• The Korean Journal of Physiology and Pharmacology
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• 제3권2호
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• pp.199-205
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• 1999

Vibrio vulnificus cytolysin caused platelet cytolysis and increased intracellular calcium concentration $([Ca^{2+}]_i)$ of rat platelets in a concentration-dependent manner. In the presence of V. vulnificus cytolysin (3 HU/ml), lactate dehydrogenase (LDH) activity was increased from $1.3{\pm}0.4%$ of control to $64.3{\pm}3.4%$ in platelet suspension buffer. In $Ca^{2+}-free$ platelet suspension buffer, however, V. vulnificus cytolysin did not induce $[Ca^{2+}]_i$ increase and LDH release. Addition of EGTA (2 mM) to suspension buffer after the initial $Ca^{2+}$ influx reversed $[Ca^{2+}]_i$ to the control level. However, a $Ca^{2+}$ channel blocker verapamil $(20\;{\mu}M)$ or mefenamic acid $(20\;{\mu}M)$ did not inhibit V. vulnificus cytolysin-induced $[Ca^{2+}]_i$ increase and LDH release. Divalent cations such as $Co^{2+},\;Cd^{2+}\;or\;Mn^{2+}$ (2 mM each) also did not alter V. vulnificus cytolysin-induced $[Ca^{2+}]_i$ increase and LDH release. V. vulnificus cytolysin (3 HU/ml)-induced calcium influx was completely blocked by lanthanum (2 mM). Lanthanum (2 mM) also completely blocked V. vulnificus cytolysin (3 HU/ml)-induced LDH release. Osmotic protectants such as, raffinose, sucrose or PEG600 (50 mM each) did not inhibit the lytic activity of V. vulnificus cytolysin. In conclusion, lanthanum sensitive $Ca^{2+}$ influx plays a significant role in Vibrio vulnificus cytolysin-induced platelet cytolysis and thrombocytopenia in V. vulnificus infection.

• 멤브레인
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• 제25권6호
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• pp.547-557
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• 2015

본 연구에서는 높은 이산화탄소 분리성능을 가지는 폴리이미드의 제조를 위해 2,2-bis(3,4-carboxylphenyl) hexafluoropropane과 두종류의 아민인 2,3,5,6-tetramethyl-1,4-phenylenediamine, Poly(ethylene glycol) bis(3-aminopropyl) terminated을 이용하여 합성을 진행하였다. 합성된 고분자를 비용매 상전이법으로 비대칭 분리막을 제조하기 위하여 고분자의 용해도 지수 추정 값과 비용매 상전이 계수 측정을 통해 용매를 선정하였고, 고분자 용액 점도 측정을 통해 분리막 제조를 위한 도프용액 중의 고분자의 함량을 결정하여 질산리튬을 첨가제로 사용하여 최종적으로 분리막을 제조하였다. 제조된 평판형 비대칭 분리막은 전자주사현미경(SEM)을 통해 질산리튬과 휘발성 용매 함량에 변화에 따른 모폴로지의 변화를 확인하였으며, 이의 변화에 따른 기체 투과도 변화를 확인하였다. 분리막 제조를 위한 도프용액 중의 휘발성 용매 함량이 작을수록 선택도 변화가 없으면서 이산화탄소 투과도가 증가하는 것을 확인하였다.

• The Korean Journal of Physiology
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• 제7권1호
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• pp.23-31
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• 1973

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.

• 한국균학회지
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• 제20권1호
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• pp.44-50
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• 1992

Rhizoctonia solani의 균사융합군(AG) 중에서 생장속도가 빠른 AG-1과 AG-4는 가장 많은 원형질체를 형성하였으며 속도가 느린 AG-3은 저조하였고 중간인 AG-2와 AG-5는 또한 중간이었다. 균핵의 형성은 AG에 따라 차이가 있었다. Cellulase "onozuka" R-l0, macerozyme R-10, ${\beta}-glucuronidase$의 효소 복합액에서 가장 많은 원형질체를 형성하였다. 5개 융합군은 30시간 배양한 3g의 균사를 0.6 M mannitol이 첨가된 효소 복합액에 $32^{\circ}C$에서 3-4시간 처리하면 가장 많은 원형질체를 형성하였다. 균핵을 형성하는 AG-1을 열처리로 불활성화시킨 후 균핵을 형성하지 않는 AG-5의 원형질체를 PEG와 ${Ca}^{2+}$로 융합시켰다. 재생배지상에서의 균총의 특징과 균핵의 형성에 의해 7 개의 원형질 융합체를 선발하고 esterase 동위효소의 zymogram 및 AG-1 과 융합체인 F150l 사이에서 일어나는 killing reaction으로 확인하였다.

