This study was carried out to examine the effects of maturation media on in vitro maturation (IVM) of porcine immature oocytes, and on subsequent in vitro fertilization (IVF) and development (IVD). Cumulus-oocyte complexes (COCs) were collected from antral follicles of porcine ovaries collected from abattoir, and were matured in vitro in modified NCSU-37 (mNCSU-37), modified NCSU-23 (mNCSU-23), or TCM-199 supplemented with 10% porcine follicular fluid (pFF). Oocytes matured in vitro, were fertilized in vitro in modified Tris-buffered medium(mTBM) with the final motile sperm concentration of 1${\times}$105 sperm/mL, and subsequently putative embryos were developed in vitro in NCSU-23. The results are as follows. 1. In the result of IVM, the rate of germinal vesicle breakdown (GVBD) and of nuclear maturation were not significantly different among the media, though numeric value of them were slightly lower in TCM-199 than in mNCSU-37 or in mNCSU-23. 2. In the result of IVF, though the rate of sperm penetration was not significantly different among the maturation media, the percentage of oocytes with male pronucleus (MPN) of ones matured in mNCSU-37 (88.0%) was significantly higher than in TCM-199 (71.1%) (p<0.05). 3. In the result of IVD, the percentage of cleaved oocytes of ones matured in mNCSU-37 (52.3%) or in mNCSU-23 (53.7%) was significantly higher than in TCM-199 (43.1%) (p<0.05), but the rate of blastocysts at day 6 was not significantly different among the maturation media, though putative embryos from oocytes matured in mNCSU-37 or in mNCSU-23 were developed more than in TCM-199. These results suggested that mNCSU-37 or mNCSU-23 was more appropriate than TCM-199 as IVM medium for porcine immature oocytes.
This study was carried out to examine the effects of epidermal growth factor (EGF) and the number of cumulus-oocyte complexes (COCs) on in vitro maturation (IVM) of porcine immature oocytes, and on subsequent in vitro fertilization (IVF) and development (IVD). COCs were collected from antral follicles of porcine ovaries collected from abattoir, and were maturated in modified NCSU-23 (mNCSU-23) with 10% pFF, 0.6 mM cysteine, 50 ${\mu}mM{\beta}-mercaptoethanol$, 1 mM dbcAMP, 10 IU/mL PMSG and 10 IU/mL hCG, which was supplemented with or without 10 ng/mL EGF and into which 50 or 15 COCs per droplet was put. Oocytes matured in vitro, were fertilized in vitro in modified Tris-buffered medium (mTBM) with the final motile sperm concentration of 1${\times}$105 sperm/mL, and subsequently putative embryos were developed in vitro in NCSU- 23. The results are as follows. 1.In the result of IVM, 10 ng/mL EGF supplement duplicated the percentage of C4 group of COCs(41% vs 81%). But the rate of germinal vesicle breakdown (GVBD) and of nuclear maturation were not significantly different between control and EGF supplemented, or between the number of COCs per culture droplet, and there was not a significant interaction between the two factors, either. 2. In the result of IVF, there was not significantly different between control and EGF supplemented, or between the number of COCs per culture droplet, or was not a significant interaction between the two factors, in the rate of sperm penetration, in the percentage of oocytes with male pronucleus (MPN), and in the rate of polyspermy. 3. In the result of IVD, there was not significantly different between control and EGF supplemented, or between the number of COCs per culture droplet in the percentage of cleaved oocytes. There was not significantly different between the number of COCs per culture droplet, but between control and EGF supplemented (p<0.01) in the percentage of blastocysts, the number of inner cell mass (ICM), trophectoderm (TC) and total cells. There was no significant interaction between the two factors anywhere. These results suggested that 10 ng/mL EGF supplement into mNCSU-23 for IVM was effective in the production of more as well as better blastocysts during IVD through increasing the number of cells in those.
