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Studies on the Determination Method of Natural Sweeteners in Foods - Licorice Extract and Erythritol (식품 중 감초추출물 및 에리스리톨 분석법에 관한 연구)

  • Hong Ki-Hyoung;Lee Tal-Soo;Jang Yaung-Mi;Park Sung-Kwan;Park Sung-Kug;Kwon Yong-Kwan;Jang Sun-Yaung;Han Ynun-Jeong;Won Hye-Jin;Hwang Hye-Shin;Kim Byung-Sub;Kim Eun-Jung;Kim Myung-Chul
    • Journal of Food Hygiene and Safety
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    • v.20 no.4
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    • pp.258-266
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    • 2005
  • Licorice Extract and Erythritol, food additives used in korea, are widely used in foods as sweetener. Its application for use in food is regulated by the standard and specification for food additives but official analytical method far determination of these sweetener in food has not been established. Accordingly, we has been carried out to set up analytical method of the glycyrrhizic acid in several foods by the way of thin layer chromatography and high performance liquid chromatography glycyrrhizic acid is qualitative anaylsis technique consists of clean-up with a sep-pak $C_{18}$ cartridge, separation of the sweeteners by Silica gel 60 F254 TLC plate using 1-butanol:4Nammonia solution:ethanol (50:20:10) as mobile solvent. Also, the quantitative analysis for glycyrrhizic acid, was performed using Capcell prk $C_{18}$ column at wavelength 254nm and DW:Acetonitrile (62:38 (pH2.5)) as mobile phase. and we has been carried out to set up analytical method of the erythritol in several foods by the way of high performance liquid chromatography. erythritol is qualitative anaylsis technique consists of clean-up with a DW and hexane. The quantitative analysis for erythritol, was performed using Asahipak NH2P-50 column, Rl and DW:Acetonitrile (25:75) as mobile phase. The glycyrrhizic acid results determined as glycyrrhizic acid in 105 items were as follows; N.D$\∼$48.7ppm for 18 items in soy sauce, N.D$\∼$5.3ppm for 12 items in sauce, N.D$\∼$988.93ppm for 15 items in health food, N.D$\∼$180.7ppm for 26 items in beverages, N.D$\∼$2.6ppm for 8 items in alcoholic beverages repectively and ND for 63 items in the ethers. The erythritol results determined as erythritol in 52 items were as follows; N.D$\∼$155.6ppm for 13 items in gm, N.D$\∼$398.1ppm for 12 items in health foods repectively and ND for 45 items in the others.

Effect of Calvarial Cell Inoculated Onto the Biodegradable Barrier Membrane on the Bone Regeneration (흡수성 차폐막에 접목된 두개관골세포의 골조직 재생에 미치는 영향)