• Membrane and Water Treatment
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• 제2권1호
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• pp.25-37
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• 2011

PES UF membranes containing silver were prepared to impart antibacterial properties for waste water treatment. Asymmetric membranes for antibacterial application were prepared from polyethersulfone (PES) and silver nitrate ($AgNO_3$) (PES/$AgNO_3$=15/2 by weight) solution in N-Methyl-2-pyrrolidone (NMP) via simple wet phase inversion technique. These membranes were characterized by polyvinylpyrrolidone (PVP) and polyethylene glycol (PEG) of different molecular weights (1000 ppm in water) at room temperature and on operating pressure of 5 bars. It was observed that the water flux of PES-$AgNO_3$ membrane is slightly lower than virgin PES but still increased linearly with the increment of pressure applied. The morphology of the resulting membranes was examined using Field-Emission Scanning Electron Microscope (FESEM) coupled with Energy Dispersive Spectroscopy (EDS). Elemental analysis using EDS proved that silver is successfully loaded on the membrane surfaces. Due to the success of loading silver on membrane surfaces, antibacterial activities were evaluated via agar diffusion method against Escherichia coli (E.coli) and Staphylococcus aureus (S.aureus) culture. By incorporating 2 wt% of silver nitrate, PES-$AgNO_3$ showed significant inhibition ring on both E.coli and S.aureus. Filtration of E.coli solution (OD 0.31) showed satisfactory rejection data with ~100% inhibition growth after 24 hours incubation at $37^{\circ}C$. Resultant membranes also exhibit better tensile strength (compared to virgin PES) up to 71% may be due to the suggested interactions. The residual silver during fabrication was measured using ICP-MS and result showed that the residual silver content of PES-$AgNO_3$ membrane was only ~1% of the original silver added in the polymer solution. These studies have shown that PES-$AgNO_3$ UF membranes are potential in improving the filtration in water treatment.

• 한국미생물·생명공학회지
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• 제16권3호
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• pp.175-181
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• 1988

섬유소 기질로부터 L-lysine을 생산할 목적으로 Cellulomonas flavigena에 Brevibacterium flavum 및 Corynebacterium glutamicum을 각각 이속간 원형질체 융합을 하여 융합조건과 융합체의 성질을 조사하였다. 각각의 parental protoplast를 동량으로 혼합한 후 30% PEG 6000으로 3$0^{\circ}C$, 30분간 융합시킨 결과 1.9$\times$$10^{-6}$~2.1$\times$$10^{-6}$의 융합빈도를 얻었고, 134주의 융합체 중 유전적 안정화가 판명되고 CMC 기질로부터 L-lysine 생성능이 인정된 FCB 3 및 FCC 19를 최종 선별하였다. FCB 3 및 FCC 19는 친주보다 DNA 함량이 높을 분 아니라 G＋C 함량도 융합전 친주의 G＋C 총함량의 약 1/2에 해당하여 반수체로서 안정화되어 있었으며, 그리고 친주와 반대로 CMC 자화능이 있어 CMC로부터 L-lysine을 배지 중에 축적하였다.

• Archives of Reconstructive Microsurgery
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• 제3권1호
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• pp.81-89
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• 1994

Firstly the wrap-around free flap from the great toe was transferred successfully by Morrison and O'Brien in 1980 for reconstruction in chronic case of amputated thumb distal to the MP joint. Since then, significant bone peg resorption and grafted bone fracture have been reported in most of cases after operation. We have performed thumb reconstructions within 2 weeks after injury in 11 cases which included acute crushing injuries or failed replantation of thumb at Guro Hospital from September, 1983 to October, 1993. The results obtained from this study are as follows: 1. There were 8 males and 3 females and the mean age was 28.8 years old ranged from 3 years old to 50 years old. 2. The most common injury mechanism was machinary injury in 10 of 11 cases. 3. We have performed thumb reconstructions by using wrap-around free flap from the great toe without bone graft in 6 cases and with bone graft in 5 cases within 2 weeks after injury. 4. During 6 years and 1 month mean follow-up period, there were bone resorptions in 3, grafted bone fracture in 1, infections in 2 and soft tissue necrosis in 3 cases. 5. We obtained satisfactable results in respect of function and cosmesis and urged to use this flap for reconstruction in acute thumb injury.

