• Title/Summary/Keyword: $CatR_I$ and $CatR_{II}$

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Function and Molecular Ecology Significance of Two Catechol-Degrading Gene Clusters in Pseudomonas putida ND6

  • Shi, Sanyuan;Yang, Liu;Yang, Chen;Li, Shanshan;Zhao, Hong;Ren, Lu;Wang, Xiaokang;Lu, Fuping;Li, Ying;Zhao, Huabing
    • Journal of Microbiology and Biotechnology
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    • v.31 no.2
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    • pp.259-271
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    • 2021
  • Many bacteria metabolize aromatic compounds via catechol as a catabolic intermediate, and possess multiple genes or clusters encoding catechol-cleavage enzymes. The presence of multiple isozyme-encoding genes is a widespread phenomenon that seems to give the carrying strains a selective advantage in the natural environment over those with only a single copy. In the naphthalene-degrading strain Pseudomonas putida ND6, catechol can be converted into intermediates of the tricarboxylic acid cycle via either the ortho- or meta-cleavage pathways. In this study, we demonstrated that the catechol ortho-cleavage pathway genes (catBICIAI and catBIICIIAII) on the chromosome play an important role. The catI and catII operons are co-transcribed, whereas catAI and catAII are under independent transcriptional regulation. We examined the binding of regulatory proteins to promoters. In the presence of cis-cis-muconate, a well-studied inducer of the cat gene cluster, CatRI and CatRII occupy an additional downstream site, designated as the activation binding site. Notably, CatRI binds to both the catI and catII promoters with high affinity, while CatRII binds weakly. This is likely caused by a T to G mutation in the G/T-N11-A motif. Specifically, we found that CatRI and CatRII regulate catBICIAI and catBIICIIAII in a cooperative manner, which provides new insights into naphthalene degradation.

Drug Resistance in Fish-Pathogenic Bacteria

  • Aoki, Takashi
    • Journal of fish pathology
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    • v.6 no.1
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    • pp.57-64
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    • 1993
  • The properties and DNA structures of R plasmids differ depending on the species of the fish-pathogens Aeromonas hydrophila, A. salmonicida, Edwardsiella tarda, Enterococcus seriolicida, Pasteurella piscicida and Vibrio anguillarum. However, some R plasmids with the same resistance markers in similar DNA structures were found in A. hydrophila and E. tarda, as well as in A. hydrophila and A. salmonicida. R plasmids from V. anguillarum were classified into three groups according to their DNA structures. The first group was detected before 1977, the second from 1980 to 1983, and the third from 1989 to 1991. R plasmids have been retained within P. piscieida having the same DNA structures and detected at various locations and times. E. seriolicida strains carrying the same R plasmids, which were encoded with resistance to macrolide antibiotics(MLs), lincomycin(LIM), and TC, and to MLs, LIM, and CP. were distributed in yellowtail farms in various districts. The chloramphenicol-resistance(cat) gene of the R plasmids of P. piscicida was classified as CAT type I. The cat of the R plasmids of E, tarda. A. salmonicida was classified as type II. The cat of R plasmids of V. anguillarum was classified into two types. One type detected before 1977, was classified as CAT IV and the other type, detected after 1980, was classified as CAT II. Tetracycline-resistance (tet) V. anguillarum, isolated before 1977 and after 1981, was classified as Tet B and Tet G, respectively. The class D tet gene was widely distributed in R plasmids from fish-pathogens A. hydrophila, E. tarda, P. piscicida, and also V. anguillarum isolated after 1989.