Journal of the korean academy of Pediatric Dentistry
/
v.27
no.1
/
pp.54-61
/
2000
Amalgam, though a widely used dental material, does not bond to the tooth substrate Therefore, retentive preparation of the cavity is necessary. Such amalgam restorations, until corrosion products form and plug the margin, will show significant marginal leakage. Unless this is prevented early on, saliva and bacteria may enter the cavity causing postoperative hypersensitivity, dissolution and collapse of the restoration, discoloration of the margin and secondary caries, leading to shortened life-span of the restoration and pulpal pathosis. Recently, a method of restoration has been introduced whereby tooth material can be preserved, cavity margin can be sealed and preventive treatment of pit and fissure can be administered while retaining all the advantages of conventional amalgam restorations. Such sealed amalgams involve removing the carious lesion without extending the cavity for prevention and using pit and fissure sealants to seal cavity margins and pit and fissures to reduce microleakage. In this study, finishing of the amalgam and sealant application were performed after different intervals following of amalgam restoration to compare the microleakage of sealed and conventional amalgam restorations. Thirty bicuspids were prepared with Class V cavity preparations on the buccal and lingual surfaces. After amalgam placement, they were divided into the following groups and treated accordingly. Group 1 : Polishing after 24 hours Group 2 : Immediate sealant application without polishing Group 3 : No polishing, but sealant applied after thermocycling 500 times After treatment, the samples were thermocycled 500 times between $5^{\circ}C$ and $55^{\circ}C$ with a dwell time of 30 seconds. After thermocycling, the samples were dipped into 1% methylene blue kept in a $37^{\circ}C$ incubator at 100% humidity for 24 hours. The teeth were then embedded in resin and cut bucco-lingually along the tooth axis and observed with a stereomicroscope to determine the degree of microleakage, The following results were obtained : 1. Group 2 showed the least microleakeage, while group 1 showed the greatest. 2. Group 1 showed significantly greater microleakage compared to group 2 (p<0.05). However, no significant differences were found between group 1 and 3(p>0.05). No significant differences in microleakage were also found between cup 2 and 3(p<0.05).
Seo, Sang-Woo;Kown, Yong-Hoon;Kim, Hyun-Jung;Nam, Soon-Hyeun;Kim, Kyo-Han;Kim, Young-Jin
Journal of the korean academy of Pediatric Dentistry
/
v.30
no.3
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pp.423-430
/
2003
The teeth bleaching with bleaching agent is widely used at recent times. Until yet the exact mechanism of the bleaching agent isn't known but it is thought that is by the complex reduction-oxidation reaction of the decomposed free radical from bleaching agent through various ways. In other words, it is supposed that the teeth are whitened by agent's changing chemical structures of stain-causing materials. The purpose of this study is to exam the change of the dentinal character by bleaching agent and to evaluate the safety of this agent. For this study, after applying 10% carbamide peroxide to enamel of human premolar for 6 hours a day for 2 weeks we examined changes of surface morphology, microhardness, composition and contents of minirals in human dentin using SEM, microhardness tester, FT-Raman spectrometer and EPMA and got following results. There was no significant difference in surface morphologic change when we examined the effect of 10% carbamide peroxide which penetrated into dentin after applied on enamel surface comparing with result from specimen in distilled water No change was shown on the surface of peritubular and intertubular dentin within the nanometeric range. The microhardness between bleached teeth and teeth stored in distilled water showed no statistically significant difference FT-Raman spectra of dentin exhibited no change of the component in human dentin. Only the least change in peaks of organic and inorganic materials were detected in Raman intencity. The total content of mineral elements in dentin with no treatment, stored only in distilled water and stored in distilled water after bleaching were $98.73{\pm}1.89,\;98.56{\pm}2.11\;and\;97.47{\pm}2.51$ respectively. Also they showed no statistically significant difference. From above results, the effect of 10% carbamide peroxide bleaching on structure of dentin is very low and the results may confirm the safety of this bleaching agent.
