• Journal of Korean Neurosurgical Society
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• 제37권4호
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• pp.287-292
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• 2005

Objective: Recent clinical studies have demonstrated that intracisternal administration of recombinant tissue plasminogen activator(rt-PA) can facilitate the normal clearing of blood from the subarachnoid space. Urokinase, a first generation fibrinolytic agent, has been used to liquify such clots with some success. Therefore, recombinant tissue plasminogen activator, a second generation fibrinolytic drug that may be safer and more effective, is studied to evaluate its dosage to lyse clots in vitro and reactivity in the brain parenchyme. Methods: Intracerebral hematomas were created by stereotactically injecting 2ml of clotted autogenous blood into the brain parenchyme of total 28 anesthetized adult cats (weighting 3.8 to 4.1 kg). The control animals (group A) received 1 ml of normal saline injected into the clots and the experimental animals received each 0.1 mg of rt-PA (group B), 0.5mg of rt-PA (group C) and 1 mg of rt-PA (group D) at 6 hours after the clot injection. Results: 1. The amount of remained clots after lysing the hematomas were as follows: $1.80{\pm}0.17ml$ in group A, $1.65{\pm}0.23ml$ in group B, $0.61{\pm}0.37ml$ in group C and $0.52{\pm}0.34$ in group D. The result indicated that hematomas in rt-PA treated groups (C & D) were lysed better than the control group. 2. At least 0.5mg of rt-PA should be required for the lysis of 2ml of hematomas. 3. Light microscopic examination revealed no histological evidence of hemorrhage in tissue sections from each brain. Conclusion: Recombinant tissue plasminogen activator may be safely and effectively employed for the lysis of intracerebral hematomas in animal model.

• Journal of the Korean Wood Science and Technology
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• 제16권4호
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• pp.3-9
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• 1988

Infrared (IR) spectroscopic techniques for the analysis of wood samples and the absorbance spectra of solid woods were presented. KBr pellets were prepared by throughly mixing approximately 300 mg of dried KBr and 1 mg of finely milled wood powder extracted with ethanol-cyclohexane previously. This mixture was made into a transparent disc by means of a pellet-making die (10 ton/$cm^2$ for 10 min). This IR techniques were applied for the analysis of archaeological wood samples. The most notable difference in the IR spectra between the recent and the archaeological waterlogged woods is that the absorption band centered at $1,730cm^{-1}$ was significantly diminished in the waterlogged ones. Total loss of absorption in $1,730cm^{-1}$ might be mainly due to the result of hemicellulose degradation. Another feature indicated by IR spectral comparision are that the degraded waterlogged wood samples showed 1) the increased intensity of the 1,600, 1,500 and $1,270cm^{-1}$ due to the residual lignin and the increased intensity at 1,470 and $1,425cm^{-1}$ due to the degradation of hemicellulose and 2) to the emergence of single band around $1,050cm^{-1}$ instead of three bands at 1,110, 1,060 and $1,040cm^{-1}$ in recent wood due to the degradation of cellulose crystalline. It was revealed from the IR examinations that the first change of wood in the waterlogged situation was the lysis of hemicellulose and the second the lysis of cellulose. It was also suggested that IR spectroscopy could serve a fast method for the investigation on the chemical characteristics of archaeological wood samples.

• Molecules and Cells
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• 제22권1호
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• pp.119-125
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• 2006

The rat is an accepted model for studying human psychiatric/neurological disorders. We provide a protocol for total soluble protein extraction using trichloroacetic acid/acetone (TCA/A) from rat (female) whole brain, 10 brain regions and the pituitary gland, and show that two-dimensional gel electrophoresis (2-DGE) using precast immobilized pH (4-7) gradient (IPG) strip gels (13 cm) in the first dimension yields clean silver nitrate stained protein profiles. Though TCA/A precipitation may not be "ideal", the important choice here is the selection of an appropriate lysis buffer (LB) for solubilizing precipitated proteins. Our results reveal enrichment of protein spots by use of individual brain regions rather than whole brain, as well as the presence of differentially expressed spots in their proteomes. Thus individual brain regions provide improved protein coverage and are better suited for differential protein detection. Moreover, using a phosphoprotein-specific dye, ingel detection of phosphoproteins was demonstrated. Representative high-resolution silver nitrate stained proteome profiles of rat whole brain total soluble protein are presented. Shortcomings apart (failure to separate membrane proteins), gel-based proteomics remains a viable option, and 2-DGE is the method of choice for generating high-resolution proteome maps of rat brain and brain regions.

