A survey of the internal parasites in 115 fishes of Ophicephalus argus was carried out in Kyungpook province during the period from January 1995 to November 1997. A species of trematode from the stomach and a species of nematode from the caeca of the fishes was discovered respectively. The trematode was identified as Azygia hwangtsinyi (Tsin, 1993) and the nematode as Pingus sinensis ($Hs\ddot{u}$, 1993). The fluke was pressed at thick of 0.1mm~0.2mm between slide glasses and fixed in 70% ethyl alcohol solution. The fluke was washed with tap water after fixation and stained with hematoxylin-carmin and mounted in balsam through routine methods. The nematode was fixed in 5% formalin solution and mounted with lactophnol or glycerine jelly. Morphology : Arygia hwantsinyi ; The fluke is elongate body with approximately parallel margins and rounded extremities in pressed preparations(Fig 1). The cuticle is unarmed. The oral sucker is ventro-subterminal in position. The ventral sucker is slightly smaller than oral sucker and positioned at 14.2% of body length from the oral sucker. The cirrus sac and genital pore is in mid-ventral line, slightly anterior to the ventral sucker. The oral sucker is succeeded by a well developed muscular pharynx. It is succeeded by a short inverted Y-shaped oesophagus. The inner wall of oesophagus is consisted of many fine folded membrane. The fine tubes of esophagus pass into the intestinal heads which are distended and presented a marked borderline between the fine tube and intestinal heads. The Intestinal heads start at level of pharynx and pass caudad in a zigzag course to end quite near caudal margin. The uterine coils occupy the intercecal area between the level of the cephalic margin of the ovary and about the level of the ventral sucker. The uterus is filled with many eggs. The testes are round or oval and placed one behind the other generally a little diagonally. The posterior testis is placed at level of 75.1% of body length. The ovary is oval, placed ahead the anterior testis. The shell gland not discernible outline is contacted with ovary ahead it. The vitelline glands are consisted of small oval vesicles. The vesicles occupy extracecal regions and between a little posterior of the ventral sucker and near caudal margin. Pingus sinensis : Female ; The cuticle is smooth and the cervical alae are well developed. The mouth opening is small, triangular, and without lips. The anterior end bears four sub-medial papillae. The oesophagus is divided into a muscular and glandular portion. The nerve ring is situated just at the junction of the muscular and glandular portions of the oesophagus. The vulva which has a prominent flap, is placed behind the middle of the body about one-third of the length of the worm from the posterior extremity. The vagina is strongly muscularised and after running anteriorly about $68{\mu}$, divides itself into two broad uterine tubes with an anterior and a posterior one. There are a few eggs in the uterine tubes. The tail is slender, straight and pointed. The ovary extends almost to the posterior extremity. Male : Male is smaller than female. The tail is pointed, curved and carrying well-developed caudal alae. A prominent muscular precloacal sucker is characteristic. The spicules are equal and short. There are four pairs of pedunculated pre-anal papillae, of which the most anterior pair are situated just at the level anterior to the muscular sucker. Two pairs of pedunculated papillae are present before the anus. There are five pairs of post-anal papillae of which the most anterior pair and the second counted from the posterior end are ventral and more marked than the ones which are situated laterally.
The cytochrome P450s (P450s) metabolizing natural products are among the most versatile biological catalysts known in plants, but knowledge of the structural basis for their broad substrate specificity has been limited. The activity of p-coumarate 3-hydroxylase (C3H) is thought to be essential for the biosynthesis of lignin and many other phenylpropanoid pathway products in plants however, all attempts to express and purify the protein corresponding C3H gene have failed. As a result, no conditions suitable for the unambiguous assay of the enzyme are known. The detailed understanding of the mechanism and substrate-specificity of C3Hdemands a method for the production of active protein on the milligram scale. We have developed a bacterial expression and purification system for the plant C3H, which allows for the quick expression and purification of active wild-type C3H via introduction of combinational mutagenesis. The modified cytochrome P450 C3H ($C3H_{mod}$) could be purified in the absence of detergent using immobilized metal affinity chromatography and size exclusion chromatography following extraction from isolated membranes in a high salt buffer and catalytically activated. This method makes the use of isotopic labeling of C3H for NMRstudies and X-ray crystallography practical, and is also applicable to other plant cytochrome P450 proteins.
