• Interdisciplinary Bio Central
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• v.1 no.3
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• pp.9.1-9.6
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• 2009

Introduction: In the mass spectrometry-based proteomics, biological samples are analyzed to identify proteins by mass spectrometer and database search. Database search is the process to select the best matches to the experimental mass spectra among the amino acid sequence database and we identify the protein as the matched sequence. The match score is defined to find the matches from the database and declare the highest scored hit as the most probable protein. According to the score definition, search result varies. In this study, the difference among search results of different search engines or different databases was investigated, in order to suggest a better way to identify more proteins with higher reliability. Materials and Methods: The protein extract of human mesenchymal stem cell was separated by several bands by one-dimensional electrophorysis. One-dimensional gel was excised one by one, digested by trypsin and analyzed by a mass spectrometer, FT LTQ. The tandem mass (MS/MS) spectra of peptide ions were applied to the database search of X!Tandem, Mascot and Sequest search engines with IPI human database and SwissProt database. The search result was filtered by several threshold probability values of the Trans-Proteomic Pipeline (TPP) of the Institute for Systems Biology. The analysis of the output which was generated from TPP was performed. Results and Discussion: For each MS/MS spectrum, the peptide sequences which were identified from different conditions such as search engines, threshold probability, and sequence database were compared. The main difference of peptide identification at high threshold probability was caused by not the difference of sequence database but the difference of the score. As the threshold probability decreases, the missed peptides appeared. Conversely, in the extremely high threshold level, we missed many true assignments. Conclusion and Prospects: The different identification result of the search engines was mainly caused by the different scoring algorithms. Usually in proteomics high-scored peptides are selected and low-scored peptides are discarded. Many of them are true negatives. By integrating the search results from different parameter and different search engines, the protein identification process can be improved.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.29 no.1
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• pp.44-50
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• 1996

Sex differentiation in the rockfish, Sebastes schlegeli, was studied by using a histological method for the appearance of primordial germ cell, formation of primitive gonad and differentiation of female and male. The primordial germ cells were buried under fibrous mesenchymal tissue between gut and mesonephric duct of pre-larva with a total length (TL) of 6.3 mm at 2 days after parturition. In juvenile of TL $5.2\~5.9cm$ at 65 days after parturition, the gonad composed of a large number of genial cell and formed of cavity along the lateral side of the gonad, differentiated to the ovary. At this time, the gonad formed seminiferous tubules by somatic cells, differentiated to the testis. In juvenile of TL $7.0\~7.2cm$ cm at 115 days after parturition, gonads divided into testis contained pigment cell and ovary absent pigment cell. S. schlegeli differentiated directly into male or female without an intermediate female phase at early indifferentiated stage. Therefore, S. schlegeli belongs to the differentiated type of gonochoristic teleosts. At 350 days after parturition, sex ratio was approximately 1 : 1(p>0.05).

• Journal of Chest Surgery
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• v.36 no.4
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• pp.219-228
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• 2003

Background:Liquid nitrogen freezing techniques have already met with widespread success in biology and medicine as a means of long-term storage for cells and tissues. The use of cryoprotectants such as glycerol and dimethylsulphoxide to prevent ice crystal formation, with carefully controlled rates of freezing and thawing, allows both structure and viability to be retained almost indefinitely. Cryopreservation of various tissues has various con-trolled rates of freezing. Material and Method: To find the optimal freezing curve and the chamber temperature, we approached the thermodynamic calculation of tissues in two ways. One is the direct calculation method. We should know the thermophysical characteristics of all components, latent heat of fusion, area, density and volume, etc. This kind of calculation is so sophisticated and some variables may not be determined. The other is the indirect calculation method. We performed the tissue freezing with already used freezing curve and we observed the actual freezing curve of that tissue. And we modified the freezing curve with several steps of calculation, polynomial regression analysis, time constant calculation, thermal response calculation and inverse calculation of chamber temperature. Result: We applied that freezing program on mesenchymal stem cell, chondrocyte, and osteoblast. The tissue temperature decreased according to the ideal freezing curve without temperature rising. We did not find any differences in survival. The reason is postulated to be that freezing material is too small and contains cellular components. We expect the significant difference in cellular viability if the freezing curve is applied on a large scale of tissues. Conclusion: This program would be helpful in finding the chamber temperature for the ideal freezing curie easily.

