• Proceedings of the Botanical Society of Korea Conference
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• 1996.07a
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• pp.48-60
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• 1996

We previously identified and characterized a predominantly pollen-expressed gene of Petunia inflata that encodes a receptor-like kinase named PRK1. The extracellular domain of PRK1 contains leucine-rich repeats which have been implicated in protein-protein interactions, and the cytoplasmic domain was found to autophosphorylate on serine and tyrosine. To investigate the function PRK1 in pollen development, we transformed P. inflata plants with a construct containing the promoter of a predominantly pollen-expressed gene of tomato, LAT52, fused to an antisense PRK1 cDNA corresponding to part of the extracellular domain of PRK1, There transgenic plants were found to each produce approximately equal amounts of normal and aborted pollen. Analysis of the inheritance of the transgene inserts in two of the transgenic plants, ASRK-13 and ASRK-20, to their progeny revealed that certain transgene inserts cosegregated with the pollen abortion phenotype. Microscopic examination of the aborted pollen grains showed that their outer wall, the exine, was essentially normal, but that their cytoplasm contained only starch-like granules. Staining of the nuclei of the microspores at different stages of uninucleate stage. However, at subsequent stages half of the microspores completed mitosis and developed into normal binucleate pollen, but the other half initially remained uninucleate, then lost their nucleio. Analysis of the amounts of PRK1 mRNA and the antisense PRK1 transcript suggested that the pollen abortion phenotype most likely resulted from down-regulation of the PRK1 gene by the antisense PRK1 transgene. These results suggest that PRK1 plays an essential role in a signal transduction pathway that mediates post-meiotic development of microspores.

• Animal cells and systems
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• v.1 no.4
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• pp.657-663
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• 1997

The CD4 and CDS coreceptors, in conjunction with the T cell receptor (TCR) , make important contributions to the differentiation of thymocytes. They have been shown to be involved in the clonal deletion and positive selection processes during T cell development in thymus. To further analyze the role of CD4 and CDS proteins during T cell differentiation, we have generated transgenic mice constitutively expressing high levels of a native CD4 and a CD4{CDSa hybrid protein. The hybrid protein is composed of CD4 extracellular domain linked to the CD8a transmembrane region and cytoplasmic tail. The transgenes were driven by human beta-actin promoter, and therefore, they were expressed in all tissues examined including thymus, spleen, and lymph nodes. The resulting CD4 and CD4{CD8${\alpha}$transgenic mice were found to express the CD4 and CD4{CD8${\alpha}$ respectively, in developing thymocytes and peripheral T cells. The expression levels of transgenic proteins were 5-10 times higher than that of endogenous CD4 in thymus. However, total surface CD4 expression (CD4 or CD4{CD8${\alpha}$ transgenic protein plus endogenous CD4) of the transgenic mice were main. tained at similar levels compared to control littermates. Surface CD4 expression on CDS T cells, however, was significantly lower than that on cells expressing endogenous CD4. These results suggest that a total avidity between developing thymocytes and thymic stromal cells is impor. tant for differentiation of thymocytes.

• Animal cells and systems
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• v.16 no.2
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• pp.104-113
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• 2012

The nucleolus is a distinct nuclear territory involved in the compartmentalization of nuclear functions. There is some evidence of a relationship between nuclear fragmentation during spermatogenesis and chromatoid body (CB) formation. The CB is a typical cytoplasmic organelle of haploid germ cells, and is involved in RNA and protein accumulation for later germ-cell differentiation. The goal of this study was to qualitatively and quantitatively describe the nucleolar cycle during the spermatogenesis of $Phrynops$ $geoffroanus$ (Reptilia Testudines), and compare this nucleolar fragmentation with CB formation in this species through the use of cytochemical and ultrastructural analysis. Qualitative analysis showed a fragmentation of the nuclear material after pachytene of the first meiotic division in the primary spermatocytes. Quantitative analysis of the nucleolar cycle revealed a significant difference in the number of nucleoli and in the size of the nucleolus between spermatogonia and early spermatids. Using ultrastructural analysis, we recorded the beginning of the CB formation process in the cytoplasm of primary spermatocytes at the same time as when nuclear fragmentation occurs. In the cytoplasm of primary spermatocytes, the CB was observed in association with mitochondrial aggregates and the Golgi complex. In the cytoplasm of early spermatids, the CB was observed in association with lipid droplets. In conclusion, our data show that the nucleolus plays a role in the CB formation process. During spermatogenesis of $P.$ $geoffroanus$, the CB is involved in some important biological processes, including acrosome formation and mitochondrial migration to the spermatozoon tail and middle piece region.

