• The Korean Journal of Physiology and Pharmacology
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• 제2권2호
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• pp.241-249
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• 1998

Platelet-activating factor(PAF) enhanced interleukin-1(IL-1) activity by the interaction with a specific receptor in rat alveolar macrophages. In this study, we investigated the role of endogenous arachidonate metabolites and intracellular calcium mobilization in the PAF-induced IL-1 activity. Alveolar macrophages were preincubated with 5-lipoxygenase and cyclooxygenase inhibitors 30 min before the addition of PAF and lipopolysaccharide(LPS). After 24h culture, IL-1 activity was measured in the supernate of sample using the thymocyte proliferation assay. Inhibition of 5-lipoxygenase by nordihydroguaiaretic acid and AA-861 completely blocked the PAF-induced enhancement of IL-1 activity with $IC_{50}\;of\;2\;{\mu}M\;and\;5\;{\mu}M$, respectively. In contrast, the inhibition of cyclooxygenase pathway by indomethacin and ibuprofen resulted in the potentiation in PAF-induced IL-1 activity with maximal effect at $1\;{\mu}M\;and\;5\;{\mu}M$, respectively. In addition, leukotriene $B_4$ and prostaglandin $E_2$ production were observed in PAF-stimulated alveolar macrophage culture. As could be expected, 5-lipoxygenase and cyclooxygenase inhibitors abolished PAF- stimulated leukotriene $B_4$ and prostaglandin $E_2$ production, respectively. The effects of PAF on intracellular calcium mobilization in alveolar macrophages were evaluated using the calcium-sensitive dye fura-2 at the single cell level. PAF at any dose between $10^{-16}\;and\;10^{-8}$ M did not increase intracellular calcium. Furthermore, there was no effective change of intracellular calcium level when PAF was added to alveolar macrophages in the presence of LPS or LPS+LTB4, and 4, 24 and 48h after treatment of these stimulants. Together, the results indicate that IL-1 activity induced by PAF is differently regulated through subsequent induction of endogenous 5-lipoxygenase and cyclooxygenase pathways, but not dependent on calcium signalling pathway.

• The Korean Journal of Physiology
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• 제28권2호
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• pp.225-231
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• 1994

The present study was undertaken to investigate the role of endogenous nitric oxide in renin release under different physiological conditions. In the first series of experiments, renin release was either inhibited by acute volume-expansion (VE) or stimulated by clipping one renal artery in the rat. VE was induced by intravenous infusion of saline (0.9% NaCl) up to 5% of the body weight over 45 min under thiopental (50 mg/kg, IP) anesthesia. VE caused a decrease of plasma renin concentration (PRC). With $N^G-nitro-L-arginine$ methyl ester $(L-NAME,\;5\;{\mu}g/kg\;per\;min)$ superadded to VE, PRC decreased further. The magnitude of increase in plasma atrial natriuretic peptide levels following VE was not affected by the L-NAME. In two-kidney, one clip rats, L-NAME-supplementation resulted in a decrease, and L-arginine-supplementation an increase of PRC. Plasma atrial natriuretic peptide levels were significantly lower in the L-arginine group than in the control. Blood pressure did not differ among the L-NAME, L-arginine, and control groups. In another series of experiments, the renin response to a blockade of NO synthesis was examined using in vitro preparations from isolated renal cortex. L-NAME significantly increased basal renin release, although it was without effect on the isoproterenol-stimulated release. These findings suggest that endogenous nitric oxide significantly contributes to the renin release. Since many factors may affect the renin release in vivo, an interaction between NO and renin under various pathophysiological states is to be further defined.

• The Korean Journal of Physiology
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• 제28권2호
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• pp.197-202
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• 1994