• Biomolecules & Therapeutics
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• v.26 no.5
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• pp.487-493
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• 2018

Cluster of differentiation 44 (CD44), a cell surface receptor for hyaluronic acid (HA), is involved in aggressive cancer phenotypes. Herein, we investigated the role of the CD44 standard isoform (CD44s) in hypoxia-inducible $factor-1{\alpha}$ ($HIF-1{\alpha}$) regulation using MCF7 overexpressing CD44s (pCD44s-MCF7). When pCD44s-MCF7 was incubated under hypoxia, levels of $HIF-1{\alpha}$, vascular endothelial growth factor, and the $HIF-1{\alpha}$ response element-derived luciferase activity were significantly increased compared to those in the control MCF7. Incubation of pCD44s-MCF7 cells with HA further increased $HIF-1{\alpha}$ accumulation, and the silencing of CD44s attenuated $HIF-1{\alpha}$ elevation, which verifies the role of CD44s in $HIF-1{\alpha}$ regulation. In addition, the levels of phosphorylated extracellular signal-regulated kinase (ERK) was higher in hypoxic pCD44s-MCF7 cells, and $HIF-1{\alpha}$ accumulation was diminished by the pharmacological inhibitors of ERK. CD44s-mediated $HIF-1{\alpha}$ augmentation resulted in two functional outcomes. First, pCD44s-MCF7 cells showed facilitated cell motility under hypoxia via the upregulation of proteins associated with epithelial-mesenchymal transition, such as SNAIL1 and ZEB1. Second, pCD44s-MCF7 cells exhibited higher levels of glycolytic proteins, such as glucose transporter-1, and produced higher levels of lactate under hypoxa. As a consequence of the enhanced glycolytic adaptation to hypoxia, pCD44s-MCF7 cells exhibited a higher rate of cell survival under hypoxia than that of the control MCF7, and glucose deprivation abolished these differential responses of the two cell lines. Taken together, these results suggest that CD44s activates hypoxia-inducible $HIF-1{\alpha}$ signaling via ERK pathway, and the $CD44s-ERK-HIF-1{\alpha}$ pathway is involved in facilitated cancer cell viability and motility under hypoxic conditions.

• Korean Journal of Veterinary Research
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• v.39 no.1
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• pp.1-11
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• 1999

This investigation was carried out to study morphological and chronological aspects of the development of the Harderian gland in the Mongolian gerbil(Meriones unguiculatus). Male and female Mongolian gerbils were sacrificed on days 1, 3, 5, 10, 30 and 60 after birth and their Harderian glands were processed for light microscopic observation. The results obtained were summarized as follows; 1. In 1-day-old Mongolian gerbil, Harderian gland was well distinguished from other tissue structures. It was composed of several immature tubules, and these tubules were separated each other by undifferentiated mesenchymal connective tissues. 2. In 3-day and 5-day-old Mongolian gerbils, the arrangement of tubules in the gland was more condensed than that of 1-day-old Mongolian gerbil. The excretory ducts started to appear in the connective tissues located between lobes. 3. In 10-day-old Mongolian gerbil, small lipid vacuoles began to be found in the cytoplasm of the secretory cells of the Harderian gland. There were some mucus-secreting cells within the epithelium of the excretory duct found in the interlobar connective tissues. 4. In 30-day-old Mongolian gerbil, there was markedly increased number of the tubules in the glands. The epithelial cells of the tubules were typically columnar in shape. Most of the columnar epithelial cells contained many small lipid vacuoles, although a few cells contained large lipid vacuoles. 5. In 60-day-old Mongolian gerbil, the Harderian gland exhibited the typical structural characteristics of the adult gland. The mature glandular structures were more distinct than those of 30-day-old animals.

• Applied Microscopy
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• v.28 no.1
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• pp.107-119
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• 1998

This study was designed to investigate the appearence and the characteristics of the apoptotic cells and the process of the joint cavity formation in mouse knee joint. Fetal mouse knee joints from 15 to 19 days of gestation were used. Paraffin-embedded serial sections, stained with H & E for light microscopic observation, Epon 812 embedded thin sections for electron microscopic observation and Lowicryl HM 20 embedded thin sections for immune-electron microscopic observation were prepared. Monoclonal antibodies to $\beta-tubulin$ and polyclonal antibodies to tissue transglutaminase were used for immune-electron microscopic study. The results obtained were as follows. 1. At 15 days of gestation, blood vessels, which have invaded in the mesenchymal cells, were present in the synovium, to form the joint cavity in the future. 2. At 16 days of gestation, the joint cleft was first appeared and several RBCs were present in the joint cleft. The invasion of blood vessels into the joint cleft was continuing, and apoptotic cells were present in the inner cell layer, adjacent to the joint cleft. Necrotic cells were also present in the outer cell layer; they were present 18 days of gestation, but apoptotic cells did not appear after 17 days of gestation. 3. In the apoptotic cells, transglutaminase were localized around vacuoles and the marginal site of the cytoplasm. 4. In the apoptotic cells, tubulin was around the endoplasmic reticulum and the marginal site of the cytoplasm. In the late stage of apoptotic cells, tubulin was localized diffusely in the cytoplasm. Tubulin was also strongly labeled around in the cytoplasm of the neighboring cell at which the apoptotic body was phagocytosed. Tubulin labeled particles were apparently increased in the seperated apoptotic bodies. On the basis of the above findings, it is proposed that during the development of the mouse knee joint, blood vessel invasion first occurs and then apoptosis and cell necrosis follow it. In the apoptotic cell, present in the synovium of the developing knee joint of the mouse. it is suggested that the redistribution of tubulin is associated with apoptotic process. And transglutaminase overexpressed in the apoptotic cell.

