Hepatic fibrosis was induced in Sprague-Dawley rats to evaluate the ultrastructural changes of fat-storing cells(Ito cells). For experimental induction of liver fibrosis, the rats were administered intraperitoneally with 0.5ml of 50% $Ccl_4$ solution per Kg body weight, twice weekly for 12 weeks. The rats were sacrified every week. The liver tissues were examined under light and eletron microscopes. And the immunohistochemical study of desmin was also performed. The results were summarized as follows : Light microscopic findings : The cellular infiltrations with inflammatory cells and Kupffer cells developed from 1 week after $Ccl_4$ injection, and were the most severe in 4 weeks. The strong immunoreactivity for desmin was also evident in 4 weeks. The centrilobular necrosis and fibrosis developed from 2 weeks after injection, and the necrosis persisted until 8 weeks. The progress of fibrosis was accompanied by decreases in cellular infiltration and reactivity for desmin, and increased gradual nodular formation was also observed. The cirrhosis was developed after 10 weeks. Electron microscopic findings : An increase in number of fat-storing cells was observed from 1 week after injection. Transitional cells characterized by a depletion of lipid droplets and a hypertrophy of the rER appeared after 2 weeks. The number of transitional cells with abundant collagen fibers in the extracellular spaces increased in 4 weeks. With progression of fibrosis the number of fat-storing cells decreased and proliferating fibroblasts with dilated rER were observed. According to these results it was revealed that there was an apparent transition from fat-storing cells to transitional cells and to fibroblasts. These cells had a few similar characteristics and may belong to the same cell population. Thus it was suggested that fat-storing cells might play an important role in hepatic fibrosis.
This study was carried out to investigate the effect of activation timing, cell cycle and passage on the development of embryos produced by cumulus cell nuclear transfer in Hanwoo (Korean cattle). Nuclear donor cumulus cells were cultured in Dulbecco's modified Eagle medium supplemented with 10% fetal bovine serum at $38.5^{\circ}C$ in a humidified atmosphere of 5% $CO_2$ in air. The 1~6 passages of serum deprived or actively dividing cumulus cells were isolated and used as donor cells. The in vitro matured oocytes were enucleated and then the isolated donor cells were introduced. One pulse of 180 volts for $15{\mu}s$ was applied to induce the fusion between karyoplast and cytoplast. The activation was done before or after the fusion. To activate, oocytes were treated with $10{\mu}M$ calcium ionophore for 5 min immediately followed by 2 mM 6-dimethylaminopurine for 3 h. The nuclear transfer embryos were cultured in $500{\mu}l$ of modified CRlaa supplemented with 3 mg/ml BSA in four well dish covered with mineral oil. After 3 days culture, culture medium was changed into modified CRlaa medium containing 1.5 mg/ml BSA and 5% FBS for 4 days. The incubation environment was 5% $CO_2$, 5% $O_2$, 90% $N_2$ at $38.5^{\circ}C$. There was no blastocyst formation when the nuclear transfer embryos were activated before the fusion, whereas, 29.9% of blastocyst formation was shown when the nuclear transfer embryos were activated after the fusion. When serum deprived and actively dividing cumulus cells were used as nuclear donor cells, the developmental rates to blastocyst were 38.5% and 40.6%, respectively. There was no significant difference between serum deprived and actively dividing cells in the developmental rates. The developmental rates to blastocyst according to 1~6 passages were 37.5~44.4%. However, there were no significant differences among passages. These results indicate that 1~6 passage cumulus cell irrespective of cell cycle could support development of nuclear transfer embryos activated after the fusion.