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Responses in Hepatic Xenobiotic Metabolizing and Antioxidant Enzymes in Javelin Goby Acanthogobius hasta Collected at Shihwa Lake (시화호에서 채집한 풀망둑 Acanthogobius hasta의 간장 약물대사효소계 및 항산화계의 반응)

  • Lee, Ji-Seon;Jeong, Jee-Hyun;Han, Chang-Hee;Shim, Won-Joon;Jeon, Joong-Kyun
    • Korean Journal of Environmental Biology
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    • v.26 no.2
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    • pp.94-101
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    • 2008
  • The aim of this study was to assess the responses of mixed function oxygenase (MFO) and antioxidative systems of feral Javelin goby, Acanthogobius hasta, caught in two sites of different pollution level in Shihwa lake, which has been a highly polluted lake by organic pollutants from nearby industrial complexes and sites. Enzymes analyzed in phase I of MFO system are cytochrome P450 (CYP), NADPH-cytochrome P450 reductase (P450R), NADH-cytochrome b5 reductase (b5R), and ethoxyresorufin deethylase (EROD). Phase II enzyme of glutathione S-transferase (GST) in MFO system was also investigated. Moreover, oxidative-enzyme system including catalase (CAT), glutathione reductase (GR) and total-glutathione peroxidase (GPX) activities and glutathione concentration in both of oxidized (GSSG) and reduced form (GSH) were determined. P450R, b5R, and GST activities of fish are relatively high in the polluted area, whereas hepatic EROD activity levels of fish in polluted area were lower than those of unpolluted area. CYP concentrations are not different between areas. These results indicated that feral Acanthogobius hasta were adaptive to highly polluted environment and exposed to oxidative stress in Shihwa lake.

Effect of Knee Joint Stimulation on the Activity of Phrenic Nerve and Inspiratory Nuron in the Cat (슬관절 자극이 횡격신경 및 흡식중추신경에 미치는 영향)

  • Cho, Dong-Ill;Han, Hee-Chul;Nahm, Sook-Hyun
    • Tuberculosis and Respiratory Diseases
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    • v.40 no.6
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    • pp.683-693
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    • 1993
  • Background: During movement the major inputs to nervous system come from firstly the muscle and joint to maintain posture and motion and secondly the chemoreceptors and baroreceptors to adjust the cardiovascular and respiratory function. Their complex relationships are generally studied for many years but the direct relation between the joint and respiratory system is not studied thoroughly until now. So this experiment was performed to determine whether the natural movement of knee joint can cause the enhancement of respiratory function by observation of the changes of respiratory rate, phrenic nerve activity and inspiratory neuron activity during the stimulation of knee joint in cat anesthetized with $\alpha$-chloralose. Method: Twenty six male adult cats were used and the extracelluar recording using bipolar platinum electrode and carbon filament electrode was done to record the changes in the activities of phrenic nerve and inspiratory neuron movement of knee joint, injection of chemicals into the joint cavity and electrical stimulation of articular nerve were done. Results: The 60 Hz. could not but 120 Hz. flexion-extension movement of knee joint increased respiratory rate(R.R.), tidal neural activity(TNA) and minute neural activity(MNA). Intra-articular injection of lactate could not increase R.R. but significantly increase TNA and MNA which represented the enhanced respiratory function. Injection of potassium chloride showed similar effects with the case of lactate but the duration of effect was shorter. The electrical stimulation of medial articular nerve with IV strength which could activate only group I and II afferents showed increased TNA and MNA during stimulation but 20 V stimulation which could activate all the afferents increased all the respiratory parameters. The changes of inspiratory neuron activity by knee joint stimulation was similar to that of phrenic nerve. Conclusion: The respiratory center could be directly stimulated by the activation of group I and II articular afferents and it seemed that the magnitude of the respiratory center enhancement is proportional to the amount of sensory information from the knee joint. These facts might suggest that the respiratory function could be enhanced even by the normal movement of knee joint.