Park, Sang Eun;Lee, Su Young;Shin, Dong Yeok;Jeong, Jin-Woo;Jin, Myung Ho;Park, Seon Young;Chung, Yoon Ho;Hwang, Hye Jin;Hong, Sang Hoon;Choi, Yung Hyun
Journal of Life Science
/
v.23
no.3
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pp.389-398
/
2013
Platycodin D is a major constituent of triterpene saponins, which is found in the root of Platycodon grandiflorum, Platycodi Radix, which is widely used in traditional Oriental medicine for the treatment of many chronic inflammatory diseases. Several pharmacological effects of this compound have been reported recently, such as anti-inflammation, immunogenicity, anti-adipogenesis, lowered cholesterol, and anti-cancer activity. However, the mechanism by which this action occurs is poorly understood. In this study, we found that platycodin D greatly increased the potential of the anti-proliferative effect in various cancer cell lines. Our data revealed that platycodin D treatment resulted in a time- and concentration-response growth inhibition of U937 cells by inducing apoptosis, as evidenced by the formation of apoptotic bodies, chromatin condensation, and the accumulation of cells in the sub-G1 phase. Apoptosis induction of U937 cells by platycodin D correlated with an increase in the Bax/Bcl-2 ratio and caused the down-regulation of IAP family members. In addition, platycodin D treatment resulted in proteolytic activation of caspase-3, the concomitant degradation of poly(ADP-ribose) polymerases, and the collapse of the mitochondria membrane potential (${\Delta}{\Psi}_m$). However, the cytotoxic effects induced by platycodin D treatment were significantly inhibited by z-DEVD-fmk, a caspase-3 inhibitor, which demonstrated the important role that caspase-3 played in the observed cytotoxic effect. These findings suggest that platycodin D may be a potential chemotherapeutic agent for use in the control of human leukemia U937 cells. These findings also provided important new insights into possible molecular mechanisms of the anti-cancer activity of platycodin D.
Kim Soo-Youn;Choi Gang Guk;Park Youn Il;Park Young Mok;Yang Young Ki;Rhee Young Ha
Korean Journal of Microbiology
/
v.41
no.1
/
pp.67-73
/
2005
Photoautotrophic bacteria are promising candidates for the production of poly(3-hydroxybutyrate) (PHB) since they can address the critical problem of substrate costs. In this study, we isolated 25 Tn5-inserted mutants of the Synechocystis sp. PCC 6803 which showed enhanced PHB accumulation compared to the wild-type strain. After 5-days cultivation under nitrogen-limited mixotrophic conditions, the intracellular levels of PHB content in these mutants reached up to $10-30\%$ of dry cell weight (DCW) comparable to $4\%$ of DCW in the wild-type strain. Using the method of inverse PCR, the affected genes of the mutants were mapped on the completely known genome sequence of Synechocystis sp. PCC 6803. As a result, the increased PHB accumulation in 5 mutants were found to be resulted from defects of genes coding for NADH-ubiquinone oxidoreductase, O-succinylbenzoic-CoA ligase, photosystem II PsbT protein or histidine kinase, which are involved in photosystem in thylakoid inner membrane of the cell. The values of $NAD(P)H/NAD(P)^+$ ratio in the cells of these mutants were much higher than that of the wild-type strain as measured by using pulse-amplitude modulated fluorometer, suggesting that PHB synthesis could be enhanced by increasing the level of cellular NAD(P)H which is a limiting substrate for NADPH-dependent acetoacetyl-CoA reductase. From these results, it is likely that NAD(P)H would be a limiting factor for PHB synthesis in Synechocystis sp. PCC 6803.
The $Na^+$-and $K^+$-induced $Ca^{++}$ release was measured isotopically by Milipore filter technique in mitochondria isolated from rabbit ventricles. The release of $Ca^{++}$ from mitochondria could be induced by 1-3 mM of $Na^+$ added in incubating medium under the presence of 0.5mM EGTA to prevent the released $Ca^{++}$ from rebinding with mitochondrial membrane. The amount of $Ca^{++}$ released was increased by increasing the concentration of $Na^+$ added. 100mM $K^+$, in itself, did not induce the $Ca^{++}$ release from cardiac mitochondria, the $Na^+$-induced $Ca^{++}$ release, however, was potentiated by the presence of $K^+$. The potentiation of $Na^+$-induced $Ca^{++}$ release by $K^+$ was proportional to the $Na^+/K^+$ ratio presented in the incubating medium. Among the monovalent cations other than $Na^+$, the release of $Ca^{++}$ from cardiac mitochondria was shared only by $Li^+$. The $Na^+$-induced $Ca^{++}$ release could be also observed in the mitochondria isolated from liver and kidney. However, the $Na^+$ sensitivity was somewhat lower in liver and kidney mitochondria than in heart mitochondria. The release of $Ca^{++}$ induced by $Na^+$ in the mitochondria isolated from the experimentally produced failured heart was not different from that in the normal heart mitochondria, and was not directly modified by $10^{-6}{\sim}10^{-5}$ M of Ouabain. From the experiments, it was suggested that the $Ca^{++}$ released from mitochondria by $Na^+$ could be used in excitation-contraction coupling process to initiate the contraction of the cardiac myofibrils. Futhermore, it appeared that the phenomenon of $Ca^{++}$ release from cardiac mitochondria by $Na^+$ and $K^+$ might be related to the inotropic effect of digitalis glycoside which could bring about the increase of $Na^+$ or the reduction of $K^+$ intracellulary through the inhibition of $Na^+$, $K^+$-ATPase.
