• Title/Summary/Keyword: intracellular fluid

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Anti-oxidant and Anti-aging Effect of Extracts from Seed of Camellia japonica L. by Supercritical Fluid Extraction (한국산 동백종자 초임계 추출물의 피부 항산화 및 항노화 연구)

  • Kim, Hee-Taek;Kim, Tae-Jun;Kim, Ee-Hwa;Kwak, Byeong-Mun
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.33 no.4
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    • pp.32-43
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    • 2020
  • Objectives : The aim of this study was to investigate the anti-oxidant and anti-aging effect of the seed of Camellia japonica L. extracted by supercritical CO2. Methods : The cell viability was performed by MTT assay. Nitric oxide (NO) production was performed by NO assay. The anti-oxidant effect was performed by beta-carotene bleaching assay and the intracellular proteome was analyzed expression of each 15 proteins by 2-D electrophoresis. Results : Fatty acid analysis of extract from Camellia japonica L. has shown oleic acid was 84%. And the antioxidant effect was about 186% compared with alpha-tocoperol(0.1%) by beta carotene bleaching assay. In 2D PAGE analysis, fifteen protein changes in five mechanisms which was collagen synthesis pathway, MMPs, ECM-cell interaction, cytokine, antioxidant enzymes were analyzed. Conclusions : On this study extract from seed of Camellia japonica L. by supercritical extraction could be used as a antioxidant and antiaging substance for the skin.

$Ca^{++}$ Polls in Isolated Rabbit and Turtle Heart (적출된 토끼와 자라심장에서의 $Ca^{++}$ Pool)

  • Kim, In-Kyo;Lee, Joong-Woo;Kang, Doo-Hee
    • The Korean Journal of Physiology
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    • v.9 no.1
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    • pp.13-22
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    • 1975
  • From the study of movements of $Ca^{++}$ in frog cardiac muscle, Niedergerke (1963) postulated that $Ca^{++}$ necessary for the cardiac contraction is stored in a specific pool. Langer et al (1967) and DeCaro (1967) also found a close relationship between the change of $Ca^{++}$ flux kinetics and the change of contractile force. According to the studies of several investigators, Ca II (Bailey and Dressel 1968) or phase I and II (Langer 1965, Langer et al 1967, 1971) in the $Ca^{++}$ washout curve was associated with cardiac contractility. This investigation was aimed to elucidate the anatomical region of the contractile active $Ca^{++}$ pool. At the same time, it was assumed in this study that $Ca^{++}$ in the sarcoplasmic reticulumn represents one of the major intracellular $Ca^{++}$ pool and cardiac contractility was also dependent on the intracellular $Ca^{++}$ concentration. Consequently, this experiment was performed at different temperatures to activate to activate inhibit the deactivating process of activated $Ca^{++}$ in the intracellular space to see if changes in the contractility decay curve existed at different temperatures. The isolated hearts of rabbits and turtles (Amyda maackii) were attached to the perfusion apparatus according to the method employed by Bailey and Dressel (1968). The isolated hearts were initally perfused with a full Ringer solution containing 2 mg/ml of inulin for 1 hr, and then $Ca^{++}$ and inulin-free Ringer solution was perfused while the isometric tension was recorded and a serial sample of perfusion fluid dripping from the cardiac apex was collected for 10 sec throughout experimental period. The above procedure was performed at $23^{\circ}C$, $30^{\circ}C$ and $38^{\circ}C$ on the rabbit heart and $10{\sim}13^{\circ}C$, $10^{\circ}C$, $25^{\circ}C$, $30^{\circ}C$ and $35^{\circ}C$ on the turtle heart. After determination of $Ca^{++}$ and inulin concentration of the samples, the $Ca^{++}$, inulin washout curve and the contractile tensin decay curve were analysed according to the method of Riggs (1963). The results were summarized as follows; 1. In the rabbit heart, there are 2 inulin compartments, 3 $Ca^{++}$ compartments and sing1e exponential decay of contractile tension. In the turtle heart, there are $1{\sim}2$ inulin compartments, $1{\sim}2$ $Ca^{++}$ compartments and $1{\sim}2$ phases of contractile tension decay. The fact that the inulin space was divided into 3 compartments in the washout curve in these hearts indicates the presence of heterogeneity in cardiac perfusion, i.e., overfused and underperfused area. 2. Ca I a9d Ca II in these hearts were found to have $Ca^{++}$ in the ECF compartments because their half times in the washout curves were far smaller than those of the inulin washout curves in the rabbit heart and similar to those of the inulin washout curves in the turtle heart. Ca III in the rabbit heart may have originated from the intracellular $Ca^{++}$ store. But no Ca III in the turtle heart was found. This may be due to the fact that the iutracellular $Ca^{++}$ pool in the turtle heart was too small to detect using this experimental procedure since sarcoplasmic reticulumn in the turtle heart is poorly developed. 3. In the rabbit heart, there were no chages in the half time of Ca I, Ca II, inulin I and inulin II at different temperatures, but the half time of Ca III was significantly prolonged at lower temperatures, and the half time of the contractile tension decay tended to be prolonged at lower temperatures but this was not significant. In the turtle heart, there were no changes in the half time of Ca I, Ca II, inulin 1, inulin II and phase I of the contractile tension decay at different temperatures, but the half time of phase II of the contractile tension decay was significantly prolonged at lower temperatures. This finding indicates that intracellu!ar $Ca^{++}$ in these hearts was also responsible particulary for maintaining the cardiac contractility at the lower temperatures. 4. The half times of contractile tension decay were shorter than those of Ca II in the $Ca^{++}$ washout curves in both animal hearts. According to the above results it was shown that $Ca^{++}$ in ECF is primarily and $Ca^{++}$ in the intracellular space is partially associated with the cardic contractility.