• 미생물학회지
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• 제21권3호
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• pp.115-126
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• 1983

The results that the effect of 6 detergents on the structural changes and biochemical composition of bacterial membrane of Escherichia coli and Bacillus cereus are as follows ; 1. Population growth of the bacteria was increased in case of the treatment with palmitoyl carnitine and sodium deoxy cholate but was increased in case of the treatment with palmitoyl carnitine and sodium deoxy cholate but was decreased by sodium dodecyl sulfate and palmitoyl choline, in E.coli and was decreased by palmitoyl carnitine and palmitoyl choline at the low concentration, in B. cereus. 2. The electron micrograph showed that cell wall lysis or cell collapse were observed in the treatment of sodium dodecyl sulfate and palmitoyl choline, and also cell wall was condensed by triton X-100 and sodium deoxy cholate, in E.coli. And in B. cereus, endospore formation of the bacteria was stimulated by palmitoyl choline, and cell lysis or structural changes of the membrane were observed in the treatment of sodium dodecyl sulfate, sodium cholate, and triton X-100, respectively. 3. As to the effect of detergent on the biochemical composition of biomembrane, the content of carnitine, in E.coli, and B.cereus, the content of structural protein and phospholipid were decreased by treatment of sodium dodecyl sulfate and structural protein was denatured by palmitoyl choline. 4. The profile of membrane protein revealed that the bacterial membrane were composed of various proteins. By dint of this result, some of membrane proteins were solubilized or changed to small molecules by the treatment of sodium dodecyl sulfate and palmitoyl choline, in E.coli and membrane protein of the biomembrane by treatment of sodium dodecyl sulfate, sodium deoxy cholate, palmitoyl choline, and palmitoyl carnitine were confirmed to be different profile as compared with those of the control, in B. cereus. Therefore, it is suggested that sodium dfodecyl sulfate and palmitoyl choline soulbilized biomembranes or inhibited membrane transport and that palmitoyl carnitine and sodium deoxy cholate were used as an energy source or stimulating the membrane transport, in E.coli. And, it is suggested that all of detergents were inhibited biomembrane synthesis, expet saponin, in B.cereus.

• 한국어병학회지
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• 제17권2호
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• pp.123-130
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• 2004

꿀벌 (Apis mellifera)의 lymph 액으로부터 정제된 항균성 펩타이드인, Apidaecin Ⅰb는 18개의 아미노산 잔기로 구성된 염기성 non-helical 펩타이드이다. 본 연구에서는 우리나라 연안의 적조 (HABs, harmful algal blooms)를 일으키는 4종의 적조생물 (Alexandrium tamarense, Chattonella marina, Cochlodinium polykrikoides 및 Gymnodinium catenatum)에 대한 Apidaecin Ⅰb의 살조효과를 조사하였다. Apidaecin Ⅰb의 4종의 적조생물에 대한 살조효과는 12.5$\mu{g}/mL$부터 50$\mu{g}/mL$에서 세포의 lysis 또는 ecdysis와 같은 형태로 현미경으로 관찰할 수 있었다. 또한 Apidaecin Ⅰb는 C. marina에 대해서 A. tamarense, C. polykrikorides 및 G. catenatum보다 더욱 강한 살조효과를 나타내었다. 이러한 HABs에 대한 Apidaecin Ⅰb의 살조효과 연구는 새로운 살조물질을 개발하기 위한 자료가 될 것으로 생각된다.

• 한국식품영양과학회지
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• 제29권6호
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• pp.1025-1029
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• 2000

방사선 조사된 쇠고기와 돼지고기의 방사선 조사 여부를 판별하는데 DNA comet assay의 활용 가능성을 검토하였다. 쇠고기와 돼지고기는 Co-60 동위 원소를 조사원으로 하여 0.1, 0.3, 0.5, 0.7, 1.0 kGy의 총흡수선량( $\pm$ 5.0%)이 되도록 조사하여, 냉동상태로 보관하였다. 시료로부터 분리 된 세포는 agarose gel과 혼합하여 슬라이드에 깔아주고, lysis 및 전기영동을 하였다. 방사선 조사된 시료의 경우 세포로부터 끌려나오는 DNA절편들은 양극을 방향으로 tail이 형성되었고, 비조사 시료의 경우 tail이 없거나, 일부에서만 작은 tail이 관찰되었다. 방사선 조사유무는 0.1 kGy부터 현미경상으로 검지가 가능하였고, 선량간 차이의 유의성 여부는 통계분석을 통하여 검지가 가능하여, 쇠고기와 돼지고기의 신속한 검지 방법으로 DNA comet assay를 활용할 수 있을 것이다.

• 한국식품과학회지
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• 제31권6호
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• pp.1628-1634
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• 1999

Listeria monocytogenes의 식품 속에서 신속하고 특이적인 검출을 위하여, listeriolysin O gene에 의한 primer, LM 1과 LM 2를 선택하여 PCR을 수행하였다. L. monocytogenes의 DNA 추출이나 cell lysis 없이 intact whole cell을 직접 이용하여 PCR을 하였으며 $10^{2-6}$ CFU 수준의 균체 배양액으로부터 L. monocytogenes에 특이적인 702 bp의 PCR 증폭 산물을 확인하였다. 우유, 닭고기, 김치 등의 식품에 L. monocytogenes를 접종하여 증균배양 전후 균의 PCR에 대한 감도와 생균수를 비교한 결과, 실험된 식품 속에서의 L. monocytogenes의 검출 감도는 순수 배양액에서의 경우에 비해 약 1/10로 둔화되었으나 역시 특이적인 검출이 가능하였다. Primer LM 1과 2를 이용한 본 실험조건에서의 L. monocytogenes의 PCR에 의한 검출은 약 4시간으로 확인이 가능하였으며, 기존의 배양 방법에 비해, 특이성이나 검출 속도 면에서 식품위생 실무에 적용하기 위한 높은 잠재력을 보여 주었다.

