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      • SCIESCOPUSKCI등재

        급성 및 만성 영양장애가 소화효소 활성 및 세포기능에 미치는 실험적 연구

        김윤수,황석귀,김명원,유용운,송정석 ( Yoon Soo Kim,Suk Kuy Whang,Myung Won Kim,Yong Wun Ryu,Chung Suk Song ) 생화학분자생물학회 1974 BMB Reports Vol.7 No.1

        A variety of digestive Enzymes of pancreatic and intestinal origin participate in the breakdown of protein, carbohydrate and lipids. It has been known that some of enzyme output of the intestinal glands can be manipulated by altering components of the diet. The incidence of patients who are suffering from gastrointestinal trouble in Korea is much higher than that of the United States. This fact might correlate with diet composition of Korean and American dishes. These facts are attractive for us to study the effect of diet composition on the digestive enzymes such as amylase, lipase, trypsin and chymotrypsin, and to decide whether the difference of diet composition affects these enzymes and whether such high incidence of gastroin testinal patients in Korea could be adequately explained by change in digestive enzyme activities of the animals fed with different diet. Our previous experiments have shown that pancreatic enzyme activites of rats fed with 5% casein diet which was prepared similarly to Korean diet in terms of protein content only were depressed as much as one fifth of that of control during two weeks feeding period. but it is true that protein is not only depleted but fat is also depleted in Korean dishes. The primary objective of the experiments described herein was to study the effect on the pancreatic enzyme activities of diet which was similarly prepared to Korean dishes and also to study the effect on the pancreatic enzyme activities of varying the proportion of fat to carbohydrate in the diet which is keeping the diet isonitrogenous and isocaloric. Data from this experiment will be discussed ire conjunction with previously published results conducted in our laboratory in order to illustrate how the high incidence of gastrointestinal patients in Korea could be adequately explained by diet prepared in similar composition to the Korean dishes. In a random distribution, male albino rats, weighing the average of 80 gm each were divided into five groups. Group I ; 5% casein : no fat : carbohydrate Group II ; 5% casein : 12% fat : carbohydrate Group III ; 18% casein : no fat : carbohydrate Group IV ; 18% casein : 12% fat : carbohydrate Group V ; 18% casein : high fat : carbohydrate (ratio of fat to carbohydrate 94 : 6) Calorie difference from protein or fat depletion was substitiuted by carbohydrate. At the end of designated time interval, the rats were sacrificed 14-15 hours after the last meal by decapitation. The pancreas was carefully romoved and placed in a cold 0. 25M sucrose solution (4℃). A 5% homogenate in 0.25M sucrose was prepared from the glandular {free of obvious lymph nodes and adipose tissue) portion of the pancreas. 5:% Homogenates were centrifuged (14,000 x g) for 15 minutes to remove cellular debris and whole cells. Enzyme analysis and protein determination were subsequently performed on the supernatant. Protein was determined by Lawry method (1951). Enzyme activity assays were performed using modification of Hepler method (1962) for amylase and the method developed by Cherry and Crandall (1932) for lipase. Chymotryptic assays were performed using a modification of method developed by Rhodes et al. (1957), as a substrate for chymotrypsinogen N-acetyl-L-tyrosine ethyl ester was used. Trypsinogen assay was similarly performed using p-toluene-sulfonyl-L-arginine methyl ester as a substrate. The assays were linear with respect to enzyme concentration over the range used under specified conditions such as temperature (37℃) and incubation period, 30 minutes for amylase, 1 hour for lipase, and for the trypsinogen and chymotrypsinogen the recording of dye disappearing of optical density was recorded 2 to 3 seconds after adding substrate-buffer-indicator The results of the experiments in which the effect of different composition of nutrient which was prepared in similar composition to Korean dishes on pancreatic enzyme acitivities are summarized as the following. When the enzyme activity

      • SCIESCOPUSKCI등재

        Cycloheximide 가 취장 소화효소 활성에 미치는 영향

        김윤수,김명원,유용운 ( Yoon Soo Kim,Myung Won Kim,Yong Wun Ryu ) 생화학분자생물학회 1974 BMB Reports Vol.7 No.2

