We estimate the fractal dimension of the ρ Ophiuchus Molecular Cloud Complex, associated with star forming regions. We selected a cube (v, l, b) database, obtained with J = 1−0 transition lines of 12CO and 13CO at a resolution of 22′′ using a multibeam receiver system on the 14-m telescope of the Five College Radio Astronomy Observatory. Using a code developed within IRAF, we identified slice-clouds with two threshold temperatures to estimate the fractal dimension. With threshold temperatures of 2.25 K (3σ) and 3.75 K (5σ), the fractal dimension of the target cloud is estimated to be D = 1.52–1.54, where P / AD/2 , which is larger than previous results. We suggest that the sampling rate (spatial resolution) of observed data must be an important parameter when estimating the fractal dimension, and that narrower or wider dispersion around an arbitrary fit line and the intercepts at NP = 100 should be checked whether they relate to rms noise level or characteristic structure of the target cloud. This issue could be investigated by analysing several high resolution databases with different quality (low or moderate sensitivity).

We present a multi-dimensional reduction method of the surveyed cube database obtained using a single- dish radio telescope in Taeduk Radio Astronomy Observatory (TRAO). The multibeam receiver system installed at the 14 m telescope in TRAO was not optimized at the initial stage, though it became more stabilized in the following season. We conducted a Galactic Plane survey using the multibeam receiver system. We show that the noise level of the first part of the survey was higher than expected, and a special reduction process seemed to be definitely required. Along with a brief review of classical methods, a multi-dimensional method of reduction is introduced; It is found that the ‘background’ task within IRAF (Image Reduction and Analysis Facility) can be applied to all three directions of the cube database. Various statistics of reduction results is tested using several IRAF tasks. The rms value of raw survey data is 0.241 K, and after primitive baseline subtraction and elimination of bad channel sections, the rms value turned out to be 0.210 K. After the one-dimensional reduction using ‘background’ task, the rms value is estimated to be 0.176 K. The average rms of the final reduced image is 0.137 K. Thus, the image quality is found to be improved about 43% using the new reduction method.

The reactive oxygen species (ROS) generated during the somatic cell transfer nuclear (SCNT) procedures may cause the mitochondrial dysfunction and DNA damage, which may result in restricts the reprogramming of SCNT embryos and play a key direct role in apoptosis. The present study was conducted to investigate the effect of antioxidant treatment during the SCNT procedures on the inhibition of mitochondria and DNA damages in bovine SCNT embryos. The reconstituted oocytes were treated with antioxidants of 25 μM β-mercaptoethanol (β-ME) or 50 μM vitamin C (Vit. C) during the SCNT procedures. In vitro fertilization (IVF) was performed for controls. Mitochondrial morphology and membrane potential (ΔΨ) were evaluated by staining the embryos with MitoTracker Red or JC-1. Apoptosis was analyzed by Caspase-3 activity assay and TUNEL assay, and DNA fragmentation was measured by comet assay at the zygote stage. Mitochondrial morphology of non-treated SCNT embryos was diffused within cytoplasm without forming clumps, while the IVF embryos and antioxidant treated SCNT embryos were formed clumps. The ΔΨ of β-ME (1.3±0.1, red/green) and Vit. C-treated (1.4±0.2, red/green) SCNT embryos were significantly higher (p<0.05) than that of non-treated SCNT embryos (0.9±0.1, red/ green), which similar to that of IVF embryos (1.3±0.1, red/green). Caspase-3 activity was not difference among the groups. TUNEL assay also revealed that little apoptosis was occurred in SCNT embryos as well as IVF embryos regardless of antioxidant treatment. Comet tail lengths of β-ME and Vit. C-treated SCNT embryos (337.8±23.5 μm and 318.7 ±27.0 μm, respectively) were shorter than that of non-treated SCNT embryos (397.4± 21.4 μm) and similar to IVF embryos (323.3±10.6 μm). These results suggest that antioxidant treatment during SCNT procedures can inhibit the mitochondrial and DNA damages of bovine SCNT embryos.

