유전자조작, 균주분리 갯메꽃 뿌리로부터 분리된 내생진균의 식물생장촉진활성

유영현 ( Young Hyun You ),윤혁준 ( Hyeok Jun Yoon ),우주리 ( Ju Ri Woo ),서영교 ( Yeong Gyo Seo ),신재호 ( Jae Ho Shin ),추연식 ( Yeon Sik Choo ),이인중 ( In Jung Lee ),김종국 ( Jong Guk Kim ) 한국미생물생명공학회 2011 한국미생물·생명공학회지 Vol.39 No.4

Eight endophytic fungal strains were isolated from the roots of Calystegia soldanella from the western coast of South Korea. The culture filtrate of the eight endophytic fungi were applied to waito-c rice seedlings in order to verify potential plant growth promotion activities. The results of bioassay indicated that the Cs-9-7 fungal strain possessed the highest plant growht promotion activity. Fungal culture filtrates were analyzed to verify secondary metabolites using gas chromatography and mass spectroscopy with selected ion monitoring (GC/MS-SIM). The culture filtrate of the Cs-9-7 fungal strain was confirmed to contain gibberellins GA3 (1.229 ng/mL), GA4 (3.535 ng/mL), GA7 (1.408 ng/mL) and GA12 (0.378 ng/mL). Polymerase chain reactions (PCR) were performed so as to determine the internal transcribed spacer (ITS) regions for the identification of isolated strains with universal primers ITS-1 and ITS-4. The Cs-9-7 fungal strain, isolated from the root of C. soldanella, has been named Aspergillus tubingensis Cs-9-7.

독도의 자생식물 뿌리에서 분리한 내생진균의 다양성과 생장촉진활성

Young-Hyun You(유영현),Hyeokjun Yoon(윤혁준),Gil Seong Lee(이길성),Ju-Ri Woo(우주리),Jae-Ho Shin(신재호),In-Jung Lee(이인중),Soon-Ok Rim(임순옥),Yeon-Sik Choo(추연식),Jong-Guk Kim(김종국) 한국생명과학회 2011 생명과학회지 Vol.21 No.7

독도에 자생하고 있는 식물의 뿌리로부터 내생진균의 분리를 시도하여, 동도로 부터 참억새, 돌피, 쇠무릎과 서도에서 갯별꽃 및 율무쑥등을 연구재료로 사용하였다. 총 21종의 내생진균을 분리하였고, universal primers ITS-1과 ITS-4를 사용하여 ITS 영역을 PCR로 증폭하여 동정하였다. 염기서열 분석결과, 동도에 자생하는 참억새에서는 Penicillium 속 균주가 75%, Aspergillus 균주가 25%, 분리되었고, 돌피에서는 Penicillium 속 균주가 55%, Aspergillus 속 균주가 30%, Zygorhynchus 속 균주가 15%가 분리되었으며, 쇠무릎에서는 Penicillium 속 균주가 50%, Aspergillus 속 균주가 12%, Gibberella 속 균주가 13%, Talaromyces 속 균주가 9%, Umbelopsis 속 균주가 8% 분리되었다. 서도에 자생하는 갯별꽃에서는 Penicillium 속 균주가 76%, Pestalotiopsis 속 균주가 24% 분리되었고, 율무쑥에서는 Penicillium 속 균주가 81%, Mucor 속 균주가 19% 분리되었다. Bioassay결과, 돌피에서 분리된 Ec-3-1균주가 식물생장촉진활성을 나타내었다. 그리고 독도의 5종류 식물에서 Penicillium 속의 내생진균이 가장 많이 존재함을 알 수 있었다. Endophytic fungi were isolated from the roots of plants growing naturally on the island of Dokdo. Plant samples, such as Miscanthus sinensis, Achyranthus japonica and Echinochloa crusgali were isolated from Dongdo, and those such as Honkenya peploides and Artemsia koidzumii were isolated from Seodo. Twenty one strains of endophytic fungi were isolated from these plants. To identify the strains, PCR (polymerase chain reaction) amplification of the partial ITS (Internal Transcribed Spacer) regions was done with universal primers ITS-1 and ITS-4 to determine the nucleotide sequence of the ITS regions. Of the strains isolated from Miscanthus sinensis, 75% were Penicillium sp. and 25% were Aspergillus sp. Fifty five percent of strains isolated from Achyranthus japonica were Penicillium sp., 30% were Aspergillus sp. and 15% were Zygorhynchus sp. Strains isolated from Echinochloa crusgali were Penicillium sp. (50%), Aspergillus sp. (12%), Giberella sp. (13%), Talaromyces sp. (9%) and Umbelopsis sp. (8%). Of the strains isolated from Honkenya peploides, 76% were Penicillium sp. and 24% were Pestalotiopsis sp. Strains isolated from Artemisia koidzumii were Penicillium sp. (81%) and Mucor sp. (19%). As a result of bioassay, Ec-3-1 strain isolated from Echinochloa crusgalli showed plant growth-promotion activity. Of all the endophytic fungi isolated, Penicillium sp. was the most abundantly distributed fungal strain in all plants used in this study.