From March 1979 through December 1986, 124 patients with early stage carcinoma of the uterine cervix received curative radiation therapy. According to FIGO classification, 35 patients were stage IB and 89 were stge II A. In stage IB, five year locoregional control, five year disease free survival, and five year overall survival was $79.0\%$, $76.4\%$ and $81.8\%$, respectively. In stage II A, five year locoregional control, five year disease free survival, and five year overall survival were $78.0\%$, $66.8\%$, and $72.1\%$, respectively. To identify prognostic factors, pretreatment parameters including age, ECOG performance status, number of pregnancies, history of diabetes mellitus and hypertension, histology, size and shape of primary tumor, CT findings and blood parameters were retrospectively analyzed in terms of locoregional control, disease free survival and overall survival using univariate analysis and multivariate analysis. In univariate analysis, tumor size on physicai examination and rectal invasion on CT significantly affected locoregional control, disease free survival and overall survival. Parametrial involvement on CT was a significant prognostic factor on locoregional control and disease free survival. Hemoglobin level affected disease free survival and overall survival. Histology and age were significant prognostic factors on locoregional control. In multivariate analysis excluding CT finding, tumor size on physical examination was a significant factor in terms of locoregioal control and overall survival. Hemoglobin level was significant in terms of disease free survival. In multivariate analysis including CT, histology was a prognostic factor on locoregional control and disease free survival. Hemoglobin level and rectal invasion on CT were significant factors on locoregional control.
Journal of the korean academy of Pediatric Dentistry
/
v.34
no.3
/
pp.359-369
/
2007
This in vitro study was performed to assess the effect of surface sealing on the microleakage of class V composite resin restorations that underwent several aging treatments. Class V cavities were prepared on the buccal surface of 100 sound extracted premolars and restored with a hybrid light-cured composite resin according to the manufacturer's instructions. They were randomly divided into two groups consisting of 50 samples: group I, without surface sealing, and group II, in which margins were etched and surface sealant was applied. After thermocycling, each group was divided into five subgroups, respectively, to represent the five aging treatments: group A = no further treatment (only thermocycling), B = toothbrushing, C = load cycling, D = toothbrushing followed by load cycling, and E = aging treatment in deionized water for six months. Microleakage was assessed by examining the penetration of 2% methylene blue dye. The following results were obtained: 1. At occlusal and cervical margins in groups without surface sealing, there was no significant difference in microleakage after the several aging treatments (p>0.05). 2. The occlusal margins of groups with surface sealing showed no significant differences after the several aging treatments (p>0.05). 3. In the cervical margins of groups with surface sealing, microleakage significantly increased after load cycling or aging in deionized water for six months (p<0.05). 4. The no-further-treatment group and the toothbrushing group with surface sealing showed less microleakage than the corresponding groups without surface sealing (p<0.05). 5. The surface-sealed groups with load cycling or aging in deionized water showed no significant difference in microleakage to the corresponding groups without surface sealing (p>0.05). In conclusion, the results of this study suggest that the surface sealant infiltrating through the gap of the cervical margin exerted a positive effect on microleakage at the initial stage, but the effect was not sufficient to overcome the stress generated by the cuspal flexure during occlusal loading and water absorption.
Kim, Yeon Sao;Kim, Seong Min;Kim, Jin Ho;Lee, Kyung Sang;Yang, Suck Chul;Yoon, Ho Joo;Shin, Dong Ho;Park, Sung Soo;Lee, Jung Hee;Choi, Yo Won;Jean, Seok Chol;Kim, Young Tae
Tuberculosis and Respiratory Diseases
/
v.43
no.4
/
pp.571-578
/
1996
Background : Long abscess is an accumulation of pus within a destroyed portion of the lung. Antibiotic therapy and postural drainge has proven to be an effective method of treatment for the majority of patients with pyogenic lung abscess. When medical therapy fails, thoracotomy and pulmonary resection are the current therapies. empyema is pus in the pleural space, and this term is deserved for effusions on which the Gram stain of the pleural fluid is positive. Initially, such collection may be drained via chest tribe. Recently, in patients who are judged to be unsuitable for surgery are in poor condition, percutaneous drainage using pig-tail catheter has been performed. We report out experience with 10 cases of lung abscess and 23 cases of empyema who were treated by percutaneous pit-tail catheter drainage. Subjects and Methods : Our study included 10 patients with lung abscess and 23 patients with empyema who were treated by percutaneous pig-tail catheter drainage, from January, 1990, to May, 1996, at Hanyang University then a pig-tail catheter was inserted into the abscess or the site of empyema under fluoroscopic and ultrasonograpic guidance. Following aspiration, the catheter was sutured into the skin, and connected to the suction tip. Catheter drainage was discontinued when the abscess of empyema was resolved in radiologically and clinically. Results : There were 2 cases of lung abscess caused by Staphylococcus aureus and Klebsiella pneumoniae and 14 cases of empyema caused by M. tuberculosis. The others were unknown. The duration of drainage was 1-2 weeks in 7 cases of lung abscess and 14 cases of empyema. In the 29 of 33 patients, percutaneous drainage were carried out successfully 20 of the 29 Gases rapidly improved. Conclusion : Percutaneous drainge is effective and relatively saute for management of lung abscess or empyema refractory to medical therapy or poor candidates for surgical treatment.