• Korean Journal of Veterinary Service
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• v.13 no.2
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• pp.162-183
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• 1990

From 1988 to 1989, 8 strains of canine parvovirus-2 (CPV-2) were isolated from the fecal specimens from the dogs that were clinically diagnosed as canine parvoviral enteritis in the veterinary hospitals located in the regions of Taejon and Chungbuk province. Studios on biological and physicochemical properties for the isolates were carried out. The results obtained by experiments are summarized as follows. 1. Among 62 fecal samples collected from the dogs with enteric diseases, 24 (38.7%) showed the haemagglutinating activity against porcine erythrocyte, ranging from 16 to 16, 384 of HA titers. 2. When 8 fecal specimens with high HA titer over 1, 000 were inocultated into CRFX cells, intranuclear inclusion bodies were obseverd in all of eight specimens, of w)lick three specimens showed cytoplasmic inclusions concurrently with the intranuclear inclusion bodies. 3. In study on species-specificity of haemagglutinating activity of the isolates, TJ-89-1 and TJ-89-2, it was found that the isolates revealed the highest haemagglutinating activity with porcine erythrocytes, showing the relatively lower haemagglutination titers with the erythrocytes from cat and rabbit. None of erythrocytes from other animals reacted with the isolates. 4. By the cross-haemagglutination inhibition test of the Isolates with reference viruses and sera, the Isolates were evidently identified as the strains of canine parvovirus-2. 5. In Physicochemical property test, it was evident that the isolates were stable in, lipid solvent, pH and heat treatment at $56^{\circ}C$ for 30 min. and contain DNA genome. 6. When seven puppies were inoculated intraorally with the isolate at HA titer of 8, 192, all of the puppies showed the symptoms of anorexia, vomiting, diarrhea and died at the 5th to 10th days post inoculation(pi). The fecal samples from all of the puppies revealed significantly high HA titers afterward the 5th days pi. Body temperature and the number of total leucocytes were slightly increased at the early stage of infection. but extremely decreased at the stage of collapse. HI titers of the sera started to increase at the 2nd to 3rd days pi reaching 512 to 1, 024 at the 4th to 5th day pi.

• Korean Journal of Veterinary Research
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• v.39 no.3
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• pp.583-592
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• 1999

We have developed the in situ hybridization(ISH) technique for rapid diagnosis of canine distemper(CD) which is the major infectious disease in dogs. In our experiment, we rapidly detected distribution of the specific canine distemper viral genome without disrupting morphology of tissues or cells. Two oligonucleotide probes for ISH were synthesized chemically and labelled 5' end with nonisotopic biotin by DNA synthesizer. The whole procedures of ISH was completed within 1~2 hours using the Microcapillary action system. On histological study, typical cytoplasmic or intranuclear inclusion bodies were observed in the trachea, bronchiole, brain, and urinary bladder with the presence of prominent red positive signals on ISH, indicating specific CDV genome from the paraffin-embedded tissues of infected 13 cases. The results showed ISH can be used as a rapid and effective diagnostic method for diagnosis of CD.