        These experiments were designed to assess the dietary-induced depletion and repletion of pancreatic enzymes mediating the adaptation of rat pancreas to the substitution of 5% casein diet for a normal diet of 18% casein and the substitution of 18% casein normal diet for a 5% casein diet which causes the pancreatic enzymes of chymotrypsinogen, trypsinogen, amylase and lipase to decrease remarkably during the 2∼4 weeks feeding period. Pancreatic chymotrypsinogen, trypsinogen and amylase of the rat`s (feeding) normal diet were severely depressed by one dose of intraperitoneal injection of cycloheximide (1㎍/gm of body weight) which is believed to be a specific inhibition of peptide bond formation. However, lipase was not affected at all by this treatment. The pancreatic enzymes of the rat fed with 5% casein diet for 4 weeks were depressed but when the 5% casein diet was substituted to a 18% casein diet the repletion of pancreatic enzymes was remakable. However, the repletion of chymotrypsinogen, trypsinogen and amylase was prevented or even more depressed when 1 ㎍/gm of body weight of cycloheximide was injected intraperitoneally, daily during the period of 18% normal casein diet feeding. The only exception was lipase which recovered to normal enzyme level in the same as noncyclohe ximide treated rats. These results indicate that the enzymatic repletion and depletion of the pancreatic enzymes in the diet are not similar to each other and are entirely different in the case of lipase, even though these pancreatic enzymes are found in identical tissues of the rat.

      • Cycloheximide가 취장 소화효소 활성에 미치는 영향

        김윤수,김명원,류용운,Kim, Yoon-Soo,Kim, Myung-Won,Ryu, Yong-Wun 생화학분자생물학회 1974 한국생화학회지 Vol.7 No.2

        쥐에서 취장소화효소 활성은 식이조성에 대단히 예민하게 반응한다. 즉 정상 18% casein 식이 대신 5% casein 식이로 대체시 또는 5% casein 식이로 2-3주 동안 기른 쥐를 정상 18% casein 식이로 대체시 취장소화효소에 미치는 영향은 대단히 민감하게 반응하여 그 효소활성이 떨어지거나 다시 회복하였다. 또한 정상식이로 기른 쥐에게 단백질합성 억제 물질인 cycloheximide($1\;{\mu}g/gm$/체중 gm)를 1회 복강내 투여로 취장소화효소 chymotrypsinogen, trypsinogen 및 amylase는 그 활성이 심하게 저하되나 lipase는 전혀 영향을 받지 않았다. 5% casein 식이에 의하여 저하된 취장효소는 정상식이로 대체시 그 효소활성 회복이 빠르다. 그러나 정상식이로 대체전에 cycloheximide($1\;{\mu}g/gm$/체중 gm)를 복강내 투여후 정상식이로 대체하면 chymotrypsinogen, trypsinogen 및 amylase는 그 회복이 전혀 되지 않거나 정상식이로 대체전보다 더 떨어졌다. lipase는 cycloheximide처치에 관계 없이 같은 시간내에 정상치로 회복하였다. 이와 같은 실험 결과는 이들 취장효소는 동일쥐의 취장세포에 존재하는 효소들이지만 cycloheximide에 의한 효소활성이 달리 반응하는 결과로 미루어 적어도 lipase 만은 그 생합성 과정이 다른 소화효소와 다름을 시사한다. These experiments were designed to assess the dietary-induced depletion and repletion of pancreatic enzymes mediating the adaptation of rat pancreas to the substitution of 5% casein diet for a normal diet of 18% casein and the substitution of 18% casein normal diet for a 5% casein diet which causes the pancreatic enzymes of chymotrypsinogen, trypsinogen, amylase and lipase to decrease remarkably during the 2~4 weeks feeding period. Pancreatic chymotrypsinogen, trypsinogen and amylase of the rat's (feeding) normal diet were severely depressed by one dose of intraperitoneal injection of cycloheximide ($1\;{\mu}g/gm$ of body weight) which is believed to be a specific inhibition of peptide bond formation. However, lipase was not affected at all by this treatment. The pancreatic enzymes of the rat fed with 5% casein diet for 4 weeks were depressed but when the 5% casein diet was substituted to a 18% casein diet the repletion of pancreatic enzymes was remakable. However, the repletion of chymotrypsinogen, trypsinogen and amylase was prevented or even more depressed when $1\;{\mu}g/gm$ of body weight of cycloheximide was injected intraperitoneally, daily during the period of 18% normal casein diet feeding. The only exception was lipase which recovered to normal enzyme level in the same as noncyclohe ximide treated rats. These results indicate that the enzymatic repletion and depletion of the pancreatic enzymes in the diet are not similar to each other and are entirely different in the case of lipase, even though these pancreatic enzymes are found in identical tissues of the rat.