Bacterial contamination reduces the semen quality, semen preservation, and cause of disease spread as well. Sperm fertility is essential factor of reproductive performance in swine. Sperm fertility is affected by semen quality such as sperm motility, abnormality, morphology, and rate of bacterial contamination. This study was conducted to determine the relationship between elapsed time after semen preservation on the changes of bacteria and semen quality. Semen was diluted with BTS extender without antibiotic for 7 days and sperm parameter and fertility were measured. Sperm motility was measured by CASA and total bacteria number was counted after 22 24 hr incubation from counting agar plate in which sperm dilute to 10 106 in 0.9% saline solution and inoculate to agar. Acrosomal integrity was measured by Chlortetracycline (CTC) staining. CTC patterns were uniform fluorescence over the whole head (pattern A), characteristic of uncapacitated acrosome-intact spermatozoa; fluorescence-free band in the post-acrosomal region (pattern B), characteristic of capacitated acrosome-intact spermatozoa; and almost no fluorescence over the whole head except for a thin band in the equatorial segment (pattern C), characteristic of acrosome reacted spermatozoa. Total number of bacteria was significantly increased (p<0.0001) 3 days after preservation. Sperm motility, viability, and morphological abnormality on elapsed time after preservation were lower from 5 (77.24±6.47, p<0.001) and 7 days (77.24±6.47, p< 0.001) after preservation compared to 1 (15.71±7.18) and 3 days(18.39±7.22) after preservation, respectively. Sperm viability was significantly lower (53.25±35.03, p<0.0001) at 7 days after preservation. Mohological abnormality of sperm was lower (p<0.001) at 1 (15.71±7.18) and 3 (18.39±7.22) days compared to (5 21.84±7.91) and 7 (22.59± 9.93) days after preservation. Acrosomal integrity and capacitation rate (pattern A) were significantly lower (p<0.001) from 5 days after preservation.

본 연구는 demecolcine 처리에 의한 탈핵과 수핵란 세포질의 세포 주기가 소 체세포 핵이식란의 발육에 미치는 영향을 검토하였다. 체외에서 16~20시간 성숙배양된 난자를 극체 방출 유무 및 MI, MII기 난자로 구분하여 0.4㎕/mL demecolcine으로 40분간 처리 후 염색체 부위가 돌출된 난자는 탈핵 후 핵이식에 공시하였다. 소의 귀 피부 세포를 탈핵란에 이식하여 전기융합과 활성화 처리(Ca-ionophore+DMAP)를 거쳐 체외 배양하였다. Demecolcine처리 후 86.2%의 난자가 염색체 부위의 돌출을 보여 이 중 98.8%가 탈핵에 성공하였다. Demecolcine은 핵이식란의 발육에 영향을 주지 않았다. 제1극체 방출란 유래 핵이식란의 배반포 발육율은 극체 미방출란 유래 핵이식란에 비하여 유의적으로 높았다(18.2% vs.4.6%, P<0.05). 한편, MI 난자 유래 핵이식란의 분할율 및 배반포 발육율은(69.4%와 5.9%) MII 난자 유래 핵이식란에 비하여 유의적으로 낮았다(96.7%와 23.9%, P<0.05). 본 연구의 결과는 demecolcine 처리가 소 난자의 탈핵에 매우 효과적이며 MII기 난자가 MI기 난자에 비하여 수핵란 세포질로 더 적절하나 극체 미방출란 및 MI기 난자도 비록 제한적이기는 하지만 핵이식란의 배반포 발육을 지원할 수 있음을 보여준다.