  • Yu, Bu-Young;Lee, Man-Sup;Kwon, Young-Hyuk;Park, Joon-Bong;Herr, Yeek
    • Journal of Periodontal and Implant Science
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    • v.29 no.3
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    • pp.483-509
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    • 1999
  • Biodegradable barrier membrane has been demonstrated to have guided bone regeneration capacity on the animal study. The purpose of this study is to evaluate the effects of cultured calvarial cell inoculated on the biodegradable barrier membrane for the regeneration of the artificial bone defect. In this experiment 35 Sprague-Dawley male rats(mean BW 150gm) were used. 30 rats were divided into 3 groups. In group I, defects were covered periosteum without membrane. In group II, defects were repaired using biodegradable barrier membrane. In group III, the defects were repaired using biodegradable barrier membrane seeded with cultured calvarial cell. Every surgical procedure were performed under the general anesthesia by using with intravenous injection of Pentobarbital sodium(30mg/Kg). After anesthesia, 5 rats were sacrificed by decapitation to obtain the calvaria for bone cell culture. Calvarial cells were cultured with Dulbecco's Modified Essential Medium contained with 10% Fetal Bovine Serum under the conventional conditions. The number of cell inoculated on the membrane were $1{\times}10^6$ Cells/ml. The membrane were inserted on the artificial bone defect after 3 days of culture. A single 3-mm diameter full-thickness artificial calvarial defect was made in each animal by using with bone trephine drill. After the every surgical intervention of animal, all of the animals were sacrificed at 1, 2, 3 weeks after surgery by using of perfusion technique. For obtaining histological section, tissues were fixed in 2.5% Glutaraldehyde (0.1M cacodylate buffer, pH 7.2) and Karnovsky's fixative solution, and decalcified with 0.1M disodium ethylene diaminetetraacetate for 3 weeks. Tissue embeding was performed in paraffin and cut parallel to the surface of calvaria. Section in 7${\mu}m$ thickness of tissue was done and stained with Hematoxylin-Eosin. All the specimens were observed under the light microscopy. The following results were obtained. 1 . During the whole period of experiment, fibrous connective tissue was revealed at 1week after surgery which meant rapid soft tissue recovery. The healing rate of defected area into new bone formation of the test group was observed more rapid tendency than other two groups. 2 . The sequence of healing rate of bone defected area was as follows ; test group, positive control, negative control group. 3 . During the experiment, an osteoclastic cell around preexisted bone was not found. New bone formation was originated from the periphery of the remaing bone wall, and gradually extended into central portion of the bone defect. 4 . The biodegradable barrier membrane was observed favorable biocompatibility during this experimental period without any other noticeable foreign body reaction. And mineralization in the newly formed osteoid tissue revealed relatively more rapid than other group since early stage of the healing process. Conclusively, the cultured bone cell inoculated onto the biodegradable barrier membrane may have an important role of regeneration of artificial bone defects of alveolar bone. This study thus demonstrates a tissue-engineering the approach to the repair of bone defects, which may have clinical applications in clinical fields of the dentistry including periodontics.

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Studies on the Factors Enhancing the Effects of Nitrogen Application of Rice Culture in Korea (수도작(水稻作)에서 시용질소효과 증대요인의 해석적(解析的) 연구)

  • Huh, Beom-Lyang
    • Korean Journal of Soil Science and Fertilizer
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    • v.16 no.2
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    • pp.131-155
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    • 1983
  • Though it has been widely known the nitrogen effects are influenced by soils, varieties, and mineral nutrients in the rice culture, few analyses in relation to the factors increasing nitrogen effect have been studied in Korea. The effects of potassium and silica on the factors increasing nitrogen effects in paddy soils were investigated in accordance with soil improvement practices and nitrogen application methods for the cultivated varieties. The results obtained are as follows. 1. For 413 paddy fields, the yield from soils without nitrogen application ranged from 200 to 850kg/10a and that from nitrogen application did 350 to 1,051kg/10a. The yield increament by nitrogen application varied 50 to 650kg/10a depending on soils. 2. Soil chemical characteristics for high yield were different between with nitrogen and without nitrogen application. In the without nitrogen application, however, contents of organic matter, phosphorous, potassium and calcium of high yield soils were lower than those of low yield, while the available silica content was higher in the former. 3. The yield increased with nitrogen application up to 22.4kg/10a and thereafter it decreased. These phenomena were supposed to be not be decrease of nitrogen uptake but by lowered silica uptake. 4. Clay soil incorporation, deep plough, and inorganic constituents control such as Ca, Mg, and $Sio_2$ were effective as soil improvement praitices. It was appeared that increases of silica content and Ca/Mg ratio were important to increase nitrogen effects. 5. For the correlation between yield and yield components, it was high between yield and panicle in low nitrogen level and so was it between grain yield and ripening rate in high nitrogen. 6. In the urea and super granule urea application plot, recovery rate of nitrogen by plant and soil was high and yield was remarkable high. 7. Regardless of fertilizer types such as ammonium sulfate and urea, the residual nitrogen was about 4kg/10a in both plots of 5.8 and 11.6kg/10a. N applied. 8. The potassium application to soil enhanced the nitrogen efficiency. It was more effective in low potassium soil. 9. Optimum pH value for gel formation in the 4% sodium silicate solution was approximately 6.6. 10. It was suggested that silica could affect to rice plant growth as the inorganic and organic chemical components.

