• Microbiology and Biotechnology Letters
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• v.37 no.3
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• pp.197-203
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• 2009

An indole oxygenase originated from Rhodococcus sp. RHA1 was cloned into the expression vector, pTrc99A, in Escherichia coli, and designated pTCAN1. The pTCAN2 was constructed from pTCAN1 by the deletion of $lacI^q$ for the constitutive expression of indole oxygenase without adding IPTG in the medium. The complete open reading frame of indole oxygenase was 1,224 bp long, which encodes a protein of 407amino acids. Crude extracts of E. coli $DH5{\alpha}$/pTCAN1 and pTCAN2, respectively, were prepared and subjected to SDS-PAGE analysis. A band corresponding to molecular mass of about 43 kDa was appeared and this result correlated with the predicted molecular mass of cloned indole oxygenase. The E. coli harboring pTCAN1 and pTCAN2, respectively, showed blue color colony in LB plate. The pigment showing blue color was prepared from E. coli $DH5{\alpha}$/pTCAN2, and identified as indigo by experiments using spectrophotometer, HPLC, and TLC. The indigo-forming activity of indole oxygenases from the whole cell of E. coli $DH5{\alpha}/pTCAN1$ cultured at LB medium added 1mM of IPTG and that of E. coli/pTCAN2 showed about 1.75nmol/min/mg DCW (dry cell weight) and 3.85 nmol/min/mg DCW, respectively. Also, the E. coli $DH5{\alpha}$/pTCAN2 produced about $236{\mu}M$ of indigo after 48 hours incubation in TB medium supplemented with 2.5 mM of tryptophan.

• Journal of Microbiology and Biotechnology
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• v.28 no.4
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• pp.588-596
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• 2018

An endo-${\beta}$-1,4-glucanase gene, cel9K, was cloned using the shot-gun method from Paenibacillus sp. X4, which was isolated from alpine soil. The gene was 2,994 bp in length, encoding a protein of 997 amino acid residues with a predicted signal peptide composed of 32 amino acid residues. Cel9K was a multimodular enzyme, and the molecular mass and theoretical pI of the mature Cel9K were 103.5 kDa and 4.81, respectively. Cel9K contains the GGxxDAGD, PHHR, GAxxGG, YxDDI, and EVxxDYN motifs found in most glycoside hydrolase family 9 (GH9) members. The protein sequence showed the highest similarity (88%) with the cellulase of Bacillus sp. BP23 in comparison with the enzymes with reported properties. The enzyme was purified by chromatography using HiTrap Q, CHT-II, and HiTrap Butyl HP. Using SDS-PAGE/activity staining, the molecular mass of Cel9K was estimated to be 93 kDa, which is a truncated form produced by the proteolytic cleavage of its C-terminus. Cel9K was optimally active at pH 5.5 and $50^{\circ}C$ and showed a half-life of 59.2 min at $50^{\circ}C$. The CMCase activity was increased to more than 150% in the presence of 2 mM $Na^+$, $K^+$, and $Ba^{2+}$, but decreased significantly to less than 50% by $Mn^{2+}$ and $Co^{2+}$. The addition of Cel9K to a commercial enzyme set (Celluclast 1.5L + Novozym 188) increased the saccharification of the pretreated reed and rice straw powders by 30.4% and 15.9%, respectively. The results suggest that Cel9K can be used to enhance the enzymatic conversion of lignocellulosic biomass to reducing sugars as an additive.

• Journal of Veterinary Clinics
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• v.16 no.2
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• pp.375-380
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• 1999

This study was conducted to develop an intrauterine inseminator (IUI) to deposit of frozen semen into uterus and to evaluate the results obtained after artificial insemination by IUI. Two Japanese spitzs (2 to 4 years of age) were used as semen donors. Semen was collected by manual masturbation into sterile glass collection tubes and separated into 3 fractions with only the sperm-rich fractions retained for further examination. Sperm motility >70%, sperm concentration of 200 to $400{\times}10^6 cells/ml$$\times$g for 5 min and poured out the suspended solution, and then diluted with 2 ml Tris-buffer which was consisted of 2.4 g Tris, 1.4 g citric acid, 0.8 g glucose, 0.1 $\mu\textrm{g}$/ml streptomycin, 100 IU/ml penicillin, 20 ml egg yolk to 100 ml mili-Q water (Ext I) or supplemented with 8 ml glycerol and 1 ml Equex STM paste to 100 rnl (Ext II). The diluted semen was cooled to 5$^{\circ}C$ in cold room, where the temperature in the sample reached 5$^{\circ}C$. Two h after beginning the cooling procedure, 2 ml of Ext II, also at 5$^{\circ}C$, was added and mixed by gently reversing the tubes several times during 1 h. The final sperm concentration for freezing was approximately $50{\times}10^6 cells/ml$. After equilibration, the semen was loaded into 0.5 ml straw and frozen on the liquid nitrogen vapour in styrofoam box. The straws were thawed at 7$0^{\circ}C$ for precisely 6 sec. After thawing of each straw, the frozen semen can survived over 50% motility. All the females were inseminated twice with 1 ml of $25{\times}10^6 cells/ml$