Numerous factors concern with the absorption of substances through the membrane of the gastrointestinal tract. To simplify the experimental condition, present work has been restricted to observe the disappearance rate of substance from the intestinal loop which was made in the jejunum, 70 cm apart from the pylorus of the adult rabbit. The purpose of the study is to clarify the absorption of urea through the jejunal wall is solely attributable to the concentration difference between the luminal fluid and plasma, and to observe the effect of adding red pepper upon the rate of absorption. The rabbits were anesthetized with nembutal, 35mg/kg I.V. Jejunal loop was made by ligating at 2 spots, 70 cm and 80cm apart from the pylorus. After rinsing with normal saline solution through the polyethylene tubing inserted from the end of the loop, 8 ml of test solution was placed through the same tubing. The test solution contained 200 mg% of urea and 150mg% of polyethylene glycol(M.W. 4,000) in normal saline solution. Right after placing the test solution the first specimen was taken through the tubing, and successive samplings were performed at 5, 10, 20, and 30 minutes. Logarithm of the difference of urea concentration between the luminal fluid and plasma was plotted against time elapsed after the onset of the experiment. If straight line is revealed, it would verify the nature of transport mechanism as diffusion, obeying the Fick's principle. The concentration of polyethylene glycol (PEG) was also measured in order to examine the change in the volume. PEG was used as the marker substance because it is not absorbable in the intestinal tract. Consequently the concentration of PEG relates inversely to the volume of the loop. Instantaneous concentration of urea in the loop times the volume will give the amount of urea remaining in the luminal fluid. The change in the amount of any substance is directly relate to the volume of the compartment and differs from the change in the concentration which is independent of the volume. After completion of the experiment without red pepper, it was added in the test solution and was centrifuged after thorough mixing. Supernatant of the mixture was placed in the loop and similar sampling were performed with the same time intervals that of previous run in order to observe the effects of the red pepper on the passive transport of the water soluble small substance, urea. The results obtained were as follows: 1. Logarithm of the concentration difference of urea between the luminal fluid and plasma was diminished exponentially as time elapsed. The decay constant in the experiment without red pepper was 0.0563/min. By adding red pepper in the test solution as much as the concentration rose to 4,000 mg% and 8,000 mg%, the decay constants were lowered to 0.0493/min and to 0.0506/min, respectively. The time interval by which the concentration difference dropped to one half of the initial value was prolonged. Without red pepper the half concentration time was 13.30 minutes, and by adding extract of red pepper, 15.31 minutes and 15.71 minutes were revealed. 2. The profile of the diminishing rate of tile amount of urea was quite different from that of the concentration because of the change in the volume of the loop during the observed period. 3. By adding the extract of red pepper, it slowed down the rate of absorption of urea in the intestinal loop, suggesting an increase in the diffusional barrier. 4. Larger dosage of red pepper brought an increase in the secretion of intestinal fluid with concomitant expansion of the luminal volume, and the retardation of the absorption of urea was noticed. This effect was largely dependent on the sensitivity of the individual animal to the red pepper, extract. The amount of urea remained after 10 minutes interval was 55.5% of the initial amount in the experiment without red pepper. On the other hand it was not consistent after administration of red pepper, showing 50.6% and 66.5% of the initial figures by adding 400 mg and 800 mg of red pepper in the test solution, respectively. It was postulated that symptom of diarrhea often encountered by taking a hot (red pepper) food might be attributable to the increase of secretion and the retardation of absorption in the intestinal tract.