      • 급성 및 만성 영양장애가 소화효소 활성 및 세포기능에 미치는 실험적 연구

        김윤수,황석귀,김명원,유용운,송정석,Kim, Yoon-Soo,Whang, Suk-Kuy,Kim, Myung-Won,Ryu, Yong-Wun,Song, Chung-Suk 생화학분자생물학회 1974 한국생화학회지 Vol.7 No.1

        저자들은 한국인 주식에 흡사한 각종 식이 조성을 달리한 인공 식이가 취장소화효소 활성에 어떻게 영향을 미치며 이들 식이 조성차이와 소화효소 활성과의 함수관계를 알아 보기 위하여 식이 조성을 달리한 각종 식이를 가지고 실험한바 다음과 같은 실험결과를 얻었다. 식이 I군(저단백, 무지방, 고함수탄소)과 식이 II군(저단백, 정상지방, 고함수탄소)으로, 4주 또는 6주동안 사육후 백쥐의 취장 소화 효소 활성을 측정하여 그 변화를 비교 관찰하였다. 정상식이(정상단백, 정상지방, 정상 함수탄소)로 해당 기간 동안 기른 쥐의 각 효소 활성치를 100%로 표시할 때 식이 I, 식이 II로 2주 동안 기른 쥐 취장 소화 효소 chymotrypsinogen은 정상치의 0~10% trypsinogen은 0~20% amylase는 10~15%, lipase 20~25%로 저하 하였다. 이와 같은 효소 활성치의 저하는 계속 4주 6주에도 유지 하였으며 식이 III군(정상단백, 무지방, 고함수탄소)에서는 정상군과 하등의 변화가 없이 소화효소 활성치를 유지하였다. 즉 소화효소 chymotrypsinogen, trypsinogen, amylase, lipase 이들 모든 효소 활성은 우리가 사용한 지방의 함량에 하등의 영향을 받지 아니 하였으며 식이중 단백질 함양유무(18%)가 효소활성 저하의 중요한 요인의 하나라는 사실을 실험적으로 입증하였다. 이와 같이 식이 조성 차이로서 저하된 각식이군의 효소 활성도는 4주, 6주에 정상식이로 대체 하였을 때 식이 I, II로 4주, 6주동안 사육한 기간에 상관없이 식이 대체 3일만에 chymotrypsinogen, trypsinogen, Iipase는 정상치로 회복하였으며 amylase는 4주에 대체한 것이 5일, 6주에 대체한 것이 7일을 요하였다. 즉 식이에 의한 amylase 효소 활성 회복 반응은 chymotrypsinogen, trypsinogen, lipase와 상이 하였다. 4, 6주후에 정상식이 대신 저단백(5%) 고지방(78%) 저함수탄소(17%)로 조성된 고 지방식이로 대체 하였을때 소화효소는 전혀 회복을 보이지 아니한 사실로 미루어 보아 소화효소 활성 회복에는 단백질함양이 절대필수요소 임을 실험적으로 입증하였으며 지방은 전혀 관계하지 않은 사실도 압증하였다. 정상단백질(18%)이 일정하고 고지방(78%) 저함수탄소(5%) 식이로 3주, 4주 동안 기른 쥐의 취장내 효소활성 chymotrypsinogen, trypsinogen, lipase는 하등의 영향을 받지 아니 하였으며 amylase 치만이 정상치 100% 보다 3주에 61.5%, 4주에 64%로 저하된 효소 활성치를 보였다. Amylase는 지방 함양보다 함수탄소양에 영향 받은 것 같다. 이상과 같은 우리의 실험적 사실은 취장 소화 효소 활성은 식이조성 차이로서 영향 받으나 지방 함양의 다소 유무에 하등의 영향을 받지 아니하고 오로지 단백질 함양 만이 그들효소의 활성 및 회복에 필수요소 임을 실험적으로 입중하였으며 amylase는 단백질 함양에도 영향 받으나 단백질 함양이 정상이고 함수탄소양을 줄인 고지방식이(함수탄소 : 지방 6 : 94)는 amylase 활성에 영향을 미친다는 사실도 구명하였다. the corresponding tissue fed with normal diet, the enzyme activities of chymotrypsinogen, trypsinogen, amylase and lipase were depressed by 0-10%, 0-20%, 10-15% and 20-25% respectively at the end of two weeks during the feeding of group I and group II and then these range of depletion of enzyme activities maintained continuously up to the end of tour and six weeks. This result suggests that digestive enzyme activities are influenced by only depletion of protein but not by various amount of fat in diet at all when carbohydrate is sufficient. Some of animals in groups I and II receiving designated diet for four and six weeks were transferred to the normal diet then pancreatic enzyme were assayed on the third, fifth and seventh