The present study was aimed to determine if endogenous L-arginine-nitric oxide (NO) pathway has central, rather than peripheral, mechanisms in blood pressure regulation. Arterial blood pressure and heart rate responses to acute inhibition of the t-arginine-NO pathway were examined in rats anesthetized with thiopental (50 mg/kg, IP). An intracerebroventricular (ICV) cannula was placed in the left lateral ventricle. The right femoral artery was cannulated to measure arterial blood pressure and the vein to serve as an infusion route. $N^G-nitro-L-arginine$ methyl ester (L-NAME) was infused either intracerebroventricularly or intravenously. ICV infusion $(1.25\;{\mu}L/min)$ of L-NAME $(20\;or\;100\;{\mu}g/kg)$ per minute for 60 min) increased the mean arterial pressure and heart rate. Plasma renin concentrations(PRC) were significantly lower in L-NAME-infused group than in the control. L-Arginine $(60\;{\mu}g/min,\;ICV)$ prevented the pressor response to ICV L-NAME. The pressor response was not affected by simultaneous intravenous infusion of saralasin, but was abolished by hexamethonium treatment. Intravenous infusion $(40\;{\mu}L/min,\;10{\sim}100\;{\mu}g/kg\;per\;minute\;for\;60\;min)$ also increased blood pressure, while it decreased heart rate. These results indicate that endogenous L-arginine-NO pathway has separate central and peripheral mechanisms in regulating the cardiovascular function. The central effect may not be mediated via activation of renin-angiotensin system, but via, at least in part, activation of the sympathetic outflow.

• 대한의생명과학회지
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• 제12권1호
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• pp.17-22
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• 2006

The baculovirus expression system is a powerful method for producing large amounts of the human erythrocyte-type glucose transport protein, heterologously. Characterization of the expressed protein is expected to show its ability to transport sugars directly. To achieve this, it is a prerequisite to know the properties of the endogenous sugar transport system in Spodoptera frugiperda Clone 21 (Sf21) cells, which are commonly employed as a host permissive cell line to support the baculovirus replication. The Sf21 cells can grow well on TC-100 medium that contains 0.1% D-glucose as the major carbon source, strongly suggesting the presence of endogenous glucose transport system. However, unlike the human glucose transport protein that has a broad substrate and inhibitor specificity, very little is known about the nature of the endogenous sugar transport system in Sf21 cells. In order to characterize further the inhibitor recognition properties of the Sf21 cell transporter, the ability of phloretin, cytochalasin B and D-fructose to inhibit 2-deoxy-D-glucose (2dGlc) transport was examined by measuring inhibition constants $(K_i)$. The $K_i's$ for reversible inhibitors were determined from plots of uptake versus inhibitor concentration. The 2dGlc transport in the Sf21 cells was very potently inhibited by phloretin, the aglucone of phlorizin with a $K_i$ similar to the value of about $2{\mu}M$ reported for inhibition of glucose transport in human erythrocytes. However, the Sf21 cell transport system was found to differ from the human transport protein in being much less sensitive to inhibition by cytochalasin B (apparent $K_i$ approximately $10\;{\mu}M$). In contrast, It is reported that the inhibitor binds the human erythrocyte counterpart with a $K_d$ of approximately $0.12\;{\mu}M$. Interestingly, the Sf21 glucose transport system also appeared to have high affinity for D-fructose with a $K_i$ of approximately 5mM, contrasting the reported $K_m$ of the human erythrocyte transport protein for the ketose of 1.5M.

• Environmental Analysis Health and Toxicology
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• 제22권2호통권57호
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• pp.147-156
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• 2007

Six week-old ICR mice were divided into four groups including NC, RC, RR and RS, then they were injected with sun ginseng (RS), red ginseng (RR) and saline (RC) intraperitoneally as an amount of 60mg/kg body weight at 1 hour, 12 hours and 36 hours before the whole body irradiation with 6.5Gy of X-ray. And the mice were sacrificed at nine days after the irradiation. The RS group were significant increase in the weight of spleen (p<0.05) and the numbers of endogenous spleen colony (p<0.001) and WBC (p<0.05) especially neutrophils, lymphocytes and monocytes in comparison with the RC group. The RR group were significant increase in the numbers of endogenous spleen colony (p<0.01) and WBC (p<0.05) especially neutrophils, lymphocytes and eosinophils in comparison with the RC group. The RS group exhibited a more increase in the weight of spleen and the number of endogenous spleen colony than the RR group. The values of ALT (alanine transaminase) and AST (aspartate transaminase) were significantly elevated (p<0.05) by radiation and they were significantly decreased (p<0.05) in the RS group to the values of the NC group. The value of TBARS(thiobarbituric acid reactive substance) was significantly elevated (p<0.01) by radiation and it was significantly decreased (p<0.05) in the RS group. And the values of SOD (superoxide dismutase) and CAT (catalase) were significantly increased (p<0.05) in the RS group in comparison with the RC group. Taken together the above results, sun ginseng demonstrated the protective effects on hepatocytes and immune and hematopoietic cells in mice exposed to medium dose of irradiation, and those radioprotective effects were a little higher or similar to the red ginseng.

