• Title/Summary/Keyword: CFU(colony-forming units)

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Ex vivo Expansion and Clonal Maintenance of CD34+ Selected Cells from Cord Blood and Peripheral Blood (제대혈 및 말포혈로부터 분리한 CD34 양성 세포의 체외 증폭 및 클론 유지)

  • Kim, Soon Ki;Ghil, Hye Yoon;Song, Sun U.;Choi, Jong Weon;Park, Sang Kyu
    • Clinical and Experimental Pediatrics
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    • v.48 no.8
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    • pp.894-900
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    • 2005
  • Purpose : Because of the unavailability of marrow transplantation, umbilical cord blood (CB) is increasingly being used. We evaluated the potential of ex vivo expansion and clonality in CD34+ cells separated from cord blood source and mobilized peripheral blood (PB) in a serum-free media. Methods : The CD34+ cells, selected from CB and mobilized PB, were expanded with hematopoietic growth factors. They were then cultured for burst-forming units of erythrocytes (BFU-E), colony-forming units of granulocytes and monocytes (CFU-GM) and colony-forming units of megakaryocytes (CFU-Mk) at culture days 0, day 4, day 7, and day 14 with various growth factors. Results : The CB-selected CD34+ cells showed significantly higher total cell expansion than those from the PB at day 7 (2 fold increase than PB). The CB-selected CD34+ cells produced more BFU-E colonies than did the PB on culture at days 7 and at day 14. Also, the CB-selected CD34+ cells produced more CFU-Mk colonies than did the PB on culture at day 4 and at day 7. Conclusion : The ex vivo expansion of the CB cells may be promising in producing total cellular expansion, CFU-Mk and BFU-E compared with PB for 7 to 14 days. The growth factors combination including megakaryocyte growth and development, flt3-ligand and interleukin-3 showed more expansion in the view of total cells and clonal maintenance compared with less combination.

A Study on Variation of Colony Forming Units of Fungi by Input Ratios of Wood Chips in Aerobic Composting of Food Wastes (음식물류폐기물의 호기성 퇴비화에 있어서 목재세편의 투입비에 따른 곰팡이의 균락형성단위의 변화에 관한 연구)

  • Park, Seok-Hwan
    • Journal of Environmental Health Sciences
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    • v.33 no.5
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    • pp.451-455
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    • 2007
  • This study was performed to evaluate the effects of input ratios of bulking material in aerobic composting of food wastes on variation of colony forming units(CFU) of fungi. Wood chips were used as a bulking material. Volume ratios of food wastes to wood chips in reactor of Control, WC-1 and WC-2 were 10/0, 10/5 and 10/10, respectively. Reactors were operated for 24 days with I hour stirring by 1rpm and 2 hours of the forced aeration rate of $80L/min{\cdot}m^3$ per day. WC-2 reached high temperature range faster than WC-1, and the maximum temperature of WC-2 was higher than that of WC-1. WC-2 reached high pH range faster than WC-1. and the maximum pH of WC-2 was higher than that of WC-1. WC-2 reached high Log(CFU/gram) range faster than WC-I, and the maximum Log(CFU/gram) of WC-2 was higher than that of WC-1. These all mean that the reaction velocity of composting of WC-2 was faster than that of WC-1. The profile of fungi changes in Log(CFU/gram) was similar to that of temperature changes (r=0.8861) not pH changes (r=0.1631).

Rapid Microbiological Assessment Method by using ATP-Bioluminescence in Ginseng Powder (ATP-Bioluminescence를 이용한 인삼분말의 미생물 신속검사법)

  • 곽이성;김천석;송용범;고셩룡
    • Journal of Ginseng Research
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    • v.25 no.3
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    • pp.127-129
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    • 2001
  • Bioluminescence technique was applied to ginseng powders. ATP bioluminescence can be used as a rapid method that can be implemented for microbiological monitoring of contaminated ginseng powders. The RLU (relative light units) of ATP was proportion to bacterial CFU (colony forming units) when in high contaminated ginseng powders ($\geq$ about 1.0$\times$10$^4$CFU/g). However, when in low contaminated ginseng powders ($\times$10$^4$CFU/g), the RLU was not proportion to CFU, respectively.

