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.
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.
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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v.13
no.3
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pp.10-16
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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.
Medical imaging modalities to image either anatomical structure or functional processes have developed along somewhat independent paths. Functional images with single photon emission computed tomography (SPECT) and positron emission tomography (PET) are playing an increasingly important role in the diagnosis and staging of malignant disease, image-guided therapy planning, and treatment monitoring. SPECT and PET complement the more conventional anatomic imaging modalities of computed tomography (CT) and magnetic resonance (MR) imaging. When the functional imaging modality was combined with the anatomic imaging modality, the multimodality can help both identify and localize functional abnormalities. Combining PET with a high-resolution anatomical imaging modality such as CT can resolve the localization issue as long as the images from the two modalities are accurately coregistered. Software-based registration techniques have difficulty accounting for differences in patient positioning and involuntary movement of internal organs, often necessitating labor-intensive nonlinear mapping that may not converge to a satisfactory result. These challenges have recently been addressed by the introduction of the combined PET/CT scanner and SPECT/CT scanner, a hardware-oriented approach to image fusion. Combined PET/CT and SPECT/CT devices are playing an increasingly important role in the diagnosis and staging of human disease. The paper will review the development of multi modality instrumentations for clinical use from conception to present-day technology and the application software.
This study was carried out to investigate effective condition for producing somatic cell nuclear transfer (SCNT) embryos of Jeju native cattle. As donor cells for SCNT, ear skin cells from Jeju native cattle were used. In experiment 1, the effect of recipient oocyte sources on the development of Jeju native cattle SCNT embryos were examined. Fusion rate of recipient oocyte and donor cell was not different between the Hanwoo and Holstein recipient oocytes (86.0% vs 89.9%). The rate of embryos developing to the blastocyst stage was significantly (p<0.05) higher in Hanwoo recipient oocytes than in Holstein recipient ones (28.2% vs 14.7%). Blastocysts derived from Hanwoo recipient oocytes contained higher numbers of total cells than those derived from Holstein ones ($115.1{\pm}40.8$ vs $101.4{\pm}33.3$), although there were no significant difference. The mean proportion of apoptotic cells in blastocyst was not different between the sources of recipient oocytes. In experiment 2, the development of Jeju native cattle and Hanwoo SCNT embryos were compared. Hanwoo oocytes were used as the recipient oocytes. Fusion rate was not different between the Jeju native cattle and Hanwoo SCNT embryos (92.1% vs 92.9%). The blastocyst rate of SCNT embryos was significantly (p<0.05) lower in Jeju native cattle than in Hanwoo (16.9% vs 31.0%). Blastocysts derived from Jeju native cattle SCNT embryos contained smaller numbers of total cells than those derived from Hanwoo ones ($136.6{\pm}33.7$ vs $149.9{\pm}39.7$), but there were no significant difference. The mean proportion of apoptotic cells in blastocyst was not different between the Jeju native cattle and Hanwoo SCNT embryos. The present study demonstrated that Hanwoo recipient oocytes were more effective in supporting production of Jeju native cattle SCNT embryos, although Jeju native cattle SCNT embryos showed reduced developmental capacity when compared to Hanwoo SCNT embryos.
M. Wang;L. Liu;L.M. Zhao;M.H. Li ;W.D. Ma;H.C. Hu ;Z.G. Wu;J.Q. Feng ;Y. Yang ;L. Zhu ;M. Chen ;T.A. Zhou;H. Jia;J. Zhang ;L. Cao ;L. Zhang ;R.R. Liang;B.J. Ding ;X.J. Zhang ;J.F. Shan;F.K. Liu ;A. Ekedahl ;M. Goniche ;J. Hillairet;L. Delpech
Nuclear Engineering and Technology
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v.54
no.11
/
pp.4102-4110
/
2022
Aiming at high-power and long-pulse operation up to 1000 s, some improvements have been made for both 2.45 GHz and 4.6 GHz lower hybrid (LH) systems during the recent 5 years. At first, the guard limiters of the LH antennas with graphite tiles were upgraded to tungsten, the most promising material for plasma facing components in nuclear fusion devices. These new guard limiters can operate at a peak power density of 12.9 MW/m2. Strong hot spots were usually observed on the old graphite limiters when 4.6 GHz system operated with power >2.0 MW [B. N. Wan et al., Nucl. Fusion 57 (2017) 102019], leading to a reduction of the maximum power capability. With the new limiters, 4.6 GHz LH system, the main current drive (CD) and electron heating tool for EAST, can be operated with power >2.5 MW routinely. Long-pulse operation up to 100 s with 4.6 GHz LH power of 2.4 MW was achieved in 2021 and the maximal temperature on the guard limiters measured by an infrared (IR) camera was about 540 ℃, much below the permissible value of tungsten material (~1200 ℃). A discharge with a duration of 1056 s was achieved and the 4.6 GHz LH energy injected into the plasma was up to 1.05 GJ. Secondly, the fully-active-multijunction (FAM) launcher of 2.45 GHz system was upgraded to a passive-active-multijunction (PAM), for which the density of optimum coupling was relatively low (below the cut-off value). Good coupling with reflection coefficient ~3% has been achieved with plasma-antenna distance up to 11 cm for the new PAM. Finally, in order to eliminate the effect of ion cyclotron range of frequencies (ICRF) wave on 4.6 GHz LH wave coupling, the location of the ICRF launcher was changed to a port that is located 157.5° toroidally from the 4.6 GHz LH system and is not magnetically connected.