• Journal of Oral Medicine and Pain
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• v.38 no.4
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• pp.291-298
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• 2013

On this study, we treated rats with restraint stress, and observed the changes with an optical microscope. Within the salivary gland tissue, we measured cell apoptosis cycle evaluation which show positive reaction on TUNEL assay, and compared within the groups. For this study, 18 rats were divided into 3 groups; 1) 2 rats of group I were selected as a normal control. 2) 2 rats of group II, as a experimental control were placed in the restraint cone for 2 hours 3) 14 rats of group III were placed in the restraint cone for 2 hours once a day. The rats were sacrificed immediately (group II, as a experimental control), 1, 2, 3, 4, 5, 6 and 7days after application of the stress and the both parotid glands were excised. The conclusions follow. 1. 5 days after giving an confining stress to the parotid gland of Rats, we can observe the hypotropy and pus and inflammation of Rat parotid gland acinar cells, and after 7 days, we can see a cell apoptosis. 2. Through the In situ DNA end labeling assay and TUNEL dye, on serous glands, benign tumor cell increased with statistically significant result after 5 days from confining stress. And the index shows maximum value on 7th days, which is same result with histological opinion. Therefore, our study shows that a cell apoptosis can be induced by restraint stress on salivary gland tissue, and we think more study should be accomplished about the cell signaling pathway in the future.

• Journal of The Korean Dental Society of Anesthesiology
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• v.7 no.1
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• pp.6-12
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• 2007

배경: 소아 진정 치료 시 서로 다른 용량의 chloral hydrate와 hydroxyzine을 복용 후 midazolam을 구강 점막 하 주사했을 때 행동 반응을 비교하였다. 방법: 총 32회 진정법을 통해 치과치료를 받은 30명의 나이 24-72개월, 체중 20 kg 미만의 미국 마취과학회 신체등급 I의 건강하지만 겁이 많고 협조가 안 되는 소아 환자를 대상으로 2개 치아이상의 보존 치료 및 발치를 필요로 하는 환자를 대상으로 하였다. 호흡기 질환이 있는 아이들은 이 연구에서 제외되었다. 연구 계획은 이대 목동 병원의 임상 실험 심사 위원회에 제출되었다. 1군은 chloral hydrate 50 mg/kg와 hydroxyzine 1 mg/kg 복용 후 점막 하 midazolam 0.2 mg/kg을 추가 투여했고 2군은 chloral hydrate 60 mg/kg와 hydroxyzine 1 mg/kg 복용 후 점막 하 midazolam 0.1 mg/kg을 주사 받았다. 50% nitrous oxide는 치료 중 두 군 모두 유지되었다. 전날 수면 시간과 약물 복용 태도를 기록하였으며 모든 치료 과정은 비디오로 촬영되었다. 맥박 산소 계측기를 이용하여 경피적 산소 포화도와 맥박수를 기록하였고 행동 반응은 Houpt scale을 이용하여 매 2분마다 40분 동안 기록되었다. 전반적인 행동 반응은 Houpt scale를 이용하여 평가되었다. 모든 자료는 SPSS 통계 프로그램을 이용하여 two sample independent t-test를 사용하였다. P 값은 0.05 미만인 경우를 통계학적으로 유의하다고 보았다. 결과: 두 군 간의 경피적 산소 포화도와 맥박수는 모두 정상 범위이며 유의한 차이가 없었다. 행동 반응 비교에서는 치료 처음 10분 동안 2군이 1군에 비해 점수가 높게 나왔으며(P < 0.05), 그 외에는 유의한 차이가 없었다. 전날 총 수면 시간과 약물을 복용하는 태도는 수면 치료 중의 행동 반응에 영향을 주지 않았다. 결론: Chloral hydrate 50 mg/kg 복용과 점막 하 midazolam 0.2 mg/kg은 chloral hydrate 60 mg/kg 복용과 점막 하 midazolam 0.1 mg/kg과 비교할 때 두 약물의 조합은 모두 소아 환자 수면 치료시 안전하고 효과적인 용량이다. Overall behavior와 Q (quiet)의 분포를 비교해 볼 때 두 군 모두 성공적인 진정효과를 기대할 수 있다.

