• Journal of Life Science
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• v.6 no.4
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• pp.241-249
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• 1996

A collagenase was isolated from the culture filtrate of Vibrio mimicus (ATCC 33658). The enzyme was purified to homogeneity by ammonium sulfate precipitation and DEAE-Sephadex A-50 chromatography, which an activity recovery of 22%. The molecular weight of the purified enzyme was estimated to be 42 kDa by SDS-polyacrylamide gel electrophoresis and gel filtration, indication a monomer structure. The optimum pH and temperature od the enzyme for insoluble collagen (Type I) were around 7.75 and 28$\circ$C, respectively. Some chelating agents and serine protease inhibitor inactivated the enzyme, but L-cysteine and histidine did not affect the activity. The amino acid composition indicated that the collagenase contained high amounts of amino acid residues of glycine and alanine. The K$_{m}$ and R$_{cat}$/K$_{m}$ values for the collagenase, using insoluble collagen (type I) as substrate, were 2.86 mg/ml and 972.28 U/mg-protein, respectively.

• Speech Sciences
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• v.15 no.2
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• pp.33-51
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• 2008

Difference of learner speech and loanword phonology is investigated in terms of Korean learners' speech and their loanword adaptation of English words with a post-vocalic word-final stop. When we compared the speech of 12 Korean learners in mid-intermediate level with that of eight English speakers, the learner speech did not reflect loanword phonology of the vowel insertion after a voiced word-final stop (e.g., rib$[\dotplus]$, bad$[\dotplus]$, gag$[\dotplus]$ vs. tip[=], cat[=], book[=]), but, instead, the target phonology of vowel lengthening before a voiced word-final stop (e.g., rib[r.I:b], CAD$[k{\ae}:d]$, bag$[b{\ae}:g]$ vs. rip[rI.p], cat$[k{\ae}t]$, back$[b{\ae}k])$. A longitudinal study of learner speech before and after instruction showed some development toward the acquisition of target phonology. The results indicate that learner speech departs from loanword phonology, and approaches to target speech in a faster rate than direct ratio. Thus, native phonology predicts loanword phonology, but lends little support to learner speech. Our results also indicate that loanword phonology is constant, while learner speech changes toward the acquisition of target phonology.

• Journal of Microbiology
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• v.44 no.6
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• pp.632-640
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• 2006

In this study, the biodegradative activities of monocyclic aromatic compounds were determined from the multi-drug resistant (MDR) Acinetobacter baumannii, which were studied in the form of clinical isolates from a hospital in Korea. These bacteria were capable of biodegrading monocyclic aromatic compounds, such as benzoate and p-hydroxybenzoate. In order to determine which pathways are available for biodegradation in these stains, we conducted proteome analyses of benzoate, and p-hydroxybenzoate-cultured A. baumannii DU202, using 2-DE/MS analysis. As genome DB of A. baumannii was not yet available, MS/MS analysis or de novo sequencing methods were employed in the identification of induced proteins. Benzoate branch enzymes [catechol 1,2-dioxygenase (CatA) and benzoate dioxygenase $\alpha$ subunit (BenA)] of the $\beta$-ketoadipate pathway were identified under benzoate culture condition and p-hydroxybenzoate branch enzymes [protocatechuate 3,4-dioxygenas $\alpha$ subunit (PcaG) and 3-carboxy-cis,cis-muconate cycloisomerase (PcaR)] of the $\beta$-ketoadipate pathway were identified under p-hydroxybenzoate culture condition, respectively, thereby suggesting that strain DU202 utilized the $\beta$-ketoadipate pathway for the biodegradation of monocyclic aromatic compounds. The sequence analysis of two purified dioxygenases (CatA and PcaGH) indicated that CatA is closely associated with the CatA of Acinetobacter radiresistance, but PcaGH is only moderately associated with the PcaGH of Acinetobacter sp. ADPI. Interestingly, the fused form of PcaD and PcaC, carboxymuconolactone decarboxylase (PcaCD), was detected on benzoate-cultured A. baumannii DU202. These results indicate that A. baumannii DU202 exploits a different $\beta$-ketoadipate pathway from other Acinetobacter species.