• Reproductive and Developmental Biology
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• 제30권3호
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• pp.189-194
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• 2006

Sperm-mediated gene transfer(SMGT) can be used to transfer exogenous DNA into the oocyte at fertilization. The main objective of this study was to assess efficiency of transferring mitochondrial DNA(mtDNA) fragment into boar spermatozoa in either presence or absence of liposome and quality of transfected spermatozoa. The mtDNA of chicken liver was isolated and purified by phenol and alkaline lysis extraction, and it was inserted to plasmid. The genome of transfected spermatozoa treated with DNase I was purified by alkaline lysis, and then amplified by the PCR analysis. After electrophoresis, DNA quantitation of each well was calculated by comparison of the band intensity with standard. As a result, exogenous DNA was composed of mtDNA fragment(1.2 kb) and plasmid(2.7 kb). On the other hand, efficiency of transfection by liposome($9.0{\pm}0.34ng/{\mu}l$) in SMGT was higher than that by DNA solution($6.9{\pm}0.53ng/{\mu}l$). However, there was no significant difference. Transfering exogenous DNA into spermatozoa was completed within 90 min of incubation. In another experiment, there were significant (p<0.05) differences between transfected spermatozoa using both DNA solution and DNA/liposome completes with unheated spermatozoa for viability ($70.8{\pm}1.80$ and $68.0{\pm}2.16%$ vs. $83.3{\pm}1.69%$, respectively) and motility($78.7{\pm}1.59$ and $79.3{\pm}2.14%$ vs. $86.7{\pm}1.59%$, respectively). This study indicates that exogenous mtDNA can be efficiently transferred into boar spermatozoa regardless of the presence of liposome, and transfected spermatozoa can also use insemination and in vitro fertilization to generate transgenic pig.

• 한국미생물·생명공학회지
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• 제13권3호
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• pp.289-296
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• 1985

본 연구는 유산간균 및 유산구균으로부터 plasmid DNA를 신속하고 간편하게 분리하기 위한 방법에 관한 것이다. 세포벽 형성 억제 인자인 glycine을 0.5% 첨가한 TCM 배지에서 유산균을 배양하였고 plasmid DNA는 mutanolysin을 처리한 cells로부터 alkaline-detergent lysis 법으로 분리되었다. 유산간균은 효소 처리 때 mutanolysin의 농도를 30$\mu\textrm{g}$/$m\ell$로 하고 37$^{\circ}C$에서 5-10분간 반응되었을 때 plasmid DNA가 아주 잘 추출되었다. 유산구균의 경우는 그 최적 조건이 조금 달랐다. 본 방법은 L. casei, L. acidophilus, L. helveticus, S. lactis, S. faecalis, S. faecium과 S. cremoris 균주로부터 plasmid DNA를 신속하게 분리하는데 사용할 수 있었다. 본 방법을 이용하여 배양액 $1.5m\ell$로부터 분리된 plasmidsDNA가 gel상에서 쉽게 확인될 수 있었다.

• 한국식품영양과학회지
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• 제12권4호
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• pp.387-394
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• 1983

zymolyase(${\beta}-1$, 3-glucanase)의 효모(酵母) 세포벽(細胞壁)에 대한 용해작용(溶解作用)을 촉진(促進)시키는 인자(因子)를 zymolyase의 조효소(粗酵素)로부터 분리(分離)하여 Sephadex G-75 gel 여과(濾過) 및 조제용(調製用) PAG-전기영동법(電氣泳動法)에 의하여 전기영동적(電氣泳動的)으로 균일(均一)하게 정제(精製)하였다. SDS-PAG 전기영동법(電氣泳動法)에 의하여 측정한 정제(精製) 인자(因子)의 분자량(分子量)은 40,500이었고, 등전점(等電點)은 pH 9.6이었다. 정제(精製)된 촉진인자(促進因子)는 395개(個)의 아미노산 잔기(殘基)로 구성(構成)되는 단일(單一) polypeptide의 염기성(鹽基性) protease임이 밝혀졌으며, 이 protease의 흡광계수(吸光係數)($E_{208,cm}^{1%}$)는 11.9, 분자흡광계수(分子吸光係數)(${\varepsilon}_{280,}mole/l$)는 $4.83{\times}10^4$이었다.