• The Korean Journal of Physiology and Pharmacology
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• v.4 no.4
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• pp.311-318
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• 2000

We cultured the rabbit inner medullary collecting duct (IMCD) cells as monolayers on collagen-coated membrane filters, and investigated distribution of the P2Y receptors by analyzing nucleotide-induced short circuit current $(I_{sc})$ responses. Exposure to different nucleotides of either the apical or basolateral surface of cell monolayers stimulated $I_{sc}.$ Dose-response relationship and cross-desensitization studies suggested that at least 3 distinct P2Y receptors are expressed asymmetrically on the apical and basolateral membranes. A $P2Y_2-like$ receptor, which responds to UTP and ATP, is expressed on both the apical and basolateral membranes. In addition, a uracil nucleotide receptor, which responds to UDP and UTP, but not ATP, is expressed predominantly on the apical membrane. In contrast, a $P2Y_1-like$ receptor, which responds to ADP and 2-methylthio-ATP, is expressed predominantly on the basolateral membrane. These nucleotides stimulated intracellular cAMP production with an asymmetrical profile, which was comparable to that in the stimulation of $I_{sc}.$ Our results suggest that the adenine and uracil nucleotides can interact with different P2Y nucleotide receptors that are expressed asymmetrically on the apical and basolateral membranes of the rabbit IMCD cells, and that both cAMP- and $Ca^{2+}-dependent$ signaling mechanisms underlie the stimulation of $I_{sc}$.

• The Korean Journal of Zoology
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• v.35 no.3
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• pp.263-270
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• 1992

Responses of immunoreactive atrial natriuretic peptide (ir-ANP) to environmental temperature were studied in the freshwater catfish, Silums usotus, acclimated at various temperatures for one week. According to increase in environmental temperature, plasma sodium and chloride concentrations, and osmolality were significantly increased, while hematocrit showed a marked decrease. When the catfishes frere acclimated at cold $(3^{\circ}C)$ or warm $(18^{\circ}C)$ temperatures, ir-ANP contents in the atrium and bulbus arteriosus were significantly lower than those at $9^{\circ}C.$ However, ventricular contents of ir-ANP urere not different among the three groups.Levels of plasma ir-ANP in the catfish increased in response to the increment of acclimated temperature. On the basis of these results, we suggest that the environmental temperature may modulate synthesis and secretion of ir-hUP in the fish. We also suggest that changes of plasma ir-ANP levels may be associated with the control of body fluid homeostasis.

• Journal of Embryo Transfer
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• v.19 no.2
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• pp.101-112
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• 2004

Chloride($Cl^-$) channels play critical roles in cell homeostasis and its specific functions such as volume regulation, differentiation, secretion, and membrane stabilization. The presence of these channels have been reported in all kinds of cells and even in frog oocytes. These essential role of $Cl^-$­ channels in cell homeostasis possibly play any role in egg homeostasis and in the early stage of development, however, there has been no report about the presence of $Cl^-$­ channel in the mammalian oocyte. This study was performed to elucidate the presence of $Cl^-$­ channels in hamster eggs. When allowing only $Cl^-$­ to pass through the channel of the egg membrane by using impermeant cation such as N-methyl-D-glucamine(NMDG), single channel currents were recorded. These channel currents showed typical long-lasted openings interrupted by rapid flickering. Mean open $time({\tau}o)$ was 43${\pm}$10.14 ms(n=9, at 50 mV). The open probability(Po) was decrease with depolarization. The current-voltage relation showed outward rectification. Outward slop conductance(32${\pm}$5.4 pS, n=22) was steeper than the inward slop conductance(10${\pm}$1.3 pS). Under the condition of symmetrical 140 mM NaCl, single channel currents were reversed at 0 mV(n=4). This reversal potential(Erev) was shifted from 0 mV at 140 mM concentration of internal NaCl(140 mM [Na+]i) to ­9.8${\pm}$0.5 mV(n=4) at 70 mM [Na+]i and 11.5${\pm}$1.9 mV at 280 mM [Na+]i(n=4) respectively, strongly suggesting that these are single $Cl^-$­ channel currents. To examine further whether this channel has pharmacological property of the $Cl^-$­ channel, specific Cl­ channel blockers, IAA-94(Indanyloxyacetic acid-94) and DIDS(4, 4'-diisothiocyan ostillben- 2-2'disulfonic acid) were applied. IAA-94 inhibited the channel current in a dose-dependent manner and revealed a rapid and flickering block. From these electrophysiological and pharmacological resluts, we found the novel $Cl^-$­ channel present in the hamster oocyte membrane. The first identification of $Cl^-$­ channel in the hamster oocyte may give a clue for the further study on the function of $Cl^-$­ channel in the fertilization and cell differentiation.

