• Title/Summary/Keyword: Cell Counting

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The Transfection of Caldesmon DNA into Primary Cultured Rat Aortic Vascular Smooth Muscle

  • Choi, Woong;Ahn, Hee-Yul
    • The Korean Journal of Physiology and Pharmacology
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    • v.3 no.6
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    • pp.597-603
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    • 1999
  • Caldesmon (CaD), one of microfilament-associated proteins, plays a key role in microfilament assembly in mitosis. We have investigated the effects of overexpression of the high molecular weight isoform of CaD (h-CaD) on the physiology of vascular smooth muscle cells (VSMCs). Rat aortic VSMCs were stably transfected with plasmids carrying a full length human h-CaD cDNA under control of cytomegalovirus promoter. The majority of the overexpressed h-CaD appears to be localized predominantly on cytoskeleton structures as determined by detergent lysis. The overexpression of h-CaD, however, does not decrease the level of endogenous low molecular weight isoform of CaD. h-CaD overexpressing VSMCs (h-CaD/VSMCs) show a decreased growth rate than that of vector-only transfected cells when determined by $[^3H]thymidine$ uptake and cell counting after fetal bovine serum (FBS) stimulation. h-CaD/VSMCs were smaller than vector-transfected cells by 18% in cell diameter. These data suggest that overexpression of h-CaD can inhibit the poliferation and the cell volume of VSMCs stimulated by growth factors and that the gene therapy with h-CaD may be helpful to prevent the conditions associated with hypertrophy and/or hyperplasia of VSMCs after arterial injuries.

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An Integrated MIN Circuit Design of DTW PE for Speech Recognition (음성인식용 DTW PE의 IC화를 위한 MIN회로의 설계)

  • 정광재;문홍진;최규훈;김종교
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.15 no.8
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    • pp.639-647
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    • 1990
  • Dynamic time warp(DTW) needs for interative calculations and the design of PE cell suitable for the operations is very important. Accordingly, this paper aims at the real time recognition design which enables large dictionary hardware realization using DTW algorithm. The DTW PE cell is seperated into three large blocks. "MIN" is the one block for counting accumulated minimum distance, "ADD" block calculates these minimum distances, and "ABS" seeks for the absolute values to the total sum of local distances. We have accomplisehd circuit design and verification for the MIN blocks, and performed MIN layout and DRC(design rule check) using 3um CMOS N-Well rule base.ing 3um CMOS N-Well rule base.

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Comparative analysis of yeast cell viability at exponential and stationary growth phases

  • An, Yejin;Jo, Nayoon;Kim, Hyeji;Nam, Dahye;Son, Woorim;Park, Jinkyu
    • Analytical Science and Technology
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    • v.35 no.4
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    • pp.181-188
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    • 2022
  • This paper describes a comparative analysis of yeast cell viability at exponential and stationary growth phases using multiple conventional techniques and statistical tools. Overall, cellular responses to various viability assays were asynchronous. Results of optical density measurement and direct cell counting were asynchronous both at exponential and stationary phases. Proliferative capacity measurement using SP-SDS indicated that cells at the end of the stationary phase were proliferative as much as exponentially growing cells. Metabolic activity assays using two different dyes concluded that the inside of cells at stationary phase is slightly less reducing compared to that of exponentially growing cells, implying that the metabolic activity imperceptibly declined as cells were aged. These results will be helpful to understand the details of yeast cell viability at exponential and stationary growth phases.

Protective Effect of Bcl-2 in NS0 Myeloma Cell Culture is Greater in More Stressful Environments

  • Tey, B.T.;Al-Rubeai, M.
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.6
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    • pp.564-570
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    • 2005
  • In the present study, the protective effects of Bcl-2 over-expression in a suspension culture (without any adaptation) and spent medium (low nutrient and high toxic metabolite conditions) were investigated. In the suspension culture without prior adaptation, the viability of the control cell line fall to 0% by day 7, whereas the Bcl-2 cell line had a viability of 65%. The difference in the viability and viable cell density between the Bcl-2 and control cell lines was more apparent in the suspension culture than the static culture, and became even more apparent on day 6. Fluorescence microscopic counting revealed that the major mechanism of cell death in the control cell line in both the static and suspension cultures was apoptosis. For the Bcl-2 cell lines, necrosis was the major mode of cell death in the static culture, but apoptosis became equally important in the suspension culture. When the NS0 6A1 cell line was cultured in spent medium taken from a 14 day batch culture, the control cell line almost completely lost its viability by day 5, whereas, the Bcl-2 still had a viability of 73%. The viable cell density and viability of the Bcl-2 cell line cultivated in fresh medium were 2.2 and 2.7 fold higher, respectively, than those of the control cultures. However, the viable cell density and viability of the Bcl-2 cultivated in the spent medium were 8.7 and 7.8 fold higher, respectively, than those of the control cultures. Most of the dead cells in the control cell line were apoptotic; whereas, the major cell death mechanisms in the Bcl-2 cell line were necrotic.