체외수정 후 체외배양48시간에 난구세포의 제거 유ㆍ무에 따라 CR/sub 1aa/ 배양액에 homoglobin을 0, 1, 5 ㎍/㎖를 첨가한 구의 상실배기 이상 체외발육성 적은 난구세포를 제거한 구에서 23.8%, 33.3% 및 26.8% 였으며, 난구세포를 제거하지 않은 구에서는 각각 39.5%, 54.8% 및 48.8%로서 난구세포를 제거하지 않은 구에 Hb를 1 ㎍/㎖를 첨가한 구가 여타구보다 통계적으로 높은 결과를 얻었다(P<0.05). 체외수정 후 체외배양 96시간 후 난구세포를 제거 유ㆍ무에 따라 Hb를 0, 1, 5 ㎍/㎖를 첨가하였을 때, 상실배기 이상 체외발육성적은 난구세포를 제거한 구에서 각각 28.6%, 46.2% 및 39.1%였으며, 난구세포를 제거하지 않은 구에서는 각각 33.9%, 67.2% 및 46.0%로서, 난구세포를 제거하지 않은 구에 Hb를 1 ㎍/㎖를 첨가구가 여타구에 비해 통계적으로 유의하게 높게 나타났다(P<0.05). CR/sub 1aa/ 배양액에 L-NAME를 0, 10, 50 및 100 mM을 첨가한 구에서 상실배 이상 발육된 체외발육성적은 각각 55.6%, 64.9%, 58.8% 및 66.7%로써 L-NAME 첨가구가 무첨가구에 비해 커다란 차이가 나타나지 않았다(P>0.05). 모든 처리구에 배반포까지 발육된 체외수정란의 세포수에는 커다란 차이가 인정되지 않았다.

본 연구는 돼지의 난포란을 체외에서 성숙, 수정시킨 체외수정란의 체외배양 체계를 확립하고 그 기작을 규명하기 위하여 체외배양액에 항산화제인 melatonin의 첨가 및 melatonin과 sodium nitroprusside(SNP)의 첨가배양이 체외수정란의 체외발육에 미치는 영향을 검토하고자 실시하였다. NCSU 23 배양액에 melatonin을 0, 1, 5 및 10nM을 첨가하여 체외배양을 실시한 결과, 배반포기까지 발육율은 17.8%, 26.1%, 20.0% 및 16.3%로서 melatonin 1nM 첨가구가 여타구에 비해 통계적으로 유의하게 높은 성적을 나타냈으며(P<0.05), 상실배기 이상 발육 성적에서도 melatonin 1 nM 첨가구가 39.1%로서 대조구 33.3%, 5 nM 첨가구의 33.3% 및 10 nM 첨가구의 27.9%보다 높은 발육율을 나타냈다(P<0.05). NCSU 23 배양액에 SNP를 0, 50 및 100 μM을 첨가하여 체외 배양한 결과, 상실배 이상 발육성적은 각각 41.9%, 25.6% 및 28.4%로서 SNP 첨가구가 대조구보다 유의적으로 낮은 성적을 나타내었다(P<0.05). NCSU 23 배양액에 대조구, SNP 50 μM, SNP 50 μM에 melatonin 1, 5 및 10nM을 혼합첨가하여 체외 발육율을 조사한 결과, 배반포기 발육율은 각각 2.5%, 1.2%, 9.9％, 5.1% 및 3.7%로서 SNP 50μM + Mel. 1nM 첨가구가 여타구 보다 높은 성적을 나타냈으며, 상실배기 이상 체외 발육율은 31.3%, 34.1%, 39.5%, 29.4% 및 39.5%로서 SNP 50μM + Mel. 1 nM 첨가구와 SNP 50 μM + Mel. 10 nM 첨가구가 여타구보다 높은 발육율을 나타냈다. 모든 처리구에서 배반포까지 발육된 체외수정란의 세포수는 커다란 차이가 인정되지 않았다.