Catecholamines에 관(關)하여 -제4편(第四編) : 심실전동발생(心室顫動發生)에 있어서의 catecholamines의 의의(意義)-

이우주(Woo Choo Lee) 대한약리학회 1983 대한약리학잡지 Vol.19 No.1

Although it has been well known that ventricular fibrillation is the most important complication during hypothermia, much investigation has failed to show the exact nature of the etiology of ventricular fibrillation. Recently, there has been considerable research on the relationship between sympathetic activity and ventricular fibrillation under hypothermia. Cardiac muscle normally contains a certain amount of norepinephrine and the dramatic effect of this catecholamines on the cardiac muscle is well documented. It is, therefore, conceivable that cardiac catecholamines might exert an influence on the susceptibility of heart muscle to tachycardia, ventricular fibrillation and arrhythmia, under hypothermia. Hypothermia itself is stress enough to increase tonus of sympatheticoadrenal system. The normal heart is supplied by an autonomic innervation and is subjected to action of circulating catecholamines which may be released from the heart. If the reaction of the heart associated with a variable amount of cardiac catecholamines is. permitted to occur in the induction of hypothermia, the action of this agent on the heart has not to be differentiated from the direct effects of cooling. The studies presented in this paper were designed to provide further information about the cardio-physiological effects of reduced body temperature, with special reference to the role of catecholamines in ventricular fibrillation. Healthy cats, weighing about 3 kg, were anesthetized with pentobarbital(30 mg/kg) intraperitoneally. The trachea was intubated and the endotracheal tube was connected to a C.F. Palmer type A.C. respirator. Hypothermia was induced by immersing the cat into a ice water tub and the rate of body temperature lowering was 1˚C per 5 to 8 min. Esophageal temperature and ECG (Lead II) were simultaneously monitored. In some cases the blood pH and serum sodium and potassium were estimated before the experiment. After the experiment the animals were killed and the hearts were excised. The catecholamines content of the cardiac muscle was measured by the method of Shore and Olin (1958). The results obtained are summarized as follows. 1) In control animal the heart rate was slowed as the temperature fell and the average pulse rates of eight animals were read 94/min at 31˚C, 70/min at 27˚C and 43/min at 23˚C if esophageal temperature. Ventricular fibrillation was occurred with no exception at a mean temperature of 20.3˚C(21-l9˚C). The electrocardiogram revealed abnormal P waves in each progressive cooling of the heart. there was, ultimately, a marked delay in the P-R interval, QRS complex and Q-T interval. Inversion of the T waves was characteristic of all animals. The catecholamines content of the heart muscle excised immediately after the occurrence of ventricular fibrillation was about thirty percent lower than that of the pre-hypothermic heart, that is, 1.0μg/g wet weight compared to the prehypothermic value of 1.41μg/g wet weight. The changes of blood pH, serum sodium and potassium concentration were not remarkable. 2) By the adrenergic receptor blocking agent, DCI(2-3 mg/kg), given intramuscularly thirty minutes before hypothermia, ventricular fibrillation did not occur in one of five animals when their body temperature was reduced even to 16˚C. These animals succumbed at that low temperature, and the changes of heart rate and loss of myocardial catecholamines after hypothermia were similar to those of normal animals. The actual effect of DCI preventing the ventricular fibrillation is not predictable. 3) Administration of reserpine(1 mg/kg, i.m.) 24 hours Prior to hypothermia disclosed reduced incidence of ventricular fibrillation, that is, six of the nine animals went into fibrillation at an average temperature of 19.6˚C. By reserpine myocardial catecholamines content dropped to 0.045μg/g wet weight.