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Use of Transgenic and Mutant Animal Models in the Study of Heterocyclic Amine-induced Mutagenesis and Carcinogenesis

  • Dashwood, Roderick H.
    • BMB Reports
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    • v.36 no.1
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    • pp.35-42
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    • 2003
  • Heterocyclic amines (HCAs) are potent mutagens generated during the cooking of meat and fish, and several of these compounds produce tumors in conventional experimental animals. During the past 5 years or so, HCAs have been tested in a number of novel in vivo murine models, including the following: lacZ, lacI, cII, c-myc/lacZ, rpsL, and $gpt{\Delta}$ transgenics, $XPA^{-/-}$, $XPC^{-/-}$, $Msh2^{+/-}$, $Msh2^{-/-}$ and $p53^{+/-}$ knock-outs, Apc mutant mice ($Apc^{{\Delta}716}$, $Apc^{1638N}$, $Apc^{min}$), and $A33^{{\Delta}N{\beta}-cat}$ knock-in mice. Several of these models have provided insights into the mutation spectra induced in vivo by HCAs in target and non-target organs for tumorigenesis, as well as demonstrating enhanced susceptibility to HCA-induced tumors and preneoplastic lesions. This review describes several of the more recent reports in which novel animal models were used to examine HCA-induced mutagenesis and carcinogenesis in vivo, including a number of studies which assessed the inhibitory activities of chemopreventive agents such as 1,2-dithiole-3-thione, conjugated linoleic acids, tea, curcumin, chlorophyllin-chitosan, and sulindac.

Role of the Salt Bridge Between Arg176 and Glu126 in the Thermal Stability of the Bacillus amyloliquefaciens ${\alpha}$-Amylase (BAA)

  • Zonouzi, Roseata;Khajeh, Khosro;Monajjemi, Majid;Ghaemi, Naser
    • Journal of Microbiology and Biotechnology
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    • v.23 no.1
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    • pp.7-14
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    • 2013
  • In the Bacillus amyloliquefaciens ${\alpha}$-amylase (BAA), the loop (residues 176-185; region I) that is the part of the calcium-binding site (CaI, II) has two more amino acid residues than the ${\alpha}$-amylase from Bacillus licheniformis (BLA). Arg176 in this region makes an ionic interaction with Glu126 from region II (residues 118-130), but this interaction is lost in BLA owing to substitution of R176Q and E126V. The goal of the present work was to quantitatively estimate the effect of ionic interaction on the overall stability of the enzyme. To clarify the functional and structural significance of the corresponding salt bridge, Glu126 was deleted (${\Delta}$E126) and converted to Val (E126V), Asp (E126D), and Lys (E126K) by site-directed mutagenesis. Kinetic constants, thermodynamic parameters, and structural changes were examined for the wild-type and mutated forms using UV-visible, atomic absoption, and fluorescence emission spectroscopy. Wild-type exhibited higher $k_{cat}$ and $K_m$ but lower catalytic efficiency than the mutant enzymes. A decreased thermostability and an increased flexibility were also found in all of the mutant enzymes when compared with the wild-type. Additionally, the calcium content of the wild-type was more than ${\Delta}E126$. Thus, it may be suggested that ionic interaction could decrease the mobility of the discussed region, prevent the diffusion of cations, and improve the thermostability of the whole enzyme. Based on these observations, the contribution of loop destabilization may be compensated by the formation of a salt bridge that has been used as an evolutionary mechanism or structural adaptation by the mesophilic enzyme.

Th e Effect of Sexuality Course on Sex - Role Stereotypes and Sexual Attitude in University Students (성 관련 강좌 이수 전후 대학생들의 성 고정관념과 성 태도의 변화)

