• Title/Summary/Keyword: Cell selection

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Association of Novel Polymorphisms in Lymphoid Enhancer Binding Factor 1 (LEF-1) Gene with Number of Teats in Different Breeds of Pig

  • Xu, Ru-Xiang;Wei, Ning;Wang, Yu;Wang, Guo-Qiang;Yang, Gong-She;Pang, Wei-Jun
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.9
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    • pp.1254-1262
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    • 2014
  • Lymphoid enhancer binding factor 1 (LEF-1) is a member of the T-cell specific factor (TCF) family, which plays a key role in the development of breast endothelial cells. Moreover, LEF-1 gene has been identified as a candidate gene for teat number trait. In the present study, we detected two novel mutations (NC_010450.3:g. 99514A>G, 119846C>T) by DNA sequencing and polymerase chain reaction-restriction fragment length polymorphism in exon 4 and intron 9 of LEF-1 in Guanzhong Black, Hanjiang Black, Bamei and Large White pigs. Furthermore, we analyzed the association between the genetic variations with teat number trait in these breeds. The 99514A>G mutation showed an extremely significant statistical relevance between different genotypes and teat number trait in Guanzhong (p<0.001) and Large White (p = 0.002), and significant relevance in Hanjiang (p = 0.017); the 119846C>T mutation suggested significant association in Guanzhong Black pigs (p = 0.042) and Large White pigs (p = 0.003). The individuals with "AG" or "GG" genotype displayed more teat numbers than those with "AA"; the individuals with "TC" or "CC" genotype showed more teat numbers than those with "TT". Our findings suggested that the 99514A>G and 119846C>T mutations of LEF-1 affected porcine teat number trait and could be used in breeding strategies to accelerate porcine teat number trait improvement of indigenous pigs breeds through molecular marker assisted selection.

Comparative Genomic and Genetic Functional Analysis of Industrial L-Leucine- and L-Valine-Producing Corynebacterium glutamicum Strains

  • Ma, Yuechao;Chen, Qixin;Cui, Yi;Du, Lihong;Shi, Tuo;Xu, Qingyang;Ma, Qian;Xie, Xixian;Chen, Ning
    • Journal of Microbiology and Biotechnology
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    • v.28 no.11
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    • pp.1916-1927
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    • 2018
  • Corynebacterium glutamicum is an excellent platform for the production of amino acids, and is widely used in the fermentation industry. Most industrial strains are traditionally obtained by repeated processes of random mutation and selection, but the genotype of these strains is often unclear owing to the absence of genomic information. As such, it is difficult to improve the growth and amino acid production of these strains via metabolic engineering. In this study, we generated a complete genome map of an industrial L-valine-producing strain, C. glutamicum XV. In order to establish the relationship between genotypes and physiological characteristics, a comparative genomic analysis was performed to explore the core genome, structural variations, and gene mutations referring to an industrial L-leucine-producing strain, C. glutamicum CP, and the widely used C. glutamicum ATCC 13032. The results indicate that a 36,349 bp repeat sequence in the CP genome contained an additional copy each of lrp and brnFE genes, which benefited the export of L-leucine. However, in XV, the kgd and panB genes were disrupted by nucleotide insertion, which increase the availability of precursors to synthesize L-valine. Moreover, the specific amino acid substitutions in key enzymes increased their activities. Additionally, a novel strategy is proposed to remodel central carbon metabolism and reduce pyruvate consumption without having a negative impact on cell growth by introducing the CP-derived mutant $H^+$/citrate symporter. These results further our understanding regarding the metabolic networks in these strains and help to elucidate the influence of different genotypes on these processes.