• The Korean Journal of Physiology and Pharmacology
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• v.24 no.4
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• pp.329-338
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• 2020

Rhinorrhea in allergic rhinitis (AR) is characterized by the secretion of electrolytes in the nasal discharge. The secretion of Cl^- and HCO₃^- is mainly regulated by cystic fibrosis transmembrane conductance regulator (CFTR) or via the calcium-activated Cl^- channel anoctamin-1 (ANO1) in nasal gland serous cells. Interleukin-4 (IL-4), which is crucial in the development of allergic inflammation, increases the expression and activity of ANO1 by stimulating histamine receptors. In this study, we investigated ANO1 as a potential therapeutic target for rhinorrhea in AR using an ANO1 inhibitor derived from a natural herb. Ethanolic extracts (30%) of Spirodela polyrhiza (SP_EtOH) and its five major flavonoids constituents were prepared. To elucidate whether the activity of human ANO1 (hANO1) was modulated by SP_EtOH and its chemical constituents, a patch clamp experiment was performed in hANO1-HEK293T cells. Luteolin, one of the major chemical constituents in SP_EtOH, significantly inhibited hANO1 activity in hANO1-HEK293T cells. Further, SP_EtOH and luteolin specifically inhibited the calcium-activated chloride current, but not CFTR current in human airway epithelial Calu-3 cells. Calu-3 cells were cultured to confluency on transwell inserts in the presence of IL-4 to measure the electrolyte transport by Ussing chamber. Luteolin also significantly inhibited the ATP-induced increase in electrolyte transport, which was increased in IL-4 sensitized Calu-3 cells. Our findings indicate that SP_EtOH and luteolin may be suitable candidates for the prevention and treatment of allergic rhinitis. SP_EtOH^- and luteolin-mediated ANO1 regulation provides a basis for the development of novel approaches for the treatment of allergic rhinitis-induced rhinorrhea.

• Journal of Life Science
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• v.19 no.6
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• pp.744-750
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• 2009

A bacterial strain producing highly viscous extracellular polysaccharide was isolated from soil. Through morphological, physiological and chemotaxonomical studies, it was identified as a Bacillus sp. and named Bacillus sp. PS-12. The extracellular polysaccharide, named PS-12 was purified by ethanol precipitation, cetylpyridinium chloride (CPC) precipitation and gel permeation chromatography. The purified polysaccharide was found to consist of glucose, mannose, galactose, and fucose, with a molar ratio of approximately 7:3.2:2:1, respectively. PS-12 was investigated for its immunostimulating activity on murine macrophage RAW264.7 cells using an ELISA assay. PS-12 stimulated the production of TNF-${\alpha}$ to a level 50 times greater than the control and also induced 1L-6 secretion in a dose-dependent manner. The cytotoxicity on RAW264.7 cells by PS-12 was relatively low with 10% cytotoxicity at 2 ${\mu}g$/ml. These results indicate that PS-12 is less cytotoxic to immune cells and possess immunomodulating activity in which it can produce cytokines including TNF-${\alpha}$ and 1L-6 from macrophages.