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Development and Expression of Porcine Embryos by Direct Injection of Sperm Treated with Exogenous DNA (외래유전자 도입정자를 이용한 돼지 체외성숙 난포란의 Intracytoplasmic Sperm Injection (ICSI) 후 후기 배로의 발달율과 외래유전자의 발현에 관한 연구)

  • 정기화;조성근
    • Journal of Embryo Transfer
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    • v.17 no.1
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    • pp.45-53
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    • 2002
  • The main goal of this study was to produce transgenic porcine embryos by direct injection of sperm-mediated exogenous DNA. Spermatozoa (6$\times$10$^{6}$ sperms of final concentration) were mixed with pcDNA LAC Z (20 ng/$\mu$l) and subjected into electroporation (300~750 volts, 25 $\mu$F, 0.4 cm electrode). After sperm injection, the oocytes were activated electrically (1.7 KV/cm, 30$\mu$sec, single pulse) in 0.3 M mannitol solution or not. The sperm injected eggs were cultured in NCSU 23 medium (0.4% BSA) at 39$^{\circ}C$, 5% $CO_2$ in air fur 144 h. The rates of cleavage and development into blastocyst stage in activation group were significantly higher than those of non-activation group (79.6% and 24.1% vs. 46.3% and 14.4%, respectively, p<0.05). Control oocytes and shame injection were developed to blastocysts low (2.5%). Sixty five (27.1%) out of 240 embryos observed in activation and non-activation groups were showed positive by X-gal staining. However, all embryos in both groups were expressed partial or mosaic pattern. These results suggested that electrical stimulation far oocytes activation after sperm injection enhances the incidence of both fertilization and development fellowing sperm injection in the pig. Our study also suggested that sperm-mediated transfer of exogenous DNA by ICSI would be used as a valuable tool for the production of transgenic porcine embryos.

The Effect of Lime and Wollastonite on an Acid Sulfate Soil (특이산성토에 대한 석회 및 규회석의 효과)

  • Park, Nae Joung;Park, Young Sun;Lee, Kyu Ha;Kim, Yung Sup
    • Korean Journal of Soil Science and Fertilizer
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    • v.5 no.1
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    • pp.25-32
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    • 1972
  • The effects of limestone and wollastonite on an acid sulfate soil were studied. In addition, the effect of wollastonite was analyzed in terms of those due to calcium and to silica in the paddy field and in the laboratory with equivalent amounts of lime and wollastonite on a calcium basis. 1. Lime and wollastonite as liming materials were equally effective in neutralizing the soil acidity. 2. Lime, however was more reactive, raising the pH up to neutralization point in three days under waterlogged conditions at $25^{\circ}C$, in the lab study, and introduced alkali damage to transplanted rice seedlings showing marked restrictions of tillering in the field even though lime was applied two weeks before transplanting. On the other hand, wollastonite reacted very slowly, taking one week to two weeks to reach neutralization even when thoroughly mixed, and did not restrict the tillering. 3. Both lime and wollastonite effectively reduced the toxic aluminium in soil as well as in the soil solution but not always in the case of ferrous iron. However the reduction effect of the toxic substances in the experimental field was not so great as expected, because typical toxic symptoms were mild only. 4. Lime considerably increased the availability of silica in soil resulting in an increase of silica content in straw. Wollastonite released extra available silica itself resulting in a greater uptake of silica. 5. Increase of silica uptake by these materials was effective in reducing rate of infection of neck blast and resulted in higher rate of ripening, and in turn increased the paddy yield. 6. Application of either one significantly diminished the effect of the other. 7. Wollastonite markedly increased tillering in the early growing stage, but decreased the rate of effective tillers finally, resulting in about the same number of ears per hill at harvesting. 8. These liming materials appear to increase the number of grains per panicle slightly.