Downregulation of $\textrm{NF}_{-{\kappa}}\textrm{B}$ activation in human keratinocytes by melanogenic inhibitors

  • Ahn, Kwang-Seok;Lee, Jinseon;Kim, Yeong-Shik
    • Proceedings of the SCSK Conference
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    • 2003.09a
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    • pp.780-803
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    • 2003
  • Exposure of skin cells, particularly keratinocytes to various nuclear factor-kappaB ($\textrm{NF}_{-{\kappa}}\textrm{B}$) activators [e.g. tumor necrosis factor-$\alpha$, interleukin-1, lipopolysaccharides, and ultraviolet light] leads to phosphorylation and degradation of the inhibitory protein, $\textrm{I}_{{\kappa}}\textrm{B}$. Liberated $\textrm{NF}_{-{\kappa}}\textrm{B}$ is translocated into the nucleus where it can change or alter expression of target genes, resulting in the secretion of extracellular signaling molecules including melanotrophic factors affecting melanocyte. In order to demonstrate the possible role of $\textrm{NF}_{-{\kappa}}\textrm{B}$ activation on the synthesis of melanotrophic factors from the keratinocytes, the activities of $\textrm{NF}_{-{\kappa}}\textrm{B}$ induced by melanogenic inhibitors (MIs) were determined in human HaCaT keratinocytes transfected with $\textrm{pNF}_{-{\kappa}}\textrm{B}$-SEAP-NPT plasmid. Transfectant cells released the secretory alkaline phosphatase (SEAP) as a transcription reporter in response to the $\textrm{NF}_{-{\kappa}}\textrm{B}$ activity and contain the neomycin phosphotransferase (NPT) gene for the dominant selection marker for geneticin resistance. MIs such as niacinamide, kojic acid, hydroquinone, resorcinol, arbutin, and glycolic acid were preincubated with transfectant HaCaT cells for 3 h and then ultraviolet B (UVB) was irradiated. $\textrm{NF}_{-{\kappa}}\textrm{B}$ activation was measured with the SEAP reporter gene assay using a fluorescence detection method. Of the Mis tested, kojic acid ($IC_{50}$/ = 60 $\mu$M) was found to be the most potent inhibitor of UVB-upregulating $\textrm{NF}_{-{\kappa}}\textrm{B}$ activation in transfectant HaCaT cells, which is followed by niacinamide ($IC_{50}$/= 540 $\mu$M). Pretreatment of the transfectant HaCaT cells with the Mis, especially kojic acid and niacinamide, effectively lowered $\textrm{NF}_{-{\kappa}}\textrm{B}$ binding measured by electrophoretic mobility shift assay. Furthermore, these two inhibitors remarkably reduced the secretion level of IL-6, one of melanotrophic factors, triggered by UV-radiation of the HaCaT cells. These observations suggest that Mis working at the in vivo level might act partially through the modulation of the synthesis of melanotrophic factors in keratinocyte.

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Lung Volume Reduction Surgery in Patients with Severe Emphysema, 7 cases (중증 폐기종 환자에서의 폐용적 감축술, 7례)

  • Jin, Ung;Lee, Sun-Hee;Kim, Si-Hoon;Wang, Young-Pil;Cho, Kyu-Do;Park, Jae-Kil;Kwak, Moon-Sub;Kim, Se-Wha
    • Journal of Chest Surgery
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    • v.32 no.6
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    • pp.543-548
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    • 1999
  • Background: These days, lung volume reduction surgery (LVRS) is used as an alternative or a bridge operation to lung transplantation in treating patients with severe emphysema. The procedure can be used in patients with pulmonary nodules combined with severe emphysema. We report the results of 21 months follow up after lung volume reduction surgery in 7 cases including 2 cases of concurrent resection of pulmonary nodules. Material and Method: Seven patients with emphysema, including 2 cases of preoperatively suspected lung cancer were operated with LVRS technique between July 1996 and June 1997. Result: Postoperative mortality was observed in a case of squamous cell carcinoma in LUL with brain metastasis, detected at postoperative 13months. Average of 21months(19-25months) follow up was done for other cases without specific events. Conclusion: LVRS is a useful operation in the treatment of patients with severe emphysema, but further evaluation should be done about the long term results and precise criteria for patient selection. Simultaneous LVRS and tumor resection could be done in patients with emphysema with marginal reserve in the hope of maximizing postoperative lung functions.