Catecholamines에 관(關)하여 -제삼편(第三編) 심장(心臟) Catecholamines에 관(關)한 실험적(實驗的) 연구(硏究)

이우주(Lee, Woo-Choo) 대한약리학회 1973 대한약리학잡지 Vol.9 No.1

During the last decade expensive studies on catecholamines have evolved new knowledge in the physiology and biochemistry of adrenergic mechanism. Cardiac muscle, receiving adrenergic fibres from the stellate, cervical and thoracic ganglia, has been repeatedly shown to have a specific capacity to uptake and to store catecholamines. The catecholamine stores in cardiac muscle have also been shown to be important sites for the action of numerous drugs. Under normal condition, a certain level of catecholamines is maintained in the stores and serves as the basis for studying the changes in the catecholamine content of the heart. Because myocardial catecholamines play such important role in the patho-physiology of the heart, it would be interesting to compare the normal level of myocardial catecholamines among various species of animals. An occasional study has dealt with myocardial catecholamines of several species and ages of animals but these have been insufficiently comprehensive to afford a basis for an understanding of the importance of these amines as related to species and ages. The present investigation was undertaken to determine whether or not there is any significance of myocardial catecholamines in the c::mrse of the evolution and development of animals. Seasonal changes, sex difference and regional and subcellular distribution of myocardial catecholamines were also examined. The concentration of cardiac catecholamines was determined by the spectrophotofluorometric procedure described by Shore and Oiin. rat, cow and pig. The ratio of norepinephrine to epinephrine also was greater as the animals progress phylogenetically. 2. Examination of the regional distribution of cardiac catecholamines in warm-blooded animals showed that the content of the auricle was generally higher than that of the septum and considerably than that of the ventricle, but the differences of contents among these regions were not so marked. 3. In the embryonic chick, cardiac catecholamines were firstly detected on the 4th day of incubation, the time before the cardiac innervation of sympathetic nerves. The concentrations of these catecholamines increased but not markedly on the 6th day of incubation, soon after the innervation of sympathetic nerves to the heart. The level of the cardiac catecholamines fluctuated throughout the remainder of embryonic development. 4. In newborn rat hearts, a considerable amount of catecholamines was present. With the development of the rats, the concentrations of myocardial catecholamines increased. The ratio of epinephrine and norepinephrine fluctuated within the range of 40 to 60 pervent. However, as development progressed, the percentage of norepinephrine continued to rise, attaining the adult value of <TEX>$80{\sim}90%$</TEX> after <TEX>$45{\sim}60$</TEX> days. In contrast, the total amount of epinephrine remained fairly constant throughout the animal s development. 5. No significant sexual differences were observed in the concentration of myocardial catecholamines in the developing rat. 6. The catecholamines in the rabbit hearts increased during the summer season (from May to August) and maintained a fairly constant level in the other seasons of the year. 7. The subcellular distribution of cardiac catecholamines was examined by differential centrifugation of homogenates of cardiac muscles in rabbits, cats and rats. The catecholamines were found to be present approximately 20% in particles of mitochondrial fraction, 45% in particles of microsomal fraction an rat, cow and pig). The ratio of norepinephrine to epinephrine also was greater as the animals progress phylogenetically. 2. Examination of the regional distribution of cardiac catecholamines in warm-blooded animals showed that the content of the auricle was generally higher than that of the septum and considerably than that of the ventricle, but the differences of contents among these regions were not so marked. 3. In the embryonic chick