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Enhancing effect of Canavalia gladiata DC semen on the hematopoietic expansion and function of stem cells (도두(刀豆)의 조혈증진 작용에 관한 연구)

  • Kim, Seung-Hyung;Kim, Kun-Hoae;Chi, Gyeong-Yup;Cho, In-Sik;Kim, Han-Young;Lee, Young-Cheol
    • The Korea Journal of Herbology
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    • v.27 no.4
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    • pp.9-16
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    • 2012
  • Objective : Canavalia gladiata DC semen (CGS) have been used to improve hematopoietic activity. In the current study, we investigated whether CGS regulate hemato-potentiating function using hematopoietic stem cells (HSCs) as a testing system. Methods : HSCs isolated from femur in mice with leukopenia and thrombocytopenia induced induced by CTX. Then, Real-time PCR was performed to measure the mRNA expression and hematopoietic related gene (EPO, IL-3, SCF, c-kit, GM-CSF), the phoaphorylation of GATA-1 and STAT-5a/b were observed by ELISA method, and the number of granulocyte erythrocyte monocyte macrophage colony-forming units (CFU-GEMM) and erythroid burst forming units (BFU-E), semisolid clonogenic assay was performed. Result : When HSCs were treated with CGS, the expression of hematopoietic related genes (EPO, IL-3, SCF, c-kit, and GM-CSF) were significantly increased at the levels of mRNA as well as production in HSCs. Additionally, CGS enhanced phosphorylation of STAT-1 and signal transducer and activator of transcription-5a/b (STAT-5a/b) in HSCs. Furthermore, CGS significantly enhanced the growth rate of granulocyte erythrocyte monocyte macrophage colony-forming units (CFU-GEMM) and erythroid burst forming units (BFU-E) in vitro. Conclusion : These result suggest that CGS has hematopoietic enhancement via hematopoietic cytokine-mediated GATA-1/STAT-5a/b pathway.

A Study on Variation of Colony Forming Units of Heterotrophic Bacteria by Input Ratios of Bulking Materials in Aerobic Composting of Food Wastes (음식물류폐기물의 호기성 퇴비화에 있어서 팽화재 투입비에 따른 타가영양세균의 균락형성단위의 변화에 관한 연구)

  • Park, Seok-Hwan
    • Journal of Environmental Health Sciences
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    • v.32 no.4 s.91
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    • pp.353-358
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    • 2006
  • This study was performed to evaluate the effects of input ratios of bulking material in aerobic composting of food wastes on variation of colony forming units of heterotrophic bacteria. Wood chips were used as a bulking material. Volume ratios of food wastes to wood chips in reactor of Control, WC-1 and WC-2 were 10/0, 10/5 and 10/10, respectively. Reactors were operated for 24 days with 1hour stirring by 1rpm and 2 hours of the forced aeration per day. WC-2 reached high temperature range faster than WC-1, and the maximum temperature of WC-2 was higher than that of WC-1. This means that the reaction velocity of composting of WC-2 was faster than that of WC-1. Judging from the profile of pH changes, composting of WC-1 proceeded slowly and continuously. Composting of WC-2 proceeded rapidly in the former half reaction period, and kept steady state of high pH in the latter half reaction period. Namely, composting of WC-2 was nearly completed in the former half reaction period. In the case of WC-1 and WC-2. the maximum temperature was followed by the rapid pH increase in 2-3 days, and this was followed by the maximum Colony Forming Units(CFU) in 3 days. But, these three items of WC-2 always appeared faster and higher than those of WC-1.

Regular Exercise Training Increases the Number of Endothelial Progenitor Cells and Decreases Homocysteine Levels in Healthy Peripheral Blood

  • Choi, Jeong Kyu;Moon, Ki Myung;Jung, Seok Yun;Kim, Ji Yong;Choi, Sung Hyun;Kim, Da Yeon;Kang, Songhwa;Chu, Chong Woo;Kwon, Sang Mo
    • The Korean Journal of Physiology and Pharmacology
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    • v.18 no.2
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    • pp.163-168
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    • 2014
  • Endothelial progenitor cells (EPCs) are known to play an important role in the repair of damaged blood vessels. We used an endothelial progenitor cell colony-forming assay (EPC-CFA) to determine whether EPC numbers could be increased in healthy individuals through regular exercise training. The number of functional EPCs obtained from human peripheral blood-derived AC133 stem cells was measured after a 28-day regular exercise training program. The number of total endothelial progenitor cell colony-forming units (EPC-CFU) was significantly increased compared to that in the control group (p=0.02, n=5). In addition, we observed a significant decrease in homocysteine levels followed by an increase in the number of EPC-CFUs (p=0.04, n=5), indicating that the 28-day regular exercise training could increase the number of EPC colonies and decrease homocysteine levels. Moreover, an inverse correlation was observed between small-endothelial progenitor cell colony-forming units (small-EPC-CFUs) and plasma homocysteine levels in healthy men (r=-0.8125, p=0.047). We found that regular exercise training could increase the number of EPC-CFUs and decrease homocysteine levels, thus decreasing the cardiovascular disease risk in men.