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Comparison of Selective Media for Isolation and Detection of Shigella spp. from Foods (식품으로부터 쉬겔라 검출을 위한 분리배지 비교)

  • In, Ye-Won;Ha, Su-Jeong;Oh, Se-Wook
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.7
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    • pp.1025-1031
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    • 2011
  • The objective of this study was to compare the performances of conventional microbiological media used in isolation of Shigella spp. from foods. Total of six selective media, including MacConkey agar (MAC), Salmonella Shigella agar (SSA), desoxycholate citrate agar (DCA), xylose lysine desoxycholate agar (XLD), hektoen enteric agar (HEA), and CHROMagar, were tested. MAC showed almost the same colony numbers as compared to tryptic soy agar (TSA) while DCA showed significantly lower colony numbers when cultivated Shigella spp. was counted in each medium. In a food recovery test with beef, pork and shrimp, S. sonnei recovered well on CHROMagar (p<0.05). With lettuce and cabbage, S. sonnei displayed significantly significant recovery (p<0.05) on SSA in comparison with other selective media. Heat-injured cells recovered well on MAC and SSA. In a specificity test using Enterobacteriaceae strains, HEA was identified as having the highest specificity among the tested media. However, Morganella spp. could not be differentiated from Shigella spp. on any of the tested selective media. Shigella spp. precluded the possibility of isolation from foods by a single 'best' selective medium. Consequently, a combination of complementary selective media or selection of appropriate media according to cell conditions must be considered for comprehensive isolation.

Selection of parental monokaryons from Korean Hypsizigus marmoreus by protoplast regeneration (원형질체 재생을 통한 느티만가닥버섯 단핵균주 선발)

  • Oh, Youn-Lee;Kong, Won-Sik;Jang, Kab-Yeul;Shin, Pyung-Gyun;Kim, Eun-Sun;Oh, Min ji;Choi, In-Geol
    • Journal of Mushroom
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    • v.13 no.3
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    • pp.270-273
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    • 2015
  • Hypsizigus marmoreus is commercially the most important edible mushroom in Japan. This mushroom is usually cultivated for a longer period (about 85~120 days) than other mushroom. In order to develop a new cultivar that has a shortened cultivation period, the genome analysis of this strain has been considered. This study aims to obtain parental monokaryotic strains reproducing 'Haemi' cultivar in Hypsizigus marmoreus for reference genome sequencing. The mycelia were cultured in MCM and MYG media for various incubation periods. Homogenized mycelia were treated with commercial cell wall degrading enzymes to maximize protoplasts production yield from Hypsizigus marmoreus. The greatest number of protoplasts was obtained from mycelia cultured in MCM media for 3 days using Novozyme enzyme. The isolated protoplasts were grown in regeneration agar media after two weeks. Regenerated colonies were picked and moved on separated dishes for microscopic observation. Neohaplonts regenerated from dikayotic strains were identified by the absence of clamp connections. We confirmed that one of monokaryotic strains is a parental strain by crossing with an original compatible strain of 'Haemi' cultivar. This parental strain will be used for reference genome sequence analysis.