• The Journal of Pediatrics of Korean Medicine
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• v.12 no.1
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• pp.183-210
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• 1998

Cheonggisan(CGS) is well known for its effect on such allergic disease as urticaria and atopic dermatitis. Gagamcheonggisan(GCGS) was formulated by subtracting several herbs from CGS and adding several herbs to CGS. Even though it is being used frequently in the clinicai medicine for the treatment of above hypersensitivity diseases, basic study to make sure the mechanism of its action is rare. In this study the author tried to know the effect of CGS and GCGS on the vascular permeability, contact dermatitis, granular secretion from mast cells and function of macrophages. The results obtained in this study are as follows : 1. Administration of CGS and GCGS decreased the vascular permeability induced by serotonin and histamine. The decrease by serotonin is more typical and dose-dependent. 2. Administration of CGS and GCGS inhibited foot-pad and ear swelling responses induced by sheep red blood cells and picryl chloride respectively, the inhibition of foot-pad swelling responses is bigger than that of ear swelling responses and both of them are not dependent on the dose3. Treatment of peritoneal mast cells with CGS and GCGS water extract decreased the histamine release triggered by compound 48／80 in a dose dependent fashion 4. Administration of CGS and GCGS increased the phagocvtic activity of peritoneal macrophages and treatment of peritoneal macrophages with CGS activated phagocytic function in a dose dependent fashion. 5. Administration of CGS and GCGS enhanced such reactive oxygen intermediates(ROIs) as superoxide and hydrogen peroxide production from peritoneal macrophages. 6. Treatment of CGS and GCGS activated peritoneal macrophages for the production of ROIs. The above results show that CGS and GCGS decreased the hypersensitivity reactions by inhibiting non-specific inflammatory mediator release and vascular permeability without affecting general immune responsiveness.

• CELLMED
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• v.4 no.2
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• pp.12.1-12.12
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• 2014

Epinephrine is a critical drug for patients at risk for anaphylaxis. Here, we suggest moxibustion as an alternative method to reduce anaphylaxis. Moxibustion was applied to the Shimen (CV5) acupoint and found to attenuate compound 48/80-induced mortality. Capsazepine, a transient receptor potential vanilloid (TRPV) 1 antagonist, significantly improved overall survival rates compared to groups treated with moxibustion or 2-aminoethoxydiphenyl borate (an activator of TRPV1, 2, and 3). Probenecid (a TRPV2 agonist) also increased survival rate and reduced histamine levels. Survival rates increased by moxibustion and probenecid were completely inhibited by ruthenium red (a TRPV2 and 3 antagonist) and gadolinium chloride (general TRPV antagonist), respectively. Passive cutaneous anaphylaxis and ear swelling were significantly reduced by moxibustion and probenecid (p < 0.05). In cardiomyocytes, TRPV2 was over-expressed by compound 48/80 and histamine but this increased TRPV2 expression decreased to baseline with moxibustion and probenecid treatment. In addition, intracellular calcium levels increased by compound 48/80 were reduced by probenecid. Overall, these findings suggest that the reduction of anaphylaxis caused by moxibustion could represent a new mechanism of moxibustion related to the regulation of TRPV2 activation and promotion of epinephrine secretion.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.5
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• pp.795-801
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• 2007

Patients with Type I diabetes mellitus have been treated with porcine insulin for several decades and pigs have recently been deemed an ideal source of microencapsulated islet cells for clinical xenotransplantation. In this study, neonatal pigs were anesthetized and sacrificed prior to a pancreatectomy. Islet cells were isolated from pancreas via collagenase digestion. Islet cells were separated and collected by hand under microscopic guidance. These cells were suspended in 1.4% sodium alginate solution and encapsulated by dropping them into 1.1% calcium chloride solution and in which the round gel in size was 250-400 ${\mu}m$ in diameter. Viability of the microencapsulated islet cells cultured in medium at $37^{\circ}C$ was assessed by MTT assay. Furthermore, insulin released in response to glucose challenge was investigated using an enzyme-linked immunosorbent assay. Secretion of insulin was low in response to the basal glucose solution (4.4 mM) in medium and was significantly higher in response to the high glucose solution (16.7 mM). The viability of microencapsulated islet cells did not differ significantly over a period of 7 days; that is, the increasing pattern of insulin concentration in the culture medium after glucose stimulation interval day was similar throughout the 7 days cultivation. In summary, experimental evidences indicated that the effects of alginate-microencapsulation prolonged survival of the neonatal porcine islets in vitro cultures and the insulin response to glucose of the islets was maintained.