5% CO₂와 5% O₂ 농도 조건 하에서 NCSU23 배양액에 aesculetin을 0, 1, 5 및 10 ㎍/㎖를 첨가한 구에서 상실배기 이상 발육된 체외배양 성적은 10 ㎍ 첨가구(35.7%)가 여타구(0 ㎍, 30.2% ; 1㎍, 29.5% ; 5㎍, 29.2%)보다 통계적으로 유의하게 높은 성적을 얻었다(P<0.05). NCSU 23 배양액에 taurine을 0, 2.5 및 5.0 mM을 첨가, 체외배양을 실시한 결과 배반포기 이상 발육된 체외발육 성적은 각각 2.8%, 2.2% 및 7.0%였으며, 상실배이상의 체외 발육성적은 26.1%, 26.9% 및 31.7%로서 taurine 5.0 mM 첨가가 체외수정란의 체외발육 성적이 유의적으로 높은 것으로 나타났다(P<0.05). NCSU 23 배앙액에 melatonin을 0, 1, 5 및 10nM을 첨가하여 체외배양을 실시한 결과, 배반포기까지 발육된 체외 발육성적은 17.8%, 26.1%, 20.0% 및 16.3%로서 melatonin 1nM 첨가구가 여타구에 비해 통계적으로 유의하게 높은 성적을 나타냈으며(P<0.05), 상실배기 이상 발육된 체외발육 성적에서는 melatonin 1nM 첨가구가 39.1%로서 대조구 33.3%, 5 nM 첨가구의 33.3% 및 10 nM 첨가구의 27.9%보다 높은 발육율을 나타냈다(P<0.05). 한편 배반포기 수정란의 세포수 조사에서는 melatonin 10 mM 첨가구가 유의하게 낮은 것으로 나타났다.

This study was performed to establish the effective culture condition for the establishment of clonal lines from porcine fetal fibroblast cells. Fibroblasts derived from a pig fetus (Day 50) were cultured and passaged two times before use. A single cell was seeded in 96-well plates, cultured in medium supplemented with different concentrations of FBS, catalase or β-mercaptoethanol (βME), and classified by cell size and morphology. Cells were passaged two times into 4-well dish before freezing. The establishment efficiencies were not different among different concentrations of FBS (0.3 to 5.1%). However, population doubling time (PDT) was significantly decreased by increasing the FBS concentration (P＜0.05). The establishment efficiency of βME-added group (10.4%) was significantly higher than those of catalase-added and control groups (3.5%, and 3.5%, respectively, p＜0.05), and PDT was significantly decreased (23.6 vs 28.1, and 25.5 h, respectively, p＜0.05). However, catalase did not show a positive effect on the establishment efficiency. Cell size and morphology did not affect the establishment efficiency and PDT of clonal lines. The result of present study shows that the establishment efficiency of clonal cell lines can be enhanced by the culture in media supplemented with 30% FBS and βME.

본 연구는 체세포의 종류, 세포 제공 개체, 계대배양 수 및 세포의 trypsin 처리시간이 소 체세포 핵이식란의 체외발육에 미치는 영향을 검토하였다. 세 종류의 체세포(피부, 근육 및 난구세포) 와 암소 3 개체를 실험에 공시하였으며, 한 개체 유래의 피부세포는 5∼30회 계대배양하였고, 핵이식 전에 1∼3분간 trypsin처리하여 핵이식에 사용하였다. 핵이식과정은 상법에 따라 전기융합법을 이용하였다. 핵이식란의 배반포 발육율은 세포의 종류(16.5∼23.9%)나 개체 간(16.4∼19.5%)에 차이가 없으나, 30회 계대 배양한 세포를 사용한 경우(5.8%)에는 5회(25.3%) 또는 15회(23.5%) 계대 배양한 세포를 사용한 경우에 비해 유의적으로 낮았다(P＜0.05). 또한, 1분간 trypsinization 한 세포를 사용한 경우(30.7%)는 3분간 trypsinization 한 경우에 비해 배반포 발육율이 유의적으로 높게 나타났다(P＜0.05). 본 연구의 결과는 소 체세포 핵이식란의 발육이 체세포의 종류 및 세포 제공 개체에 따라서는 영향을 받지 않으나, passage 수 및 체세포의 trypsinization 시간에 따라서는 영향을 받을 수 있음을 시사한다.