• Korean Journal of Fisheries and Aquatic Sciences
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• v.7 no.1
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• pp.15-27
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• 1974

Since the summer of the year 1972 ulcer disease of common carp, colour carp ana goldfish had suddenly spread widely and caused a serious mortality at fish farms especially around Gim-hae and Yang-san, near Busan. The present study aimed to find out the causative organisms, histopathological changes and the way of treatment, and the results are summarized as follows : Two kinds of pathogenic bacteria, i. e, Chondrococcus columnaris and Aeromonas sp. were isolated from the mucus of the lesion. According to the macroscopic findings, these symptoms began with the hemorrhagic spots under scales which fell out, after the collapse of the dermis, which was followed by lesion to form ulcer, and then the muscle was exposed. The fin was eroded mostly from the distal part, but sometimes from the basal part. Gills showed grey colour, and this part of the tissue collapsed. According to the histopathological findings, the dermis was exposed after the collapsed of the epithelium of the skin, and the necrosis of the musculature occurred, the muscular fibre being destroyed. The epithelial cells of gill tissue proliferated, thus gill filaments were conglutinated and collapsed. Fatty degeneration happened at the liver but the other organs seem to be normal. Treatment with the following mixtures were effective the water temperature of $22\~25^{\circ}C$, but not effective when the temperature was under as low as $13\~15^{\circ}C$. Mixture 1. Aivet (HB-115.HCI) 0.3ppm Malachite green 0.2ppm. Dipterex 0.4ppm Mixture 2. Furanace (P-7138) 0.05ppm Malachite green 0.2ppm Dipterex 0.4ppm When lesion healed, the epithelium, dermis, and the muscular fibre were regenerated.

• Journal of Korean Ophthalmic Optics Society
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• v.17 no.1
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• pp.91-97
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• 2012

Purpose: This purpose of study is to investigate the effect of UV-A-blocking or brown-tinted ophthalmic lens against A2E photooxidation which known as one of the etiologies of AMD(Age-related macular degenaration). Methods: The photooxidation of A2E, synthetic product of two molecules of all-trans-retinal and ethanolamine, was induced by the exposure to blue light (420~470 nm, $94mW/cm^2$) for 3 minutes. The inhibitory effect of UVblocking or brown-tinted ophthalmic lens against A2E photooxidation was evaluated by UV absorbance and HPLC analysis of remained A2E after the exposure to blue light. Results: UV-blocking ophthalmic lens could not inhibit A2E photooxidation induced by blue light irradiation. There was no difference in A2E photooxidation in the presence of brown-tinted ophthalmic lens to block 15% of visible ray, however, those lenses blocking 55% or 86% of visible ray showed the inhibitory effect of A2E photooxidation as 9.98% and 16.55%, respectively. By HPLC analysis, the amount of residual A2E which was not blocked by any lens was $199.29{\pm}26.53{\mu}M$, however, the inhibitory effect against A2E photooxidation was shown in the presence of brown-tinted lens. The remained A2Es were $264.58{\pm}31.91{\mu}M$ and $402.93{\pm}28.68{\mu}M$ in brown-tinted lenses of 55% and 86% blocking visible ray, respectively. However, there was no inhibitory effect against A2E photooxidation in the case of UV-blocking lens by HPLC analysis. Conclusions: In this study, brown-tinted ophthalmic lens was confirmed to have the inhibitory effect against the photooxidation of A2E, a causing substance of AMD onset.

• Radiation Oncology Journal
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• v.23 no.3
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• pp.161-168
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• 2005

Purpose: The changed expressions of $TGF-{\beta}1$, as a key cytokine in the fibrotic process, due to melatonin with potent antioxidative effects, were investigated in the irradiated lung using fibrosis-sensitive C57BL/6 mice. Materials and Methods: Female C57BL/6 mice were divided into control irradiation-only, and melatonin (300 mg/kg i.p. 1 hr before irradiation) pretreatment groups. The thoraces of the mice were irradiated with a single dose of 12 Gy. The mRNA expressions of $TGF-{\beta}1$ in the lung tissue 2 and 4 weeks after irradiation were quantified using semiquantitive RT-PCR, and the cellular origin and expression levels of $TGF-{\beta}1$ protein were identified using immunohistochemical staining. Results: The relative mRNA expression levels in the irradiation-only and melatonin pretreatment groups 2 and 4 weeks after irradiation were 1.92- and 1.80-fold (p=0.064) and 2.38- and 1.94-fold (p=0.004) Increased, respectively compared to those in the control group. increased expressions of $TGF-{\beta}1$ protein were prominently detected in regions of histopathologicai radiation injury, with alveolar macrophages and septal epithelial cells serving as important sources of $TGF-{\beta}1$ expression. At 2 and 4 weeks after irradiation, the expression levels of protein were $15.8\%\;vs.\;16.9\%$ (p=0.565) and $36.1\%\;vs.\;25.7\%$ (p=0.009), respectively. Conclusion: The mRNA and protein expressions of $TGF-{\beta}1$ in the lung tissue following thoracic irradiation with 12 Gy were significantly decreased by melatonin pretreatment at 4 weeks. These results indicate that melatonin may have a possible application as an antifibrotic agent in radiation-induced lung injury.