Twenty-four cannulated Small-tailed Han${\times}$Poll Dorset wethers (BW $47.5{\pm}2.1kg$) were used to determine the effects of forage to concentrate ratio (40:60 vs. 70:30), monensin supplementation (0, 15 or 30 ppm, DM basis) and interactions of these two factors on cis-9, trans-11 conjugated linoleic acid (cis-9, trans-11 CLA) and trans-11 octadecenoic acid (trans11-$C_{18:1}$) concentrations in ruminal contents and plasma in sheep. The experiment was designed as a $2{\times}3$ factorial. The diet contained Chinese wild rye grass hay (Aneurolepidium Chinese), cracked corn, soybean meal, NaCl, limestone and trace mineral premix. Dietary crude fat and linoleic acid ($C_{18:2n-6}$) were adjusted with soybean oil to about 7.0% and 24.0 mg/g (DM basis), respectively. High forage diets increased (p<0.001) the concentrations of trans11-$C_{18:1}$ and cis-9, trans-11 CLA in ruminal contents and plasma. Monensin supplementation increased (p<0.001) the concentration of trans11-$C_{18:1}$ in ruminal contents, but had no effect on that of cis-9, trans-11 CLA. Concentrations of trans11-$C_{18:1}$ (p<0.019) and cis-9, trans-11 CLA (p<0.022) in plasma increased with dietary monensin levels. Interactions of forage: concentrate ratio and monensin level tended to affect the concentrations of trans11-$C_{18:1}$ (p<0.091) and $C_{18:2n-6}$ (p<0.083) in ruminal contents. Increasing forage levels increased the concentrations of trans11-$C_{18:1}$ and cis-9, trans-11 CLA in the rumen. Supplementing with monensin increased the ruminal production of trans11-$C_{18:1}$ and concentrations of trans11-$C_{18:1}$ and cis-9, trans-11 CLA in plasma.
Because liquids with high molecular weight such as mineral oil have low vapor pressure at room temperature, it is generally thought to be difficult to lose them to evaporation. However, when they are dispersed into air in small droplets during application in machining processes, their surface area becomes considerably higher. To determine the potential for metalworking fluids (MWF) filter losses, MWF mist was generated and collected on polyvinyl chloride (PVC) filters in test chamber. After collected MWF was exposed to clean air during designated period (range 10~240 minutes) and the filters were desiccated, losses were evaluated. As duration of clean air passing through PVC filter increased, loss of MWF gradually increased. MWF lost after 10 minutes ranged form 12.4 % to 21.8 % of the original loading mass, on average 53.3 % of the total loss. These results indicate that significant mass of MWF collected on PVC filters can be lost at the beginning of air sampling. Loss of MWF collected on PVC filter also occurred during desiccation without active airflow. In multiple regression to identify which factors influence the loss of MWF collected on PVC filter, both duration of air passing through PVC filter and MWF age (fresh vs. used) were significant predictor (p=0.0001). Therefore, workers' exposure to MWF measured method 0500, may underestimate true concentration. Further study is needed to develop a new method to quantify the workers' exposure to airborne MWF mist accurately.
Park, Kee-Sang;Kim, Ju-Hwan;Lee, Taek-Hoo;Song, Hai-Bum;Chun, Sang-Sik
Clinical and Experimental Reproductive Medicine
/
v.28
no.2
/
pp.79-86
/
2001
Objective: Mouse pre-antral follicles require the addition of gonadotropins (Gns) to complete maturation and ovulation of oocyte and antrum formation in vitro. However, we tried examination of in vitro growth of mouse pre-antral follicles in medium without Gns and/or phygiological factors. And also, pre-antral follicles were isolated from ovaries by mechanical method. Our present studies were conducted to evaluate on the growth of follicles and intra-follicular oocytes and antrum formation in vitro of mouse pre-antral follicles in two different media. Methods: Pre-antral follicles ($91{\sim}120{\mu}m$) were isolated mechanically by fine 30G needles not using enzymes from ovaries of 3-6 week-old female ICR mice. Isolated pre-antral follicles