• The Korean Journal of Pharmacology
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• v.12 no.1
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• pp.45-55
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• 1976

The effect of positive inotropic agents on the contractile properties of myocardial muscle were studied in the cat papillary muscle preparation. For the purpose, the effects of ouabain $(1{\times}10^{-6}g/ml)$, norepinephrine (0.05r/m1) and Aconiti tuber butanol fraction (AF(5), $1{\times}10^{-4}$, $5{\times}10^{-4}$, $1{\times}10^{-3}$, $2{\times}10^{-3}g/ml$) on the contractile dynamics of the papillary muscle preparation isolated from right ventricle of cat were observed in terms of the characteristics of isometric twitch and the lengh-tension relation, the force-velocity relation and the load-extension relation of the series elastic component of contractile model of A.V. Hill. All the studied inotropic drugs similary increased the rate and the intensity of the developed isometric tension, while shortened the time from onset of contraction to peak tension and the total duration of contraction. In the afterloaded simultaneous isotonic and isometric contraction, they also similary increased the maximal velocity of shortening accompanied with the increasing the maximum developed force. In the load-extension relation all the drugs, however, had no appreciable influence on the properties of the series elastic component. Increasing the concentration, Aconiti tuber butanol fraction produced more pronounced effect on all the studied parameters of isometric and isotonic contraction of cat papillary muscle preparation. From the aspect of contractile dynamics, it seemed that the positive inotropic effect of ouabain, norepinephrine and Aconiti tuber butanol fraction are similary achieved through an influence on the behavior of the contractile component only.

• Parasites, Hosts and Diseases
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• v.60 no.2
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• pp.127-131
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• 2022

Feline hemotropic mycoplasmosis (hemoplasmosis) is an infection of the red blood cells caused by the Mycoplasma haemofelis (Mhf), Candidatus Mycoplasma haemominutum (CMhm), and Candidatus Mycoplasma turicensis (CMt). The existence of Mhf, CMhm, and CMt has been demonstrated in feral cats in Korea using molecular methods, but no clinical cases have yet been reported. This study reports 2 clinical cases of hemotropic mycoplasmosis caused by CMhm and CMt in 2 anemic cats. The first case was a client-owned intact female domestic shorthair cat that presented with fever, pale mucous membranes, and normocytic normochromic non-regenerative anemia. Prior to referral, an immunosuppressive prednisolone dose was administered at the local veterinary clinic for 1 month. The cat was diagnosed with high-grade alimentary lymphoma. Organisms were found on the surface of the red blood cells on blood smear examination. The second case was of a rescued cat that presented with dehydration and fever. The cat had normocytic normochromic non-regenerative anemia. Necropsy revealed concurrent feline infectious peritonitis. Polymerase chain reaction assay targeting 16S rRNA revealed CMhm infection in case 1 and dual infection of CMhm and CMt in case 2. Normocytic normochromic non-regenerative anemia was observed in both cats before and during the management of the systemic inflammation. This is the first clinical case report in Korea to demonstrate CMhm and CMt infections in symptomatic cats.

• Parasites, Hosts and Diseases
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• v.49 no.3
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• pp.207-212
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• 2011

Rapid serodiagnostic methods for Toxoplasma gondii infection in cats are urgently needed for effective control of transmission routes toward human infections. In this work, 4 recombinant T. gondii antigens (SAG1, SAG2, GRA3, and GRA6) were produced and tested for the development of rapid diagnostic test (RDT). The proteins were expressed in Escherichia coli, affinity-purified, and applied onto the nitrocellulose membrane of the test strip. The recombinant SAG1 (rSAG1) showed the strongest antigenic activity and highest specificity among them. We also performed clinical evaluation of the rSAG1-loaded RDT in 182 cat sera (55 household and 127 stray cats). The kit showed 0.88 of kappa value comparing with a commercialized ELISA kit, which indicated a significant correlation between rSAG1-loaded RDT and the ELISA kit. The overall sensitivity and specificity of the RDT were 100% (23/23) and 99.4% (158/159), respectively. The rSAG1-loaded RDT is rapid, easy to use, and highly accurate. Thus, it would be a suitable diagnostic tool for rapid detection of antibodies in T. gondii-infected cats under field conditions.

• Biomolecules & Therapeutics
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• v.23 no.2
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• pp.189-194
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• 2015