Kim, Jong-Uk;Choi, Dong-Soon;Joo, Hyun;Min, Churl-K.
KSBB Journal
/
v.23
no.3
/
pp.231-238
/
2008
The second family member of tissue inhibitors of matrix metalloproteinases, TIMP-2, is a 21kDa protein which inhibits matrix metalloproteinases 2 (MMP-2). Expression of mammalian proteins in E. coli often forms inclusion bodies that are made up of mis-folded or insoluble protein aggregates. The requirement for the formation of 6 disulfide bonds in the process of the TIMP-2 folding is likely to be incompatible with the reducing environment of E. coli. However, this incompatibility can be often overcome by introducing a mutagenesis that could lead to enhancement of the protein solubility. In this reason, we have attempted to express the soluble TIMP-2 in E. coli by applying a modified staggered extension process (StEP), one of the in vitro PCR-based recombinant mutagenesis methods, and error-prone PCR. C-terminally located CAT fusion protein with respect to mutated TIMP-2 proteins enables us to differentiate the soluble TIMP-2 from the insoluble in E. coli by virtue of chloramphenicol resistance. According to this scheme, E. coli harboring properly-folded CAT fused to TIMP-2 protein was selected, and some of the resulting colonies exhibited an enhanced, soluble expression of TIMP-2 compared to the wild type, implying (i) the StEP technique is successfully employed to enhance the proper folding thereby increasing the solubility of TIMP-2, and (ii) the CAT dependent screening may be a simple and effective method to differentiate the soluble protein expression in E. coli.
Journal of the Korean Society of Food Science and Nutrition
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v.40
no.2
/
pp.235-244
/
2011
The physicochemical properties and quality changes in salted and dried brown croaker products using deep seawater salt were investigated for preparation with different pretreatment, salting and drying conditions. Optimum salt concentration, salting time and drying time for product were 20% of body weight, 12 hrs at room temperature ($25{\pm}2^{\circ}C$), and 24 hrs at $30{\pm}2^{\circ}C$, respectively. The volatile basic nitrogen (VBN), peroxide value (POV), viable cell count and acid value (AV) of viscera and blade removed products were 18.9~22.4 mg%, 21.0~32.5 meq/L, and 2.3~4.4 mg/mL, $2.0\sim3.5{\times}10^3$ CFU/g, respectively and only viscera removed products were 31.2~38.1 mg%, 40~55.2 meq/L, 7.0~11.5 mg/mL, $4.1\sim5.5{\times}10^3$ CFU/g, respectively. Changes in quality of salted and dried brown croaker products were investigated during storage at room temperature and $5{\pm}2^{\circ}C$. The POV, AV and viable cell counts of salted and dried brown croaker products increased in commercial salt used and only viscera removed products compared with deep seawater salt used and viscera and blade removed products during storage time. Results in this study showed that the deep seawater salt was very effective for quality control of salted and dried brown croaker products and pretreatment method was a very important factor for preparation products. The optimum conditions for preparation of salted and dried brown croaker product were 20% of body weight for salt concentration, 12 hrs at $25{\pm}2^{\circ}C$ for salting time, and 24 hrs at $30{\pm}2^{\circ}C$ for drying time. Optimum storage time for quality maintenance was 14 days at $5{\pm}2^{\circ}C$.
Mutual growth limitation was observation when the two antagonistic fungi was come in contact with each other. Brown line was formed 2day after contact with Trichoderma spp., and then, green spores formed overnight. The laccase activity of L. edodes was stimulated when this fungus wsa co-incubated with Trichoderma spp. for a few days in liquid media. In sawdust-rice bran nixtures, outstanding broun line developed when the two antagonistic fungi co-cultured. The pH of the substrates changed from 5.5 to 4.5 after overgrowth, suggesting a difference in the degradation ability and the preference of the two fungi for the lignocellulose material.
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