Park H. S.;Kim T. S.;Lee Y. H.;Jung S. Y.;Lee M. Y.;Jin J. I.;Park J. K.;Lee J. S.;Kim C. H.
Reproductive and Developmental Biology
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v.28
no.3
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pp.181-185
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2004
This study was conducted to examine whether activation treatments, source of oocytes and culture conditions affect in vitro developmental ability of caprine oocytes. Mature Korean native goats were pretreated with intravaginal CIDR for 10 days. The goats were then treated with a single intramuscular injection of 1,000 IU PMSG on Day 8 or twice daily injection of a total of 70 mg FSH for 3 days from Day 8 of CIDR insertion for superovulation. All the goats were injected with 10 mg PGF/sub 2a/ on Day 8 and 400 IU hCG on Day 10 of CIDR. Oocytes were surgically collected by oviduct flushing(in vivo maturation) or direct follicle aspiration(in vitro maturation) through mid-ventral incision at 35 h after hCG injection. Fifteen to twenty oocytes were placed in TCM-199 medium containing 25 mM Hepes and hormones under mineral oil at 39℃ in a humudified atmosphere of 5% CO₂ in air for 22 to 24 h. After maturation, the oocytes were activated by electric stimulation or ionomycin + 6-DMAP. The activated oocytes were then cultured in M16, TCM-199 and mSOF media supplemented with proteins at 39℃ for 6 to 7 days. Activation treatments did not affect cleavage of the oocytes. The cleavage rates were 64.1% (41/64) in oocytes activated by electric stimulation and 76.5% (218/285) in oocytes activated by ionomycin + 6-DMAP. The proportion of development to blastocyst was 15.6% (34/218) in oocytes activated by ionomycin + 6-DMAP, but activation by electric stimulation did not support embryos developed beyond morula stage. There were no differences in the cleavage rates of activated oocytes experiencing in vivo (86.8%, 66/76) and in vitro maturation (69.0%, 127/184). However, the development rate to blastocyst stage was significantly (P<0.05) higher for oocytes matured in vivo (50.0%, 33/66) compared to in vitro (0.8%, 1/127). Culture conditions did not affect the cleavage of -activated oocytes. The cleavage rates were 51.6% (49/95) in M16, 64.3% (18/28) in TCM-199 and 81.0% (145/179) in mSOF, respectively. By contrast, the development rate of activated oocytes to stage was greater (P<0.05) for oocytes cultured in mSOF medium (23.4%, 34/145) than in M16 or TCM-199 (0.0%). Our results suggest that source of oocytes and culture conditions are major factors affecting in vitro development of caprine parthenogenetic oocytes.
This experiment was conducted to select optimum minor cereal crop and variety for the marginal agricultural lands so that they would be used to increase the agricultural land use rate in mountainous areas. This experiment used three different marginal agricultural lands, that is, slope land, barren land and gravelly land. Soil pH of control field was 7.85, slightly alkali, whereas gravelly land showed strong acid(pH 5.1). EC of marginal agricultural lands was generally lower compared to the control, especially barren land showed 0.05dS $m^{-1}$ which was the lowest among marginal agricultural lands. Foxtail millet showed no significant difference of heading and ripening date among marginal agricultural lands. Foxtail millet of slope land showed the highest culm length(169.5 cm) and grains per ear(4913.9 grains) among marginal agricultural lands except control field. Growth duration of three varieties was varied from 112 to 113days, which was not different statistically. In grains per ear of common millet, slope land showed 787.1 grains which was the highest among marginal agricultural lands except control field. Among varieties, Byorukgijang showed the highest grains per ear(827.2 g/ear), whereas 1000 grain weight was 5.5 g which was lower compared to that of Hwanggijang(6.2 g). In sorghum, slope land showed the highest grains(2563.8 grains) per ear and ears per 3.3 $m^2$(26.0 ears) except control field. Growth duration of three sorghum varieties almost the same from 122 to 123 days, whereas the highest grains per ear was obtained from Moktaksusu(2357.6 grains). Jangmoksusu showed the highest 1000 grain weight(23.8 g) among three sorghum varieties. In yield of three foxtail millets, the highest yield was obtained from slope land(282.0 kg/10a) among marginal agricultural lands and Mongdangjo showed the highest yield(252.3 kg/10a) among three varieties. In yield of three common millets, the highest yield was obtained from slope land(196.0 kg/10a) among marginal agricultural lands and Byorukgijang showed the highest yield(173.8 kg/10a) in three varieties. In three sorghum varieties, the highest yield was obtained from slope land(301.7 kg/10a) among marginal agricultural lands and Moktaksusu showed the highest yield(236.5 kg/10a) among three varieties.
