• Title/Summary/Keyword: Unfused

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Fertilization by Microinjection of Mouse Round Spermatid (생쥐 원형정자세포의 미세주입에 의한 수정)

  • 이상민;백청순;구덕본;김묘경;김진회;박흠대;이훈택;정길생
    • Korean Journal of Animal Reproduction
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    • v.19 no.3
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    • pp.171-179
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    • 1995
  • This study was carried out to investigate the fertilizing ability of round spematids isolated from seminiferous tubules. A round spermatid was introduced into the perivitelline space of a mature oocyte using Leitz micromanipulators and then subjected to electrofusion. Electrofusion was induced by applying a single DC pulse of 90V with a duration of 60$\mu$sec using Model 611 Square Wave Stimulator(Phipps and Bird, U.S.A) in 0.3 M sucrose fusion medium containing 0.05mM CaCl2 and 0.1mM MgSO4, Oocyte pre-activation was conducted by exposure to a single DC(80V, 80$\mu$sec) pulse in electrofusion medium at 1 hour before electrofusion. The incidence of fusion with pre-activated oocytes(23.8%, 57/239) was higher than that with nonactivated oocytes(6.7%, 3/45). The most of electro-stimulated mouse oocytes cleaved regardless of the success or failure of fusion. Karyotyping of embryos that developed into blastocysts after exposure to the fusion pulse were performe. We found that blastocysts from the fused oocytes were diploid whereas blastocysts from the unfused oocytes were haploid. About 11.7 and 11.5% of fused and unfused oocytes were developmental potentials of fused and unfused oocytes. Therefore, these results suggest that the mouse mture oocyte can be fertilized by fusion with a round spermtid and subsequently developed normally.

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Improvement of the Protection Algorithm Based on Voltage Difference Method for Detecting Arcing Faults within 22.9kV Shunt Capacitor Banks (22.9kV급 병렬 커패시터 뱅크 내부의 아크 고장 판별을 위한 전압차동 보호 알고리즘의 개선 방안)

  • Lim Jung-Uk;Kwon Young-Jin;Kang Sang-Hee;Yuk Yoo-Kyoung
    • The Transactions of the Korean Institute of Electrical Engineers A
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    • v.54 no.2
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    • pp.61-66
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    • 2005
  • This paper presents a refined protection algorithm of the unfused 22.9kV shunt capacitor banks in grounded wye connection to improve the existing algorithm using the voltage difference method. It is difficult to detect ground faults with arc near the input points or ground faults near the grounding point by the existing algorithm using only the voltage balanced relay. This paper shows that ground faults with arc near the input point can be detected by harmonics analysis of the differential voltage and that it has no impact of harmonics out of nonlinear loads which have the quantitative influence on capacitor banks. Thus the proposed method using harmonics analysis can be a proper detection method. In case of ground faults near the grounding point, an OVGR is being added recently and its validity is verified in this paper. The proposed method is applied to a 22.9kV example system and is verified that the proposed algorithm can detect clearly faults which are not easy to detect by the existing method.

On the Possible Fusion-Promoting Factor Secreted from Cultured Myoblasts (培養 鷄胚 筋原細胞로부터 분비된 細胞融合 촉진 물질에 관한 연구)

  • Park, Hye-Gyeong;Park, Young-Chul;Lee, Chung-Choo;Ha, Doo-Bong
    • The Korean Journal of Zoology
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    • v.29 no.4
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    • pp.294-306
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    • 1986
  • In order to find out whether myoblast cells release into the culture medium any substances that induce or promote the fusion of myoblasts, chick embryonic myoblasts were cultured and the cultured medium (muscle-conditioned medium, MCM) was collected. The MCM was then added to the newly cultured myoblasts to examine if it has fusion-promoting activity. The MCM was also analyzed for its protein content before and after its addition to the second culture. The MCM apparently showed fusion-promoting activity when applied to unfused young myoblasts, suggesting that it contained substances that promote the fusion and that had been released from cells fo the previous culture. Analysis of proteins in the myoblasts and in the MCM suggested that the released protein was absorbed by or tightly bound to myoblasts of the second culture. One of the released proteins of about 175 kilodalton was degraded to a polypeptide of approximately 145 kilodalton, which appeared to act upon the membrane proteins of unfused myoblasts so as to stimulate their membrane to fuse with neighboring cells.