배양액이 마우스 핵이식 재구축배의 초기발생에 미치는 영향

  • 심보웅;권오용;이은화;김순희;서정선
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.76-76
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    • 2002
  • 본 실험은 Piezo-미세조작기(PrimeTech Ltd., Japan)를 사용하여 마우스 핵이식 후 재구축배를 CZB와 KSOM 두가지 배양액을 사용하여 체외배양성적을 비교 검토하였다. MII의 미수정란은 성숙한 4~5주령 B6D2Fl에 hCG 주사 후 14시간째에 과적 방법을 통해 난관의 팽대부로 부터 회수하였고, metaphase II chromosome-spindle complex와 최소량의 세포질을 내경이 10$\mu\textrm{m}$인 피펫으로 흡입하여 탈핵하였다. 핵이식에 사용된 난구세포(8-l0$\mu\textrm{m}$)는 3시간동안 12% PVP 에처리 하여 piezo-미세조작기를 이용하여 세포질에 세포의 핵을 직접 미세주입 하였다. 핵이식 후 생존한 재구축배는 2시간동안 배양한 후 10mM SrC1$_2$와 5$\mu\textrm{g}$/$m\ell$의 cytochalasin B가 첨가된 $Ca^{2+}$-free CZB에서 6시간 활성화 처리하였다, 활성화 처리 후 위전핵이 관찰된 재구축란을 CZB 와 KSOM 배지에서 배양하면서 발달률을 비교하였고, 상실배 및 배반포배로 발달한 재구축배를 day 3 대리모에 이식하였다. 표 1에서 보는 바와 같이 재구축배의 2-cell로의 발달률에 있어서 KSOM이 CZB에 비하여 유의적으로 높게 나타났으며(P<0.05), 또한 4-cell과 상실배/배반 포배로의 발달률에 있어서도 KSOM이 CZB에 비하여 유의적으로 높은 발달률을 나타내었다(P<0.01). 또한 KSOM 배지에서 배양된 상실배/배반포배를 대리모에 이식한 경우에 11.5 d.p.c에 생존한 태아가 관찰되었다. 이상의 결과로 핵이식 재구축배의 활성화 처리 후의 발생에는 KSOM 배지가 CZB 배지에 비하여 유효함을 확인 할 수 있었다.그와 같은 배양 기술을 이용하여 외래유전자를 도입한 일련의 결과에 관하여 보고 하고자한다., 이것은 세포내 유전자가 transfection되지 않은 세포도 neo selection에서 선발된다는 것을 제시하고 있다. 따라서 체세포를 이용한 형질전환동물 생산을 위해서는 세포내 유전자 도입과 선발 과정에서 나타난 colony에 대하여 보다 엄격한 screen을 하는 것이 필요한 것으로 생각된다.로 우점하였다. 여름철 식물플랑크톤 대발생에 영향은 수온과 직산염이 중요하였으나, 부유물질 크게 기여하지 못하였다.애를 확인하고 지도 관점을 파악하는 것을 포함한다. 그러나 본 논문은 역사발생적 수학 학습-지도 원리의 실제적인 적용에 관하여는 기초적인 연구에 지나지 않기 때문에, 역사발생적 원리를 학교수학에 실제적으로 적용하기 위해서는 각각의 내용에 대한 철저한 역사적 분석을 바탕으로 하는 후속 연구가 필요하다./TEX>구성교육${\lrcorner}$이 조선총독부의 관리하에서 실행되었다는 것을, 당시의 사범학교를 중심으로 한 교육조직을 기술한 문헌에 의해 규명시켰다.nd of letter design which represents -natural objects and was popular at the time of Yukjo Dynasty, and there are some documents of that period left both in Japan and Korea. "Hyojedo" in Korea is supposed to have been influenced by the letter design. Asite- is also considered to have been "Japanese Letter Jobcheso." Therefore, the purpose of this study is to look into the origin of the letter designs in t

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Development of Liquid Crystal Optic Modulation Based X-ray Dosimeter by Using CdS Sensor (CdS 센서를 이용한 액정 광변조 X-선 검출 시스템 개발)

  • Noh, Si-Cheol;Kang, Sang-Sik;Jung, Bong-Jae;Choi, Il-Hong;Kim, Hyun-Hee;Cho, Chang-Hoon;Park, Jun-Hong;Park, Ji-Koon
    • Journal of the Korean Society of Radiology
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    • v.5 no.6
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    • pp.357-361
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    • 2011
  • In this study, the liquid-crystal optical modulation X-ray detection system using a CdS which is a family of II-IV compound semiconductor was proposed. The system consist of the detector, the signal processing part, the liquid-crystal driving parts, microcontroller, and I/O parts, and was designed to be suitable for miniaturization and portable. In addition, the system can measure a wide range X-ray by using the detecting range selection. In order to evaluate the performance of the proposed system, the CdS sensor's output characteristics were confirmed in accordance with changes of dose, and excellent correlation was determined. And also, the optical penetration ratio was discussed in accordance with changes of the applied voltage by measuring the change of the liquid-crystal in accordance with changes of the applied voltage. Through these results, the characteristics of the liquid-crystal optical modulation system such as the excellent reproducibility and the noise immunity were confirmed. And we considered that the CdS cell-based liquid-crystal optical modulated portable X-ray detection system could be applied to compact, low-cost, portable system.

