• Journal of Microbiology and Biotechnology
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• v.17 no.12
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• pp.1930-1936
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• 2007

A sequential optimization strategy based on statistical experimental designs was employed to enhance the production of cyclosporin A (CyA) by Tolypocladium inflatum DSMZ 915 in a submerged culture. A 2-level Plackett-Burman design was used to screen the bioprocess parameters significantly influencing CyA production. Among the 11 variables tested, sucrose, ammonium sulfate, and soluble starch were selected, owing to their significant positive effect on CyA production. A response surface methodology (RSM) involving a 3-level Box-Behnken design was adopted to acquire the best process conditions. Thus, a polynomial model was created to correlate the relationship between the three variables and the CyA yield, and the optimal combination of the major media constituents for cyclosporin A production, evaluated using the nonlinear optimization algorithm of EXCEL-Solver, was as follows (g/l): sucrose, 20; starch, 20; and ammonium sulfate, 10. The predicted optimum CyA yield was 113 mg/l, which was 2-fold the amount obtained with the basal medium. Experimental verification of the predicted model resulted in a CyA yield of 110 mg/l, representing 97% of the theoretically calculated yield.

• Journal of Periodontal and Implant Science
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• v.33 no.4
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• pp.643-650
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• 2003

Cyclosporin-A(CsA)는 장기와 조직 이식에 따른 거부반응을 조절하기 위해 사용되는 면역억제제로, 이식의학의 발달과 더불어 사용량이 증가하고 있다. CsA의 부작용중의 하나인 치은과증식은 30-50%의 빈도로 발발하고 있다. 최근 macrolide 계열의 항생제인 azithromycin을 이용하여 이런 부작용을 억제시킨다는 임상 보고가 있어서, 이를 실험적으로 확인하고자 하였다. 이를 위해 CsA를 투여한 적이 없는 환자에서 정상 치은조직을 채취, 치은섬유아세포를 배양하였다. 우선 CsA에 대한 치은섬유아세포의 반응을 보기 위해 다양한 농도($10^{-8}-10^{-10}$g/ml)로 처치하여, 세포 증식량과 교원질 합성량을 MTT assay와 Sirol Collagen Assay를 이용하여 측정하였다. 이에 반응을 보인 조건과 세포를 대상으로 다양한 농도($10^{-8}-10^{-10}$g/ml)의 azithromycin을 CsA와 동시 처치하여 아래와 같은 결과를 얻었다. 1. CsA는 일부 치은섬유모세포의 증식을 증가시켰다. 그러나 Collagen 합성능에는 변화를 주지 않았다. 2. Azithromycin은 정상 치은섬유아세포의 증식능에 영향을 미치지 않았다. 3. Azithromycin은 CsA 에 반응을 보인 세포의 증식을 감소시켰으며, 이는 정상 수준과 유사하였다. 이상의 결과에서 azithromycin이 CsA에 의한 치은과증식 치료에 유익하다고 사료된다.

• Journal of Pharmaceutical Investigation
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• v.39 no.1
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• pp.59-63
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• 2009

Soybean phosphatidylcholine microparticles loaded with cyclosporin A (CsA) were prepared by the modified emulsion solvent diffusion and ionic gelation method, in which chitosan on the surface of the microparticles was crosslinked with various concentrations of tripolyphosphate (TPP). The morphology of the particles was characterized by scanning electron microscopy (SEM). The change of particle size and zeta-potential by chitosan on the surface of the lipid microparticles were systematically observed. The encapsulation efficiency and loading capacity of CsA in the particles were determined by high performance liquid chromatography (HPLC). In vitro release kinetics was studied using the dialysis method. In the results, the mean particle size and the zeta-potential of lipid microparticles increased when the attached chitosan was cross-linked (from 2.5 to 6.2 ${\mu}m$ and from -37.0 to +93.0 mV, respectively). The cyclosporin A-loaded lipid microparticles appeared discrete and spherical particles with smooth surfaces. The encapsulation efficiency of CsA was between 79% and 90% while the loading capacity was between 41% and 56%. In vitro release study showed that the crosslinkage of chitosan by TPP significantly delayed the release of CsA from the particles in a concentration-dependent manner. Thus, the release of CsA from the lipid microparticles could be controlled by tripolyphosphate used as a cross-linking agent.