Purpose: The HSV1-tk reporter gene system is the most widely used system because of its advantage that direct monitoring is possible without the introduction of a separate reporter gene in case of HSV1-tk suicide gene therapy. In this study, we investigate the usefulness of the reporter probe (substrate), $9-(4-[^{18}F]Fluoro-3-hydroxymethylbutyl)$guanine ($[^{18}F]FHBG$) for non-invasive reporter gene imaging using PET in HSV1-tk expressing hepatoma model. Materials and Methods: Radiolabeled FHBG was prepared in 8 steps from a commercially available triester. The labeling reaction was carried out by NCA nucleophilic substitution with $K[^{18}F]/K2.2.2.$ in acetonitrile using N2-monomethoxytrityl-9-14-(tosyl)-3-monomethoxytritylmethylbutyl]guanine as a precursor, followed by deprotection with 1 N HCl. Preliminary biological properties of the probe were evaluated with MCA cells and MCA-tk cells transduced with HSV1-tk reporter gene. In vitro uptake and release-out studies of $[^{18}F]FHBG$ were performed, and was analyzed correlation between $[^{18}F]FHBG$ uptake ratio according to increasing numeric count of MCA-tk cells and degree of gene expression. MicroPET scan image was obtained with MCA and MCA-tk tumor bearing Balb/c-nude mouse model. Results: $[^{18}F]FHBG$ was purified by reverse phase semi-HPLC system and collected at around 16-18 min. Radiothemical yield was about 20-25%) (corrected for decay), radiochemical purity was >95% and specific activity was around >55.5 $GBq/{\mu}\;mol$. Specific accumulation of $[^{18}F]FHBG$ was observed in HSV1-tk gene transduced MCA-tk cells but not in MCA cells, and consecutive 1 hour release-out results showed more than 86% of uptaked $[^{18}F]FHBG$ was retained inside of cells. The uptake of $[^{18}F]FHBG$ was showed a highly significant linear correlation ($R^2=0.995$) with increasing percentage of MCA-tk numeric cell count. In microPET scan images, remarkable difference of accumulation was observed for the two type of tumors. Conclusion: $[^{18}F]FHBG$ appears to be a useful as non-invasive PET imaging substrate in HSV1-tk expressing hepatoma model.
Background: Lung clearance of inhaled $^{99m}Tc$-DTPA reflects alveolar epithelial permeability and it had been reported as more sensitive than conventional pulmonary function tests in detecting lung epithelial damage. However, measuring lung clearance of inhaled $^{99m}Tc$-DTPA by gamma camera may not always reflect alveolar epithelial permeability exactly because it is influenced by mucociliary clearance depending on the site of particle deposition. Moreover, this method takes much time and patient's effort because he has to sit or lie still in front of the camera for a prolonged period. Most of the absorbed DTPA is excreted in urine within 24 hours and the amount of excreted DTPA in urine during the first few hours after inhalation is influenced by absorption rate which is correlated with the alveolar-epithelial permeability suggesting that the urinary excretion, especially in first few hours, may be an alternate index for lung clearance. The purpose of this study was to evaluate the usefulness of ratio of excreted $^{99m}Tc$-DTPA in 2 hour and 24 hour urine as an index of alveolar-epithelial damage. Methods: Pulmonary function tests including diffusing capacity and lung clearance of $^{99m}Tc$-DTPA measured by gama camera ($T_{1/2}$) and 2hr/24hr urine excretion ratio (Ratio) of inhaled $^{99m}Tc$-DTPA in 8 normal subjects and 14 patients with diffuse interstitial lung disease were compared. Results: 1) In the normal control, there was significant negative correlation between the $T_{1/2}$ and the Ratio (r=-0.77, p<0.05). In patients with diffuse interstitial lung disease, there also was significant negative correlation between $T_{1/2}$ and Ratio(r=-0.63, p<0.05). 2) In diffuse interstitial lung disease patients, the $T_{1/2}$ was $38.65{\pm}11.63$ min which was significantly lower than that of normal control, $55.53{\pm}11.15$ min and the Ratio was $52.15{\pm}10.07%$ also signifantly higher than that of the normal control, $40.43{\pm}5.53%$ (p<0.05). 3) There was no significant correlations between $T_{1/2}$ or Ratio and diffusing capactiy of lung in both patients and controls (p>0.05). Conclusion: These results suggests that 2hr/24hr urine excretion ratio of inhaled $^{99m}Tc$-DTPA is a useful simple bedside test in assessing alveolar epithelial permeability and that it may be used as an additive follow-up test in patients with diffuse interstitial lung disease complementing conventional pulmonary function tests.