The purpose of this study was to fine out the general physical status of the neonates, and to identify the risk factors of the mothers and the neonates which were significantly related to the neonatal diseases during hospitalization. The data were obtained from clinical records of 1098 neonates born in Seoul Red cross Hospital between January 1st of 1984 and December 31th of 1986. The results of this study were summarized as follows: 1. General characteristics of the maternal group. 1) The average of maternal age was 26.6 years, the $91.7\%$ of the mothers de liveried at the age of 20-34 years old. 2) The distribution of the types of delivey were as follows : spontaneous delivery $39.9\%$, cesarean section $32.4\%$, vaccum extraction $25.7\%$, and breech delivery$2.0\%$. 3) The $40.3\%$ of the total de liveried mother had experienced abortion. 4) The $42.3\%$ of the total deliveried mother had one or more obstetric risk factors. 2. General characteristics of the neonatal group. 1) In the distribution of sex, male was $49.4\%$, female $50.6\%$. 2) The average of birth weights was 3,020gm. The distribution of birth weight were as follows; nomal weight $85.5\%$, low birth weight $12.7\%$ and high birth weight $2.5\%$. 3) The average of gestational age was 39.2 weeks. The distribution of gestational age were as follows; full term $77.4\%$, preterm $13.7\%$, and postterm $8.9\%$. 4) The average of Apgar Score was 9.0 at one minute and 9.6 at five minutes. 5) The $5.7\%$ of the neonates had one or more neonatal risk symptoms and signs at birth. 3. Apgar Score by the maternal and neonatal factors. In Apgar Score at one minute, normal group was higher than that of abnormal group. Apgar Score at five minutes was slightly higher than that at one minute. 4. The distribution of the maternal risk factors and the neonatal risk factors. 1) The total numbers of the maternal risk factors were 1376. The distribution of the maternal risk factors were as follows: obstetric factor $33.7\%$, abortion $32.2\%$, breech and cesarean section delivery $27.5\%$ and maternal age under 19 years and over 35 years $6.6\%$. 2) The total numbers of the neonatal risk factors were 517. The distribution of the neonatal risk factors were as follows: gestational age under 37 weeks and over 42 weeks $48.0\%$, birth weight under 2500gm and over 4000gm $12.2\%$, Apgar score under 4 at one munute $6.4\%$ and Apgar score at five munutes $2.7\%$. 3) The total numbers of the obstetric risk factors were 661. The types of the obstetric risk factors were meconium stained amniotic fluid $22.0\%$, premature rupture of membrane $17.5\%$. absence prenatal care $14.1\%$, unmarried pregnancy $10.3\%$, placenta problem $9.0\%$, toxemia $8.0\%$. 4) The total numbers of the neonatal risk symptoms and signs at birth were 83. The types of the neonatal risk symptoms and signs were respiratory distress $65.1\%$, neonatal apnea $14.4\%$, convulsion $13.3%$, meconium aspiration syndrome $4.8\%$, cyanosis $2.4\%$. 5. The relationship between the maternal risk factors and the neonatal risk factors. 1) Maternal age under 19 years or over 35 years was significantly related to Apgar Score under 4 at 5 minutes. 2) Breech delivery or cesarean section was significantly related to neonatal risk factor at birth such as birth weight, gestational age, Apgar Score at one minute and at five minutes. and neonatal risk symptoms and signs. 3) Obstetric risk factors were significantly related to the neonatal risk factors at birth. 4) Abortion was not related to the neonatal risk factors. 6. The relationship between neonatal diseases during hosptalization and the maternal or the neonatal risk factors. 1) The total numbers of neonatal diseases during hospitalization were 281. The distribution of neonatal diseases were as follows: birth trauma $38.1\%$, infectious disease $31.3\%$, hematologic disease $21.4\%$, respiratory disease $6.0\%$, neurologic disease $2.5\%$. cardiovascular disease $0.7\%$. 3) Most maternal risk factors except abortion were significantly related to neonatal diseases. 4) Most neonatal risk factors at birth were significantly related to neonatal diseases.
Ha, Ji Hoon;Lee, Hye Mi;Kwon, Soon Sik;Kim, Hae Soo;Kim, Moon Jin;Jeon, So Ha;Jeong, Yoo Min;Hwang, Jun Pil;Park, Jong-Ho;Choi, Yung-Key;Park, Jino;Park, Soo Nam;Park, Dong-Sik
Journal of the Society of Cosmetic Scientists of Korea
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v.39
no.4
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pp.259-269
/
2013
In this work, licorice extracts were prepared using various extraction conditions such as extraction solvent, temperature, and time from Glycyrrhiza uralensis (G. uralensis) produced in Korea and China and Glycyrrhiza glabra (G. glabra) in Uzbekistan. The optimum extraction condition was selected from the extraction yields and antioxidative activities of extracts. Korea licorice extracts showed the highest free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity (46.05%) under the extraction condition of 85% ethanol at $60^{\circ}C$ for 6 hours. The prominent ROS (reactive oxygen species) scavenging activity using luminol-dependent chemiluminescence assay and the cellular protective effect against $^1O_2$ induced cellular membrane damage were also shown from the extracts obtained from the same condition. Especially, Korea G. uralensis extracts exhibited the higher prominent protective effect (${\tau}_{50}$ = 116.4 min) than (+)-(+)-${\alpha}$-tocopherol (${\tau}_{50}$ = 28.5 min) and the extraction yield of Korea licorice extract was 18.75%, which is 1.2 times and 2.5 times higher than that of Uzbekistan and China, respectively. These results indicate that the extraction condition of 85% ethanol at $60^{\circ}C$ for 6 hours is optimal to prepare licorice extracts, which can be applicable as anti-oxidative cosmetic materials.
