• Title/Summary/Keyword: viable cells

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A Study on the Induction of Sister-Chromatid Exchanges in Chinese Hamster Ovary $K_1$ Cells by Exposure to Cadmium (Chinese Hamster Ovary $K_1$ 세포의 자매염색분체교환에 미치는 카드뮴의 영향에 관한 연구)

  • Hwang, In-Kyung;Kim, Don-Kyoun
    • Journal of Preventive Medicine and Public Health
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    • v.23 no.2 s.30
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    • pp.178-184
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    • 1990
  • In order to examine the mutagenicity of cadmium dichloride the author studied the induction of sister chromatid exchanges in chinese hamster ovary $K_1$ cells which treated with cadmium dichloride at various concentrations. The results obtained were as follows : 1. In cells treated with $10^{-4}M$ cadmium dichloride, a small number of cells were viable but no mitosis was bound. 2. The frequencies of sister chromatid exchanges in cells treated with $10^{-5}M\;and\;10^{-6}M$ cadmium dichloride as $10.7{\pm}1.9\;and\;8.3{\pm}2.1$, respectively, were significantly increased for control ($6.0{\pm}2.3$). (P<0.05). 3. There were dose-dependent relationship between the concentration of cadmium dichloride and frequency of sister chromatid exchanges in cells treated with cadmium dichloride at concentration ranging from $10^{-5}\;to\;10^{-7}M$.

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Mechanisms of Glucose Uptake in Cancer Tissue (악성종양의 포도당 섭취 기전)

  • Chung, June-Key
    • The Korean Journal of Nuclear Medicine
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    • v.33 no.1
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    • pp.1-10
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    • 1999
  • Cancer cells are known to show increased rates of glycolysis metabolism. Based on this, PET studies using F-18-fluorodeoxyglucose have been used for the detection of primary and metastatic tumors. To account for this increased glucose uptake, a variety of mechanisms has been proposed. Glucose influx across the cell membrane is mediated by a family of structurally related proteins known as glucose transporters (Gluts). Among 6 isoforms of Gluts, Glut-1 and/or Glut-3 have been reported to show increased expression in various tumors. Increased level of Glut mRNA transcription is supposed to be the basic mechanism of Glut overexpression at the protein level. Some oncogens such as src or ras intensely stimulate Glut-1 by means of increased Glut-1 mRNA levels. Hexokinase activity is another important factor in glucose uptake in cancer cells. Especially hexokinase type II is considered to be involved in glycolysis of cancer cells. Much of the hexokinase of tumor cells is bound to outer membrane of mitochondria by the porin, a hexokinase receptor. Through this interaction, hexokinase may gain preferred access to ATP synthesized via oxidative phosphorylation in the inner mitochondria compartment. Other biologic factors such as tumor blood flow, blood volume, hypoxia, and infiltrating cells in tumor tissue are involved. Relative hypoxia may activate the anaerobic glycotytic pathway. Surrounding macrophages and newly formed granulation tissue in tumor showed greater glucose uptake than did viable cancer cells. To expand the application of FDG PET in oncology, it is important for nuclear medicine physicians to understand the related mechanisms of glucose uptake in cancer tissue.

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USP44 Promotes the Tumorigenesis of Prostate Cancer Cells through EZH2 Protein Stabilization

  • Park, Jae Min;Lee, Jae Eun;Park, Chan Mi;Kim, Jung Hwa
    • Molecules and Cells
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    • v.42 no.1
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    • pp.17-27
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    • 2019
  • Ubiquitin-specific protease 44 (USP44) has been implicated in tumor progression and metastasis across various tumors. However, the function of USP44 in prostate cancers and regulatory mechanism of histone-modifying enzymes by USP44 in tumors is not well-understood. Here, we found that enhancer of zeste homolog 2 (EZH2), a histone H3 lysine 27 methyltransferase, is regulated by USP44. We showed that EZH2 is a novel target of USP44 and that the protein stability of EZH2 is upregulated by USP44-mediated deubiquitination. In USP44 knockdown prostate cancer cells, the EZH2 protein level and its gene silencing activity were decreased. Furthermore, USP44 knockdown inhibited the tumorigenic characteristics and cancer stem cell-like behaviors of prostate cancer cells. Inhibition of tumorigenesis caused by USP44 knockdown was recovered by ectopic introduction of EZH2. Additionally, USP44 regulates the protein stability of oncogenic EZH2 mutants. Taken together, our results suggest that USP44 promotes the tumorigenesis of prostate cancer cells partly by stabilizing EZH2 and that USP44 is a viable therapeutic target for treating EZH2-dependent cancers.