The Glucosinolate and Sulforaphane Contents of Land Race Radish and Wild Race Radish Extracts and Their Inhibititory Effects on Cancer Cell Lines (재래종 무와 갯무 추출물의 암세포주 증식 저해 활성 및 Glucosinolate와 Sulforaphane의 함량)

  • Choi, Sun-Ju;Choi, A-Reum;Cho, Eun-Hye;Kim, So-Young;Lee, Gun-Soon;Lee, Soo-Seong;Chae, Hee-Jeong
    • Journal of the East Asian Society of Dietary Life
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    • v.19 no.4
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    • pp.558-563
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    • 2009
  • The inhibitory effects of land race radish (LRR) and wild race radish (WRR) extracts on cancer cell lines were investigated. A and their glucosinolate and sulforaphane contents were analyzed. The anticancer activitiesy of the LRR and WRR extracts on the breast cancer cell line MCF-7 were determined by a CCK (cell counting kit) assay, in which WWR showed higher inhibition rates than LRR. The sulforaphane content of WRR was higher than that of LRR. In the lung cancer cell line, A-549, WRR showed higher inhibition rates and a higher total glucosinolate content than LRR. The glucosinolate contents of the radishes were analyzed by the Pd-quicktest method, showing that WRR contained more glucosinolate than LRR in both the trunk and root. In conclusion, these results indicate that wild race radish could be used for the quality improvement of radishes.

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Effect of Adefovir Dipivoxil on the Inhibition of Osteogenic Differentiation of Mesenchymal Stem Cells and Osteoblasts (아데포비어가 중간엽 줄기세포와 조골세포의 골형성 분화 억제에 미치는 영향)

  • Ho PARK
    • Korean Journal of Clinical Laboratory Science
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    • v.55 no.4
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    • pp.284-290
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    • 2023
  • Adefovir dipivoxil (ADV) is used for the treatment of hepatitis and acquired immunodeficiency syndrome, but long-term use can cause osteoporosis. In this study, the effect of ADV on the osteocyte maturation process was evaluated at the level of undifferentiated cells using mesenchymal stem cells (MSCs) and osteoblasts (MG63). First, MSCs and MG63 cells were treated with ADV at different concentrations, and then a Cell Counting Kit-8 analysis was performed to determine the effect on the proliferation of each cell. Additionally, crystal violet and Hoechst staining were performed for the morphological analysis of each cell and nucleus. To determine the cause of cell hypertrophy, the transforming growth factor-beta (TGF-β) expression was investigated, and alkaline phosphatase (ALP) staining and activity were measured to determine the degree of differentiation of the MSCs and MG63 cells into mature osteocytes. The results confirmed that the ADV increases the expression of TGF-β in MSCs and MG63 cells, causing cellular and nuclear hypertrophy, and can cause osteoporosis by inhibiting cell proliferation and affecting the differentiation of mature osteocytes. Therefore, it is believed that these results can be used as a basis for understanding the adverse effects of ADV at a cytological level in basic medicine and clinical research.

Exosomes Secreted by Toxoplasma gondii-Infected L6 Cells: Their Effects on Host Cell Proliferation and Cell Cycle Changes

  • Kim, Min Jae;Jung, Bong-Kwang;Cho, Jaeeun;Song, Hyemi;Pyo, Kyung-Ho;Lee, Ji Min;Kim, Min-Kyung;Chai, Jong-Yil
    • Parasites, Hosts and Diseases
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    • v.54 no.2
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    • pp.147-154
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    • 2016
  • Toxoplasma gondii infection induces alteration of the host cell cycle and cell proliferation. These changes are not only seen in directly invaded host cells but also in neighboring cells. We tried to identify whether this alteration can be mediated by exosomes secreted by T. gondii-infected host cells. L6 cells, a rat myoblast cell line, and RH strain of T. gondii were selected for this study. L6 cells were infected with or without T. gondii to isolate exosomes. The cellular growth patterns were identified by cell counting with trypan blue under confocal microscopy, and cell cycle changes were investigated by flow cytometry. L6 cells infected with T. gondii showed decreased proliferation compared to uninfected L6 cells and revealed a tendency to stay at S or G2/M cell phase. The treatment of exosomes isolated from T. gondii-infected cells showed attenuation of cell proliferation and slight enhancement of S phase in L6 cells. The cell cycle alteration was not as obvious as reduction of the cell proliferation by the exosome treatment. These changes were transient and disappeared at 48 hr after the exosome treatment. Microarray analysis and web-based tools indicated that various exosomal miRNAs were crucial for the regulation of target genes related to cell proliferation. Collectively, our study demonstrated that the exosomes originating from T. gondii could change the host cell proliferation and alter the host cell cycle.