Catecholamines에 관(關)하여 -제5편(第五編) : 자궁(子宮) catecholamines에 관한 실험적(實驗的) 연구(硏究)-

이우주,Lee, Woo-Choo 대한약리학회 1983 대한약리학잡지 Vol.19 No.1

The uterus receives adrenergic terminals from the mesenteric ganglia and considerably large amount of catecholamines have been shown to be contained in this organ. On the other hand, the activities of epinephrine, norepinephrine or adrenergic nerve on uterine motility is so complicated that many controversial results have been reporter. Recently, a large number of reports concerning the changes of uterine catecholamines content have appeared, but little is known about the role of uterine catecholamines in their activities on uterine motility. The present experiments were undertaken to determine the significance of the intrinsic uterine catecholamines in the physiology of uterus. Female albino rabbits weighing approximately 2 kg were employed in this experiment. uterine strip3 were prepared and suspended in a constant temperature $bath(38^{\circ}C)$ containing 100 ml of Locke's solution aerated with 95% oxygen and 5% carbon dioxide. Spontaneous motility was recorded on a smoked drum with an isotonic lever. The catecholamines concentration of the uterus was determined according to the Procedure described of Shore and Olin (1958). Human uterus obtained from patients was also used to determine the catecholam ines content of myometrium. Followings are summarized results. 1) On the non-pregnant rabbit uterine strips, epinephrine and norepinephrine significantly elevated the tonus and stimulated the spontaneous motility. Pretreatment with dichloroisoproterenol(DCI), an adrenergic beta-receptor blocker, enhanced the stimulatory activity of epinephrine or norepinephrine. On the other hand, pretreatment with dibenamine, an adrenergic alpha-receptor blocker, rendered the uterine muscle to exhibit inhibition after the administration of epinephrine or norepinephrine. Following the treatment with both DCI and dibenamine, epinephrine or norepinephrine produced no appreciable effects on the spontaneous motility of the uterus. These results suggest there exist both alpha and beta-adrenergic receptors in the uterine muscle and the response to epinephrine of the former is predominant over that of latter in the non-pregnant uterus of rabbits. The total catecholamines concentration of the non-pregnant uterus was $351\;m{\mu}g/g$ and the fractional concentrations of epinephrine and norepinephrine were $125\;m{\mu}g/g(35.7%)$ and $226\;m{\mu}g/g$ respectively. It is interesting to note that the catecholamines content of uterus was characterized by a high fractional corcentration of epinephrine relative to norepinephrine. 2) On the pregnant rabbit uterine strips, the effects of epinephrine and norepinephrine varied according to the period of pregnancy. The response to epinephrine of adrenergic beta receptor of uterus increased during pregnancy, and the effect of catecholamine was inhibitory in the early pregnancy but became stimulatory as the pregnancy progressed. This stimulating action on the uterine motility was found to occur through the action of norepinephrine. The uterine catecholamines concentration was markedly reduced during pregnancy. The catecholamines concentration was started to decrease in the early pregnancy, reached the lowest level in the mid-pregnancy and then started to increaae again in the late pregnancy when the total catecholamines content became the highest level of all. This increase of catefholamines in late pregnancy was chiefly due to the increase of norepinephrine. These results suggest that the uterine motility may be related to the catecholamines content, especially norepinephrine content in the uterus. 3) Bilateral oophorectomy of rabbits results in a marked shrink of the uterus in size. The spontaneous motility of the uterine segment of these animals was very weak and irregular. Norepinephrine produced inhibitory effect, whereas epinephrine was stimulatory or inhibitory effect on the uterine segment. The total catecholamines tontent in whole uterus was markedly reduced. The

최신 이뇨제

이우주,Lee Woo Choo 대한간호협회 1965 대한간호 Vol.4 No.2

생약진피(生藥陳皮)의 심혈관계(心血管系)에 대한 약리작용(藥理作用)

이병학(Lee, Byung-Hak),조태순(Cho, Tai-Soon),이우주(Lee, Woo-Choo) 대한약리학회 1974 대한약리학잡지 Vol.10 No.2

Citrus unshiu Marc., a family of Rutaceae, has been used as antitussives, antiemetics, expectorants, spasmolytics, stomachics and digestants in oriental medicine. But there is few reports on the pharmacological effects of Citrus unshiu pericarpium. Present experiment was performed to observe the effects of the water extract of Citrus unshiu on isolated auricle and intestine in rabbits and blood pressure in rats and also to study on the active component on the cardiovascular system by means of thin layer chlomatography and spectrophotometer. 1. The water extract showed positive inotropic and positive chronotropic effect in the concentration of 1 X 10^-4g/ml, 5 X 10^-4g/ml, and 1 X 10^-3g/ml. 2. The motility of isolated rabbit intestine was significantly depressed by the water extract. 3. The blood pressure was markedly increased by the water extract. 4. The blood vessel of rabbit ear was constricted by perfusion of the ether extract. 5. The purified alkaloid of Citri pericarpium by ion exchange resin showed the same Rf value with l-synephrine on thin layer chromatography. 6. The purified alkaloid of Citri pericarpium by thin layer chromatography showed the same absorption curve with l-synephrine in the ultraviolet scanning.

