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.
Angiotensin converting enzyme(ACE) belongs to the class of zinc protease and plays an important role in the regulation of blood pressure. In this experiment, we investigated the inhibitory activities of ninety four plant extracts on ACE. The extracts were prepared by water and refluxing with 70% and 100% methanol. Among the extracts, two plant extracts such as Cassia tora, Persicaria cochinchinensis Kitagawa showed more than 60% inhibitory activities, and Foeniculum vulgare Gaertner, Scutellaria baicalensis Georgl, Caragana sinica (Buchoz) Rehder, Inula britannica var. chinensis showed $45.2{\sim}49.7%$ inhibitory activities. Twenty eight plant extracts such as Hemerocallis fulva L, Camptotheca acuminata Decne, Inula britannica var. chinensis, Xanthium strumarium, Polygonatum odoratum, Phellodendron amurense Rupr, Coix lachryma-jobi var. mayuen, Prunus ansu, Hibiscus mutabilis L, Thchosanthes kirilowii, Helianthus annuus, Juglans sinensis showed $30.3{\sim}39.7%$ Inhibitory activities. These results suggest that plant extracts which contain high ACE inhibitory activities may be useful as anti-hypertension agents and to the treatment of hypertension.
The present study was conducted to examine some factors affecting in vitro development and fecundity of embryos recloned with somatic cell nuclear transfer (SCNT). Fibroblast cells retrieved from the ear of a 3-week-old, cloned Korean goat (Jinsoonny) were used as karyoplast donors and serum-starvation was conducted in tissue culture medium (TCM)-199 supplemented with 0.5% FBS. Recipient oocytes were surgically collected by flushing the oviducts 35 h after hCG injection following FSH priming. The zonae pellucidae of the oocytes were partially perforated with a laser drill and a donor cell was transferred into an enucleated oocyte. The couplets were electrically fused and activated by ionomycin (5 min) and 6-DMAP (4 h). The reconstructed embryos were cultured in mSOF medium containing 0.8% BSA at $39^{\circ}C$ in an atmosphere of 5% $CO_2$, 5% $%O_2$, 90% $N_2$ for 12 to 15 h. Re-cloned embryos (2- to 4-cell stages) were surgically transferred into the oviducts of the recipients and pregnancy was subsequently diagnosed by progesterone assay and ultrasound on Days 21 and 63 of pregnancy. The fusion rate following 1st fusion pulse was higher (p<0.05) in 2nd cloning (65.9%) compared to 1st cloning (51.0%), but it was not different in the other groups. The rate of cleavage after fusion was significantly higher (p<0.05) in 1st (77.7%) than in 2nd cloning (56.0%). A total of 175 re-cloned embryos were transferred into 28 recipients. On day 21 and 60 after transfer, 11 (39.3%) and 4 recipients (17.4%) were pregnancy, respectively. In comparison of pregnancy rate by estrous synchronization, a total of 66 and 109 re-cloned embryos were transferred into 11 recipients in natural estrus and 17 recipients in induced estrus, respectively. Five (45.4%) and 2 recipients (18.2%) in natural estrus were pregnant on days 21 and 63 while 6 (35.3%) and 2 (11.8%) recipients in induced estrus were pregnant, respectively. These results show that recloning of goat can be achieved by SCNT and estrous synchronization between donor and recipient animals may be one of the major factors affecting success rate.
Since 1966, practical use of active heavy analogs of the hormones has been also worked out as an insecticide and brought the features of it to the light as cocoon producer. On the other hand, it is expected to afford the increase of silk productivity resulted from control of the fifth larval period by delaying normal development. With these regards, some of analogs have been tried to apply practically to the silkworm for an increase of cocoon productivity. One of the synthesized juvenile hormone available is "Manina". And it is presently used for the increase of silk productivity in Korea. For the practical use, it is very essential the varietal differences in the increase of silk productivity by topical application was tested and the obtained results are summarized as follows. 1. It was evident that the fifty larval period was extended by topical application after 48 hrs. of the last ecdysis, ranging from 8 hrs. to one day, as compared to the control. 2. In pupal rates, there is no significance between control and treatments. It proved that there was no toxicity to silkworm by topical application in general, except jam 120. With regards to an increase of cocoon yield in Japanese, it was resulted from 17∼24% of cocoon yield from 10,000 larvae, as compared to that of control. In case of Chinese, the incrasing rates were varied from 15∼26% of cocoon yield, 17.8kg of it with 26% increase for Jam 122 and 16.7kg of it with 25% increase for Jam 118. In case of all hybrids, an increase of the cocoon yield took places from 20% to 31% and the weight of cocoon layer for the Japanese increased by 6 to 14%, those for the Chinese by 4 to 7% and those for the hybrids ranged from 21 to 29% increase. 3. It was recognized that the hybrid vigor rate took places with the hybrids between high responsing parents to juvenile hormones.