• Biomaterials and Biomechanics in Bioengineering
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• 제2권1호
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• pp.23-32
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• 2015

Facile immobilization of growth factors in hyaluronic acid (HA) hydrogels using dual enzymes is reported in the paper. The hydrogels were formed by using horseradish peroxidase (HRP) and hydrogen peroxide ($H_2O_2$) and transforming growth factor-${\beta}3$ (TGF-${\beta}3$) was covalently conjugated on the hydrogels in situ using tyrosinase (Ty) without any modifications. For the preparation of hydrogels, HA was grafted with poly(ethylene glycol) (PEG), which was modified with a tyrosine. The gelation times of the HA hydrogels were ranging from 415 to 17 s and the storage moduli was dependent on the concentration of $H_2O_2$ and Ty (470-1600 Pa). A native TGF-${\beta}3$ (200 ng/mL) was readily encapsulated in the HA hydrogels and 17% of the TGF-${\beta}3$ was released over 1 month at the Ty concentration of 0.5 KU/mL, while the release was faster when 0.3 KU/mL of Ty was used for the encapsulation (27%). It can be suggested that the growth factors resident in the hydrogels for a long period of time may lead cells proliferating and differentiating, whereas the growth factors that are initially released from the hydrogels can induce the ingrowth of cells into the matrices. Therefore, the dual enzymatic methods as facile gel forming and loading of various native growth factors or therapeutic proteins could be highly promising for tissue regenerative medicines.

• Journal of Microbiology
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• 제42권3호
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• pp.228-232
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• 2004

In this paper, we investigated the development of a simplified and rapid primary capture step for the recovery of M13 bacteriophage from particulate-containing feedstock. M13 bacteriophage, carrying an insert, was propagated and subsequently purified by the application of both conventional multiple steps and expanded bed anion exchange chromatography. In the conventional method, precipitation was conducted with PEG/NaCl, and centrifugation was also performed. In the single step expanded bed anion exchange adsorption, UpFront FastLine$\_$TM/20 (20mm i.d.) from UpFront Chromatography was used as the contactor, while 54$m\ell$ (H$\_$o/=15cm) of STREAMLINE DEAE (p=1.2 g/㎤) from Amersham Pharmacia Biotechnology was used as the anion exchanger. The performance of the two methods were evaluated, analysed, and compared. It was demonstrated that the purification of the M13 bacteriophage, using expanded bed anion exchange adsorption, yielded the higher recovery percentage, at 82.86％. The conventional multiple step method yielded the lower recovery percentage, 36.07％. The generic application of this integrated technique has also been assessed.

• 한국균학회지
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• 제21권4호
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• pp.323-331
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• 1993

The protoplast formation and intergeneric protoplast fusion between Gliocladium virens and Trichoderma harzianum were attempted to obtain fusants. Protoplast formation was the most effective when the strains were treated with concentration of 5 mg/ml of Novozyme 234 and Cellulase at $25^{\circ}C$ for 3 hours in phosphate buffer, pH 6.5, supplemented with 0.6 M sorbitol as osmotic stabilizer. Auxotrophic mutants of G. virens G88 did not grow in minimal medium and benomyl resistant T. harzianum T95 from wild types, however, was selected by treatment with UV light as genetic marker to isolate fusants. When the intergeneric protoplast fusion between G. virens G88 and T. harzianum T95 was carried out using 30% PEG 4000 containing 10 mM $CaCl_{2}$, and 50 mM glycine (pH 8.5) as fusogenic agent at $25^{\circ}C$ for 10-15 min, the fusion frequency was $0.8{\times}10^{-4}$. Fusants obtained from intergeneric protoplast fusion were spontaneously segregated into va rious strains by continous culture on complete medium. Several intergeneric hybrids were classified into three types: parent-like hybrids, segregants, and recombinants.