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An Electron Microscopic Study on the Effect of Calcium-free Reperfusion in Isolated Perfused Guinea Pig Heart after Global Ischemia (허혈후 칼슘 결핍 용액의 재관류가 적출 관류 기니픽 심근 세포에 미치는 영향에 관한 전자현미경적 관찰)

  • Oh, Seung-Hwan;Kim, Ho-Dirk;Rah, Bong-Jin
    • Applied Microscopy
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    • v.20 no.1
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    • pp.65-76
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    • 1990
  • The effect of calcium-free reperfusion for 5, 10, and 15 minutes, respectively, followed by continuous reperfusion with normal Tyrode solution containing 1.0mM calcium chloride, after global ischemia in the isolated perfused guinea pig heart by Langendorff techniques was examined with transmission electron microscope. Compared to the nomal Tyrode solution-perfused control hearts, the 5 minute calcium-free-reperfused hearts showed loss or thickening of Z lines, focal sarcolemmal disruption, mitochondrial swelling, clumping of chroma-tin, intracellular fluid accumulation, and some separation of cell junctions, especially the fasciae adherentes. These changes became more severe in the hearts of 10 minute calcium-free reperfusion. Subsarcolemmal larger bleb and near complete separation of cell junctions were noticed. In the 15 minute calcium-free-reperfused hearts, irreversible ultrastructural changes including contraction bands, biazrre mitochondria, and sarcolemmal destruction were widely distributed. The severity of myocardial changes were in accordance with the duration of calcium-free reperfusion. These changes indicate that calcium-free reperfusion regardless of its duration could not salvage the post-ischemic myocardium probably due to development of calcium paradox.

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INTRACELLULAR AMINO ACID PROFILE OF RUMEN BACTERIA AS INFLUENCED BY UREA FEEDING AND ITS DURATION

  • Kobayashi, Y.;Wakita, M.;Hoshino, S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.6 no.4
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    • pp.619-622
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    • 1993
  • Rumen bacterial amino acids in sheep on urea diet were monitored to assess a possible change in amino acid synthesis as a long term response to high rumen ammonia environment. A sheep was fed a semipurified diet with soybean meal, followed by a diet with urea as a main nitrogen source. Mixed rumen bacteria were harvested from ruminal fluid taken 3 h after feeding (twice in soybean meal feeding and 6 times in urea feeding) and fractionated as cell wall, proteins and protein-free cell supernatant of monitor amino acids in each fraction. Ruminal ammonia concentration at the sampling ranged from 5.7 to 39.5 mgN/dl. Cell wall and protein fractions of mixed rumen bacteria were stable in their amino acid composition regardless of nitrogen sources of diet and the feeding duration. However, protein-free cell supernatant fraction showed a higher alanine proportion with urea feeding (18.6 and 28.2 molar % of alanine for samples from sheep fed soybean meal and urea, respectively) and its duration (20.6 and 32.9 molar % for samples from sheep on urea diet for 1 and 65 days, respectively). Total free amino acid level of bacteria was depressed in the initial period of urea feeding but restored on 65th day of the feeding. These results suggest that an alanine synthesizing system may develop in rumen bacteria as urea feeding becomes longer.