days. The recovery of these enzymes is remarkable, and apparently the rats fed protein-and lipid-depleted diet were able to respond immediately to the stimulus of diet. Complete recovery of chymotrypsinogen, trypsinogen and lipase was observed after three days feeding regardless rats fed with protein- and lipid-depleted diet for four or six weeks except amylase which was recovered after five days feeding in the group of rats fed diet I and II for four weeks and seven days feeding in the group of rats fed diet I and II for six weeks. No difference in the recovery rate of enzyme was observed between diets I and II and also between the group of the rats when the rats in groups I and II shifted to the normal diet at four weeks and six weeks. Rats fed protein- and lipid-depleted for four and six weeks were not able to respond to the stimulus of diet, at all, which consists of high lipid. low protein, and low carbohydrate. This result illustrates that the essential factor for stimulus of enzyme activity is protein in diet but not carbohydrate and fat. The effect of high proportion of fat to carbohydrate (94: 6) in isonitrogenous, isocaloric diet on pancreatic enznyme activities also was studied in rats and compared to the effect of normal isocaloric diet. The diet which provided 57 cal/day and the experiment lasted for four weeks, no effect of this diet on pancreatic chymotrypsinogen, trypsinogen and lipase was observed. However, pancreatic amylase was reduced to 61.5% at the end of three weeks and 64% at the end of four weeks. This result indicates that pancreatic amylase activity may be related when carbohydrate is restricted but protein is normal in diet. These results indicate that when various dietary composition is related to the pancreatic enzyme activity, one of main influential factor in diet on pancreatic enzyme is protein, but not fat at all. However pancreatic amylase acivity may be related when carbohydrate is resticted but protein is normal in diet. Our experimental evidence tends to indicate that enzyme synthesis in the pancreas is regulated, at least in part by the composition of amino acid pool(s) in the acinar cells of pancreas. This may be due to the decreased amount of amino acid in the dietary protein, but not composition of fat. It can be postulated that the decrease in the dietary protein affects synthesis and secretion of pancreatic digestive enzymes, and then the intracellular structure of the cell, resulting in decrease of cell activity and tissue debility.