• Journal of the korean academy of Pediatric Dentistry
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• v.32 no.1
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• pp.75-88
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• 2005

Ameloblastoma is the most commonly occurring odontogenic tumor in oral cavity. Although most are benign epithelial neoplasm, they are generally considered to be locally aggressive and destructive, exhibiting a high rate of recurrence. The biological behavior of this neoplasm is a slowly growing, locally invasive tumor without metastasis, therefore malignant neoplasm, changed its histological appearance to carcinoma or showed distant metastasis, is only defined clinically. In this study, we identified the differentially expressed genes(DEGs) in stages under benign or malignant ameloblastoma compared with normal patient using ordered differential display(ODD) reverse transcription(RT)-PCR and $GeneFishing^{TM}$ technology. ODD RT-PCR is rather effective when the investigation of samples containing very small amounts of total RNA must be accomplished. ODD RT-PCR used the means of amplification with anchored T-primer and adaptor specific primer. bearing definite two bases at their 3' ends and so this method could display differential 3'-expressed sequence taqs(ESTs) patterns without using full-length cDNAs. Compared with standard differential display, ODD RT-PCR is more simple and have enough sensitivity to search for molecular markers by comparing gene expression profiles, However, this method required much effort and skill to perform. $GeneFishing^{TM}$ modified from DD-PCR is an improved method for detecting differentially expressed genes in two or more related samples. This two step RT-PCR method uses a constant reverse primer(anchor ACP-T) to prime the RT reaction and arbitrary primer pairs(annealing control primers, ACPs) during PCR. Because of high annealing specificity of ACPs than ODD RT-PCR, the application of $GeneFishing^{TM}$ to DEG discovery generates reproducible, authentic, and long(100bp to 2kb) PCR products that are detectable on agarose gels. Consequently, various DEGs observed differential expression levels on agarose gels were isolated from normal, benign, and malignant tissues using these methods. The expression patterns of the some isolated DEGs through ODD RT-PCR and $GeneFishing^{TM}$ were confirmed by Northern blot analysis and RT-PCR. The results showed that these identified DEGs were implicated in ameloblastoma neoplasm processes. Therefore, the identified DEGs will be further studied in order to be applied in candidate selection for marker as an early diagnosis during ameloblastoma neoplasm processes.

• Tuberculosis and Respiratory Diseases
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• v.66 no.3
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• pp.205-210
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• 2009

Background: CXCL10 and CXCL11, which are family of CXCR3 ligands, are expressed by lymphocytes and even by bronchial epithelial cells if the cellular immunity is activated. This study evaluated the potential utility of CXCL10 and CXCL11 in the serum for active pulmonary tuberculosis in comparison with lung cancer, which activates the cellular immunity, and benign lung diseases. Methods: Patients who newly visited Pusan National University Hospital from January 2007 to December 2007 and were suspected of having lung cancer or tuberculosis were enrolled prospectively. The patients were classified pathologically and clinically into three groups, 47 with lung cancer, 18 with active pulmonary tuberculosis and 38 control patients with benign pulmonary disease. ELISA was used to determine the levels of CXCL10 and CXCL11 were determined in the serum. Results: The level of CXCL10 and CXCL11 were significantly higher in the active pulmonary tuberculosis group than in the lung cancer and benign lung disease groups (p<0.001, Kruskal-Wallis). The level of CXCL11 was significantly higher in the lung cancer group than in the benign pulmonary disease group, but there was no significant difference in level of CXCL10 between the three groups (p<0.001, p=0.655, respectively, Mann-Whitney U). The level of CXCL10 in patients with stage III+IV lung cancer was significantly higher than those with stage I+II, but there was no significant difference in the level of CXCL11 between the groups (p<0.001, p=0.07, respectively, Mann-Whitney U). There was no significant difference in the level of CXCL10 and CXCL11 between those with the presence and absence of lung cancer metastasis. There was a significant correlation between the level of CXCL10 and CXCL11 (r=0.223, p<0.001). Conclusion: CXCL10 and CXCL11 may be a potential useful markers for active pulmonary tuberculosis if used alongside other diagnostic methods.