were cultured in $20{\mu}l$ droplets of TCM (n=17; follicles: $107.8{\pm}1.58{\mu}m$; oocytes: $57.9{\pm}1.2{\mu}m$) or MEM (n=12; follicles: $109.3{\pm}2.53{\mu}m$; oocytes: $55.4{\pm}1.6{\mu}m$) under mineral oil on the 60 mm culture dish. All experimental media was supplemented with 10% FBS without Gns and/or physiological factors. Pre antral follicles were individually cultured for 8 days. Antram formation and growth of pre-antral follicles and intra-follicular oocytes were evaluated using precalibrated ocular micrometer at X200 magnifications during in vitro culture. Results were analyzed using combination of Student's t-test and Chi-square, and considered statistically significant when p<0.05. Results: Antrum formation had started in two culture media on day 2. On day 8, antrum formation had occurred in 58.3% of pre-antral follicles cultured in DMEM, but only in 23.5% of those cultured in TCM (p=0.0364). Growth of pre-antral follicles and intra-follicular oocytes were observed on day 4 and 8. On day 4, follicular diameter was similar (p=0.1338) in TCM ($119.4{\pm}2.58{\mu}m$) and MEM ($125.4{\pm}4.52{\mu}m$). However, on day 8, diameters of pre-antral follicles cultured in MEM ($168.9{\pm}17.29{\mu}m$) were significantly bigger (p=0.0248) than that in TCM ($126.7{\pm}4.28{\mu}m$). On day 4 and 8, diameters of intra-follicular oocytes were similar in TCM ($67.1{\pm}1.3$ and $72.4{\pm}0.9{\mu}m$) and MEM ($65.2{\pm}1.7$ and $73.3{\pm}1.5{\mu}m$), respectively. Conclusion: We can conform that medium without Gns and/or physiological factors can be used for in vitro antrum formation and growth of pre-antral follicles and intra-follicular oocytes in mouse. In conclusion, MEM supplemented with FBS can be used for growth in vitro of mouse pre-antral follicles isolated mechanically.
Go, In Hee;Jeong, Hye Young;Park, Ju Hyeon;Jeong, Sir Lin;Jo, A Hyeon
Journal of Conservation Science
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v.31
no.4
/
pp.331-339
/
2015
This study were conducted the particle-size analysis on 57 blue pigments to understand the step distribution characteristics of blue pigments made out of Azurite by using Malvern's Mastersizer 2000. As the result, most of the conventional blue pigments in Korea, Japan, and Chinese showed good granularity step separation except for few, and the smaller the particle, the more the Span value increased and the wider the granularity distribution range. On the basis of Friedman and Sanders's Grain size, most of the pigments were sand size to silt size. 72.2% of B-100 was clay size and 2.5% of A-14 was gravel size. Even the same components can differ by the grain size directly affecting the important property such as color formation, oil absorption, specific gravity, usability, etc. so the information about the granularity distribution would be used for basic data to deal with natural pigments.
Kim, Chaeyeon;Park, Ki-Su;Kang, Sung-Min;Kim, Jongmin;Song, YoungShin;Lee, Chang-Soo
Korean Chemical Engineering Research
/
v.53
no.6
/
pp.740-745
/
2015
This study investigated the effect of different collection methods on physical properties of pectin hydrogels in microfluidic synthetic approach. The pectin hydrogels were simply produced by the incorporation of calcium ions dissolved in continuous mineral oil. Then, different collection methods, pipetting, tubing, and settling, for harvesting pectin hydrogels were applied. The settling method showed most uniform and monodispersed hydrogels. In the case of settling, a coefficient of variation was 3.46 which was lower than pipetting method (18.60) and tubing method (14.76). Under the settling method, we could control the size of hydrogels, ranging from $30{\mu}m$ to $180{\mu}m$, by simple manipulation of the viscosity of pectin and volumetric flow rate of dispersed and continuous phase. Finally, according to the characteristics of simple encapsulation of biological materials, we envision that the pectin hydrogels can be applied to drug delivery, food, and biocompatible materials.