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Availability of Heavy Metals in Soils with Different Characteristics and Controversial Points for Analytical Methods of Soil Contamination in Korea (토양특성별 중금속 유효도와 토양오염 평가방법의 개선점)

  • Jung, Goo-Bok;Kim, Won-Il;Lee, Jong-Sik;Shin, Joung-Du;Kim, Jin-Ho;Yun, Sun-Gang
    • Korean Journal of Environmental Agriculture
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    • v.24 no.2
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    • pp.106-116
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    • 2005
  • This experiment was conducted to investigate available extraction capacity and potential mobility of heavy metal according to the distribution property and contamination level of heavy metals in soils and to suggest a reform measure of soil environment assessment methodology applied with soil quality and the official soil heavy metal test methods in domestic and foreign countries. The soils were collected from the natural forest paddy with long-term application of same type fertilizer, and paddies near metal mine and industrial complex. The post-treatment methods of soil were partial extraction, acid digestion and sequential extraction methods. For the heavy metal contents with different soil properties, it was shown that their natural forest and paddy soil were slightly low and similar to the general paddy soil, while their paddies near metal mine and industrial complex were higher than the standard level of Soil Environment Protection Act. Heavy metal concentrations in the soils with different soil properties had difference between $HNO_3\;and\;HNO_3+HCl$ extractant by US-EPA 3051a method. There were highly significant positive relationships in both two methods. It was appeared that the higher extractable concentration ratio with 0.1N-HCl to total heavy metal content with $HNO_3+HCl$ extractant the greater total heavy metal content. There were highly significant positive correlationship between total heavy metal content and extractable content with 0.1N-HCl. For extractable capacity of soil extractable solution compared to the total heavy metal content it was appeared that it extractable method with 0.1N-HCl was higher than those with EDTA and DTPA. In extractable ratio with 0.1N-HCl in the contaminated paddy soils near mine and industrial complex, it was shown that the lower soil pH, the higher total heavy metal content. The order of a potential mobility coefficient by distribution of heavy metal content with ie different typies in the soil was Cd>Ni>Zn>Cu>Pb. It could be known that contamination characteristics of heavy metals with different types of soils were affected by different heavy metal components, contamination degree and soil chemical properties, and heavy metal concentration with different extractable methods had great variations with adjacent environment. To be compared with assessment methodology of soil environment impact at domestic and foreign countries with our results, it might be considered that there was necessary to make a single analysis method based on total heavy metal content with environmental overloading concept because of various analysis methods for total heavy metal content and present analysis method with great variation according to soil environment. In spite of showing higher concentration of heavy metal with acidic digestion than the extractable method, it might be considered that there is need to be adjusted the national standard of soil heavy metal contamination.

Bioequivalence of Burophil Capsule to Surfolase Capsule (Acebrophylline 100 mg) (설포라제 캡슐(아세브로필린 100 mg)에 대한 부로필 캡슐의 생물학적 동등성)

  • Cho, Hea-Young;Park, Eun-Ja;Kang, Hyun-Ah;Kim, Se-Mi;Park, Chan-Ho;Oh, In-Joon;Lim, Dong-Koo;Lee, Myung-Hee;Lee, Yong-Bok
    • Journal of Pharmaceutical Investigation
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    • v.35 no.3
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    • pp.179-185
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    • 2005
  • Acebrophylline is a compound produced by salifying ambroxol with theophylline-7 -acetic acid. After acebrophylline administration, the salt splits into these two components which feature a peculiar pharmacokinetic behavior, an adequate ambroxol and a low theophylline-7-acetic acid serum levels. The purpose of the present study was to evaluate the bioequivalence of two acebrophylline capsules, Surfolase (Hyundai Pharm. lnd. Co., Ltd.) and Burophil (Kuhnil Pharm. Co., Ltd.), according to the guidelines of the Korea Food and Drug Administration (KFDA). The release of ambroxol from the two acebrophylline formulations in vitro was tested using KP VIII Apparatus II method with various dissolution media (pH 1.2, 4.0, 6.8 buffer solution and water). Twenty eight healthy male subjects, $23.25{\pm}1.43$ years in age and $64.82{\pm}6.77$ kg in body weight, were divided into two groups and a randomized $2{\times}2$ cross-over study was employed. After two capsules containing 100 mg as acebrophylline were orally administered, blood was taken at predetermined time intervals and the concentrations of ambroxol in serum were determined using HPLC with electrochemical detector (ECD). The dissolution profiles of two formulations were similar at all dissolution media. In addition, the pharmacokinetic parameters such as $AUC_t$, $C_{max}$ and $T_{max}$ were calculated and ANOVA test was utilized for the statistical analysis of the parameters using logarithmically transformed $AUC_t$, $C_{max}$ and untransformed $T_{max}$. The results showed that the differences between two formulations based on the reference drug Surfolase, were -1.64, -3.33 and -0.92% for $AUC_t$, $C_{max}$ and $T_{max}$, respectively. There were no sequence effects between two formulations in these parameters. The 90% confidence intervals using logarithmically transformed data were within the acceptance range of log 0.8 to log 1.25 $(e.g., \;log\;0.93{\sim}log\;1.05\;and\;log\;0.88{\sim}log\;1.05$ for $AUC_t$, and $C_{max}$, respectively). Thus, the criteria of the KFDA bioequivalence guideline were satisfied, indicating Burophil capsule was bioequivalent to Surfolase capsule.