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Morphologic Diversities of Sacral Canal in Children;Three-Dimensional Computed Tomographic Study

  • Kim, Dae Wook;Lee, Seung Jun;Choi, Eun Joo;Lee, Pyung Bok;Jo, Young Hyun;Nahm, Francis Sahngun
    • The Korean Journal of Pain
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    • v.27 no.3
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    • pp.253-259
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    • 2014
  • Background: Caudal block is a common technique in children for reducing postoperative pain, and there have been several reports on the variations of the sacral canal in children. However, previous studies have mainly focused on the needle trajectory for caudal block, and there is limited information on the structural variations of the sacrum in children. The purpose of this study was to analyze the anatomic variations of sacral canals in children. Methods: Three-dimensional computed tomographic images were analyzed. The data from the images included (1) fusion of the sacral vertebral laminae and the sacral intervertebral space (2) existence of the sacral cornua and (3) the types of sacral hiatus. The types of sacral hiatus were classified into 3 groups: group I (fusion of S3 or S4 vertebral laminae), group II (unfused vertebral arch with the distance of the S3 and S4 vertebral laminae < 50% of the distance between the cornua), and group III (unfused vertebral arch with the distance of the S3 or S4 vertebral laminae ${\geq}50%$ of the distance between the cornua). Results: A total of 143 children were included in this study. All of the sacral vertebral arches were not fused in 22 children (15.4%). Cornua were not identified bilaterally in 5 (3.5%) and unilaterally in 6 (4.2%) children. In the sacral hiatus, group II and group III were identified in 22 (15.4%) and 31 (21.7%) children, respectively. Conclusions: The sacral canal has various anatomical variations in children. Careful attention must be paid to identify the correct anatomic landmark.

The strain of the spring ligament complex at different arthrodesis of the hindfoot for treatment of the flatfoot (평편족의 치료로 이용한 관절고정술시 스프링인대군 (spring ligament complex)에서 strain의 변화)

  • Lee, Kyung-Tai;Bae, Joon-Woo
    • Journal of Korean Foot and Ankle Society
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    • v.1 no.1
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    • pp.38-42
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    • 1997
  • It was well known that arthrodesis of the tarsal joint is an exellent procedure to correct the flatfoot deformity for relieving pain. Recently, concept of the selective tarsal joint fusion instead of the triple fusion was developedto preserve the joint motion. To investigate and compare the effect of the each different tarsal fusion, we measured the strain at the spring ligament, medial roof of the medial longitudianl arch. Five fresh frozen cadevar foot specimens, with distal half of the tibia were utilized. The superomedial portion of the spring ligament complex was dissected from the origin to the insertion. For each specimen, a calibrated open liquid metal strain guage was secured along the length of the superomedial portion. Under the specially devised test rig, measurement of the strain was taken at each test condition from the tare weight 18.2 lb followed by 38.2 lb., 82.2 lb and a maximum loads of 134.6 lb. : 1) unfused condition, 2) isolated subtalar fusion 3) isolated talonavicular fusion 4) combined subtalar and talonavicular fusion 5) triple fusion. Statistics showed that siginificant reduction in strain following the triple fusion, and from the subtalar fusion to triple fusion.

