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Establishment of Spermatogonial Stem Cells using Total Testicular Cell Culture System in Mouse (정소세포의 체외 혼합배양 방법을 이용한 생쥐 정원 줄기세포 확립)

  • Lee, Won Young;Kim, Hee Chan;Kim, Dong Hoon;Chung, Hak Jae;Park, Jin Ki;Song, Hyuk
    • Reproductive and Developmental Biology
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    • v.37 no.3
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    • pp.143-148
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    • 2013
  • Spermatogenesis is initiated from spermatogonial stem cells (SSCs) that has an ability of self-renewal and unipotency to generate differentiating germ cells. The objective of this study is to develop the simple method for derivation of SSCs using non-sorting of both spermatogonia and feeder cells. Simply uncapsulated mouse testes were treated with enzymes followed by surgical mincing, and single cells were cultured in stempro-$34^{TM}$ cell culture media at $37^{\circ}C$. After 5 days of culture, aciniform of SSC colony was observed, and showed a strong alkaline phosphatase activity. Molecular characterization of mouse SSCs showed that most of the mouse SSC markers such as integrin ${\alpha}6$ and ${\beta}1$, CD9 and Stra8. In addition, pluripotency embryonic stem cell (ESC) marker Oct4 were expressed, however Sox2 expression was lowered. Interestingly, expression of SSC markers such as Vasa, Dazl and PLZF were stronger than mouse ESC (mESC). This data suggest that generated mouse SSCs (mSSCs) in this study has at least similar biomarkers expression to mESC and mSSCs derived from other study. Immunocytochemistry using whole mSSC colony also confirmed that mSSCs generated from this study expressed SSC specific biomarkers such as c-kit, Thy1, Vasa and Dazl. In conclusion, mSSCs from 5 days old mouse testes were successfully established without sorting of spermatogonia, and this cells expressed both mESC and SSC specific biomarkers. This simple derivation method for mSSCs may facilitate the study of spermatogenesis.

Study on Cosmeceutical Activities from Fermented Ginseng Berry Extracts (진생베리 발효추출물의 화장품 약리활성)

  • Kim, Il-Chool
    • Journal of the Korean Applied Science and Technology
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    • v.37 no.1
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    • pp.28-37
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    • 2020
  • Ginseng berry contains a large amount of Ginsenoside Re and has anti-inflammatory, anticancer, hypoglycemic and whitening effects. In this study, Rhizopus Oligosporus strain was used to establish ginseng berry fermentation process and cosmetic pharmacological activity of ginseng berry fermented product was analyzed.. The electron donating ability of ginseng berry extract by fermentation shown 81% at 1,000 ㎍/mL concentration. The ABTS+ radical scavenging ability of shown 100.2% at 1,000 ㎍/mL concentration. The tyrosinase inhibitory effect which is related to skin-whitening, was 57% at the concentration of 1,000 ㎍/mL. The elastase inhibitory effect which is related to skin-wrinkle, was 47% at 1,000 ㎍/mL concentration. Also, the collagenase inhibition effect was 33% at 1,000 ㎍/mL concentration. From these results, ginseng berry extracts by fermentation is considered to have anti-inflammatory, anti-wrinkle effect and whitening effect. Therefor, ginseng berry fermented product is expected to be very useful as an anti-inflammatory and anti-aging cosmetic raw material.

Comparison of Quantitative Endotoxin against 5 Species of Enterobacteriaceae (장내세균 5종의 Endotoxin 정량 비교)

  • Kwon, Pil Seung
    • Korean Journal of Clinical Laboratory Science
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    • v.48 no.2
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    • pp.124-129
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    • 2016
  • Endotoxin, also known as lipopolysaccharide (LPS) produced by the cell wall of gram negative bacteria can be present in any liquid or on any biomaterial. Endotoxin in blood can cause fever and inflammation. In this study, we compared bacterial endotoxin using Escherichia coli O157:H7, Klebsiella oxytoca, Salmonella Typhi, Shigella sonnei and Morganella morganii. Bacteria were cultured for use in the experiment, and diluted to $1.5{\times}10^8CFU/mL$. A check marked sensitivity confirmatory test of the Limulus amebocyte lysate (LAL) reagent was performed to examine the validity. The end point reaction to each bacteria sample was confirmed with 10 fold dilution and then the final reaction end point was confirmed by 2 fold dilution between the dilution step and the upper dilution step. According to the results, in detection of endotoxins in more than 0.015 EU/mL, E. coli O157 was 75~37.5 CFU/mL, K. oxytoca 37.5~18.75 CFU/mL, M. morganii and S. Typhi 3.75~1.875 CFU/mL, and S. sonnei 7.5~3.75 CFU/mL. The resulting value was finally ensured by a confirmation test for the inhibitory factor. Based on this study, conduct of further research on bacterial endotoxin is encouraged.