P450 1A2 is responsible for the metabolism of clinically important drugs and the metabolic activation of environmental chemicals. Genetic variations of P450 1A2 can influence its ability to perform these functions, and thus, this study aimed to characterize the functional significance of three P450 1A2 allelic variants containing nonsynonymous single nucleotide polymorphisms (P450 $1A2^*8$, R456H; $^*15$, P42R; $^*16$, R377Q). Variants containing these SNPs were constructed and the recombinant enzymes were expressed and purified in Escherichia coli. Only the P42R variant displayed the typical CO-binding spectrum indicating a P450 holoenzyme with an expression level of ~ 170 nmol per liter culture, but no P450 spectra were observed for the two other variants. Western blot analysis revealed that the level of expression for the P42R variant was lower than that of the wild type, however the expression of variants R456H and R377Q was not detected. Enzyme kinetic analyses indicated that the P42R mutation in P450 1A2 resulted in significant changes in catalytic activities. The P42R variant displayed an increased catalytic turnover numbers ($k_{cat}$) in both of methoxyresorufin O-demethylation and phenacetin O-deethylation. In the case of phenacetin O-deethylation analysis, the overall catalytic efficiency ($k_{cat}/K_m$) increased up to 2.5 fold with a slight increase of its $K_m$ value. This study indicated that the substitution P42R in the N-terminal proline-rich region of P450 contributed to the improvement of catalytic activity albeit the reduction of P450 structural stability or the decrease of substrate affinity. Characterization of these polymorphisms should be carefully examined in terms of the metabolism of many clinical drugs and environmental chemicals.

• Korean Journal of Plant Resources
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• v.26 no.1
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• pp.60-67
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• 2013

A laboratory experiment was conducted to determine the content of phenolics and flavonoids, antioxidant activity and antioxidant enzyme activity for the extracts from cowpea seed and sprouts. Plant length and weight of cowpea sprouts were significantly increased until 7 days after seeding. Total phenolics level [mg chlorogenic acid equivalents (CAE) $kg^{-1}$ DW] was highest in dry seed (DS) extracts of cowpea ($63.9mg\;kg^{-1}$), followed by imbibed seed (IS) ($56.8mg\;kg^{-1}$) and 1-day-old sprout (1DOS) extracts ($46.4mg\;kg^{-1}$), and significantly reduced with increase of sprout age (p < 0.05). The antioxidant activity of the methanol extracts from all the samples showed same tendency to the results of total phenolics level, and dose-dependently increased. DPPH (1,1-diphenyl-2-picryl hydrazyl radical) free radical scavenging activity was higher in DS (87.3%) and IS (41.2%) than in cowpea sprouts from 1DOS to 7DOS, ranging from 17.1 to 30.4%. Antioxidant enzymes, APX, POX, and POX activities were highest in 7DOS and lowest in DS. SOD activity showed much higher activity in sprouts and in seeds. Correlation coefficient between physiological-active substance and the activity was highest between APX and CAT activities ($r^2$= 0.9574). Especially, total phenolics content was more highly correlated with antioxidant or with antioxidant enzyme activities than was total flavonoid level.

• Microbiology and Biotechnology Letters
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• v.39 no.1
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• pp.93-96
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• 2011

It has been shown that a nucleotide substitution at position 770 in Escherichia coli 16S rRNA, which is implicated in forming the evolutionary conserved B2c intersubunit bridge, has a detrimental effect on ribosome function. In order to isolate second-site revertants that complement ribosomes containing C770G, we performed a random mutagenesis of the 16S rRNA gene and selected clones that could produce more CAT protein translated by specialized ribosome. One of the clones contained two nucleotide substitutions at positions 569 and 904 (C569G and U904C) and these mutations partially complemented the loss of protein-synthesis ability caused by C770G. Further studies using the isolated revertant will provide information about which part of 16S rRNA is interacting with C770 and the consequence of the structure formed by these interactions in the process of protein synthesis.

• Parasites, Hosts and Diseases
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• v.49 no.4
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• pp.445-448
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• 2011

A survey was performed to find out the intermediate hosts of Gnathostoma nipponicum in Jeju-do (Province), the Republic of Korea. In August 2009 and 2010, a total of 82 tadpoles, 23 black-spotted pond frogs (Rana nigromaculata), 7 tiger keelback snakes (Rhabdophis tigrinus tigrinus), 6 red-tongue viper snakes (Agkistrodon ussuriensis), and 2 cat snakes (Elaphe dione) were collected in Jeju-do and examined by the pepsin-HCl digestion method. Total 5 gnathostome larvae were detected in 3 (50%) of 6 A. ussuriensis, 70 larvae in 3 of 7 (42.9%) R. tigrinus tigrinus, and 2 larvae in 2 of 82 (8.7%) frogs. No gnathostome larvae were detected in tadpoles and cat snakes. The larvae detected were a single species, and $2.17{\times}0.22mm$ in average size. They had characteristic head bulbs, muscular esophagus, and 4 cervical sacs. Three rows of hooklets were arranged in the head bulbs, and the number of hooklets in each row was 29, 33, and 36 posteriorly. All these characters were consistent with the advanced third-stage larvae of G. nipponicum. It has been first confirmed in Jeju-do that R. nigromaculata, A. ussuriensis, and R. tigrinus tigrinus play a role for intermediate and/or paratenic hosts for G. nipponicum.