The present study was attempted to investigate the effects of total ginseng saponin (G75), panaxadiol-type (PDS) and panaxatriol-type saponin (PTS) on contractile responses of vasoconstrictors in aortic smooth muscle stripes of normotensive (NR) and spontaneous hypertensive rats (SHR). Phenylephrine (an adrenergic $\alpha$$\_$1/-receptor agonist) and high potassium (a membrane depolarizing agent) caused greatly contractile responses in both NR and AHR aorta, respectively. Phenylephrine- and high potassium-induced contractile responses were greater in NA than those in SHR aortic smooth muscle stripes. In NR, the contractile responses of high potassium (5.6$\times$10$\^$-2/ M) were not affected in the presence of GTS (300 $\mu$g/ml), PDS (300 $\mu$g/ml), and PTS (300 $\mu$g/ml), respectively whereas phenylephrine (10$\^$-6/ M)-induced contractile responses were markedly inhibited. In SHR, the contractile responses of high potassium (5.6$\times$10$\^$-2/ M) were not affected in the presence of GTS (300 $\mu$g/ml), PDS (300 $\mu$g/ml), and moderate doses of PTS (150-300 $\mu$g/ml), respectively but greatly blocked by high concentration of PTS (600 $\mu$g/ml). Phenylephrine (10$\^$-6/ M)-induced contractile responses were inhibited in a dose dependent fashion (150-600 $\mu$g/ml) by the pretreatment with PTS while not altered in the presence of GTS (300 $\mu$g/ml) and PDS (300 $\mu$g/ml), respectively. Taken together, these experimental results suggest that ginseng saponins cause vascular relaxation through blockade of adrenergic $\alpha$$\_$1/-receptors and some unknown mechanisms, and that there is some difference in sensitivity of vascular smooth muscle between NR and SHR in responses to ginseng saponins. It seems that panaxatriol type of some ginseng saponins has the greatest potency in vascular relaxation.
Kim, Sun-Q;Shin, Mi-Kyoung;Auh, Q-Schick;Lee, Jin-Yong;Hong, Jung-Pyo;Chun, Yang-Hyun
Journal of Oral Medicine and Pain
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v.32
no.2
/
pp.137-150
/
2007
Trees emit phytoncide into atmosphere to protect them from predation. Phytoncide from different trees has its own unique fragrance that is referred to as forest bath. Phytoncide, which is essential oil of trees, has microbicidal, insecticidal, acaricidal, and deodorizing effect. The present study was performed to examine the effect of phytoncide on Porphyromonas gingivalis, which is one of the most important causative agents of periodontitis and halitosis. P. gingivalis 2561 was incubated with or without phytoncide extracted from Hinoki (Chamaecyparis obtusa Sieb. et Zucc.; Japanese cypress) and then changes were observed in its cell viability, antibiotic sensitivity, morphology, and biochemical/molecular biological pattern. The results were as follows: 1. The phytoncide appeared to have a strong antibacterial effect on P. gingivalis. MIC of phytoncide for the bacterium was determined to be 0.008%. The antibacterial effect was attributed to bactericidal activity against P. gingivalis. It almost completely suppressed the bacterial cell viability (>99.9%) at the concentration of 0.01%, which is the MBC for the bacterium. 2. The phytoncide failed to enhance the bacterial susceptibility to ampicillin, cefotaxime, penicillin, and tetracycline but did increase the susceptibility to amoxicillin. 3. Numbers of electron dense granules, ghost cell, and vesicles increased with increasing concentration of the phytoncide, 4. RT-PCR analysis revealed that expression of superoxide dismutase was increased in the bacterium incubated with the phytoncide. 5. No distinct difference in protein profile between the bacterium incubated with or without the phytoncide was observed as determined by SDS-PAGE and immunoblot. Overall results suggest that the phytoncide is a strong antibacterial agent that has a bactericidal action against P. gingivalis. The phytoncide does not seem to affect much the profile of the major outer membrane proteins but interferes with antioxidant activity of the bacterium. Along with this, yet unknown mechanism may cause changes in cell morphology and eventually cell death.
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