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Survivin protects fused cancer cells from cell death

  • Do, Mihyang;Kwak, In-Hae;Ahn, Ju-Hyun;Lee, In Jeong;Lee, Jae-Ho
    • BMB Reports
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    • v.50 no.7
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    • pp.361-366
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    • 2017
  • Tetraploidy, a potential precursor of cancer-associated aneuploidy, is produced either by cell fusion or failure of cytokinesis. In this study, low p53-expressing HeLa cells were used to address the fate of cancer cells after fusion. We found that massive cell death or growth arrest occurred a few days after fusion. Interestingly, cells with larger nuclei preferentially died after fusion, suggesting that a larger deviation of DNA content is a strong inducer of apoptosis. Notably, a fraction of cells escaped cell death. Also, the stability of survivin increased, and its localization changed preferentially to the cytosol in fused cells. Knockdown of survivin decreased the survival of fused cells, more than observed in unfused cells, showing increased dependency of fused cells on survivin. Collectively, after cancer cell fusion, some fused cells avoid the apoptotic crisis partly owing to survivin, and continue to proliferate, a process that contributes to human cancer progression.

Antimicrobial Activity of Bacteriophage Endolysin Produced in Nicotiana benthamiana Plants

  • Kovalskaya, Natalia;Foster-Frey, Juli;Donovan, David M.;Bauchan, Gary;Hammond, Rosemarie W.
    • Journal of Microbiology and Biotechnology
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    • v.26 no.1
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    • pp.160-170
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    • 2016
  • The increasing spread of antibiotic-resistant pathogens has raised the interest in alternative antimicrobial treatments. In our study, the functionally active gram-negative bacterium bacteriophage CP933 endolysin was produced in Nicotiana benthamiana plants by a combination of transient expression and vacuole targeting strategies, and its antimicrobial activity was investigated. Expression of the cp933 gene in E. coli led to growth inhibition and lysis of the host cells or production of trace amounts of CP933. Cytoplasmic expression of the cp933 gene in plants using Potato virus X-based transient expression vectors (pP2C2S and pGR107) resulted in death of the apical portion of experimental plants. To protect plants against the toxic effects of the CP933 protein, the cp933 coding region was fused at its Nterminus to an N-terminal signal peptide from the potato proteinase inhibitor I to direct CP933 to the delta-type vacuoles. Plants producing the CP933 fusion protein did not exhibit the severe toxic effects seen with the unfused protein and the level of expression was 0.16 mg/g of plant tissue. Antimicrobial assays revealed that, in contrast to gram-negative bacterium E. coli (BL21(DE3)), the gram-positive plant pathogenic bacterium Clavibacter michiganensis was more susceptible to the plant-produced CP933, showing 18% growth inhibition. The results of our experiments demonstrate that the combination of transient expression and protein targeting to the delta vacuoles is a promising approach to produce functionally active proteins that exhibit toxicity when expressed in plant cells.

Oil Pipeline Weld Defect Identification System Based on Convolutional Neural Network

  • Shang, Jiaze;An, Weipeng;Liu, Yu;Han, Bang;Guo, Yaodan
    • KSII Transactions on Internet and Information Systems (TIIS)
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    • v.14 no.3
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    • pp.1086-1103
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    • 2020
  • The automatic identification and classification of image-based weld defects is a difficult task due to the complex texture of the X-ray images of the weld defect. Several depth learning methods for automatically identifying welds were proposed and tested. In this work, four different depth convolutional neural networks were evaluated and compared on the 1631 image set. The concavity, undercut, bar defects, circular defects, unfused defects and incomplete penetration in the weld image 6 different types of defects are classified. Another contribution of this paper is to train a CNN model "RayNet" for the dataset from scratch. In the experiment part, the parameters of convolution operation are compared and analyzed, in which the experimental part performs a comparative analysis of various parameters in the convolution operation, compares the size of the input image, gives the classification results for each defect, and finally shows the partial feature map during feature extraction with the classification accuracy reaching 96.5%, which is 6.6% higher than the classification accuracy of other existing fine-tuned models, and even improves the classification accuracy compared with the traditional image processing methods, and also proves that the model trained from scratch also has a good performance on small-scale data sets. Our proposed method can assist the evaluators in classifying pipeline welding defects.