Design of High-Performance Motion Estimation Circuit for H.264/AVC Video CODEC (H.264/AVC 동영상 코덱용 고성능 움직임 추정 회로 설계)

  • Lee, Seon-Young;Cho, Kyeong-Soon
    • Journal of the Institute of Electronics Engineers of Korea SD
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    • v.46 no.7
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    • pp.53-60
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    • 2009
  • Motion estimation for H.264/AVC video CODEC is very complex and requires a huge amount of computational efforts because it uses multiple reference frames and variable block sizes. We propose the architecture of high-performance integer-pixel motion estimation circuit based on fast algorithms for multiple reference frame selection, block matching, block mode decision and motion vector estimation. We also propose the architecture of high-performance interpolation circuit for sub-pixel motion estimation. We described the RTL circuit in Verilog HDL and synthesized the gate-level circuit using 130nm standard cell library. The integer-pixel motion estimation circuit consists of 77,600 logic gates and four $32\times8\times32$-bit dual-port SRAM's. It has tile maximum operating frequency of 161MHz and can process up to 51 D1 (720$\times$480) color in go frames per second. The fractional motion estimation circuit consists of 22,478 logic gates. It has the maximum operating frequency of 200MHz and can process up to 69 1080HD (1,920$\times$1,088) color image frames per second.

Further Evidence of Linkage at the tva and tvc Loci in the Layer Lines and a Possibility of Polyallelism at the tvc Locus

  • Ghosh, A.K.;Pani, P.K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.5
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    • pp.601-605
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    • 2005
  • Three lines of White Leghorn (WL) chickens (IWJ, IWG and IWC) maintained at Central Avian Research Institute, Izatnagar (UP), were used for chorioallantoic membrane (CAM) and liver tumour (LT) assay. Eleven-day-old embryos of each line were partitioned into three groups and inoculated with 0.2 ml of subgroup A, subgroup C and an equal mixture of subgroup A and C Rous sarcoma virus (RSV). Subgroup virus receptor on the cell surface membrane for subgroup A is coded for by tumour virus a (tva) locus and for subgroup C by tumour virus c (tvc) locus. The random association of the genes at the tva and tvc loci in IWJ and IWC line was assessed and the $x^2$-values for phenotypic classes were found to be significant, indicating the linkage between the tva and tvc loci. The linkage value was estimated to be 0.09 on pooled sex and pooled line basis. On the basis of four subclass tumour phenotypes a 4-allele model was proposed for tva locus having $a^{s1}$, $a^{s2}$, $a^{r1}$ and $a^{r2}$ alleles and the frequencies were calculated as 0.47, 0.13, 0.13 and 0.27 for IWJ line, 0.31, 0.33, 0.14 and 0.22 for IWG line and 0.44, 0.11, 0.21 and 0.24 for IWC line, respectively. Similarly, for tvc locus the frequencies of four alleles i.e. $c^{s1}$, $c^{s2}$, $c^{r1}$ and $c^{r2}$ were calculated as 0.42, 0.20, 0.21 and 0.17 for IWJ line, 0.42, 0.17, 0.27 and 0.14 for IWG line and 0.30, 0.21, 0.16 and 0.33 for IWC line, respectively. The $x^2$-values for all classes of observations were not significant (p>0.05), indicating a good fit to the 4-allele model for the occurrence of 4-subclass tumour phenotypes for tva and tvc loci. On the basis of the 2-allele model both tva and tvc locus carries three genotypes each. But, on the basis of the 4-allele model tva and tvc locus carries 10 genotypes each. The interaction between A-resistance and C-resistance (both CAM and LT death) was ascertained by taking the 10 genotypes of tva locus and 3 genotypes of tvc locus by pooling the lines and partitioning the observations into 3 classes. The $x^2$-values for the genotypic classes of CAM (-) LT (+) and CAM (-) LT (-) phenotypes to mixed virus (A+C) infection were found to be highly significant (p<0.01), indicating increased resistance, which indicates the joint segregation of $a^r$ and $c^r$ genes, suggesting the existence of close linkage between the tva and tvc loci. Therefore, an indirect selection approach using subgroup C viruses can be employed to generate stocks resistant to subgroup A LLV, obviating contamination with the most common agent causing LL in field condition.