      • Flayin Analogue가 백서 간장내 포도당 및 지방 대사에 미치는 영향

        김원용,황석귀,김윤수,Kim, Won-Yong,Whang, Suk-Kuy,Kim, Yoon-Soo 생화학분자생물학회 1978 한국생화학회지 Vol.11 No.1

        Lambooy (1961) has shown that, while several analogues of riboflavin are able to replace the vitamins in the metabolism of some microorganism, 7-ethyl-8-methyl flavin is able to serve as the sale flavin in the metabolism of the rat. This flavin is able to serve as a completely adequate replacement for riboflavin in the metabolism of Wistar (Lambooy, 1961) and Sprague-Dawley rats (Yoon S. Kim et al, 1966) in terms of growth, survival, optimal physical appearance, and efficient utilization of food. Administration of 7-ethyl-8-methyl flavin to weanling rats as their only source of flavin permits a normal growth with a concomitant rapid and extensive loss of succinic dehydrogenase activity (SDH) from the liver, heart and kidney to within 24, 48, and 67% of the normal level, respectively, on day 42 following initiation of a 7-ethyl-8-methyl flavin diet. This result was essentially identical with that previously reported (Y.S. Kim and Lambooy, 1967). The purpose of the experiments described here is to study whether the decreased SDH activity in the tissue of rats is related to the metabolism of glucose and de novo biosynthesis of lipid from glucose in the liver of rats when D-glucose-$^{14}C$(U) is administered intraperitoneally to the rat maintaining SDH activity from liver, heart and kidney at 24, 48 and 67% of the normal level. This is also to study whether the total triglyceride and the total lipid content in the liver of rats is influenced by the loss of SDH activity in the liver of rats which have received 7-ethyl-8-methyl flavin. The present study shows that there was no significant difference in the total amount of $^{14}CO_2$ expired from D-glucose-$^{14}C$(U) through the respiration of rats which had received 7-ethyl-8-methyl flavin compared to control rats which received riboflavin, indicating that the matabolic rate of glucose in the tissues of rat receiving 7-ethyl-8-methyl flavin is similar to that of the control rats. However, compared to the control level only 50% of $^{14}C$ total lipids were incorporated by de novo biosynthesis from D-glucose-$^{14}C$(U) in the liver of rats receiving 7-ethyl-8-methyl flavin but the total content of triglyceride and total lipids in the liver of rats receiving 7-ethyl-8-methyl flavin were increased by 29 and 30% over the normal level. This may be due to the increased of lipid transport from the peripheral adipose tissue due to the decreased SDH activity in the liver of rats. 7-Ethyl-8-methyl flavin이 SDH 효소 활성에 미치는 영향에 대하여 Kim and Lambooy (1967)에 의해 보고된 바 있다. 즉 이 flavin 투여후 30일에 쥐 간장에서 SDH 효소 활성은 정상군의 해당조직 효소 활성치를 100%로 할때 23%로 떨어져 100일 까지 계속 20% 내외로 유지하여, 심장에서는 투여 30일에 64% 50일에 43% 100일에서 31% 내외였고, 신장에서는 30일에 72% 50일에 68% 100일에 56% 내외를 유지한다고 보고 하였다. 저자는 본 실험을 시작하기전에 Kim and Lambooy (1967)방법에 따라 쥐에 이 flavin을 투여한 후 42일에 SDH 효소 활성을 측정한바 간장에서 24% 심장에서 48% 신장에서 76%로 떨어졌으나, 그 쥐의 성장, 체중의 증가, 식이 섭취량 및 외모등이 riboflavin 투여군과 동일한 사실을 재확인 하였으며 이런 SDH 효소 활성을 유지하는 쥐에다 D-glucose $^{14}C$ (U)를 마리당 $55{\times}10^5$ dpm 씩 복강내 주입하고 이 쥐를 metabolic cage에 넣어 호흡을 통하여 나오는 총 $^{14}CO_2$의 배설량을 methylcellosolve:ethanolamine 용액 (2:1)에 포착시켜 정량한바 riboflavin 투여군에서는 평균 $257192{\pm}8630\;cpm$/3hrs/gm of body wt. 7-ethhyl-8-methyl flavin 군은 평균 $255426{\pm}1160\;cpm$/3hrs/gm of body wt. 로 두군의 총 $^{14}CO_2$ 배설량은 서로 같았다. 7-Ethyl-8-methyl flavin에 의하여 SDH 활성이 떨어졌음에도 총 $^{14}CO_2$ 배설량에 하등의 변동이 없는 이 두군에서 성장 및 체중증가, 식이 섭취량, 외모 등의 동일성으로 미루어보아 체내에 전체적으로 영향을 주는 대사장애는 없는 것으로 사려되나, 투여한 D-glucose $^{14}C$ (U)로부터 간장내 지방 de novo 생합성은 평균 $13080{\pm}420$ cpm per gm에서 $6595{\pm}400$ cpm per gm 으로 7-ethyl-8-methyl flavin 투여군에서 50%나 현저히 감소하였다. 간장지방내 $^{14}C$ 는 모두투여한 $^{14}C$ (U)로부터 합성된 것임으로 이 결과는 간장 조직내에서 SD H 활성 저하가 지방 de novo 생합성과 밀접한 관계가 있음을 시사해준다. 즉 SD H 활성이 떨어지면 Krebs cycle 저하를 초래하고 이것은 glucose로 부터 지방의 de novo 생합성을 떨어뜨리는 결과로 사려된다. 간장내 총 triglyceride 함량은 정상군에서 평균 $1.93{\pm}0.083gm%$이나 7-ethyl-8-methyl flavin 투여군에서는 $2.49{\pm}0.085gm%$로 29%가 증가하였으며 간장내 총지방함량 역시 정상군에서 평균 $4.53{\pm}0.299gm%$인데 비해 7-ethyl-8-methyl flavin 투여군에서는 $5.90{\pm}0.157gm%$로 정상군보다 30% 증가하였다. 이상 결과로 미루어 보아 7-ethyl-8-methyl flavin 투여로 간장에서 SDH 합성이 24%로 떨어지면 간장내 지방의 de novo 생합성은 50% 감소되나 총 triglyceride 함량은 정상군보다 29%, 총지방함량은 30% 증가하였다. 이는 아마도 체내 SDH 활성저하가 말초 지방조직으로부터 간장조직으로 지방산의 이동을 증가시키는 결과로 사려된다.

