen, and kidney, except for the abdominal fat, showed no significant difference. The weight of abdominal fat was the highest in T1, but there were no significant difference among C, T2, and T3. The HMG-CoA reductase activity was the highest in T1 followed by T3, C but T2 was lowest. The daily fecal excretions of the total sterol, neutral sterol and acid sterol was highest in T2 but lowest in T1. The results of this study show that the egg consumption reduces the blood lipid through facilitation of fecal excretions of sterols and inhibition of enzyme activity in cholesterol biosynthesis, in the liver of animal and human.

Effects of YK-209 Mulberry Leaves on Antioxidative Defense System of Liver in Streptozotocin-Induced Diabetic Rats (YK-209 뽕잎이 Streptozotocin 유발 당뇨쥐 간조직의 항산화계에 미치는 영향)

  • 유수경;이순재
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.31 no.6
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    • pp.1065-1070
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    • 2002
  • The purpose of this study was investigated the effects of YK-209 mulberry leaves on antioxidative defense system of liver in diabetic rats induced with streptozotocin (STZ). Male Sprague-Dawley rats weighing 100$\pm$10 g were randomly assigned to one normal and four STZ-induced diabetic groups; YK-209 mulberry leaves free diet (DM group),0.1% YK-209 mulberry leaves diet (DM-0.1Y group),0.2% YK-209 mulberry leaves diet (DM-0.2Y group) and 0.4% YK-209 mulberry leaves diet (DM-0.4Y group). Diabetes was induced by intravenous Injection of 55 mg/kg body weight of STZ in sodium citrate buffer (pH 4.3) via tail vein after 4 weeks feeding of experimental diets. Rats were sacrificed at the 9th day of diabetic states. Liver weight in all four diabetic groups were higher than normal group, but YK-209 mulberry supplementation groups were lower than DM group. Hepatic superoxide dismutase (SOD) activity was significantly decreased in all diabetic groups, compared with normal group. Hepatic glutathione peroxidase (GSHpx) activity was 7.3% decreased in DM group, compared with normal group, but those of DM-0.1Y and DM-0.2Y groups were maintained the normal level. The hepatic thiobarbituric acid reactive substances was markedly increased by 144% in DM group, compared with normal group, but those of DM-0. 1Y, DM-0.2Y groups were maintained the normal level. The contents of lipofuscin in liver were increased by 100% in DM group compared with normal group, but those of DM-0. 1Y, DM-0.2Y and DM-0.4Y groups were decreased to 42% 43% and 44%, respectively, compared with DM group. The hepatic superoxide radical (0$^2$-) contents in DM group were increased to 81%, compared with normal group, but those of DM-0.1Y and DM-0.4Y groups were similar to those of normal group. The present result indicate that YK-209 mulberry leaves regarded to suppress lipid peroxidation as an free radical scavenger system by the inhibition of oxidative stress.

Biochemical and Nutritional Effects of Rice, Barley, and Millet on Rats (백미대맥(白米大麥) 및 속(粟) 사료(飼料)가 실험용(實驗用) 백서(白鼠)에 미치는 영양학적(營養學的) 및 생물화학적(生物化學的) 연구(硏究))

  • Haw, Kum
    • Journal of Nutrition and Health
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    • v.1 no.1
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    • pp.9-18
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    • 1968
  • Dividing into six(6) experimental groups; Stock Diet(SD) Group, Rice Diet(RD) Group, Rice Diet supplemented with Barley(RD-BD) Group, Barley Diet(BD) Group, Rice Diet supplemented with Millet(RD-MD) Group, and Millet Diet(MD) Group, biochemical and nutritional effects of rice, barley, and millet on rats are studied. The grains used in this experiment are polished ones and purchased from market. Six(6) weaning male albino rats are housed into the individual cages in each group and fed with each experimental diet and water ad libitum for 21 weeks. The composition of the experimental diets are shown in Table 1. At the end of experiment, the appearences of rats are observed and the bloods are sampled by heart puncture method following ether anesthesia for determinations of serum protein and serum cholesterol. And the livers are sampled immediately right after heart after heart puncture for determinations of liver weight, liver nitrogen, liver fat, and liver glycogen. And also liver tissues are observed histologically. The result of growth experiment is shown in Table 2 and Figure 7. The results of blood analysis are shown in Table 3 and the results of liver analysis are shown in Table 4. And the histological observations of liver tissue are shown in Figures 1, 2, 3, 4, 5, and 6. According to the above results, it could be summarized as follows; 1) The normal appearences are observed at SD Group and BD Group. Alopecia is observed at RD, RD-­BD, RD-MD, and MD groups and the wet appearences of hair are observed at RD-MD and MD groups. The color of hair is changed to yellowish white at RD, RD-MD groups. The maximum growth inhibition, is observed at MD group and the growths of RD, RD-BD, BD, and RD-MD groups are significantly lower than that of SD group. The growth of RD group is lower than those of RD-BD and BD groups, though this difference is not significant. 2) At the blood analysis, the highest serum protein content is shown at SD group and the lowest serum protein is observed at MD group. The serum cholesterol contents are not significantly different among the six(6) experimental groups. 3) The highest liver weight is observed at SD group and the lowest weight is shown at MD group. However, oppositely the highest liver weight per 100 grams of body weight at MD group and the lowest liver weight per 100 grams of body weight at SD group are observed as shown in Table 6. 4) The differences of liver nitrogen contents of RD, RB-BD, BD, RD-MD, and MD Groups are not significant except SD group showing the highest content. 5) The significant highest content of liver fat is observed at RD group and the lowest content of liver fat is shown at SD group. And no significant differences in contents of liver glycogen are observed among the six(6) experimental groups. 6) No significant abnormal histological changes of liver tissue are observed at SD, RD-BD, and BD groups. However, the fat metamorphosis at peripheral zone of liver tissue is observed at RD Group and the some histological abnormal changes are observed at RD-MD and MD groups. According to the above summaries, the followings might be estimated and recommended; 1) The BD group shows histological normality of liver tissue, improvement of liver fat and liver nitrogen, normality of appearence, and better growth compared with RD Group. This could be indicated that barley is nutritionally and biochemically better than rice as the feeding diet on rat. And it might be considered that there might be the species speciality for nutritionary evaluation of food. 2) Recently, Marshall indicated that the high consumption of carbohydrate causes fatty liver and Kondo reported that the higher consumption of rice causes the more heart disease and the shorter life span. Considering these reports, the higher consumption of rice as a main dish in our country should be considered as an important national nutritional problem. And this might be improved by supplementing with barley or other grains to rice.

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