• Journal of Periodontal and Implant Science
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• v.39 no.2
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• pp.119-128
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• 2009

Purpose: The aim of this study was to evaluate the stemness of cells from canine dental tissues and bone marrow. Methods: Canine periodontal ligament stem cells (PDLSC), alveolar bone stem cells (ABSC) and bone marrow stem cells(BMSC) were isolated and cultured. Cell differentiations (osteogenic, adipogenic and chondrogenic) and surface antigens (CD146, STRO-1, CD44, CD90, CD45, CD34) were evaluated in vitro. The cells were transplanted into the subcutaneous space of nude mice to assess capacity for ectopic bone formation at 8 weeks after implantation. Results: PDLSC, ABSC and BMSC differentiated into osteoblasts, adipocytes and chondrocytes under defined condition. The cells expressed the mesenchymal stem cell markers differently. When transplanted into athymic nude mice, these three kinds of cells with hydroxyapatite /${\beta}$- tricalcium phosphate (HA/TCP) carrier showed ectopic bone formation. Conclusions: This study demonstrated that canine dental stem cells have stemness like bone marrow stem cells. Transplantation of these cells might be used as a therapeutic approach for dental stem cell-mediated periodontal tissue regeneration.

• Journal of Aquaculture
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• v.20 no.1
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• pp.47-50
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• 2007

The primordial germ cells of the oblong rockfish, Sebastes oblongus were buried under fibrous mesenchymal tissues between gut and mesonephric duct of pre-larva with 4.0 mm total length (TL) at 1 day after the parturition. In the juvenile of 22.0 mm TL at 71 days after the parturition, the gonad composed of a large number of gonial cell and formed of cavity along the lateral side of the gonad, differentiated to the ovary. At this time, the gonad formed seminiferous tubules by somatic cells, was differentiated to the testis. The smallest oblong rockfish that possessed primary oocytes was about 42.1 mm TL at 141 days. Spermatogonia remained quiescent until most fish were over 42.1 mm TL at 141 days. The oblong rockfish the differentiated directly into male or female without an intermediate female phase at the early indifferentiated stage. Therefore, the oblong rockfish belongs to the differentiated type of gonochoristic teleosts.

• Tuberculosis and Respiratory Diseases
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• v.46 no.1
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• pp.129-135
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• 1999

We describe unusual manifestations of congenital cystic adenomatoid malformation (C.C.A.M.) of the lung, such as movable fungal ball-like intracystic blood clots and hemoptysis, which were detected in previously healthy 23 years-old woman. We identified blood clots only after left upper lobectomy and could not distinguish from fungal ball with radiographic methods. CCAM of the lung, rare and lethal form of congenital pulmonary cystic disease, was initially introduced by Ch, in and Tang in 1949. The histogenesis of this lesion is characterized by polypoid glandular tissue proliferation and overgrowth of mesenchymal elements due to cessation of bronchiolar maturation which occurred in after 16weeks intrauterine period. In 80-95% of reported cases, the lesion was confined to a single lobe and there was no lobe and right and left lung predilection The clinical presentation may be widely variable, ranging from intrauterine fetal death to late discovery in childhood with recurrent pulmonary infection. But there,s no reports which were misdiagnosed with intracystic fungal ball. The treatment choice is lobectomy of affected lobe. There,s a few case reports with rhabdomyosarcoma, bronchiolar cell carcinoma and myxosarcoma arising in CCAM patients. Therefore, early resection is recommended even if asymtomatic cases. We experienced a rare case of CCAM of the lung in 23 years old female, and there were intracystic fungal ball-like movable blood clots in lower portion of left lung. After left upper lobectomy was performed, now she is discharged and followed up without any complications.

• Tuberculosis and Respiratory Diseases
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• v.65 no.4
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• pp.334-338
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• 2008

Solitary fibrous tumors of the pleura (SFTPs) are relatively rare tumors that originate from the mesenchymal cells of the submesothelial tissue of the pleura. Patients with SFTPs are often asymptomatic, while some patients experience pleuritic chest pain, cough and/or dyspnea. We report here on a case of SFTP, and the patient presented with septic shock and respiratory failure that required mechanical ventilation. A 68-year-old woman was admitted for the evaluation of her dyspnea and generalized edema. Chest imaging studies showed an 18 cm-sized voluminous mass occupying the right thoracic cavity with anterior displacement of hilar structures and atelectasis of the right lung. Immediately after admission, she developed pneumonia and septic shock that required antibiotics and mechanical ventilation. She displayed a partial response to medical treatment, and then complete excision of the tumor was performed and the pathologic examination revealed benign SFTP. Afterward, she fully improved without evidence of recurrence until now.