Improving Soluble Expression of β-Galactosidase in Escherichia coli by Fusion with Thioredoxin

  • Nam, E.S.;Jung, H.J.;Ahn, J.K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.12
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    • pp.1751-1757
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    • 2004
  • Recombinant heterologous proteins can be produced as insoluble aggregates partially or perfectly inactive in Escherichia coli. One of the strateges to improve the solubility of recombinant proteins is fusion with a partner that is excellent in producing soluble fusion proteins. To improve the production of soluble $\beta$-galactosidase, the gene of Thermus thermophilus KNOUC112 $\beta$-galactosidase (KNOUC112 $\beta$-gal) was fused with thioredoxin gene, and optimization of its expression in E. coli TOP10 was performed. KNOUC112 $\beta$-gal in pET-5b was isolated out, fused with thioredoxin gene in pThioHis C, and transformed to E. coli TOP10. The $\beta$-galactosidase fused with thioredoxin was produced in E. coli TOP10 as dimer and trimer. The productivity of fusion $\beta$ -galactosidase expressed via pThioHis C at 37$^{\circ}C$ was about 5 times higher than that of unfused $\beta$-galactosidase expressed via pET-5b at 37$^{\circ}C$. Inclusion body of $\beta$-galactosidase was formed highly, regardless of the induction by IPTG when KNOUC112 $\beta$ -gal was expressed via pET-5b at 37$^{\circ}C$. Fusion $\beta$ -galactosidase expressed at 37$^{\circ}C$ via pThioHis C without the induction by IPTG was soluble, but the induction by IPTG promoted the formation of inclusion body. Lowering the incubation temperature for the expression of fusion gene under 25$^{\circ}C$ prevented the formation of inclusion body, optimally at 25$^{\circ}C$. 0.07 mM of IPTG was sufficient for the soluble expression of fusion gene at 25$^{\circ}C$. The soluble production of Thermus thermophilus KNOUC112 $\beta$-galactosidase could be increased about 10 times by fusion with thioredoxin, and optimization of incubation temperature and IPTG concentration for induction.

A Study on the production of Monoclonal Antibodies against Rhodotrodula rubra (효모양진균의 단크론 생성)

  • Ryeom K.;Kim S. C.;Lee J. H.
    • Journal of environmental and Sanitary engineering
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    • v.4 no.1 s.6
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    • pp.17-28
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    • 1989
  • Having studied the production of monoclonal antibodies for developing a diagnosis medicine which shall be detected by a high-sensitivity test by using Rhodotorula rubra as a fungi-host which had been extracted through biochemical tests and follow-up examinations on Yeast-like fungi obtained from pulmonary tissues of pulmonary tuberculosis patients who had been in Kong ju National Tuberculosis Hospital from Jun. to Dec. in 1987, I. have gained such results as follows: 1. The fusion rate was influenced by feeder cell layers, cell density and time required to the cell fusion with cells in myelona subculture. 2. The fusion rate did not show any significant difference when the cell was applyed with two molecular weights, i.e., 1500 and 4000, of polyethylene glycol. 3. Fused cells after the addition of HAT selection media were bright and round, whereas unfused myelona cells and spleen cells were shrunk and granulated. 4. The cell fusion rate turned out to be about $57.2\%$(150 wells / 264 wells). 5. $10\%$(15 wells / 150 wells) of the positive reaction was detected in monoclonal antibody screening. 6. The titer which had reacted positively to Rhodotorula rubra fungal-host was 800 times in density after the gradual dilution of the produced monoclonal antibodies with Indirect ELISA method. 7. The Strongest specific reaction came out after the peroxidase labelled anti-human Immunogobulin had been applyed to Rhodotorula rubra for activating its nature after making drift with Carbonate-bicarbonate buffer (pH 9.6) and drying completely.

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