Biokinetics of Carbohydrate and Lipid Metabolism in Normal Laying Hen [Part 1] -Determination of Turnover of Glucose- (정상산란계(正常産卵鷄)에 있어서 탄수화물(炭水化物)과 지질대사(脂質代謝)의 생동역학(生動力學) 제1보[第一報] -포도당 대사회전(代謝回轉)의 측정(測定)-)

  • Chiang, Y.H.;Riis, P.M.
    • Applied Biological Chemistry
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    • v.20 no.2
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    • pp.205-209
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    • 1977
  • The pool size of plasma glucose, turnover rate and other concerned items for glucose metabolism in normal laying hen were investigated by a single-injection method using $U-C^{14}-glucose$. The 11.6 nCi of pure dose was injected to a hen normally fed through the wing vein. The glucose concentration in plasma sample taken at 5 minutes after injections was 214mgper 100ml. From the plottings of logarithmic standard specific activities of plasma taken from 5 to 120 minutes against the time after injection and from the regresion analysis, metabolic states were determined. The pool size was 1.07g, turnover rate was 0.024 per minute, turnover time was 41 minutes, utilization rate was 26mg/min. (0.83 g/hr/kg B.W. 3/4) and glucose space(extracellular fluid volume) was 25.3 per cent of body weight. The values obtained from. 10-50 minutes samples were similar to those described above, which we from 5-120 minutes samples.

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Enzymatic Hydrolysis of Yellowfin Sole Skin Gelatin in a Continuous Hollow Fiber Membrane Reactor (연속식 중공사막 반응기를 이용한 각시가자미피 젤라틴의 가수분해)

  • KIM Se-Kwon;BYUN Hee-Guk;KANG Tae-Jung;SONG Dae-Jin
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.26 no.2
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    • pp.120-132
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    • 1993
  • A continuous hollow fiber membrane reactor(CHFMR) was developed and optimized for the production of yellowfin sole(Limanda aspera) skin gelatin hydrolysates using trypsin. The results were summerized as follows: The $K_m$ value of the CHFMR was 2.4 times higher than that of the batch reactor, indicating reduced enzyme affinity for the substrate. The $K_2$ value of the CHFMR was 8.5 times lower than that of the batch process, showing a significant reduction in trypsin activity in the CHFMR. The optimum operating conditions for the CHFMR process were $55^{\circ}C$, pH 9.0, flux 7.79 ml/min, residence time 77min, and trypsin to substrate ratio, 0.01(w/w) After operating for 60min under the above conditions, $79\%$ of the total amount of initial gelatin was hydrolysed. Enzyme leakage was observed through the 10,000 MWCO membrane after the 20min of reactor operation, while none occurred after 5hr. Total enzyme leakage was about $12.95\%$ at $55^{\circ}C$ for 5hrs. However, there was no apparent correlation between enzyme leakage and substrate hydrolysis. The membrane has a significant effect on trypsin activity loss for 60min of the CHFMR operation. The CHFMR operating with the membrane lost $34\%$ of the initial activity versus a $23\%$ loss of activity after 3hr in the continuous reactor lacking the hollow fiber membrane. The measurement of fouling property showed that relative flux reduction was $91\%$ and flux recover rate was $92\%$ at $10\%$ substrate solution. The productivity(378.85mg product/mg enzyme) of the CHFMR was more than 4 times higher than that of the batch reactor at $55^{\circ}C$.