• The Korean Journal of Physiology
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• v.20 no.1
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• pp.103-124
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• 1986

Since it has been suggested that atrial receptor may be involved in the mechanism of extracellular volume regulation, it was shown that the granularity of atrial cardiocytes can be changed by water and salt depletion, and that an extract of cardiac atrial tissue, when injected intravenously into anesthetized rats, was shown to cause a large and rapid increase in renal excretion of sodium. Various natriuretic peptides were isolated and synthetized, and the effects were investigated by many workers. Most studies, however, have been carried out under anesthesia and there have teen some controversies over direct effect of the factor on the renal function. Therefore, it was attempted in this study to access the effects of an atrial extract and a synthetic natriuretic factor in unanesthetized rabbits. Intrarenal arterial infusion of atrial extract caused a rapid increase of urinary volume and excretion of sodium. Glomerular filtration rate and renal plasma flow were both increased with no change in filtration fraction. The ventricular extract produced no change in urinary excretion of electrolytes, nor in renal hemodynamics. Intrarenal infusion of synthetic atrial natriuretic factor caused increases of renal excretory rate of sodium, chloride and potassium, and $FE_{Na}$. Glomerular filtration rate, renal plasma flow increased. And free water clearance also increased. Accentuated excretory function correlated well with increased glomerular filtration rate and renal plasma flow during infusion and for 10 minutes following the cessation of the infusion. Renin secretion rate decreased during constant infusion of atrial natriuretic factor. However, no correlation was found with the changes in glomerular filtration rate, renal plasma flow, or urinary excretion of sodium. These results suggest that atrial extract or atrial natriuretic factor induces changes in renal hemodynamics, as in excretion of electrolytes either indirectly through hemodynamic changes or directly by inhibiting tubular reabsorption. At the same time, renin secretory function is affected by the factor possibly through an unknown mechanism.

• International Journal of Oral Biology
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• v.43 no.4
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• pp.217-222
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• 2018

Phagocytosis is a fundamental process in which phagocytes capture and ingest foreign particles including pathogenic bacteria. Several oral pathogens have anti-phagocytic strategies, which allow them to escape from and survive in phagocytes. Impaired bacteria phagocytosis increases inflammation and contributes to inflammatory diseases. The purpose of this study is to investigate the influences of various agents on oral pathogenic phagocytosis. To determine phagocytosis, Streptococcus mutans, Fusobacterium nucleatum, Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis were stained with 5-(and-6)-carboxyfluorescein diacetate succinimidyl ester (CFSE), and was measured using flowcytometery and confocal microscopy. The influencing factors on phagocytosis were evaluated through the pretreatment of ROS inhibitor (N-acetyl-L-cysteine (NAC)), lysozyme, potassium chloride (KCI) and adenosine triphosphate (ATP) in THP-1 cells. Expression of pro-inflammatory cytokines was determined by enzyme-linked immunosorbent assay (ELISA). The phagocytosis of various bacteria increased in a MOI-dependent manner. Among the tested bacteria, phagocytosis of P. gingivalis showed the highest fluorescent intensity at same infection time. Among the tested inhibitors, the NAC treatment significantly inhibited phagocytosis in all tested bacteria. In addition, NAC treatment indicated a similar pattern under the confocal microscopy. Moreover, NAC treatment significantly increased the bacteria-induced secretion of $IL-1{\beta}$ among the tested inhibitors. Taken together, we conclude that the phagocytosis occurs differently depending on each bacterium. Down-regulation by ROS production inhibited phagocytosis and lead increased of oral pathogens-associated inflammation.