• Clinical and Experimental Pediatrics
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• v.53 no.4
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• pp.525-531
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• 2010

Purpose : Expression levels of tumor necrosis factor (TNF)-${\alpha}$ expression on the mucosa of the small intestine is increased in patients with villous atrophy in food protein-induced enterocolitis syndrome (FPIES). TNF-${\alpha}$ has been reported to induce apoptotic cell death in the epithelial cells. We studied the TNF family and TNF-receptor family apoptosis on the duodenal mucosa to investigate their roles in the pathogenesis of FPIES. Methods : Fifteen infants diagnosed as having FPIES using standard oral challenge test and 5 controls were included. Terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) staining was performed to identify the apoptotic cell death bodies. Immunohistochemical staining of TNF-${\alpha}$, Fas ligand (FasL) for TNF family and TNF-related apoptosis-including ligand (TRAIL) receptor 1 (DR4), TRAIL receptor 2 (DR5), and Fas for TNF-receptor family were performed to determine the apoptotic mechanisms. Results : $TUNEL^+$ was significantly more highly expressed in the duodenal mucosa of FPIES patients than in controls ($P$-0.043). TNF-${\alpha}$ ($P$=0.0001) and DR4 ($P$=0.003) were significantly more highly expressed in FPIES patients than in controls. Expression levels of FasL, Fas, and DR5 were low in both groups and were not significantly different between the 2 groups. Conclusion : These results suggest that FPIES pathogenesis is induced by apoptosis, and that TNF-${\alpha}$ expression and DR4 pathway may have an important role in apoptosis.

• Reproductive and Developmental Biology
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• v.32 no.3
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• pp.153-158
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• 2008

Tissue-specific and temporal regulation of milk protein gene expression is advantageous when creating transgenic animal that produces foreign protein into milk. Gene expression, i.e. protein production, is regulated not only by promoter strength but also mRNA stability. Especially, poly A tail length by polyadenylation affects in vivo and in vitro mRNA stability and translation efficiency of the target gene. In the present study, nucleotide sequence of 3'-UTR was analyzed to evaluate the effects of mRNA stability on the target gene expression. Based on the poly A signal of 3' -untranslated region (UTR), nucleotide sequences of putative cytoplasmic polyadenylation elements (CPEs) and downstream elements (DSEs: U-rich, G-rich, GU-rich) were analyzed and used to construct 15 luciferase reporter vectors. Each vector was transfected to HC11 and porcine mammary gland cell (PMGC) and measured for dual luciferase expression levels after 48 hours of incubation. Luciferase expression was significantly higher in construct #6 (with CPE 2, 3 and DSE 1 of exon 9) and #11 (with CPE 2, 3 and DSE 1, 2 and 3 of exon 9) than construct #1 in the PMGC. These results suggest that expression of target genes in PMGC may be effectively expressed by using the construct #6 and #11 on production of transgenic pig.

• Childhood Kidney Diseases
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• v.14 no.2
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• pp.143-153
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• 2010

Purpose: Recently, massive proteinuria has been observed in some transplant patients after switching cyclosporine A (CsA) to sirolimus. To evaluate the pathogenesis of sirolimus-associated proteinuria, we investigated the early changes in slit diaphragm molecules by various administrative conditions of sirolimus and CsA. Methods: In vitro-Mouse podocytes were incubated with buffer (C), sirolimus ($10\;{\mu}g/mL$) after CsA ($10\;{\mu}g/mL$) (C-S), sirolimus only (S) and CsA and sirolimus simultaneously (C+S) for 12, 24, and 48 hours. In vivo- twenty four SPF female Wistar rats were divided into 4 groups buffer (C), sirolimus after 2 weeks of CsA (C-S), sirolimus only (S) and CsA and sirolimus simultaneously (C+S). All groups were treated by intraperitoneal injection every other day for 4 weeks (CsA: 25 mg/kg, sirolimus: 0.5 mg/kg). The changes in mRNA of slit diaphragm molecules were examined by RT-PCR. Results: The mRNA of nephrin was significantly decreased in group C-S and C+S in vitro. In vivo, the mRNA of nephrin in all groups using sirolimus and the mRNA of podocin in group C-S and C+S were decreased. Microscopically, group C-S and C+S showed small vacuolization and calcification in proximal tubular epithelial cells. Immunohistochemistry using nephrin and podocin antibodies did not show remarkable decrease of staining along the glomerular capillaries. Electron-microscopically, focal fusion of foot processes was seen in group C-S and C+S. Conclusion: This study suggests the decrease of slit diaphragm molecules (nephrin and podocin) in podocyte may be one of the causes of sirolimus associated proteinuria, and podocyte injury by sirolimus may need a primary hit by CsA to develop the proteinuria.