Kim, Bong-Su;Pyo, Sung-Jai;Cho, Yong-Gyi;Shin, Chai-Ho;Cho, Jin-Woo;Kim, Chang-Ho
The Korean Journal of Nuclear Medicine Technology
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제13권3호
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pp.10-16
/
2009
Purpose: As the number of patients has increased since the installation of a PET/CT, we are now examining about 2500-3000 annually. We have realized that if we properly adjust a pitch under the same condition of a CT during a PET/CT exam, radiation quantity that reaches the patient can change. In order to reduce the exposure dose of a patient, the research examines a method of reducing the exposure dose of a patient by controlling the pitch during a PET/CT exam, viewing whether the adjustment of the pitch influences CT image and PET SUV. Methods: The equipment used is a Biograph Positron Emission Tomography (PET) Scanner (CT type: TRCT-240-130 (WCT-240-130)) of Siemens company. For the evaluation of exposure dose of a patient, we measured radiation quantities using a PTW-DIADOS 11003/1383, which is a CT radiation measurement instrument used by Siemens. We measured and analyzed the space resolutions of CT images caused by the change of pitches using an AAPM Standard Phantom in order to see how the adjustment of pitches influenced the CT images. In addition, in order to obtain SUVs caused by each change of pitches using a PET source made with a solid radioactive cylinder phantom, we confirmed whether the SUVs changed in the PET/CT images by calculating the SUVs of the fusion images caused by the change of pitches after obtaining CT and PET images and finishing the test. Results: 2slice CT scanner showed that radiation quantities largely dropped when pitches ranged from 0.7 to 1.3 and that the reduction of radiation quantities were smaller when pitches ranged from 1.5 to 1.9. That is, we found that the bigger pitch values are the smaller the radiation quantities of a patient are. Moreover, we realized that there is no change of SUVs caused by the increase of pitches and that pitch values do not influence PET SUVs and the quality of CT images. It is judged that using 1.5 as a pitch value contributes to the reduction of exposure dose of a patient as long as there is no problem in the quality of an image. Conclusions: When seeing the result of the research, hospital using a PET/CT should make an effort to reduce the exposure dose of a patient seeking pitch values appropriate for their hospital within the range in which there is no image distortion and PET SUVs are not influenced from pitches. We think that the research can apply to all multi-detectors having a CT scanner and that such a research will be needed for other equipments in the future.