Photodynamic Inactivation of Moraxella catarrhalis (Moraxella catarrhalis의 광역학적 비활성화)

  • Hong, Seong-No;Kwon, Pil-Seung;Kim, Dae-Sik
    • Korean Journal of Clinical Laboratory Science
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    • v.39 no.1
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    • pp.19-24
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    • 2007
  • The aim of this study was to evaluate the bacterial effects of Moraxella catarrhalis in otitis media with effusion (OME) by photodynamic therapy (PDT). Bacterial suspensions (10000 CFU/mL) were prepared. The colony forming units (CFU) of Moraxella catarrhalis have been measured after an application of photogem plus 632 nm diode laser irradiation. One ml of the bacterial suspensions have been incubated in the dark for 3h with various concentrations of photogem ($0.625{\sim}5.0_{\mu}g/mL$) and then irradiated with 632 nm diode laser ($15J/cm^2$). After, the PDT Moraxella catarrhalis suspensions ($50{\mu}L$) were inoculated on chocolate agar plate and cultured in the dark at $37^{\circ}C$, 5% $CO_2$ condition for 18h. The colony forming units off the bacteria were measured. Also transmission electron microscopy (TEM) was employed to evaluate the effect of otitis media pathogens by PDT. The nucleus of Moraxella catarrhalis was stained using green fluorescent nucleic acid dye thiazole orange and the fluorescence intensity of the nucleus was measured by flow cytometry. The PDT was effective in killing Moraxella catarrhalis at the photogem dose of $5.0_{\mu}g/mL$, respectively, As assessed by flow cytometry analysis the fluorescence intensity of the nucleus got lower after PDT. TEM result appeared to able to cause damage to the bacterial membranes. On the basis of these findings, bacterial photodynamic therapy with photogem can be considered to be a promising new therapeutic approach for OME.

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Studies on the regulation of Hematopoietic enhancement of Brassica campestris var narinosa., Canavalia gladiata DC semen and their combinational prescription via Jak2/STAT5/GATA1 Pathway in Sca-1+ hematopoietic stem cells (Sca-1+골수조혈세포에서 JAK2/STAT5/GATA-1 신호전달 경로를 통한 다채, 도두 그리고 두 조합물에 의한 조혈증진 조절에 관한 연구)

  • Kim, Kunhoae;Kim, Seung-Hyung;Cho, In-Sik;Kim, Han-Young;Kim, Dong-Seon;Lee, Young-Cheol
    • The Korea Journal of Herbology
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    • v.28 no.4
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    • pp.7-16
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    • 2013
  • Objectives : Brassica campestris var narinosa (BCN), Canavalia gladiata DC semen (CGD) and their combinational prescription (BCN+CGD) have been use to demonstrate to regulate hematopoiesis. In the current study, we investigated whether Brassica campestris var narinosa, Canavalia gladiata DC semen and their combinational prescription is related to hemato-potentiating function using Sca-$1^+$ hematopoietic stem cells (Sca-$1^+HSCs$) as a testing system. Methods : Sca-$1^+HSCs$ isolated from femur in C57bl/6 mice with leukopenia and thrombocytopenia induced by cyclophosphamide (CTX). Then, Real-time PCR was performed to measure the mRNA expression, ELISA and haematopoiesis-related gene (EPO, TPO, IL-3, SCF, c-kit, GM-CSF), the phosphorylation of JAK2, GATA-1 and STAT-5a/b were observed by western blot, and the numbers of $CD117^+/Sca-1^+$ cell and the number of granulocyte erythrocyte monocyte macrophage colony-forming units (CFU-GEMM) and erythroid burst forming units (BFU-E), semisolid clonogenic assay was performed. Result : When Sca-$1^+HSCs$ were treated with Brassica campestris var narinosa, Canavalia gladiata DC semen and their combinational prescription with rIL-3/rSCF, the expression of haematopoiesis-related (EPO, TPO, IL-3, SCF, c-kit, and GM-CSF) were significantly increased at the levels of mRNA as well as production in Sca-$1^+HSCs$. Additionally, CGS enhanced phosphorylation of JAK2, GATA-1, and signal transducer and activator of transcription-5a/b (STAT-5a/b) in Sca-$1^+HSCs$. Furthermore, their combinational prescription (BCN+CGD) significantly enhanced the growth rate of granulocyte erythrocyte monocyte macrophage colony-forming units (CFU-GEMM) and erythroid burst forming units (BFU-E) in vitro. Conclusion : These result suggest that Brassica campestris var narinosa (BCN) and Canavalia gladiata DC have hematopoietic enhancement via hematopoietic cytokine-mediated JAK2/GATA-1/STAT-5a/b pathway, and their combinational prescription (BCN+CGD) has superior hematopoietic enhancement to those of individual extracts.