Status of High Risk Group Fabry Disease Screening in Korea by Measuring Globotriacocylceramide in Body Fluid using Electrospray-MS/MS (탠덤매스에의한 체액 중 Globotriaocylceramide(Gb-3)의 측정을 이용한 한국인 고 위험도군에서의 파브리병 스크리닝)

  • Yoon, Hye-Ran
    • YAKHAK HOEJI
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    • v.55 no.1
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    • pp.56-63
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    • 2011
  • Fabry disease (FD) is an X-linked inborn error of glycoshpingolipid metabolism resulting from mutation in the enzyme ${\alpha}$-galactosidase A gene. The disease is an X-linked lipid storage disorder and the lack of ${\alpha}$-Gal A causes an intracellular accumulation of glycosphingolipids, mainly globotriaosylceramide (Gb-3). Measurement of Gb-3 in plasma has clinical importance for monitoring after enzyme replacement therapy for confirmed FD patients. Using electrospray ionization MS/MS we had developed, a simple, rapid, and highly sensitive analytical method for Gb-3 in plasma was used for the purpose of screening FD among high risk groups in Korean population. To date, no comprehensive results for FD screening have been performed and reported in Korea. We screened 1,100 outpatients from 13 hospitals (including clinics) to assess the incidence of FD among patients in high risk groups. For patients with borderline level amount of Gb-3, we repeated Gb-3 or performing complementary or confirmative assay with ${\alpha}$-Gal A activity and DNA mutaion analysis for confirmation diagnosis. Of 1,100 we diagnosed 3 FD with 2 classical type and 1 carrier (0.27%).

Pulmonary Function Test and Body Composition Analysis in Obese Children (비만 소아에서 폐기능 검사와 체성분 분석에 대한 연구)

  • Shin, Jee Seon;Park, Ji Hye;Kim, Ji Young;Kim, Su Jung;Hong, Young Mi
    • Clinical and Experimental Pediatrics
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    • v.48 no.6
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    • pp.588-593
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    • 2005
  • Purpose : Obesity is associated with disturbances of ventilatory functions in adults. But few studies have evaluated the pulmonary complications of obesity in the pediatric population. The purpose of this study is to clarify the effects of obesity on pulmonary function and body composition in obese children. Methods : Forty seven obese children whose ages ranged from nine to twelve years were evaluated for their body composition(intracellular fluid, extracellular fluid, protein mass, mineral mass, soft lean mass, fat mass, percent body fat, fat distribution) by bioelectrical impedance analysis. Hemoglobin, serum glucose, aspartate aminotransferase(AST), alanine aminotransferase(ALT), total cholesterol and triglycerides were measured. Pulmonary function test was performed by spirometer. Results : Intracellular fluid, protein mass, fat mass, percent body fat and fat distribution were significantly higher in severely obese children with an obesity index of more than 150 percent compared with those with an index of less than 150 percent. Peak expiratory flow rate(PEFR) was significantly lower in severely obese children with obesity index of more than 150 percent compared with those with less than 150 percent($241.7{\pm}14.6L/sec$ vs $276.8{\pm}64.3L/sec$). PEFR, forced expiratory flow 25 percent($FEF_{25}$), mid expiratory flow rate(MEFR), forced expiratory flow 50 percent($FEF_{50}$), forced expiratory volume in 1st second($FEV_1$) and forced vital capacity(FVC) were decreased in 37.0 percent, 14.8 percent, 14.8 percent, 11.1 percent, 3.7 percent and 3.7 percent of obese children, respectively. Conclusion : PEFR was significantly decreased in obese children. Pulmonary function test must be performed in severely obese children and more extended study is needed in other age groups.