Journal of the Korean Association of Oral and Maxillofacial Surgeons
/
v.26
no.6
/
pp.581-590
/
2000
Alterations in the cellular genome affecting the expression or function of genes controlling cell growth and differentiation are considered to be the main cause of cancer. Over 30 oncogenes can be activated by insertional mutagenesis, single point mutations, chromosomal translocations and gene amplification. The ras oncogenes have been detected in $15{\sim}20%$ of human tumors that include some of the most common forms of human neoplasia and are known to acquire their transforming properties by single point mutations in two domains of their coding sequences, most commonly in codons 12 and 61. The ras gene family consists of three functional genes, N-ras, K-ras and H-ras which encode highly similar proteins of 188 or 189 amino acid residues generically known as P21. ras proteins have been shown to bind GTP and GTP, and possess intrinsic GTPase activity. Experimental study was performed to observe the mutational change of the ras gene family and apply the results to the clinical activity. 36 Golden Syrian Hamster each weighing $60{\sim}80g$ were used and painted with 0.5% DMBA by 3 times weekly on the right buccal cheek(experimental side) for 6, 8, 10, 12, 14 and 16 weeks. Left buccal cheek (control side) was treated with mineral oil as the same manner of the right side. The hamsters were sacrificed on the 6, 8, 10, 12, 14 & 16 weeks. Normal and tumor tissues from paraffin block were completely dissected by microdissection and DNA from both tissue were isolated by proteinase K/phenol/chloroform extraction. Segments of the K-ras and H-ras gene were amplified by PCR using the oligonucleotide primers corresponding to the homologous region (codon 12 and 61) of the hamster gene, and then confirmational change of ras genes was observed by SSCP and autosequencing analysis. The results were as follows : 1. Malignant lesion could be found in the experimental side from the experimental six weeks. 2. One hamster among six showed point mutation of the H-ras codon 12($G{\rightarrow}A$ transition) at the experimental 10 and 14 weeks. 3. One of six at 6 weeks, two of six at 8 weeks and one of six at 12 weeks revealed the confirmational change of the H-ras codon 61($A{\rightarrow}T$ transversion). 4. The incidence of point mutation of H-ras codon 12 and 61 were 5.5%(2 of 36) and 11%(4 of 36) respectively. 5. Point mutation of the K-ras could not be seen during the whole experimental period. Form the above results, these findings strongly support the concept that H-ras oncogenes may have the influence of the DMBA induced carcinoma of hamster buccal pouch.
The data acquired from seismic exploration can be used to detect the existence of oil and gas resources through appropriate processing and interpretation. The seismic attributes indicating the existence of resources are extracted from amplitude information, where the Q-factor representing intrinsic attenuation plays an useful role of hydrocarbon indicator. So, the accuracy of Q-factor estimation is very important to investigate the existence of resources. In this study, we calculated the Q-factor and analyzed the error rate through a numerical example. To mimic real data, random noise was added to the synthetic data. With the noise-added data, the Q-factor was estimated and the error rate was analyzed by using the spectral ratio method (SRM) and peak frequency shift method (PFSM). Both methods provided a relatively accurate Q-factor when the signal-to-noise ratio was 90 dB. However, the peak frequency shift method (PFSM) produced better results than the spectral ratio method (SRM) as the level of random noise increased.
Proceedings of the Korean Society of Embryo Transfer Conference
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2002.11a
/
pp.69-69
/
2002
Transgenic animals production tools have been valuable for research and purpose. The current methods of gene transfer, microinjection and nuclear transfer, which are widely used in transgenic animal production, but all most methods has only had limited success in production of larger species. Here, we report the possibility of a sperm-mediated gene transfer method in porcine embryos. Oocytes were collected from ovaries harvested at a local slaughterhouse were matured in 500${mu}ell$ drops of TCM-199 under mineral oil at 38.5$^{\circ}C$ in a humidified atmosphere of 5%CO2 in air. After 42-43h of in vitro maturation oocytes were denuded. for sperm injection into the cytoplasm of the porcine oocytes, sperm suspension in NIM medium are subjected extraction with TritonX-100 before mixing with a green fluorescent gene (GFP). Sperm with Tritonx-100 were prepared by adding TritonX-100 to a final volume of 0.05% in the sperm suspension and mixing by trituration for 60s before two wishes in NIM medium at 2$^{\circ}C$. A(ter wishing, sperm were mixed with TritonX-100 at $25^{\circ}C$ followed by washes at 2$^{\circ}C$. Sperm were resuspended in ice cold NIM to a final volume of 400${mu}ell$ and 2-20ng/${mu}ell$ DNA were triturated on ice for 60s. All microinjection was performed in HEPES-buffered CZB medium at room temperature within 2h. After culture in NCSU-23 for 72h, percent of porcine embryos transfected GFP gene are 20.7%(6/29) in 20ng/${mu}ell$ sperm-DNA mixed group and other groups were 3.7 %(2/54)and 4.7%(3/67). These data suggests that sperm-mediated gene transfer method should be used to the production tool of transgenic pig efficiently.
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