      • 비단열 모세관의 형상 변경에 따른 소형 냉동 사이클 성능 해석

        남기원(Ki-Won Nam),이대용(Dae Yong Yi),박상구(Sang-Goo Park),정지환(Ji Hawn Jeong),김윤수(Lyun-Su Kim) 대한설비공학회 2009 대한설비공학회 학술발표대회논문집 Vol.2009 No.-

        The present simulation focuses on the effect of the shape of capillary tube-suction line heat exchangers(SLHX), The results in the three cases of the SLHX types show that both of the location and length of heat exchange section influence the coefficient of performance(COP) and cooling capacity. Simulation shows the COP may be improved by 4.6% and the cooling capacity may be improved by 13.6% in the Lateral type.

      • SCIESCOPUSKCI등재

        Flavin Analogue 가 백서 간장내 포도당 및 지방 대사에 미치는 영향

        김원용,황석귀,김윤수 ( Won Yong Kim,Suk Kuy Whang,Yoon Soo Kim ) 생화학분자생물학회 1978 BMB Reports Vol.11 No.1

        Lambooy (1961) has shown that, while several analogues of riboflavin are able to replace the vitamins in the metabolism of some microorganism, 7-ethyl-8-methyl flavin is able to serve as the sole flavin in the metabolism of the rat. This flavin is able to serve as a completely adequate replacement for riboflavin in the metabolism of Wistar (Lambooy, 1961) and Sprague-Dawley rats (Yoon S. Kim et al, 1966) in terms of growth, survival, optimal physical appearance, and efficient utilization of food. Administration of 7-ethyl-8-methyl flavin to weanling rats as their only source of flavin permits a normal growth with a concomitant rapid and extensive loss of succinic dehydrogenase activity (SDH) from the liver, heart and kidney to within 24, 48, and 67% of the normal level, respectively, on day 42 following initiation of a 7-ethyl-8-methyl flavin diet. This result was essentially identical with that previously reported (Y.S. Kim and Lambooy, 1967). The purpose of the experiments described here is to study whether the decreased SDH activity in the tissue of rats is related to the metabolism of glucose and de novo biosynthesis of lipid from glucose in the liver of rats when D-glucose-^(14)C(U) is administered intraperitoneally to the rat maintaining SDH activity from liver, heart and kidney at 24, 48 and 67% of the normal level. This is also to study whether the total triglyceride and the total lipid content in the liver of rats is influenced by the loss of SDH activity in the liver of rats which have received 7-ethyl8-methyl flavin. The present study shows that there wa.s no significant difference in the total amount of ^(14)CO₂ expired from D-glucose-^(14)C(U) through the respiration of rats which had received 7-ethyl-8-methyl flavin compared to control rats which received riboflavin, indicating that the metabolic rate of glucose in the tissues of rat receiving 7-ethyl-8-methyl flavin is similar to that of the control rats. However, compared to the control level only 50% of ^(14)C total lipids were incorporated by de novo biosynthesis from D-glucose-^(14)C(U) in the liver of rats receiving 7-ethyl-8-methyl flavin but the total content of triglyceride and total lipids in the liver of rats receiving 7-ethyl-8-methyl flavin were increased by 29 and 30% over the normal level. This may be due to the increased of lipid transport from the peripheral adipose tissue due to the decreased SDH activity in the liver of rats.

      • SCIESCOPUSKCI등재

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