• Journal of Microbiology and Biotechnology
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• v.15 no.5
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• pp.1047-1053
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• 2005

Cyclosporin A binding protein type-19 kDa peptidyl-prolyl cis/trans isomerase (PPIases, EC 5.2.1.8) of Euglena gracilis was purified and some of its biochemical characters were elucidated. Purification of the PPIase was achieved by employing a series of steps involving ammonium sulfate precipitation, Superdex G-75 gel filtration chromatography, Mono­Q anion and Mono-S cation exchange chromatographies, and Superdex S-200 gel filtration chromatography on FPLC. Purified PPIase had a specific activity of 8,250 units/mg, showing a 27-fold increase compared with that of cell-free extract of Euglena gracilis. The enzyme consisted of a single polypeptide chain with a molecular mass of 19 kDa. It showed high substrate specificity to succinyl-Ala-Ala-Pro-Phe-p-nitroanilide, and $k_{car}/K_{m}$, for this substrate was found to be $61.19{\times}10^5/sec$. The isomer distributions were investigated at an equilibrium of seven different peptide substrates, varying Xaa in Suc-Ala-Xaa-Pro-Phe-p-nitroanilide in dimethylsulfoxide. The cis/trans equilibrium constants were estimated to be from 0.14 (Ile) to 0.63 (Gly), which correspond to $12.00\%\;to\;38.52\%$ of the cis population, respectively, under experimental condition. The enzyme was highly sensitive to the immunosuppressive ligand cyclosporin A, but not to other immunosuppressants such as FK506 and rapamycin. Thus, it appears to belong to the class of cyclophilin.

• The Journal of Dong Guk Oriental Medicine
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• v.2 no.2
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• pp.1-15
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• 1993

The purpose of this study is to observe the effect of Woogyuyem on serum reaction and hepatic tissue in Cyclosporin A treated rats. In this study, the experimental rats divided two group(Control group and Sample group). Under the same condition, both control group and sample group were injected to abdomen with Cyclosporin A for 7days. And then, control group were administered water, sample group were administered Woogy-uyeum for 300days. The change of transaminase and bile acid and SOD activity in blood serum, hepatic tissue were measured at 3rd, 7th, 15th, 30th day. The obtained results are summarised as follows : 1. In the change of SGOT contents, as compared with control group, sample group was significantly decreased at 7th and 15th day. 2. In the. change of SGPT contents, as compared with control group, sample group was significantly decreased at 15th day. 3. In the change of serum bile acid contents, as compared with control group, sample group was significantly decreased at 7th and 15th day. 4. In the change of serum SOD activity, as compared with control group, sample group was significantly increased at 15th and 30th day. 5. In the change of hepatic tissue, as compared with control group, the vacuolation of hepatocytes in the sample group was reduced, the liver plates were reconstructed, and the epitheriai cell wasn't destructed.

• KSBB Journal
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• v.11 no.5
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• pp.613-621
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• 1996

The effects of important medium components such as carbon, nitrogen sources and amino acids on the production of cyclosporin A(CyA) were investigated in an immobilized fungal cell fermentation using Tolypocladium inflatum. As carbon sources in the synthetic medium, fructose and maltose stimulated CyA production remarkably compared to glucose when ammonium sulfate was supplemented as a nitrogen source. In the absence of ammonium sulfate in the medium, however, CyA biosynthesis was reduced considerably without regard to C-sources tested. Ammonium sulfate was found to be the best N-source, and also ammonium phosphate and ammonium citrate showed some positive effects on CyA production. Optimum concentration of ammonium sulfate was 10g/L, and supplementation of ammonium sulfate at the start of fermentation was found to be the most efficacious for maximal production of CyA. Among the constituent amino acids of cyclic peptide, CyA, L-valine had the most significant effect on the biosynthesis of CyA, and maximum CyA production was observed when 10 g/L of L-valine was initially added.