Kim, Jong Yeop;Kim, Cheol Hong;Shin, Hyun Won;Chae, Young Je;Choi, Chul Young;Shin, Tae Rim;Park, Yong Bum;Lee, Jae Young;Bahn, Joon-Woo;Park, Sang Myeon;Kim, Dong-Gyu;Lee, Myung Goo;Hyun, In-Gyu;Jung, Ki-Suck
Tuberculosis and Respiratory Diseases
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v.60
no.6
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pp.653-662
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2006
Background: The changes in the pulmonary function observed in burn patients with an inhalation injury are probably the result of a combination of airway inflammation, chest wall and muscular abnormalities, and scar formation. In addition, it appears that prolonged ventilatory support and an episode of pneumonia contribute to the findings. This study investigated the changes in the pulmonary function in patients with inhalation injury at the early and late post-burn periods. Methods: From August 1, 2002, to August 30, 2005, surviving burn patients who had an inhalation injury were enrolled prospectively. An inhalation injury was identified by bronchoscopy within 48hours after admission. Spirometry was performed at the early phase during admission and the recovery phase after discharge, and the changes in the pulmonary function were compared. Results: 37 patients (M=28, F=9) with a total burn surface area (% TBSA), ranging from 0 to 18%, were included. The initial $PaO_2/$FiO_2$ratio and COHb were $286.4{\pm}129.6mmHg$ and $7.8{\pm}6.6%$. Nine cases (24.3%) underwent endotracheal intubation and 3 cases (8.1%) underwent mechanical ventilation. The initial X-ray findings revealed abnormalities in, 18 cases (48.6%) with 15 (83.3%) of these being completely resolved. However, 3 (16.7%) of these had residual sequela. The initial pulmonary function test, showed an obstructive pattern in 9 (24.3%) with 4 (44.4%) of these showing a positive bronchodilator response, A restrictive pattern was also observed in 9 (24.3%) patients. A lower DLco was observed in only 4 (17.4%) patients of which 23 had undergone DLco. In the follow-up study, an obstructive and restrictive pattern was observed in only one (2.7%) case each. All the decreased DLco returned to mormal. Conclusions: Most surviving burn patients with an inhalation injury but with a small burn size showed initial derangements in the pulmonary function test that was restored to a normal lung function during the follow up period.
Purpose: factor analysis and independent component analysis (ICA) has been used for handling dynamic image sequences. Theoretical advantages of a newly suggested ICA method, ensemble ICA, leaded us to consider applying this method to the analysis of dynamic myocardial $H_2^{15}O$ PET data. In this study, we quantified patients' blood flow using the ensemble ICA method. Materials and Methods: Twenty subjects underwent $H_2^{15}O$ PET scans using ECAT EXACT 47 scanner and myocardial perfusion SPECT using Vertex scanner. After transmission scanning, dynamic emission scans were initiated simultaneously with the injection of $555{\sim}740$ MBq $H_2^{15}O$. Hidden independent components can be extracted from the observed mixed data (PET image) by means of ICA algorithms. Ensemble learning is a variational Bayesian method that provides an analytical approximation to the parameter posterior using a tractable distribution. Variational approximation forms a lower bound on the ensemble likelihood and the maximization of the lower bound is achieved through minimizing the Kullback-Leibler divergence between the true posterior and the variational posterior. In this study, posterior pdf was approximated by a rectified Gaussian distribution to incorporate non-negativity constraint, which is suitable to dynamic images in nuclear medicine. Blood flow was measured in 9 regions - apex, four areas in mid wall, and four areas in base wall. Myocardial perfusion SPECT score and angiography results were compared with the regional blood flow. Results: Major cardiac components were separated successfully by the ensemble ICA method and blood flow could be estimated in 15 among 20 patients. Mean myocardial blood flow was $1.2{\pm}0.40$ ml/min/g in rest, $1.85{\pm}1.12$ ml/min/g in stress state. Blood flow values obtained by an operator in two different occasion were highly correlated (r=0.99). In myocardium component image, the image contrast between left ventricle and myocardium was 1:2.7 in average. Perfusion reserve was significantly different between the regions with and without stenosis detected by the coronary angiography (P<0.01). In 66 segment with stenosis confirmed by angiography, the segments with reversible perfusion decrease in perfusion SPECT showed lower perfusion reserve values in $H_2^{15}O$ PET. Conclusions: Myocardial blood flow could be estimated using an ICA method with ensemble learning. We suggest that the ensemble ICA incorporating non-negative constraint is a feasible method to handle dynamic image sequence obtained by the nuclear medicine techniques.