Relationship between water and coffee intake on Health-related Physical Fitness and obesity of women's university student (여대생의 건강체력과 비만에 대한 물과 커피 섭취량과의 관계)

  • Lee, Lu-Ry;Ko, You-Sun
    • Journal of the Korean Applied Science and Technology
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    • v.37 no.4
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    • pp.649-658
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    • 2020
  • The purpose of this study is to verify the difference in health and obesity according to water and coffee intake. The study subject is 900 female college students. Physical fitness factors measured in this study were physical strength(grip strength, back muscle strength), muscle endurance (sit-up), Flexibility(Sit & reach) and cardiopulmonary endurance(harvard step). The factors of body composition (intracellular fluid, extracellular fluid, protein, mineral, body water content, muscle mass, lean body mass, body fat mass) and obesity (BMI, body fat percentage, abdominal fat percentage, obesity) were measured with the Inbody equipment. SPSS program was used for all data analysis. Group differences were verified by two-way ANOVA and one-way ANOVA. The results are as follows. There was no interaction effect between water and coffee intake on health and obesity. Therefore, the difference of the dependent variables according to each independent variable was verified. There was no significant difference in physical fitness according to water intake and coffee intake, but there was a significant difference in body composition (intracellular fluid, protein, minerals, body water, muscle mass and lean body mass). Water drinking group more than 7 cups had significantly higher body composition than the 1-4 cups water drinking group. In the case of coffee, the body composition of the 3 cup intake group was significantly higher than that of the none intake group. Obesity was significantly higher than the none intake and 1-2 cup intake groups. Based on the study results, water intake was not related to obesity, but coffee was related. For further studies, it would be better to consider gender and age differences in drinking various beverages such as water and coffee and that would be used to suggest a desirable way of water intake according to individual characteristics of the better health.

Effect of Equine Chorionic Gonadotropin and Porcine Follicle-Stimulating Hormone on Oocyte Maturation and Embryonic Development after Parthenogenesis and Nuclear Transfer in Pigs (체외성숙 배양액에 첨가된 eCG 및 돼지 FSH가 돼지 미성숙 난자의 체외성숙과 단위 발생 및 핵이식 난자의 체외발육에 미치는 영향)

  • You, Jin-Young;Jeong, Chan-Woo;Kim, Jin-Young;Lee, Eun-Song
    • Journal of Embryo Transfer
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    • v.24 no.3
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    • pp.213-220
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    • 2009
  • The objective of this study was to examine the effect of eCG and various concentrations (20, 40, and 80 ${\mu}g/ml$) of porcine FSH on nuclear maturation and intracellular glutathione (GSH) level of oocytes, and embryonic development after parthenogenetic activation (PA) and somatic cell nuclear transfer (SCNT) in pigs. Immature pig oocytes were matured in TCM-199 supplemented with porcine follicular fluid, cysteine, pyruvate, EGF, insulin, and hormones (10 IU/ml hCG and 10 IU/ml eCG or $20{\sim}80{\mu}g/ml$ FSH) for the first 22 h and then further cultured in hormone-tree medium for an additional 22 h. Nuclear maturation of oocytes ($85{\sim}89%$) was not influencem foreCG and various concentrations FSH. Embryonic development to the cleavage stage ($86{\sim}94%$) and mean number of cells in blastocyst ($33{\sim}37$ cells) after PA were not altered but blastocyst formation e-treignificaddlor(p<0.05) improvem forthe supplementation eith 80 ${\mu}g/ml$ FSHr(64%) compared to 47%, io8%, iand 47% in oocytes that were treated with eCG, 20,i and 40 ${\mu}g/ml$ FSH,i numectivelo. In SCNT, fusion ($78{\sim}83%$) of cell-cytoplast couplets and siosequent embryo cleavage ($82{\sim}88%$) were not influencem fordifferent gonadotropins but blastocyst formation tended to increase forthe supplementation eith 80 ${\mu}g/ml$ FSHr(25% vs. $11{\sim}18%$). Our nuults demonstrated that oocyte maturation and embryonic development after PA and SCNT e-frinfluencem fortype of gcem fortype of gits concentration. In this study, supplementation of maturation medium eith 80 ${\mu}g/ml$ FSHrimproved preimplantation development of PA and SCNT pig embryos, probably by increasing intracellular GSH concentration of matured oocytes.