• Korean Journal of Animal Reproduction
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• v.21 no.3
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• pp.293-302
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• 1997

Follicular oocytes of Grade I and II were collected from 2~6 mm ovarian follicles and matured in vitro (IVM) for 24 hrs in TCM-199 su, pp.emented with 35$\mu\textrm{g}$/ml FSH, 10$\mu\textrm{g}$/ml LH, and 1$\mu\textrm{g}$/ml estradiol-17$\beta$ at 39$^{\circ}C$ under 5% CO2 in air. They were fretilized in vitro (IVF) by epididymal spermatozoa capacitated with heparin for 12 hrs. The zygotes were then co-cultured in vitro with bovine oviducted epithelial cells (BOEC) for 7 to 9 days. The optimal time for IVM, the successful enucleation of IVM oocytes by micromanipulation at different oocyte ages after IVM, and the ideal culture system for IVM for effective IVF and in vitro development of IVM-IVF embryos was examined for in vitro production of nuclear recipient oocytes and nuclear donor embryos. To improve the efficiency of nuclear transplantation (NT) of IVF embryo into IVM follicular oocytes, this study evaluated the optimal electric condition and oocytes age for activation of IVM oocytes and in vitro development of NT embryos. In vitro development of NT embryos with preactivation or non-preactivation in enucleation oocytes, cell number of IVN-IVF embryos, and NT embryos wre also examined. The results obtained were as follows; 1. The most suitable enucleation time was at 24 hpm (83.3%) rather than that of 28 hpm(69.6%) and 32 hpm(50.0%). 2. There was no difference among the fusion rates of NT embryos at the voltages of 0.75, 1.0 and 1.5 kV/cm, but the in vitro development rates to morule and blastocyst were significantly (P<0.05) higher at the voltage of 0.75(12.5%) and 1.0kV/cm (12.6%) compared to 1.5kV/cm(0%). 3. No significant difference in activation rates were seen in NT embryos stimulated for 30, 60 and 120 $\mu$sec (71.7, 85.2 and 71.9%, respectively), but the in vitro development rates to morulae and blastocyst were significantly (P<0.05) higher in the oocytes stimulated for 30 $\mu$sec (11.6%) and 60 $\mu$sec(10.7%) than 120 $\mu$sec(0.0%). 4. The fusion rates (71.0 and 87.3%) and the in vitro development rates (9.1 and 12.7%) to morula and blastocyst were seen in the NT embryos stimulated at 28 and 32 hpm under the condition of 1.0 kV/ml, 60 $\mu$sec. However, at 24 hpm the fusion rates were 64.8% and the in vitro development to morula and blastocyst were not seen. 5. The fusion rates between the 8~12, 13~17 and 18~22-cell stage of IVM-IVF embryos were not significantly different. The in vitro development rates of the fused embryos to morula and blastocyst which were received from a blastomere of 8~12, 13~17 and 18~22-cell stages of IVM-IVF embryos were 14.9, 8.3 and 6.5%, respectively. 6. The in vitro development rate of the enucleated recipient oocytes with preactivation (24.2%) to morula and blastocyst was significantly (P<0.05) higher than that of non-preactivation (12.8%). 7. The cell numbers of NT blastocyst and IVM-IVF blastocyst cultured during 7~9 days were 63$\pm$11 and 119$\pm$23, and then their the mean cell cycle number were 5.98 and 6.89, respectively.

• Korean Journal of Environmental Biology
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• v.32 no.3
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• pp.216-224
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• 2014

Decrease in dissolved oxygen concentrations associated with temperature fluctuation is an important criteria to evaluate the mortality rate of the species. Based on this parameter, we investigated the survival rate, physiological response and histological change of warty sea squirt. It was found that the survival rate of the warty sea squirt species was 63.3% at $23^{\circ}C$ and 56.6% at $26^{\circ}C$ respectively. However, exposure of six days at $29^{\circ}C$ caused deaths among species, which indicated the 6day-$LT^{50}$ of the tested species to be $24.58^{\circ}C$ ($19.48{\sim}35.48^{\circ}C$). Further, after 11 day of exposure, the dissolved oxygen concentration has been found to decrease, with the survival rate of 20% at $4.0mg\;L^{-1}$ and deaths at $2.0mg\;L^{-1}$, thus 11day-$LC^{50}$ calculated to be $3.88mg\;L^{-1}$ ($3.29{\sim}4.57mg\;L^{-1}$). In addition, decrease in rate of oxygen consumption and excretion of ammonia was also noted at this critical water temperature and dissolve doxygen concentration. Moreover, there has been common histopathological changes were observed in warty sea squirt's gill pouch, digestive tract, and tunic as follows such as: proliferation of epithelial cells, condensation and necrosis, permeation of phagocyte and blood cell, loss of cilium and muscular fiber degeneration. Based on our study results, we suggest that these parameters can also be useful to evaluate the survival rate and physiological response in other species.