• KSBB Journal
• /
• v.19 no.4
• /
• pp.308-314
• /
• 2004

A degenerate set of PCR primers based on two conserved regions (heme binding region and oxygen ligand pocket) were designed and successfully applied to amplify DNA fragments of cytochrome P450 hydroxylase (CYP) genes from a rare actinomycetes, S. benihana. The PCR amplified products were employed as a DNA probe to clone the entire CYP genes from S. benihana genomic library. Five different CYP-positive cosmids were isolated by colony hybridization as well as PCR confirmation. The complete nucleotide sequencing of five different CYP genes revealed that each unique CYP showed a significant amino acid homology to previously-known CYP genes involved in streptomycetes secondary metabolism. In addition, four CYP genes (CYP502, CYP503, CYP504, CYP506) were found to be linked to ferredoxin genes in the chromosome, and the CYP503 gene showed the high degree of amino acid similarity to the previously well-characterized CYP105 family in streptomycetes.

• Childhood Kidney Diseases
• /
• v.6 no.1
• /
• pp.120-122
• /
• 2002

A 9-year-old boy of B blood group with end-stage renal disease due to IgA nephropathy received group O kidney transplantation from his father On day 9, he developed intravascular hemolysis, and anti-B autoantibody formation was confirmed. We diagnosed as immune hemolytic anemia due to passenger lymphocyte from donor, and cyclosporine withdrawl was done. Anemia resolved spontaneously, but on day 18, graft dysfunction developed, and graft biopsy revealed acute allograft rejection. Although hemolysis due to autoantibody is very rare and often mild, and the role of hemoglobinuria on acute rejection in this case is not certain, we recommend consideration of aggressive management on severe hemolysis after minor mismatched kidney transplantation. (J Korean Soc Pediatr Nephrol 2002 ; 6 : 120-2)

• Biomolecules & Therapeutics
• /
• v.6 no.3
• /
• pp.296-302
• /
• 1998

The bioequivalence of two cyclosporin A products was evaluated in 26 normal male volunteers (age 25 ~33 yr, body weight 56~84 kg) following single oral administration. Test product was a hard capsule containing the granule of cyclosporin A (Chong Kun Dang Corp., Korea) and reference product, Sandimmun", was a soft capsule containing surfactant, oil, alcohol and cyclosporin A (Sandoz, Swiss). Both products contain 100 mg of cyclosporin A. Four capsules of the test and the reference product were administered to the volunteers, respectively, by randomized two period cross-over study (2$\times$2 Latin square method). Average drug concentrations at each sampling time and pharmacokinetic parameters were not significantly different between two products (p>0.05); the area under the concentration-time curve to last sampling time (24 hr) (AU $Co_{24}$) (5034.8$\pm$ 1760.6 vs 4635.4$\pm$ 1158.9 ng . h/ml), maximum plasma concentration ( $C_{max}$) (1002.7$\pm$353.1 vs 980. 4$\pm$ 171.7 ng/71), and mean residence time (MRT) (6.16$\pm$0.81 vs 5.64$\pm$0.50 h). The differences of mean AUC 7-,4,7~, T_ and MRT between the two products (7.93,2.22,16 and 8.39%, respectively) were less than 20% given as a guideline. The power (1-$\beta$) and treatment difference ($\Delta$) for AU $Co_{24}$, $C_{max}$ and MRT were more than 0.8 and less than 0.2, respectively. Although $T_{max}$ of the two products was significantly different each other (p<0.05), $T_{max}$ might be an insignificant parameter because cyclosporin A generally requires long-term administration. From these results, the two products are bioequivalent.alent.t.

• Microbiology and Biotechnology Letters
• /
• v.38 no.4
• /
• pp.475-480
• /
• 2010

Actinomycetes are Gram-positive soil bacteria and one of the most important industrial microorganisms due to superior biosynthetic capabilities of many valuable secondary metabolites as well as production of various valuable bioconversion enzymes. Among them are cytochrome P450 hydroxylase (CYP), which are hemoproteins encoded by a super family of genes, are universally distributed in most of the organisms from all biological kingdoms. Actinomycetes are a rich source of soluble CYP enzymes, which play critical roles in the bioactivation and detoxification of a wide variety of metabolite biosynthesis and xenobiotic transformation. Cyclosporin A (CyA), one of the most commonly-prescribed immunosuppressive drugs, was previously reported to be hydroxylated at the position of 4th N-methyl leucine by a rare actinomycetes called Sebekia benihana, leading to display different biological activity spectrum such as loss of immunosuppressive activities yet retaining hair growth-stimulating side effect. In order to improve this regio-selective CyA hydroxylation in S. benihana, previously-identified several secondary metabolite up-regulatory genes from Streptomyces coelicolor and S. avermitilis were heterologously overexpressed in S. benihana using an $ermE^*$ promoter-containing Streptomyces integrative expression vector. Among tested, SCO4967 encoding a conserved hypothetical protein significantly stimulated region-specific CyA hydroxylation in S. benihana, implying that some common regulatory systems functioning in both biosynthesis and bioconversion of secondary metabolite might be present in different actinomycetes species.