Purpose: Scintimammography is one of the screening tests for the early diagnosis of breast cancer. It has been widely accepted as very useful in assessing masses that have not been detected in breast scanning. This method is highly sensitive and specific with respect to the diagnosis of primary and relapsing breast cancer. It has some difficulties, however, in detecting tumors sized 1 cm and below due to the radioactivity around the breast and the geometrical structure of the equipment. The recent introduction of high-resolution Breast-specific Gamma Imaging (BSGI) has made it possible to more accurately discriminate between malignant and benign tumors than with any other test method. Thus, the possibility of an unnecessary biopsy being performed has decreased. The purpose of this study was to examine the diagnostic capacity of mammography, breast sonography, and scintimammography, which are used for the early diagnosis of known breast cancer, and of BSGI, and to evaluate the skillfulness of radiologists. Materials and Methods: The 53 volunteers participants who had no clinical manifestation of breast cancer underwent the BSGI in February 2009. In the BSGI procedure, scanning images were obtained from the craniocaudal projection (CC) and the mediolateral Oblique projection (MLO), as well as from the additional $80{\times}80$-matrix-sized views at various angles in the Present Time method, 10 minutes after the 25 mCi $^{99m}Tc$-MIBI was injected. Results: The results of the BSGI showed that two participants had masses in their breast tissue. As the results of the diagnosis of four participants were not clear, they were retested and the results of the second test were negative. The results of the clinical screening test for breast cancer showed that the sensitivity of BSGI, scintimammography, mammography, and breast sonography was 86.5%, 77.8%, 85~90%, and 66.7%, respectively, and that their specificity was 92.4%, 84.2%, 20~42%, and 68%, respectively. Conclusion: The autodiagnosis and breast cancer screening test are needed for the early diagnosis of breast cancer. It was not easy, however, to accurately determine the presence of a mass in the breast using the existing breast cancer screening test. The patients with unclear test findings were made to undergo a histologic biopsy for a more accurate diagnosis. It is expected that the BSGI can provide useful information for the early diagnosis of breast cancer and of primary breast cancer, and will reduce the performance of unnecessary biopsies because of its higher sensitivity and specificity than existing breast cancer screening tests.
Two experiments were conducted to investigate the effect of dietary selenium sources on performance and selenium retention in broiler chickens and laying hens. In experiment 1, the effects of dietary selenium sources and levels on the weight gain, feed intake, feed conversion, and selenium retention of meat in broiler chickens were investigated. for each growth phase, the basal diet was supplemented with 0 (control), 0.12 and 0.24 ppm Se from sodium selenite (SS) and 0.12, 0.24 and 0.60 ppm Se from selenium yeast(SY). Weight gain was significantly increased(P<0.05) in supplemental 0.24 and 0.60 ppm SY compared to the 0.24 ppm SS by diet during day 1 to 35, but feed intake and feed conversion were not affected by the source or the level of Se. Selenium concentrations of breast and leg muscle were significantly increased(P<0.05) in supplemental SS and SY compared to the control, and linearly increased(P<0.05) as dietary. Se level increased by SY, but there was no difference in supplemental 0.12 ppm SS compared to 0.24 ppm SS. In experiment 2, 12-week-experiment using Hy-Line laying hens(31 wk of age) was conducted to compare the effects of selenium sources and levels on egg production, egg weight, daily egg mass, feed intake, feed conversion, egg quality, and selenium retention of egg in laying hens. A corn-soybean meal basal diet was supplemented with 0 (control), 0.06 and 0.12 ppm Se from sodium selenite (SS) and 0.06, 0.12 and 0.30 ppm Se from selenium yeast(SY). Feed conversion was significantly improved(P<0.05) in supplemental 0.06 ppm SS compared to the control, but egg production, egg weight, daily egg mass, and feed intake were not affected by source and level of Se. Haugh unit was not affected by source or level of Se. Yolk color was significantly(P<0.05) higher in supplemental 0.3 ppm SY compared to the control and other supplement in week 12. Eggshell breaking strength was significantly(P<0.05) higher in supplemental 0.06 ppm SY(P<0.05). Thickness of eggshell was not affected by source or level of Se. Se concentrations of egg was significantly improved(P<0.05) in supplemental SS and SY compared to the control, and was significantly increased(P<0.05) as dietary Se level increased by SS and SY, especially SY more effective compared to the SS.
Changes in the fine structure of testicular Leydig cell from hatching to adulthood were studied in Korean native chickens of 1, 2, 4, 6, 8, 10, 12, 14, 16, 18, 21, 24, 28, 32, 44, 52 and 64 weeks (n=13 chickens per group) of age. The objective of this study were to elucidate Leydig cell ultrastructure during testicular development. Testes of chickens were fixed by whole body perfusion using a fixative containing 2.5% glutaraldehyde in cacodylate buffer, processed and embedded in Epon-araldite. Using $1{\mu}m$ sections stained with methylene blue-azure II, qualitative and quantitative(stereological) morphological studies were performed. The ultrastructural changes of the Leydig cell were investigated by ultrathin section with the transmission electron microscope. The stages of the Leydig cell development described focus on mitochondria, endoplasmic reticulum, and lipid droplets which are involved in androgens as fullows. 1) Approaching puberty. The closely packed Leydig cells and sparse intercellular space. The nucleus occupied a large portion of the Leydig cell volume. The population of Leydig cells contained two types of cells that differed in the appearance of their nuclei which were either highly electron-opaque or relatively electron-lucid. The cytoplasm was characterized by large amounts of lipid droplets, relatively few spherical mitochondria, and sparse smooth endoplasmic reticulum. 2) Puberty to adult. The Leydig cells which display features compatible with significant androgen synthesis: large volume of cytoplasm containing extended smooth endoplasmic reticulum, abundant mitochondria, and reduction of lipid droplets.