  • Lee, Kye-Eun;Kim, Nam-Sun
    • Women's Health Nursing
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    • v.9 no.1
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    • pp.5-17
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    • 2003
  • Purpose : The purpose of this study was 1) to classify university student's attitudes toward sex, 2) to confirm the effects of sexuality course on sex-role stereotypes and sexual attitude in university student s. Method : The subjects in this study were 212 K university students in Kangwon Province between 3/7/2001 $\sim$ 6/13/2001. The instruments used for this study were the general characteristics, sexual attitude and sex-role stereotypes. The data was analyzed by frequency, percentage, mean, standard deviation, paired t-test, ANOVA, Pearson correlation coefficients, Factor analysis and Cronbach's $\alpha$ using the SPSS program. Result : 1. University students' attitude toward sex are divided into three types. Type I are called open mined : 16, 33, 28, 10, 18, 2, 30, 25, 26, 27, 22, 19, 29, 21, 9, 5 items. Type II are supporters of virginal purity : 8, 6, 31, 17, 14, 33 items. Type III express love through sexual intercourse : 20, 24, 7, 3, 15, 32 items. 2. There was a significant difference in the appearance and occupational characteristics of sex-role stereotypes before and after the sexuality course (t=2.562, p<.05). 3. In Type III, there was a significant difference in sexual attitude before and after the sexuality course (t=3.576, p<.0001). 4. The data showed the relationships between type III of sexual attitude and sex-role stereotypes (r=-.3 15, p<.0001). 5. Sex-role stereotypes according to the demographic characteristics before sexuality course were significantly different by age, gender, experience of military service, experience of sex edu cat ion an d experience of sexual intercourse. Sex-role stereotypes according to the demographic characteristics after sexuality course were significantly different by gender and experience of sexual intercourse. 6. Sexual attitude according to the demographic characteristics before sexuality course were significantly different by maj or (type I), age, grade, experience of military service and experience of sexual intercourse (type II), age, grade, gender, experience of military service, experience of sex education and experience of sexual intercourse. Sexual attitude according to the demographic characteristics after sexuality course were significantly different by religion and major(type I), age and grade(type II), age, grade, gender, religion and experience of sexual intercourse. Conclusion : This study showed that a sexuality course was effect ive in changing the sex-role stereotypes and sexual attitude of university students.

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Role of Catecholamines in Ventricular Fibrillation (Catecholamines에 관(關)하여 -제4편(第四編) : 심실전동발생(心室顫動發生)에 있어서의 catecholamines의 의의(意義)-)

  • Lee, Woo-Choo
    • The Korean Journal of Pharmacology
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    • v.19 no.1
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    • pp.15-35
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    • 1983
  • 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 $1^{\circ}C$ 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 $31^{\circ}C$, 70/min at $27^{\circ}C$ and 43/min at $23^{\circ}C$ if esophageal temperature. Ventricular fibrillation was occurred with no exception at a mean temperature of $20.3^{\circ}C(21-l9^{\circ}C)$. 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, $1.0\;{\mu}g/g$ wet weight compared to the prehypothermic value of $1.41\;{\mu}g/g$ 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 $16^{\circ}C$. 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 $19.6^{\circ}C$. By reserpine myocardial catecholamines content dropped to $0.045\;{\mu}g/g$ wet weight. 4) Bretylium pretreatment(20 mg/kg, i.m.), which blocks the release of catecholamines, Prevented the ventricular fibrillation under hypothermia in four of the eight cats. The pulse rate, however, was approximately the same as control and in some cases was rather slower. 5) Six cats treated with norepinephrine(2 mg/kg, i.m.) or DOPA(50 mg/kg) and tranylcypromine(10 mg/kg), which tab teen proved to cause significant increase in the catecholamines content of the heart muscle, showed ventricular fibrillation in all animals under hypothermia at average temperature of $21.6^{\circ}C$ and the pulse rate increased remarkably as compared with that of normal. Catecholamines content of cardiac muscle of these animals markedly decreased after hypothermia but higher than control animals. 6) The functional refractory periods of isolated rabbit atria, determined by the paired stimulus technique, was markedly shortened by administration of epinephrine, norepinephrine and isoproterenol. 7) Adrenergic beta-blocking agents, such as pronethalol, propranolol and sotalol(MJ-1999), inhibited completely the shortening of refractory period induced by norepinephrine. 8) Pretreatment with either phenoxftenbamine or phentolamine, an adrenergic alphatlocking agent, did not modify the decrease in refractory period induced by norepinephrine. From the above experiment it is possible to conclude that catecholamines play an important role in producing ventricular fibrillation under hypothermia. The shortening of the refractorf period of cardiac muscle induced by catecholamines mar be considered as a partial factor in producing ventriculr fibrillaton and to be mediated by beta-adrenergic receptor.

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