황금(黃芩)의 심장(心臟)에 대한 약리작용(藥理作用)

노재열(Jai-Youl Ro),이우주(Woo-Choo Lee) 대한약리학회 1975 대한약리학잡지 Vol.11 No.2

The adrenergic blocking activity and refractory period of cardiac muscle on isolated rabbit atria were measured after administration of Scutellaria. In rabbits and cats the antiarrhythmic action of Scutellaria on atrial and ventricular arrhythmias produced by epinephrine or ouabain was examined and also compared with that of propranolol and quinidine. The alcoholic extract of Scutellaria produced a marked decrease in heart rate and contractile amplitude of the isolated rabbit atria. Pretreatment with Scutellaria rendered the atria to fail to respond to epinephrine, indicating that this crude drug possesses an adrenergic blocking activity. The extract produced a marked prolongation of the refractory period of atrial muscle. The extract effectively abolished the spontaneous arrhythmia occurring in the isolated rabbit atria. As propranolol and quinidine it also suppressed the atrial arrhythmia induced by ouabain. The extract prevented, as propranolol and quinidine, the induction of ventricular arrhythmia arising from excessive dose of epinephrine in anesthetized rabbits and cats. With regard to the ventricular arrhythmia induced by a continuous infusion of ouabain, the alcoholic extract of Scutellaria exerted some suppressive effect in anesthetized rabbits but no effect on cats. From the above results, it may be concluded that Scutellaria is effective against atrial and ventricular arrhythmias. The antiarrhythmic effects of this drug may be the result of adrenergic beta receptor blocking and cardiac depressive activities including prolongation of the refractory period of cardiac muscle.

고양이에서 체표냉각에 따른 심혈관계 변동에 관한 연구

안영수(Young Soo Ahn),고창만(Chang Mann Ko),이우주(Woo Choo Lee) 대한약리학회 1983 대한약리학잡지 Vol.19 No.1

Hypothermia is an essential preparatory procedure for cardiac surgery, which lows the metabolic rate and myocardial oxygen demand. However, hypothermia itself is a stress enough to change the tonus of sympathoadrenal system, especially the cardiovascular responses to the catecholamines. It is reported that the positive chronotropic and inotropic response of catecholamines is exaggerated during hypothermia because of decreased norepinephrine uptake at the junctional cleft or decreased catecholamine metabolism. On the other hand, there are evidences of diminished catecholamines responses in low temperature ana further, interconversion of adrenergic receptors is also suggested. Present investigation was planned to observe the cardiovascular changes and its responses to catecholamines during surface hypothermia in cat. Healthy mongrel cats, weighing 2 ~ 3 kg, anesthetized with secobarbital(30 mg/kg), were permitted to hypothermia by external cooling technic. Esophageal temperature, ECG (lead II), heart rate, left ventricular pressure with dP/dt, carotid artery pressure and left ventricular contractile force were monitored with Polygragh (Model 7, Grass), and the respiration was maintained with artificial respirator (V 5 KG, Narco). Followings are summarized results. 1) Surface cooling caused progressive decrease of body temperature and reached l8.8 ± 0.8˚C and 16.9 ± 0.6˚C in 120 and 150 min respectively, after immersion into ice water, and ventricular fibrillation was developed at 20.4 ± 0.65˚C. 2) Heart rate, blood pressure and myocardial contractility were decreased after initial increase as the body temperature falls. 3) Systolic and diastolicdd P/dt of left ventricular pressure were decreased and that the decrement of diastolic dP/dt was more marked. 4) On ECG, ST depression, Twave inversion and prolongation of PR interval were prominent in hypothermia, and moreover, the prolongation of PR interval was marked just prior to the development of ventricular fibrillation. 5) The cardiovascular responses to catecholamines, especially to isoproterenol, were suppressed under hypothermia.

麻藥의 藥理學的 作用

李宇柱 대한의학협회 1965 대한의학협회지 Vol.8 No.9