Effect of Potassium Iodide on Erythrosine-Mediated Photodynamic Therapy on Streptococcus Mutans Biofilms (Streptococcus mutans 바이오필름에 대한 에리스로신 매개 광역동 치료 시 potassium iodide의 효과)

  • Yongsoon, Kim;Howon, Park;Juhyun, Lee;Haeni, Kim;Siyoung, Lee
    • Journal of the korean academy of Pediatric Dentistry
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    • v.49 no.3
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    • pp.321-328
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    • 2022
  • The aim of this in-vitro study is to evaluate the effect of potassium iodide (KI) on erythrosine-mediated photodynamic therapy (PDT) against Streptococcus mutans biofilms. S. mutans ATCC 25175 was cultured to form a biofilm on a hydroxyapatite disk. After diluting erythrosine to 20 μM and KI to 10, 50, and 100 mM, respectively, PDT was performed. The number of surviving bacteria was calculated as colony forming units (CFU)/mL and the statistical significance of the difference between groups was confirmed by Bonferroni post-hoc analysis. Cell viability was visually evaluated using confocal laser scanning microscopy (CLSM). As a result of the experiment, a significant decrease (p < 0.05) in CFU was observed in the experimental groups in which PDT was performed after applying KI regardless of the concentration of KI. In addition, a significant reduction (p < 0.05) in CFU was observed in the experimental group to which 100 mM KI was applied compared to 10 mM KI. The same results were confirmed when observing CLSM. KI significantly improved the efficacy of erythrosine-mediated PDT on S. mutans biofilms at all concentrations. This may compensate for the low sensitivity of PDT to biofilm-state bacteria strains, but it is necessary to establish an optimal clinical protocol through further research.

Comparison of Optimal Storage Temperature and Collection Reagents for Living Bacterial Cells in Swab Samples (면봉시료에서 세균의 보존을 위한 최적 보관 온도와 채취 시약의 비교)

  • Lee, Yeong Ju;You, Hee Sang;Lee, Song Hee;Lee, So Lip;Lee, Han;Sung, Ho Joong;Kang, Hee Gyoo;Hyun, Sung Hee
    • Korean Journal of Clinical Laboratory Science
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    • v.53 no.4
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    • pp.326-332
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    • 2021
  • Swabs are useful and common sampling tools in various research fields, such as medicine, ecology, biotechnology, forensic medicine, and pollutant monitoring systems. Collection reagents are one of the essential components in sampling. It is important to develop a sample collection kit and designate an appropriate storage temperature because samples need to be stored for a long time. The purpose of this study was to identify the effects of three collection reagents and three storage temperatures on the recovery of living bacteria without media. We selected Escherichia coli and Staphylococcus aureus as representative environmental bacteria. Distilled water (DW), phosphate buffered saline (PBS), and Tris-EDTA (TE) buffer were used as collection reagents and stored at 22℃, 4℃, and -70℃ after sampling. The results of using each collection reagent and storage temperature on the bacteria were compared using relative light units (RLU) and the number of colony forming units (CFU). When using -70℃ storage temperature and the TE buffer, the number of living bacteria and the RLU values remained constant. It is therefore recommended that the sample be stored at -70℃ immediately after collection and a TE buffer solution be used as the collection reagent.