• The Journal of Korean Institute of Electromagnetic Engineering and Science
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• v.21 no.10
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• pp.1169-1176
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• 2010

The RF front-end for L1/L2 dual-band Global Positioning System(GPS) receiver is presented in this paper. The RF front-end(down-converter) using low IF architecture consists of a wideband low noise amplifier(LNA), a current mode logic(CML) frequency divider and a I/Q down-conversion mixer with a poly-phase filter for image rejection. The current bleeding technique is used in the LNA and mixer to obtain the high gain and solve the head-room problem. The common drain feedback is adopted for low noise amplifier to achieve the wideband input matching without inductors. The fabricated RF front-end using $0.18{\mu}m$ CMOS process shows a gain of 38 dB for L1 and 41 dB for L2 band. The measured IIP3 is -29 dBm in L1 band and -33 dBm in L2 band, The input return loss is less than -10 dB from 50 MHz to 3 GHz. The measured noise figure(NF) is 3.81 dB for L1 band and 3.71 dB for L2 band. The image rejection ratio is 36.5 dB. The chip size of RF front end is $1.2{\times}1.35mm^2$.

• Korean Journal of Poultry Science
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• v.33 no.4
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• pp.317-321
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• 2006

The effects of dietary humic substances (HS) on egg Production and egg Quality were studied using 252(55-wk old) ISA brown laying hens. laying were divided into 21 groups of 12 hens each and seven groups (experimental units) were assigned to 1) CON (basal diet), 2) HS5 (basal diet 4- 5% humic substances) or 3) HS10 (basal diet +10% humic substances) in a completely randomized block design. Hens had free access to diets and water fur 6 wk. Egg Production and egg quality were monitored over the 6-wk Period. Results showed that 10% dietary HS decreased egg Production and yolk diameter (P<0.05) compared to CON. Egg weight and yolk cole. were improved (P<0.05) in HS10 compared to CON. Egg shell breaking strength was increased significantly (P<0.05) when hens were fed HS5 diet compared to the others. There were no effects of treatments on egg shell thickness, yolk index, albumen height and Haugh nit. The results suggest that the dietary supplementation of HS at 5% or 10% decreases egg Production, but HS at 5% can increase egg shell breaking strength. Hens fed 10% HS could increase egg weight and yolk color and decrease yolk diameter.

• Neonatal Medicine
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• v.17 no.2
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• pp.254-261
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• 2010

Little is known about neonatal mitochondrial disease, though mitochondrial metabolic disorders may often present in the neonatal period because of the high energy requirement of neonate. In newborn period, common presentations are not specific and the disease course may be rapid and fatal. In this study, we report three cases of neonatal mitochondrial disease. The first case was strongly suspected because of sudden seizure and mental change with severe lactic acidosis, and multiorgan failure. Plasma lactate/pyruvate (L/P) ratio was increased to 55.6 with marked lactic aciduria and increased plasma alanin up to 2,237 nmol/mL. In the second patient, a peritoneal dialysis was performed for acute adrenal and renal failure, but metabolic acidosis persisted. Plasma L/P ratio was increased to 23.9, and MRC I (mitochondrial respiratory chain defect) was diagnosed through the enzymatic analysis of the muscles. The third case showed repetitive episode of lactic acidosis during the first two months of life, hypotonia, failure to thrive and feeding difficulties. We found markedly increased cerebrospinal fluid L/P ratio up to 57 though plasma L/P ratio(19.4) was borderline with increased plasma lactate. The lactate peak was prominent in brain magnetic resonance spectroscopy (MRS). MRC II was confirmed through muscle biopsy. Plasma lactate level and lactate peak of brain MRS were normalized after conservative treatment.