Phosphorylation of AQP4 Water Channel Regulates Water Permeability (Aquaporin 4 water channel 인산화에 의한 수분 투과도의 조절)

  • 박권희;정동근;정진섭;이재숙;예운해;서덕준;배혜란
    • Journal of Life Science
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    • v.10 no.5
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    • pp.456-466
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    • 2000
  • Aquaperin 4 (AQP4) is the mercurial water channel expressed abundantly in brain, especially the region related with cerebrospinal fluid reabsorption and osmoregulation. The primary structure of AQP4 water channel was elucidated but the molecular mechanism of AQP4 channel regulation is still unknown. To investigate the possible regulation of AQP4 water channel by phosphorylation via various protein kinases, osmotic water permeability of AQP4 expressed in Xenopus oocytes was measured by videomicroscopy technique. Forskolin (10 $\mu$M) did not affect osmotic water permeability of oocytes injected with AQP4 cRNA, excluding the regulation of AQP4 water cnannel by protein kinase A. Osmotic water permeability (P아래첨자) of AQP4-expressed oocytes was ingibited by the pretreatmeat of BAPTA/AM (up to 500$\mu$M), an intracellular Ca윗첨자 chelator, and calmidazolium (100$\mu$M), a specific Ca윗첨자/calmodulin antagonist, in a dose-dependent manner. The inhibition of osmotic water permeability (P아래첨자) by the calmidazolium treatment was completely reversed by the addition of calyculin A (0.1$\mu$M), a nonspecific phosphatase inhibitor. Phorbol 12-myristate 13-acetate (PMA), a protein kinase C activator, had biphasic effects on osmotic water permeability in AQP4 cRNA injected oocytes depending on its concentration; 21% increase by 100 nM PMA, 35% decrease by 1$\mu$M PMA. These effects were reversed with 2$\mu$M staurosporine, a nonspecific PKC inhibitor. These results suggest that phosphorylation of AQP4 water channel by Ca윗첨자/calmodulin kinase and protein kinase C might regulate the osmotic water permeability.

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Intracellular Messenger Role of Cyclic Nucleotides in Exocrine Secretion of Guinea Pig Pancreas (취외분비에 미치는 cyclic nucleotides의 역할)

  • Lee, H.W.;Kim, W.J.;Hong, S.S.
    • The Korean Journal of Pharmacology
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    • v.13 no.2
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    • pp.41-48
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    • 1977
  • In 1968, Case et al. first studied the importance of cyclic AMP as an intermediate in the action of secretin and cholecystokinin-pancreozymin and they suggested that the action of secretin, not that of cholecystokinin-pancreozymin, may be mediated through cyclic AMP. Recently Albano et al. reported that in the exocrine pancreas each of the two major physiological functions is modulated a specific cyclic nucleotide, enzyme secretion by cyclic GMP, and fluid and ionic secretion by cyclic AMP. But in pancreas still conflicting results have been reported on the role of cyclic nucleotides in enzyme and electrolyte secretion. In these study, the role of cyclic nucleotides in the exocrine pancreatic secretion was examined. The results are as follows. 1) Very strong stimulation on amylase release from guinea pig pancreatic slice was produced by 1 unit of cholecystokinin-pancreozymin but as compared to that of cholecystokinin-pancreozymin very weak response was observed by 1 unit of secretion or $1\;{\mu}g$ of VIP. 2) Both cholecystokinin-pancreozymin and acetylcholine produced a rapid and marked rise in cyclic GMP as well as cyclic AMP in isolated pancreatic tissue. However, both secretin and VIP failed to alter significantly the basal level of cyclic GMP in pancreatic fragments. 3) Atropine inhibited acetylcholine mediated amylase release, but did not affect the cholecystokinin-pancreozymin response. Furthermore, atropine pretreatment produced a marked inhibitory effect on the increase of tissue cyclic nucleotides induced by cholecystokinin-pancreozymin and acetylcholine. In summary, these results suggest that whereas the pancreatic secretion produced by secretin and VIP is modulated by the formation of cyclic AMP, the pancreatic enzyme secretion in response to cholecystokinin-pancreozymin and acetylcholine is triggered by both cyclic AMP and cyclic GMP.

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