• The Korean Journal of Physiology and Pharmacology
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• 제21권3호
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• pp.317-325
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• 2017

Non-alcoholic fatty liver disease (NAFLD) has become the most prevalent liver disease in parallel with worldwide epidemic of obesity. Reactive oxygen species (ROS) contributes to the development and progression of NAFLD. Peroxisomes play an important role in fatty acid oxidation and ROS homeostasis, and catalase is an antioxidant exclusively expressed in peroxisome. The present study examined the role of endogenous catalase in early stage of NAFLD. 8-week-old male catalase knock-out (CKO) and age-matched C57BL/6J wild type (WT) mice were fed either a normal diet (ND: 18% of total calories from fat) or a high fat diet (HFD: 60% of total calories from fat) for 2 weeks. CKO mice gained body weight faster than WT mice at early period of HFD feeding. Plasma triglyceride and ALT, fasting plasma insulin, as well as liver lipid accumulation, inflammation (F4/80 staining), and oxidative stress (8-oxo-dG staining and nitrotyrosine level) were significantly increased in CKO but not in WT mice at 2 weeks of HFD feeding. While phosphorylation of Akt (Ser473) and $PGC1{\alpha}$ mRNA expression were decreased in both CKO and WT mice at HFD feeding, $GSK3{\beta}$ phosphorylation and Cox4-il mRNA expression in the liver were decreased only in CKO-HF mice. Taken together, the present data demonstrated that endogenous catalase exerted beneficial effects in protecting liver injury including lipid accumulation and inflammation through maintaining liver redox balance from the early stage of HFD-induced metabolic stress.

• 한국조경학회지
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• 제18권4호
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• pp.127-140
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• 1991

The Primary purpose of this study is to investigate factors and variables which have significant effects on user satisfaction with recreational facilities in Taejong-Dae recreational complex, thereby establishing indices of planning and development of urban parks and open space. To test the causal models of this research, the date were gathered by self-administered questionnaires from 967 households in Pusan City which were selected by the multi-stage probability sampling methood. The analysis of the multi-stage primarily consists of two phase : The first analysis dealt exploratory factor analysis which identified major factors involved in satisfaction with recreational activities and facilities in Taejong-Dae recreational complex and the second analysis tested the fit of the causal models of this research by employing LISREL methodology. There are three advantages of using LISREL over other multivariate analysis methods : First, measurement error is allowed and calculated in LISREL, otherwise there is a risk of seriously misleading estimates of coefficients ; Second, LISREL deals with latent variables or unmeasured variables ; Third, it enables to test causal relations among variables. The factors analysis identified that five factors are involved in satisfaction with recreational facilities. The five factors of satisfaction with recreational facilities are space for repose and relaxation, active recreation facilities such as pool and zoo, physical exercise facility, convenience and maintenance facility, and linear facility, and linear facility for walking. The second phase analysis tested the fit of the causal models for satisfaction with recreational facilities to the data and identified statistically significant causal linkage among overall satisfaction with Taejong-Dae recreational complex, other endogenous factors and exogenous variables. Overall fits of both causal models were very good. Among endogenous factors, facility for repose and relaxation. linear facility for walking, active recreation facility, facility for convenience and maintenance were identified as having significant effects on overall satisfaction. Exogenous variables which have significant effects on endogenous variables wer also identified. These significant relationships indicate important factors and variables that should be considered in planning and development of the recreational complex. On the basis of these significant causal relationships, implications for planning and the delovepment of Taejong-Dae recreational complex were suggested.

• 한국동물학회지
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• 제7권2호
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• pp.25-36
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• 1964