Activation Mechanism of Protease in the Germination of Mulberry Seeds (뽕나무 종자 발아시의 Protease 발현기구)

  • 배계선
    • Journal of Sericultural and Entomological Science
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    • v.35 no.1
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    • pp.1-6
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    • 1993
  • The activity change of mulberry seeds protease was compared during germination for 5 days at 28$^{\circ}C$ in the dark place after daily hormone injection of different concentration. The protease from germinated mulberry seeds for 4 days was partially purified and the enzyme characteristics was investigated. The protease activity of mulberry seeds treated by hormone was highest with 10 $\mu$m GA3 followed by 10 $\mu$M zeatin and 10 $\mu$M kinetin. The protease activity of mulberry seeds was increased by 14% with 10ml agar culture that control at 4th day of germination. The protease from mulberry seeds was purified 313 fold by DEAE-Toyo-pearl 650M, Butyl-Toyopearl, Hydrozylapatite and Toyopearl HW 55M. After purification, the specific activity of the enzyme was 175 units/mg. Optimum pH and temperature of protease from mulberry seeds was 5.0 and 37$^{\circ}C$, respectively. The protease was stable below 37$^{\circ}C$ and the enzyme activity was decreased by 50%, when incubated at 52$^{\circ}C$ for 10minutes. The protease activity of mulberry seeds was inhibited by metal ions such as mercury, iron, zinc, copper, but activited by magnesium, choromium, aluminium ions. The Km value of the protease was 0.89mM with azocasein as a subscribe.

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Electrical Explorations in and around the Nanjido Waste Landfill (난지도 폐기물 매립장과 그 주변 지역에서의 전기탐사)

  • Lee, Kiehwa;Yoon, Jong-Ryeol
    • Journal of the Korean Society of Groundwater Environment
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    • v.2 no.2
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    • pp.64-71
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    • 1995
  • Electrical soundings were conducted in and around the Nanjido waste landfill in August, 1994 and February, 1995. Schlumberger array was adopted and 37 and 22 soundings were performed around and in the Nanjido landfill, respectively. Besides, self potentials were measured at 50 points, in front of the right Nanjido landfill. Interpretations of the sounding data show low resistivity zones of about 10 Ω-m at depth ranging from 10 to 80 m from the surface in front of the landfill and of about 6 Ω-m at depth ranging from 37 m to 130 m in the landfill. It appears that these low resistivity zones are contaminated by or saturated with leachate, and their depths are deeper than those of boring data by 20∼30 m. These results indicate the possibility of contamination of weathered zone and the upper part of the bed rock in these areas. But sounding data obtained at the back of the landfill reveal more resistive and thinner low resistivity zones than those in and in front of the landfill. Thus it is concluded that the degree of contamination by leachate in and in front of the landfill is greater than that at the back of the landfill.

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Estimation of optimal pumping rate, well efficiency and radius of influence using step-drawdown tests (단계양수시험을 이용한 최적 양수량, 우물효율 및 영향반경 산정)

  • Choi, Hyun-Mi;Lee, Jin-Yong;Cheon, Jeong-Yong;Jun, Seong-Chun;Kwon, Hyung-Pyo
    • The Journal of Engineering Geology
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    • v.20 no.2
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    • pp.127-136
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    • 2010
  • Optimal pumping rate, well efficiency and radius of influence were estimated using field step-drawdown tests. According to the analysis results, optimal pumping rates were estimated as 9.37, 16.20 $m^3/day$ for KDPW 1 and 8.11, 14.10 $m^3/day$ for KDPW 2. The well efficiency was calculated as 72.02~90.73% for KDPW 1 while it was 70.62~88.52% for KDPW 2. In the meanwhile, the steady-state analysis yielded the radius of influence (ROI) of 3.50~31.92 m in case of pumping at KDPW 1 and the ROI of 0.14~37.43 m in case of pumping at KDPW 2. In addition, the transient analysis produced the ROI of 0.02~8.34 m for KDPW 1 pumping and the ROI of 0.24~9.68 m for KDPW 2 pumping. The methodology used in this study can be usefully applied in the pump and treat remediation design for contaminated groundwater.

Bfl-1/A1 Molecules are Induced in Mycobacterium Infected THP-1 Cells in the Early Time Points

  • Park, Sang-Jung;Cho, Jang-Eun;Kim, Yoon-Suk;Cho, Sang-Nae;Lee, Hye-Young
    • Biomedical Science Letters
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    • v.18 no.3
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    • pp.201-209
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    • 2012
  • Apoptosis is a physiological programmed cell death process. Tubercle bacilli inhibit apoptosis of alveolar macrophages and phagolysosome fusion. We investigated whether the Bcl-2 family anti-apoptotic member, Bfl-1/A1, plays an important role in the anti-apoptotic process during mycobacterial infection. PMA-treated human monocytoid THP-1 cells were infected with mycobacteria (H37Rv, BCG, and K-strain) at a multiplicity of infection (MOI) of 10 for 0, 1.5, 3, 6, 9, 12, 18, 24, 48, or 72 h. In addition, PMA-treated THP-1 cells were pretreated with specific inhibitors for 45 min before stimulation with mycobacteria at an MOI of 10 for 4 h. After the indicated time, the cells were subject to reverse transcription-polymerase chain reaction (RT-PCR) analysis, and a Bfl-1/A1-specific Western blot was performed. In PMA-differentiated THP-1 cells, the expression level of Bfl-1/A1 mRNA was increased by Mycobacterium tuberculosis (MTB) H37Rv infection. The mRNA level of Bfl-1/A1 peaked 3 h after MTB infection, then declined gradually until 9 h. However, Bfl-1/A1 mRNA induction gradually re-increased from 24 h to 72 h after MTB infection. No difference in Bfl-1/A1 expression was detected following infection with MTB H37Rv, K-strain, or M. bovis BCG. These results were not dependent on mycobacterial virulence. Moreover, mRNA levels of other anti-apoptotic molecules (Mcl-1, Bcl-2, and Bcl-xL) were not increased after MTB H37Rv or K-strain infection. These results suggest that mycobacteria induce the innate immune host defense mechanisms that utilize Bfl-1/A1 molecules at early time points, regardless of virulence.