The study was conducted from May to September in 1994 to investigate applicability of the Hearing Handicap Inventory for the Elderly-Screening version(HHIE-S) in parallel with the pure-tone audiometer to the initial screening test of noise-induced hearing loss(NIHL) in some noise-exposed workers. Subjects were selected by systemic sampling that took every 10th person from 6, 700 workers taking the annual occupational health examination by the department of Health Maintenance of Dongsan Hospital Keimyung University in Taegu. The authors administered the pure-tone audiometric test and self-reported questionnaire of HHIE-S including items of sociodemographic and job-related variables concurrently. The final subjects analysed were 1,019(488 males and 531 females) excluding fourteen persons who had many missing values in their questionnaires. The reliability coefficients of HHIE-S scale by Cronbach's alpha were 0.84. In the univariate analysis of hearing handicap measured by the HHIE-S, work duration, military service and the hearing threshold loss at 1KHz and 4KHz by the initial audiometer were significant in males while age, work duration and hearing threshold loss at 1KHz and 4KHz were significant in females. In the stepwise linear regression analysis, hearing threshold loss at 1KHz and 4KHz, was the only selected variable explaining the hearing handicap in males and hearing threshold loss at 1KHz and 4KHz, age, and work duration were selected in females. In ROC curves for HHIE-S scores against NIHL as gold standard which was defined by the follow-up audiogram as more than 30dB of the average of 0.5/1/2KHz and 50dB at 4KHz, the optimal cutoff for the parallel HHIE-S appeared to be 8. The results suggest that HHIE-S appeared to have some reliability and validity in this data and might be used in screening NIHL in parallel with pure-tone audiometer in noise-exposed workers.
Back J. J.;Park C. K.;Yang B. K.;Kim C. I.;Cheong H. T.
Reproductive and Developmental Biology
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제29권3호
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pp.175-180
/
2005
This study was conducted to examine the effects of demecolcine-assisted enucleation and recipient cell cycle stage on the development of bovine somatic cell nuclear transfer (NT) embryos. In vitro cultured oocytes for $16\~20$ h were classified by first polar body (1st PB) extrusion and cell cycle stage (MI and MII) and treated $0.4\;{\mu}L/mL$ demecolcine for 40 min before enucleation. Enucleated oocytes were fused electrically with bovine ear skin cells, activated by Ca-ionophore+DMAP, and cultured in vitro. Most of eggs ($86.2\%$) treated with demecolcine protruded a chromosome mass and enucleated efficiently ($98.8\%$, (P<0.05). Demecolcine did not have a deteriorative effect on the development of NT embryos. Developmental rate of NT embryos reconstituted with oocytes extruded 1st PB significantly higher than that of NT embryos produced by oocytes without 1st PB ($18.2\%\;vs.\;4.6\%\cdot$, P<0.05). Cleavage and blastocyst formation rate of embryos reconstituted with MI oocytes ($69.4\%\;and\;5.9\%$, respectively) were significantly lower than those of embryos reconstituted with MII oocytes ($96.7\%\;and\;23.9\%$, respectively, P<0.05). From the present result, it is suggested that domecolcine is useful for the enucleation of recipient oocytes in bovine NT procedures, and MII oocytes rather than MI oocytes are more appropriate for recipient cytoplasm. Although, the potential to develop into blastocysts of NT embryos produced by 1st PB-nonextruded and MI oocytes was very low, these oorytes could be used for NT.
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