The present paper deals with the comparative study on phylogenic difference in the patterns of energy metabolism of brain slices of several vertebrate species by measuring oxygen consumptionwith glucose-6-phosphate, glucose-1-phosphate, glyceraldehyde-3-phosphate or glutamate as respiratory substrate employing Warburg's manometric method, by determination of the utilization rate of glucose using glucose-1-C14 by analyzing patterns of free amino acid distribution , and by histochemical determination using glucose-1-C14 by analyzing patterns of free amino acid distribution acid distribution , and by histochemical determination of glycogen contents. 1. Glucose enhances the oxygen consumption of brain slices of animals belinging to reptile, aves and mammalia while it shows a tendency to decrease that of animals belonging to pisces and amphibia. 2. Glucose-6--phosphate increase oxygen consumption more than glucose in every species examined, while glucose-1-phosphate and glyceraldehyde-3-phosphate increase that of Rana nigromaculata only . In general m, it appears that phosphosugars are more effective as a respiratory substrate to those species which have less endogenous respiration than to those having larger endogenous respiration. 3. Similar patterns of free amino acid distribution and the relative amount are found among the species and in every species examined glutamic acid is detected in the larges amount . ${\gamma}$-Amino butyric acid, glycine, alanine and aspartic acid are found in every species. 4. Ophicephalus showed less oxygen consumption than endogenous respiration when glutamate was added to the medium. When sodium fluoride was added, the oxygen consumption was some what increased . Such phenomenon wasnot found in the frog. 5. The result of histochemcial analysis of the brain showed that glycogen was abundantly present in the fish , amphibia , and especially in the reptile and that no distinctive grains of glycogen were found in the bird and mammal . From these facts, it may be supposed that anaerobic glycolysis as energy source dominates in fish and amphibia and aerobic respiration through the oxidation of glucose dominates in bird and mamal , the reptile occupying transitional position between these two categories. The way of obtaining energy for brain activity by the oxidation of glucose supplied from the circulating blood is seemed to be first acquired by reptile and the function is completed both in aves and mammal.

• Journal of Nutrition and Health
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• 제30권4호
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• pp.406-414
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• 1997

It has been known that dietary phytate decreases the absorption of body zinc pool which is composed of the dietary and endogenous zinc in the body. The purpose of this study was to examine the effect of phytate on the absorption of total bodyzinc in Zn-depleted rats. Rats were Zn-depleted with either low(0.8%) or high(1.6%) Ca diet containing sodium phytate for 4 weeks. After zinc depletion, rats were assigned into phytate or non-phytate dietary groups within each low-or high-Ca dietary group. ant feces were collected for 2 weeks of the initial collection and 1 week after dietary crossover, during which the phytate and the non-phytate diet was switched over within the same Ca group. The content of Zn and Ca measured by atomic absorption spectrophotometer and phytate content was analyzed. food intake was higher in the high Ca group than in the low Ca group(p <0.0001), and was also higher in the non-phytate group than in the phytate group(p <0.0001). Food intake and phytate level affected body weight gain in rats(p <0.0001). Zinc excretion in the total feces was higher in the phytate group than in the non-phytate group at both low and high Ca level(p <0.0001), except during the crossover collection period in high Ca group. Calcium, however, didn't show any synergistic effect on phytate effect(p <0.05). This study showed that phytate decreased the absorption of total body zinc at both low and high Ca levels in Zn-depleted rats. A large portion of total body zinc originated from the endogenous zinc pool in these rats. The results of the present study showed the same effect of phytate on the endogenous zinc in Zn-depleted rats as in a previous study, confirming that phytate adversely affects zinc bioavailability, especially under marginal and poor zinc nutrition.

• Biomolecules & Therapeutics
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• 제25권4호
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• pp.396-403
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• 2017

Under normal, non-stressed conditions, intracellular p53 is continually ubiquitinated by MDM2 and targeted for degradation. However, in response to severe genotoxic stress, p53 protein levels are markedly increased and apoptotic cell death is triggered. Inhibiting the ubiquitination of p53 under conditions where DNA damage has occurred is therefore crucial for preventing the development of cancer, because if cells with severely damaged genomes are not removed from the population, uncontrolled growth can result. However, questions remain about the cellular mechanisms underlying the regulation of p53 stability. In this study, we show that p53-inducible gene 3 (PIG3), which is a transcriptional target of p53, regulates p53 stability. Overexpression of PIG3 stabilized both endogenous and transfected wild-type p53, whereas a knockdown of PIG3 lead to a reduction in both endogenous and UV-induced p53 levels in p53-proficient human cancer cells. Using both in vivo and in vitro ubiquitination assays, we found that PIG3 suppressed both ubiquitination- and MDM2-dependent proteasomal degradation of p53. Notably, we demonstrate that PIG3 interacts directly with MDM2 and promoted MDM2 ubiquitination. Moreover, elimination of endogenous PIG3 in p53-proficient HCT116 cells decreased p53 phosphorylation in response to UV irradiation. These results suggest an important role for PIG3 in regulating intracellular p53 levels through the inhibition of p53 ubiquitination.