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Effects of Adenosine on the Action Potentials of Rabbit SA Nodal Cells (동방결절 활동전압에 대한 아데노신 효과)

  • Kim, Ki-Whan;Ho, Won-Kyung
    • The Korean Journal of Physiology
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    • v.18 no.1
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    • pp.19-35
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    • 1984
  • Since the first report of Drury and $Szent-Gy{\ddot{o}}rgyi$ in 1929, the inhibitory influences of adenosine on the heart have repeatedly been described by many investigators. These studies have shown that adenosine and adenine nucleotides have overall depressant effects, similar to those of acetylcholine. Heart beats become slow and weak. It is also well known that adenosine is a potent endogenous coronary vasodilator. Many investigations on the working mechanisms of adenosine have been focused mainly on the effects of the coronary blood flow. However, the cellular mechanisms underlying the inhibitory action of adenosine on sinus node are not well understood yet. Thus, this study was undertaken to examine the behavior of rabbit SA node under influence of adenosine. In these series of experiments three kinds of preparations were used: whole atrial pair, left atrial strip, and isolated SA node preparations. The electrical activity of SA node was recorded with conventional glass microelectrodes 30 to 50 $M{\Omega}$. The preparations were superfused with bicarbonate-buffered Tyrode solution of pH 7.35 and aerated with a gas mixture of $3%\;CO_2-97%\;O_2$ at $35^{\circ}C$. In whole atrial pair, adenosine suppressed sinoatrial rhythm in a dose-dependent manner. Effect of adenosine on atrial rate appeared at the concentration of $10^{-5}M$ and was enhanced in parallel with the increase in adenosine concentration. Inhibitory action of adenosine on pacemaker activity was more prominent in the preparation pretreated with norepinephrine, which can steepen the slope of pacemaker potential by increasing permeability of $Ca^{+2}$. Calcium ions in perfusate slowly produced a marked change in sinoatrial rhythm. Elevation of the calcium concentration from 0.3 to 8 mM increased the atrial rate from 132 to 174 beats/min, but over 10 mM $Ca^{+2}$ decreased. The inhibitory effect of adenosine on sinoatrial rhythm developed very rapidly. Atrial rate was recovered promptly from the adenosine-induced suppression by the addition of norepinephrine, but extra $Ca^{+2}$ was less suitable to restore the suppression of atrial rate. Adenosine suppressed also atrial contractility in the same dosage range that restricted pacemaker activity, even in the reserpinized preparation. In isolated SA node preparation, spontaneous firing rate of SA node at $35^{\circ}C$(mean{\pm}SEM, n=16) was $154{\pm}3.3\;beats/min. The parameters of action potentials were: maximum diastolic potential(MDP), $-73{\pm}1.7\;mV: overshoot(OS), $9{\pm}1.4\;mV: slope of pacemaker potential(SPP), $94{\pm}3.0\;mV/sec. Adenosine suppressed the firing rate of SA node in a dose-dependent manner. This inhibitory effect appeared at the concentration of $10^{-6}M$ and was in parallel with the increase in adenosine concentration. Changes in action potential by adenosine were dose-dependent increase of MDP and decrease of SPP until $10^{-4}M$. Above this concentration, however, the amplitude of action potential decreased markedly due to the simultaneous decrease of both MDP and OS. All these effects of adenosine were not affected by pretreatment of atropine and propranolol. Lowering extra $Ca^{2+}$ irom 2 mM to 0.3 mM resulted in a marked decrease of OS and SPP, but almost no change of MDP. However, increase of perfusate $Ca^{2+}$ from 2 mM to 6 or 8 mM produced a prominent decrease of MDP and a slight increase of OS and SPP. Dipyridamole(DPM), which is known to block the adenosine transport across the cell membrane, definately potentiated the action of adenosine. The results of this experiment suggest that adenosine suppressed pacemaker activity and atrial contractility simultaneously and directly, by decreasing $Ca^{2+}-permeability$ of nodal and atrial cell membranes.

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