• The Korean Journal of Nuclear Medicine
• /
• v.34 no.4
• /
• pp.360-365
• /
• 2000

We experienced a case of cerebral hypoperfusion due to cyclosporine neurotoxocity confirmed only by Tc-99m ECD brain SPECT. A 53-year-old female had received allogenic peripheral blood stem cell transplantation due to refractory plasmacytoid lymphoma. Cyclosporine and steroid had been administrated to prevent graft versus host disease. Twenty days after transplantation, she became delirious and suffered from generalized tonic-clonic seizure. Immediately, brain MRI and MR angiography were performed and these studies did not show any abnormal findings. However, Tc-99m ECD brain SPECT showed diffuse hypoperfusion in the left cerebral hemisphere and blood cyclosporine level was 962.6 ng/ml. Cyclosporine administration was stopped and discontinuation of cyclosporine resulted in disappearance of all neurological symptoms. The same neurological symptoms recurred with cyclosporine re-administration for management of exacerbated graft versus host disease. In this case, Tc-99m ECD brain SPECT proved very helpful in the diagnosis of cycloporine neurotoxicity.

• Journal of Chest Surgery
• /
• v.32 no.1
• /
• pp.1-4
• /
• 1999

Background: The transplantation of organs between phylogenetically disparate or harmonious species has invariably failed due to the occurrence of hyperacute rejection or accerelated acute rejection. But, concordant cardiac xenograft offer us an opportunity to study xenotransplantation in the absence of hyperacute rejection. Current therapeutics for the prolongation of survival of rodent concordant xenotransplantation are not ideal with many regimens having a high mortality rate. Cyclosporine A & Mycophenolate Mofetil are new immunosuppresive agent which has been shown to be effective at prolonging survival of allograft, as purine synthesis inhibitor. Material and Method: We used white mongrel rats as recipient and mice as donor, divided 4 groups(n=6), control group(Group 1) has no medication or pretreatment, Group 2 has splenectomy as pretreatment 7∼10 days before transplantation, Group 3 has Cyclosporine A treatment group, Group 4 has combined treatment of Cyclosporine A & Mycophenolate Mofetil(RS 61443). We compared survival time. Reuslt: We can't find significant difference of survival time between each groups. Conclusion: We concluded that rejection of cardiac xenograft was different from rejection of allograft, and new immunossuppresive Agent(Mycophenolate Mofetil, Cyclosporine A) was not effective for prolongation of survival time after cardiac xenograft.

• Journal of Chest Surgery
• /
• v.30 no.9
• /
• pp.876-884
• /
• 1997

Fourteen patients underwent orthotopic heart transplantation between March 1994 and May 1996 in Seoul National University Hospital. There were 9 male and 5 female patients, and the mean age was 40.8 $\pm$ 12.4 years ranged from 12 to 56 years. All patient were in NYHA Fc III or IV preoperatively. The underlying heart diseases were dilated cardiomyopathy in 11 and restrictive cardiomyopathy in 3. The mean age of donors was 24.9$\pm$ 10.2 years and the causes of the brain death were head trauma by traffic accidents in 8, subarachnoid hemorrhage in 2, 1 asphyxia, 1 fall down injury, 1 brain tumo , and 1 drowning, respectively The blood type was identical in 11, compatible in 2, and incompatible in 1 patient. The direct bicaval anastomosis technique was used in 11 cases, and standard right atrial anastomosis was done in the remaining 3 cases. The graft ischemic time was 158$\pm$44 minutes ranged 94 to 220 minutes. There were two hospital deaths(14.3%). The causes of deaths were 1 right ventricular failure followed by suspected cyclosporine induced hemolytic uremic syndrome and rejection, and 1 delayed massive bleeding, probably from rupture of the anastomotic pseudoaneurysm, respectively. The follow-up duration was 16$\pm$9 months(3 to 28 months). There was one late death(8.3%). All the other patients were in NYHA Fc I except one patient who was in hospital because of the acute rejection. The actuarial survival rates including hospital deaths were 93.7% at 1 month, 86.9% at 6 months, and 77$\pm$12% at 2 years. Conclusively, heart transplantation is the good strategy for the management of end stage heart disease with acceptable operative mortality and early follow-up results.

• Journal of Chest Surgery
• /
• v.42 no.2
• /
• pp.135-140
• /
• 2009

Background: Chronic rejection after a cardiac allograft usually occurs about six months after the operation. Vasculopathy due to chronic rejection causes atherosclerosis in the coronary artery of the transplanted heart and then this causes myocardial injury. We intended to discover and document those findings that occur in a transplanted ascending aorta. Material and Method: In rats weighting $200{\sim}300gm$ (Spraque-Dawley rat), we carried out heterotopic heart allo-transplantation with the modified Ono-Lindsey method and then the rats were administrated cyclosporine (10mg/kg/day). After three months survival, we acquired biopsy materials from the native ascending aorta and the allo-transplanted ascending aorta and we compared them. We classified each severity of 1) intimal thickening, 2) medial hyperplasia, 3) medial calcification, 4) medial inflammation and 5) chondroid metaplasia, which are specific biopsy findings for chronic rejection after a cardiac allograft. Each severity was classified, according to the opinion of one pathologist, in the native ascending aorta biopsies (n=9) and the allo-transplanted ascending aorta biopsies (n=13). The data of the control group and the study group were statistically analyzed with using the Mann-Whitney test (SPSS version 12.0 window). Result: The important changes of the allo-transplanted aorta were intimal thickening (p<0.0001), medial calcification (p=0.045), medial inflammation (p<0.0001) and chondroid metaplasia (p=0.045), but not medial hyperplasia (p=0.36). Conclusion: Cardiac allograft vasculopathy was seen in the transplanted ascending aorta, the same as was seen in the coronary artery, after allograft cardiac transplantation. We have reached the conclusion that chronic rejection also progresses in the aorta.