• Journal of Life Science
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• v.14 no.5
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• pp.847-854
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• 2004

Allergy is a multi-factorial disease influenced by genetic and environmental factors. As the number of allergy-affected people is increasing in developed countries, there is an increasing interest in genetic predisposition to the allergy. A number of genes and chromosomal region have been identified to be linked to allergy including rhinitis, asthma and atopy. In order to understand the genetic background for the allergy-affected people, we investigated genetic predisposition among students enrolled in Busan Science Academy. Among 138 students, about 30% students had some allergy-related disorder including rhinitis, asthma and atopy. We analyzed several single nucleotide polymorphisms (SNPs) within two genes, Inter-leukin-4(IL-4) and Interleukin-4 receptor(IL-4R), which are involved in the induction of allergy reaction with the Th2 immunity. For 96 samples obtained from students, we analyzed 9 SNPs including -590 C/T and -34 C/T in IL-4, and I75V, Q576R, E375A, e406R, 5411L, S761P and S727A in IL-4R. From the analysis, these SNPs showed slight differences among normal and allergy-affected students, but these differences was not enough to predict the predisposition to the allergy. In contrast to previous reports, we could not find SNP(s) related with allergy. These results suggest that genetic tests recently performed in Korea widely have to be reassessed for its validity of genetic predisposition. [Supported by grants from MOST]

• Journal of Plant Biology
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• v.13 no.1
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• pp.33-46
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• 1970

Pinus densiflora stands are common secondary forest communities on infertile soils in Korea. The stands are results of long severe past biotic pressure such as cutting, burning and grazing. These could be regarded as biotic climax in Korea. Because of their prevalent occurrence, relatively simple species and age composition, and their domestic economic importance, study of their distributional patterns may give some basic knowledge for better utilization of land resources in Korea. To detect distributional patterns and interspecific associations ten pine stands, each of which was homogenious with respect to topography and physiognomy, were subjectively selected from pine stands in Kyunggi Province near Seoul in 1969 and were made object of this study. Four contiguous systematic samples of count for trees, shrubs and seedlings from belt transects were collected from homogeneous areas within ten natural pine stands. The belt transect was 64m or 128m in length, and 1m, 2m or 4m in width. Basic units within the transect ranged from 64 to 256. The data from the contiguous transects were analysed in terms of multiple split-plot experiment. Departure from randomness of stem distribution, i.e., pattern, was tested in terms of variance mean ratio. For the detection of association between species, correlation coefficient was calculated for different block sizes. The values of ${\gamma}$ were tested by the usual t-test. Fine trees within one of the stands showed significant regular distribution through out the blocks. Within other eight stands pines were randomly distributed at basic unit with 4$\times$4m, 2$\times$2m, 2$\times$1m and 1$\times$1m. One significantly clumped distribution at basic unit 2$\times$2m, however, was observed from one of the pine stands. These randomly distributed groups were themselves significantly regularly distributed throughout the blocks for four pine stands. For the other four pine stands, in addition to the random distribution at the basie unit(the primary random group), randomly distributed groups with 32m dimension(the secondary random groups) were also observed. Both the primary and the secondary random groups were significantly regularly distributed at the rest of blocks. Pine seedlings were not distributed randomly thoughout the blocks. Within three of the ten stands they were contagiously distributed. Important shrub species underneath pines such as Querus serrata, Q. acutissima, Leapedeza intermedia, Rhododendron Yedoense var. poukhanenae, Juniperus utilis, Rhododendron mucronulatum var. ciliatum shnwed consistently similar distributional pattern with the pine at each stand. The shrub species pairs; Rhododendron Yedoense var. poukhanenae/Quercus serrata, Rhododendron mucronulatum var. ciliatum/Lespedeza intermedia were significantly negatively associated from 1m to 4m dimensional block sizes but became significantly positively associated from 8m sized blocks on. On the other hand the shrub species pairs; Lespedeza intermedia/Robinia Pseudoacacia, and Lespedeza bicolor var, japonica/Lespedeza intermedia were also significnalty negatively associated from 1m to 8m sized blocks but became significantly positively associated from 16m sized blocks on. The associational pattern between Rhododendron mucronul tun var. poukhanenae and Lespedeza intermedia was not consistent throughout the stands. In some stands negative associations were observed throughout the blocks except NS 32. From these observatons micro-edaphic variation within the pine stands seems not to be great enough to cause distributinal difference of pine trees within the ten pine stands. Among each species and pine seedings, however, the edaphic variation within the pine stands may be great enought to cause distributional variation.