• KSBB Journal
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• v.11 no.1
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• pp.30-36
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• 1996

Based on fermentation data for cyclosporin A production, simple Monod kinetics was proposed for both immobilized and suspended cultures. Higher value of $\mu$mas and lower value of Km suggest better catalytic activity of the immobilized cells than the parallel suspended cells. Furthermore, lower Km value in the immobilized cell system indicates higher affinity of the immobilized cells for carbon substrate as compared with the suspended cells. For immobilized cell cultures, these parameters were also utilized for the estimation of effectiveness factor, an indicator for intraparticle mass transfer resistance. Based on simulation studies, optimum radius of celite beads was turned out $100 ~ 500{\mu}m$In this simulation work, we examined the influence of biosupport size and immobilized biomass density on diffusional resistance of substrate inside the bead matrix. In order to maintain uniformly distributed cell activities in biosupport, it was essential to determine optimum slze of particle, and then to estimate the most economic loaded biomass content.

• Tuberculosis and Respiratory Diseases
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• v.55 no.6
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• pp.631-635
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• 2003

Idiopathic interstitial pneumonia is a group of lung disease classified by clinical, radiological and pathologic findings. Steroid is described in many reports as an effective treatment for the idiopathic interstitial pneumonitis. For the treatment of steroid non-responders, immuno-suppressive or cytotoxic agents are used as second line drug of choice. We experienced a case of nonspecific interstitial pneumonitis who was treated initially with glucocorticoid and antibiotics without effects, but later did respond to cyclosporin treatment. This 63 year-old patient was admitted due to dyspnea, cough, and sputum production. He was started on glucocorticoid and antibiotics after he was diagnosed as having nonspecific interstitial pneumonitis by open lung biopsy. The initial steroid treatment was not effective, however, after cyclosprin treatment, the patient showed clinical and radiologic improvements.

• Journal of Periodontal and Implant Science
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• v.30 no.4
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• pp.747-757
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• 2000

Cyclosporin A(CsA) is an immunosuppressive agent widely used for preventing graft rejecting response in organ transplantation. The basic properties of CsA to osteoblast has not been well known yet. A better understanding of the mechanisms of CsA function on bone could provide valuable information regarding basic properties of bone remodeling, pharmacotherapeutic intervention in metabolic bone disease, and the consequences of immunosuppression in bone physiology. The purpose of this study was to investigate the effect of CsA on osteoblast by evaluating parameters of proliferation, collagen synthetic activity, alkaline phosphatase activity, and ALP mRNA expression in mouse calvarial cell. 1. CsA ($3{\mu}g/m{\ell}$) treated mouse calvarial cell showed statistically significant increase in cell proliferation.(P<0.05) 2. CsA($1,\; 3{\mu}g/m{\ell}$) treated MC3T3 cell line showed statistically significant increase in cell proliferation. 3. The amount of collagen of CsA($3{\mu}g/m{\ell}$) treated mouse calvarial cell was decreased statistically significantly. 4. Alkaline phosphatase activity was increased statistically significantly in CsA treated group($1{\mu}g/m{\ell}$). 5. mRNA expression of ALP was increased in CsA treated group These results suggest that CsA could affect bone remodeling by modulating proliferation & differentiation of osteoblast.

• KSBB Journal
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• v.9 no.2
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• pp.200-210
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• 1994

In batch bioreactor fermentations for cyclosporin A (CyA) production, good potential for bioprocess improvement was demonstrated in the immobilized cell system, providing appreciably better utilization of the catalytic activity of the biomass than the freely suspended cells, especially during the exponential phase. When concentrated nutrient medium was added pulsely during the exponential phase of cell growth(at hour 139 of fermentation), reactivation and regermination in both immobilized and suspended cell cultures were observed to contribute to the longevity of CyA production, maintaining maximum CyA titre until 250 hours of fermentation. Contrarily, simple batch fermentations without any supplement of medium in both systems showed repid decrease in CyA concentrations during the late stationary phase. Notably, the CyA yield coefficient $(Y_p/x)$ for the immobilized cell system was maintained quite high even after the pulse addition of the concentrated full medium, reaching almost 80% of the level attained during the exponential phase. This is in sharp contrast when compared with the corresponding value of 58% in the case of parallel-suspended cells. This pattern of CyA production resulted in considerably enhanced CyA production in the immobilized cell system, reaching almost 2 time higher maximum CyA production in comparison with the free cell system.