• Korean Journal of Veterinary Research
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• v.39 no.3
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• pp.425-434
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• 1999

The morphological studies on the cecal development in the 60-, 90-, and 120-day-old fetuses and the newborns of Korean native goats were investigated by scanning and transmission electron microscopy. The results were summarized as follows ; Scanning electron microscopic studies : 1. In the 60-day-old fetuses, fold-like shapes protrusion on the cecal mucosa surface appeared. In the 90-day-old fetuses, the cecal villi appeared to be columnar shapes. In the 120-day-old fetuses, the cecal villi showed various tongue-like or columnar shapes. In the newborns, only the rudimental trace of the villi and the intestinal glands were observed. Transmission electron microscopic studies : 2. In the 60-day-old fetuses, the cecal epithelia were simple columnar in some areas and stratified columnar in others, and the epithelial cells contained nuclei, nucleoli, ER, mitochondria, Golgi complexes, zonula occludens, desmosomes, digitiform intercellular junctions, and large masses of the glycogen granules. 3. In the 90-day-old fetuses, the cecal epithelia were simple columnar in some area and stratified columnar in other. The microvilli of the cecal epithelia became much larger and longer than those in the 60-day-old fetuses, and intercellular junctions were developed, and increased numbers of ER, mitochondria, Golgi complexes were observed and the goblet cells contained a lot of the secretory granules. 4. In the 120-day-old fetuses, the cecal epithelia were only simple columnar in all areas. Microvilli and cytoplasmic organelles were well developed and the irregular annular nuclei were observed. 5. In the newborns, the cecal epithelia were covered with extensive microvilli, and the goblet cells with secretory granules were protruded into the lumen. And some goblet cells secreted the secretory granules into the lumen.

• Korean Journal of Veterinary Research
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• v.37 no.1
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• pp.41-57
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• 1997

This study was carried out to investigate the effect of selenium on the adriamycininduced renal lesions in male Sprague Dawley rats. A total of 60 Sprague-Dawley male rats were divided into 2 control groups(C1: saline, C2: selenium) and 2 treatment groups(T1: adriamycin, T2: adriamycin+selenium). The rats of the C1 and T1 groups were given normal saline(0.15ml/rat), the rats of the C2 and T2 groups were given sodium selenite(0.5mg/kg) intraperitoneally three days a week for 4 weeks. The treatment groups were dosed intraperitoneally with adriamycin(2mg/kg/day) five days at the second week. Animals were sacrificed at the 1st week, 2nd week and 3rd week after dosing with adriamycin. The morphologic abnormalities of the glomeruli and tubules in the kidney of male rats were examined histopathologically and electron microscopically.The results obtained were as follows : The mean body weight of adriamycin dosed group was significantly decreased as compared with that of control group at 4th week(p<0.05). In adriamycin and selenium dosed group, the mean body weight was decreased until the end of 2nd week but gradually increased from 3rd to 4th week. The histopathological findings of the renal corpuscle in adriamycin dosed group were parietal epithelial cell proliferation, vacuolization of glomerulus, and thickened basement membrane of the parietal epithelium. Proximal convoluted tubules were significantly dilated and the lumens were filled with renal cast. These lesions were generally not very significant in the rats given adriamycin and selenium. The electron microscopical findings of the renal glomerulus in the adriamycin dosed group were focal loss and fusion of the pedicels of the podocyte, and some vacuoles in the cytoplasm of the podocytes. There were numerous cytoplasmic vacuoles in the proximal and distal convoluted tubular cells. However, these ultrastructural changes were not significantly observed in the renal tubules of the rats of adriamycin and selenium dosed group. These results suggest that selenium may act as an inhibitor of the renal lesions induced by adriamycin in male rats.

• Molecules and Cells
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• v.38 no.4
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• pp.349-355
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• 2015