An Experimental Study on Bacterial Adhesion onto Activated Carbon and Ceramic (활성탄 및 세라믹 재질에의 세균 부착성에 대한 연구)

  • Kwon Sung-Hyun;Cho Dae-Chul;Rhee In-Hyoung
    • Journal of Environmental Science International
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    • v.14 no.12
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    • pp.1163-1170
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    • 2005
  • The microbial adsorption characteristics of two different media for biological treatment were studied using attached diverse microbes onto activated carbon and ceramic. The results in the experiments of the characteristics of physical adhesion on two different media with addition of high and low concentrated substrate in the culture were observed that the efficient of adhesion onto F-400 activated carbon was higher over that of ceramic due to the surface area of media. The irradiation treatment by ultrasonication with 400 W power and 3 min retention time on the media without addition substrate conditions and subsequent mixing throughly the culture showed the highest efficiency of cell detachment on the media. Three different microbes, P. ovalis, A calcoaceticus, and B. subtillis were used for the study of the characteristics of microbial adhesion on the media. p ovalis showed the highest adhesion capability while B. subtillis showed the lowest capability adhesion onto media either addition of substrate in the culture. The mixed bacterial culture showed $10\%$ lower removal efficiency of DOC in the low concentrated substrate culture compared to the single pure culture. Whileas, it did not show significant difference between two cultures at high concentrated substrate. It was also observed same population density of microorganism by counting of microbes adhered to microbial media with an ultrasound treatment.

Quantitation of Antigen-Antibody Reaction Condition for Development of Fluorescence Image-based CD4 Rapid Test (형광 영상 기반 CD4 신속 검사법 개발을 위한 항원-항체 반응 조건 정량화)

  • Kim, Subin;Kim, Jung Kyung
    • Journal of the Korean Society of Visualization
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    • v.13 no.1
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    • pp.35-42
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    • 2015
  • CD4+ T-cell count determines the effectiveness for antiretroviral therapy (ART) in patients with human immunodeficiency virus (HIV). Although ART slows the progression of HIV to AIDS, rapid counting of CD4+ T lymphocytes with a drop of patient's blood sample is urgently needed to ensure timely ART treatment in rural areas. Recently point-of-care CD4 testing devices have been developed by using non-flow based imaging cytometer incorporated with a sample cartridge where CD4+ T cells are reacted with fluorescently tagged specific antibodies. Here we conducted an experimental study using a conventional fluorescence microscope-based imaging system to quantitate the interaction of CD4 antibodies with CD4+ T cells at different reaction conditions. We demonstrated that a fast and affordable point-of-care CD4 test is feasible with a far less amount of antibodies and a shorter incubation time compared with a conventional sample preparation protocol for flow cytometry. We also proposed a general method to evaluate and compare the detection limit across different CD4 counting platforms by using fluorescently labelled microbeads for intensity calibration.

Selective Media Containing Antibiotics for Counting Bifidobacteria (항생제를 이용한 Bifidobacteria의 선택배지)

  • Lee, Jeong-Jun;Shin, Myeong-Su;Na, Seog-Hwan;Bae, Hyoung-Suk;Baek, Young-Jin
    • Microbiology and Biotechnology Letters
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    • v.22 no.3
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    • pp.309-315
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    • 1994
  • Selective agar media were constructed for the counting of bifidobacteria in dairy produ- cts containing bifidobacteria, lactobacilli and streptococci. This media containing antibiotics inhibited the growth of lactobacilli and streptococci at less than 10$^{5}$ cfu/ml, but had no influence on the recovery of bifidobacteria. In order to inhibit the growth of 10$^{5}$ cfu/ml of lactobacilli and streptoco- cci, the addition of 1.0~2.0 $\mu$g/ml of tetracycline in BL agar medium was needed. When 25 $\mu$g/ml of neomycin and paromomycin were mixed with 1.0 $\mu$g/ml of tetracycline in BL agar medium, it was able to inhibit the growth of 10$^{6}$ cfu/ml of lactobacilli and streptococci but had a little negative effect on the recovery of colonies of bifidobacteria. The results revealed that the BL agar medium containing 1.0 $\mu$g/ml of tetracycline was suitable to count the cell number of bifidobacteria selecti- vely in the presence of a 1 to 10$^{5}$-fold excess of L. casei, L. acidophilus and S. salivarius subsp.thermophilus.

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