The purpose of this study was to evaluate the levels of blood lead and cadmium in urban and rural populations. Blood samples were collected from 100 urban (Busan) residents and 150 rural residents (Jinju-84, Gijang-66) from July 1 to August 30, 2007. The blood lead and cadmium levels were analyzed using flameless atomic absorption spectrophotometry and were compared by age, gender and smoking status. The mean levels of blood lead in urban-Busan, rural-Jinju and rural-Gijang residents were $6.38{\pm}2.86{\mu}g/dl$, $5.41{\pm}2.16{\mu}g/dl$ and $4.50{\pm}2.87{\mu}g/dl$, respectively. There was a significant difference in the level of blood lead between urban-Busan residents and rural-Gijang residents. The mean levels of blood cadmium in urban-Busan, rural-Jinju and rural-Gijang residents were $0.85{\pm}0.44{\mu}g/l$, $1.57{\pm}0.78{\mu}g/l$ and $0.95{\pm}0.54{\mu}g/l$, respectively. There was a significant difference in the level of blood cadmium between urban-Busan residents and rural-Jinju residents. This study showed that the levels of blood lead and cadmium were significantly different between urban and rural populations. The level of blood lead was highest in urban-Busan residents, but the level of blood cadmium was highest in rural-Jinju residents. Further studies are needed to define the cause of high levels of blood lead and cadmium related to area of residence and personal habits.
A lab-scale MF membrane reactor was installed to investigate the membrane permeability, characteristics of membrane fouling at each conditions, and quality of permeate (liquid livestock manure) in the separation of solid-matters using membrane. Experiment was divided three filtration type such as follows; continuous filtration, gravity filtration, and intermittent filtration. As a result of experiment, flux 1 LMH was maintained for 7days, and trans-membrane pressure(TMP) was increased gradually under 10cmHg, but it was increased immediately after 10cmHg, respectively. However, the flux was increased, the Tmax was decreased exponential more and more. During the pure-flux test, most of the fouling of membrane was reversible. At the gravity filtration, permeate could be obtained as 1.75 LMH for 3.5days without any other electronic pressure. As an investigation of membrane surface, this study could be decided that the reason of fouling at the lower flux (Run 1 and 2) was attached matters in membrane surface, but at the higher flux (Run 4-6) was concentration polarization.
For investigating transfer factors of $^{137}Cs$ in the arable land of Korea, mature crop plants and topsoils were collected from paddy and upland fields located at 33 areas of the country and $^{137}Cs$ concentrations were measured by ${\gamma}-spectrometry$. The $^{137}Cs$ concentrations in soil were in the range of $0.7{\sim}17.7$ Bq/kg-dry in the paddy fields and $1.2{\sim}27.8$ Bq/kg-dry in the upland fields. The $^{137}Cs$concentrations in hulled seed, detected for 12 areas only, were in the range of $0.019{\sim}0.111$ Bq/kg-dry and those in Chinese cabbage, detected also for 12 areas only, were in the range of $0.012{\sim}0.066$ Bq/kg-fresh. Soil-to-plant transfer factors of $^{137}Cs$ were in the range of $1.2{\times}10^{-3}{\sim}1.1{\times}10^{-2}$ for hulled seed and $6.8{\times}10^{-4}{\sim}1.7{\times}10^{-2}$ for Chinese cabbage. Inboth plant stuffs, the factor tended to decrease with increasing soil organic matter or cation exchange capacity and, in hulled seed, it tended to increase with increasing soil clay content. No statistical significance was, however, found in all those relationships. Present results can be utilized for estimating radiation risk resulting from the food consumption by Korean people and deciding agronomical counter-measures at the time of an nuclear accident.
This study was conducted to examine in vitro developmental ability of porcine embryos after somatic cell nuclear transfer. The porcine ear fell was cultured in vitro for confluency in serum-starvation condition(TCM-199 + 0.5% FBS) far 3~6 days of cell confluency. The zona pellucida of IVM oocytes were partially drilled using laser system. Single somatic cell was individually transferred into enucleated oocytes. And the reconstructed embryos were electrically fused(single DC 1.9kv/cm, 30$\mu$ sec) with 0.3M mannitol. After electrofusion, embryos were activated(single AC 5v/mm, 5sec) and cultured in HCSU-23 medium containing 10% FBS at 39$^{\circ}C$, 5% $CO_2$ in air for 6 to 8 days. The fusion rate of donor cells was 45.6, 36.8 and 46.1% in 3~4, 5~6 days of serum starvation and non serum starvation(N-S), and were 52.7. 53.0 and 51.7% in 1~2. 5~6 and 13~14 passages of donor cell culture, respectively. No significant difference was found in the fusion rate of donor cells by the duration of serum starvation treatment or the number of donor cell passages. By the size of donor cells, however, the fusion rate was significantly higher(P<0.05) for reconstructed embryos derived from 25r $\mu$m $\geq$ site of donor cells (65.3%) than that of 25~30$\mu$ m(42.5%) or 30$\mu$ m(45.5%)$\leq$ cells. The cleavage rate was significantly (P<0.05) higher in 3~4 darts of serum starvation treatment(67.1%) than that in N-S (50.7%) or 5~6 days of starvation(57.1%). The activation rate by the size of donor cells in fused oocytes was 56.5, 68.8 and 58.5%, respectively, and was not significant.
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