EphA7 has been implicated in the regulation of apoptotic cell death in neural epithelial cells. In this report, we provide evidence that EphA7 interacts with caspase-8 to induce apoptotic cell signaling. First, a pull-down assay using biotinylated ephrinA5-Fc showed that EphA7 co-precipitated with wild type caspase-8 or catalytically inactive caspase-8 mutant. Second, co-transfection of EphA7 with caspase-8 significantly increased the number of cleaved caspase-3 positive apoptotic cells under an experimental condition where transfection of EphA7 or caspase-8 alone did not affect cell viability or apoptosis. EphA4 also had a causative role in inducing apoptotic cell death with caspase-8, whereas EphA8 did not. Third, caspase-8 catalytic activity was essential for the apoptotic signaling cascade, whereas tyrosine kinase activity of the EphA4 receptor was not. Interestingly, we found that kinase-inactive EphA4 was well co-localized at the plasma membrane with catalytically inactive caspase-8, suggesting that an interaction between these mutant proteins was more stable. Finally, we observed that the extracellular region of the EphA7 receptor was critical for interacting with caspase-8, whereas the cytoplasmic region of EphA7 was not. Therefore, we propose that Eph receptors physically associate with a transmembrane protein to form an apoptotic signaling complex and that this unidentified receptor-like protein acts as a biochemical linker between the Eph receptor and caspase-8.

• Molecules and Cells
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• v.39 no.3
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• pp.266-279
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• 2016

The mechanism by which 12-O-tetradecanoylphorbol-13-acetate (TPA) bypasses cellular senescence was investigated using human diploid fibroblast (HDF) cell replicative senescence as a model. Upon TPA treatment, protein kinase C (PKC) ${\alpha}$ and $PKC{\beta}1$ exerted differential effects on the nuclear translocation of cytoplasmic pErk1/2, a protein which maintains senescence. $PKC{\alpha}$ accompanied pErk1/2 to the nucleus after freeing it from $PEA-15pS^{104}$ via $PKC{\beta}1$ and then was rapidly ubiquitinated and degraded within the nucleus. Mitogen-activated protein kinase docking motif and kinase activity of $PKC{\alpha}$ were both required for pErk1/2 transport to the nucleus. Repetitive exposure of mouse skin to TPA downregulated $PKC{\alpha}$ expression and increased epidermal and hair follicle cell proliferation. Thus, $PKC{\alpha}$ downregulation is accompanied by in vivo cell proliferation, as evidenced in 7, 12-dimethylbenz(a)anthracene (DMBA)-TPA-mediated carcinogenesis. The ability of TPA to reverse senescence was further demonstrated in old HDF cells using RNA-sequencing analyses in which TPA-induced nuclear $PKC{\alpha}$ degradation freed nuclear pErk1/2 to induce cell proliferation and facilitated the recovery of mitochondrial energy metabolism. Our data indicate that TPA-induced senescence reversal and carcinogenesis promotion share the same molecular pathway. Loss of $PKC{\alpha}$ expression following TPA treatment reduces pErk1/2-activated SP1 biding to the $p21^{WAF1}$ gene promoter, thus preventing senescence onset and overcoming G1/S cell cycle arrest in senescent cells.

• The Journal of Korean Medicine
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• v.20 no.3 s.39
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• pp.27-35
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• 1999

The purpose of this study was to investigate the histochemical effect of Jojoongikgi-tang (JIT) on hairless mice induced by DNCB. For the study, DNCB was applied on the infrascapular region of the mices skin and then JIT was orally administered. As a result, the following histochemical changes of the dermis were observed through light and electron microscopes, and statistical data. The results obtained were as follows: 1. In the 2 days experimental group, more histamine mast cells dermis occurred than in the control or normal group when observed under light microscope. When an electron microscope was used, histochemical reactive cells in the dermis were found as mast cells that contained more cytoplasmic granules than in the control or normal group. 2. In the 5 days experimental group, the number of mast cells were decreased over the control or normal group when observed under light microscope. When an electron microscope was used, mast cells in the control contained secretory granules with higher electron density than those of the experimental group. 3. As a result of statistical analysis, the mean value of mast cells in the normal 2 days control and experimental groups was not significantly different (p<0.05). 4. However, the mean values of it in the normal ($19.1000{\pm}6.3154$), 5 day control ($103.4500{\pm}42.2704$) and 5 day experimental groups ($35.9500{\pm}8.5746$) were significantly different (p<0.05). From the above results, it is concluded that JIT is efficient against the purpura induced by DNCB.