This study was conducted to examine the viability of Korean native striped cattle (Bos namadicus Falconer, Chikso) clone embryos after embryo transfer. Chikso somatic cell nuclear transfer (SCNT) embryos were produced by fusion of ear skin cells derived from a female Chikso with enucleated oocytes matured in vitro for 18-24 hr. After in vitro culture of SCNT embryos for 7 to 8 days, fresh or vitrified blastocysts derived from SCNT were transferred into a uterine horn of recipient cows. Fifteen of total 43 recipients were pregnant at Day 50 and 4 recipients were maintained to term. Three IVF-derived calves and 1 clone Chikso calf were born. Pregnancy rate was higher when fresh embryos were transferred to recipients compared to vitrified embryos, but development to term was not different between both groups. The clone Chikso calf died at 5 days after birth due to the fullness of amniotic fluid in rumen and the infection of umbilical cord. The result of the present study shows that clone Chikso calf can produced from the embryo transfer of SCNT embryos, however, solution of abortion problem is necessary to improve the cloning efficiency.
Jwa, Mi-Ri;Shin, Se-Jeong;Albright, Charles F.;Song, Ki-Won
BMB Reports
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v.32
no.1
/
pp.92-97
/
1999
Cytokinesis and septation should be coordinated to nuclear division in the cell division cycle for precise transmission of the genome into daughter cells. byr4, an essential gene in fission yeast Schizosaccharomyces pombe, regulates the timing of cytokinesis and septation in a dosage-dependent manner. We examined the intracellular localization of the Byr4 protein by expressing byr4 as a fusion of green fluorescence protein (GFP). The Byr4 protein localizes as a single dot on the nuclear periphery of interphase cells, duplicates before mitosis, and the duplicated dots segregate with the nuclei in anaphase. The behavior of Byr4p throughout the cell cycle strongly suggests that Byr4p is localized to the spindle pole body (SPB), a microtubule organizing center (MTOC) in yeast. The presence of the Byr4 protein in the SPB is consistent with its function to coordinate mitosis and cytokinesis. We also mapped the domains of Byr4p for its proper localization to SPB by expressing various byr4 deletion mutants as GFP fusions. Analyses of the diverse byr4 deletion mutants suggest that the indirect repeats and the regions homologous to the open reading frame (ORF) YJR053W of S. cerevisiae in its C-terminus are essential for its localization to the SPB.
PET/CT fused image with anatomical and functional information have improved medical diagnosis and interpretation. This fusion has resulted in more precise localization and characterization of sites of radio-tracer uptake. However, a motion during whole-body imaging has been recognized as a source of image quality degradation and reduced the quantitative accuracy of PET/CT study. The respiratory motion problem is more challenging in combined PET/CT imaging. In combined PET/CT, CT is used to localize tumors and to correct for attenuation in the PET images. An accurate spatial registration of PET and CT image sets is a prerequisite for accurate diagnosis and SUV measurement. Correcting for the spatial mismatch caused by motion represents a particular challenge for the requisite registration accuracy as a result of differences in PET/CT image. This paper provides a brief summary of the materials and methods involved in multiple investigations of the correction for respiratory motion in PET/CT imaging, with the goal of improving image quality and quantitative accuracy.
Proceedings of the Korean Institute of Information and Commucation Sciences Conference
/
2013.05a
/
pp.938-940
/
2013
According to the increasing concern for the safety management of nuclear power plants, the growing need for the development of a monitoring system for the management. Smart revolution sparked the evolution of wireless networks and the spread of smart phones, smart pads and mobile-centric, as well as the change of life, injury, industrial growth, and the Internet economy. In this paper, a fusion of the latest smart technology to meet the traditional nuclear industry in the era of the smart revolution, radiation safety management system is proposed to Content Basic imaging and dose output data in real time, and provides basic monitoring functions required to manage the radiation safety.
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