Development of a Functional Mixed-Starter Culture for Kefir Fermentation (Kefir 배양용 기능성 복합 Starter 개발)

  • Lee, Bomee;Yi, Hae-Chang;Moon, Yong-II;Oh, Sejong
    • Journal of Dairy Science and Biotechnology
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    • v.36 no.3
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    • pp.178-185
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    • 2018
  • Kefir, which originates in the Caucasian mountains, is a cultured milk beverage produced by a combination of acidic and alcoholic fermentation. Kefir products are commonly used as food vehicles to deliver health-promoting materials including kefran and lactic acid bacteria to consumers. The aim of this study was to develop a freeze-dried starter culture without yeast and assess the suitability of kefir-like dairy products for the growth of lactic acid bacteria and the acidification of milk. Pasteurized whole milk (SNF 8.5%) stored at $25^{\circ}C$ was aseptically inoculated with starter cultures (0.002% w/v); it was kept at $25^{\circ}C$ until the pH attained a value of 4.6. Ten grams of the kefir-like product sample was diluted with 90 mL of 0.15% peptone water diluent in a milk dilution bottle, followed by uniform mixing for 1 min. Viable cells of Lactobacillus species were enumerated on modified-MRS agar (pH 5.2), with incubation at $37^{\circ}C$ for 48 h. Viable cells of Lactococcus species were enumerated on M17-lactose agar, with incubation at $32^{\circ}C$ for 48 h. The pH attained a value of 4.6 after fermentation for 9 h 30 min (Starter 1), 9 h 45 min (Starter 2), and 12 h (Starter 3). The viable cell count of Lactobacillus sp. and Lactococcus sp. was initially $10^5{\sim}10^6CFU/g$; it increased significantly to $10^9CFU/g$ after 12 h of incubation. During the storage of the kefir-like products at $4^{\circ}C$ for 1 4 days, the total viable cell numbers were unchanged, but the pH decreased slightly. The consistency of the kefir products increased gradually during the storage. The organoleptic properties of the kefir products fermented using the new starter culture are more desirable than those of commercial kefir. These results suggest that the newly developed starter culture without yeast could be suitable for kefir fermentation.

Characterization of exopolysaccharide-producing lactic acid bacteria from Taiwanese ropy fermented milk and their application in low-fat fermented milk

  • Ng, Ker-Sin;Chang, Yu-Chun;Chen, Yen-Po;Lo, Ya-Hsuan;Wang, Sheng-Yao;Chen, Ming-Ju
    • Animal Bioscience
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    • v.35 no.2
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    • pp.281-289
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    • 2022
  • Objective: The aim of this study was to characterize the exopolysaccharides (EPS)-producing lactic acid bacteria from Taiwanese ropy fermented milk (TRFM) for developing a clean label low-fat fermented milk. Methods: Potential isolates from TRFM were selected based on the Gram staining test and observation of turbid suspension in the culture broth. Random amplified polymorphic DNA-polymerase chain reaction, 16S rRNA gene sequencing, and API CHL 50 test were used for strain identification. After evaluation of EPS concentration, target strains were introduced to low-fat milk fermentation for 24 h. Fermentation characters were checked: pH value, acidity, viable count, syneresis, and viscosity. Sensory evaluation of fermented products was carried out by 30 volunteers, while the storage test was performed for 21 days at 4℃. Results: Two EPS-producing strains (APL15 and APL16) were isolated from TRFM and identified as Lactococcus (Lc.) lactis subsp. cremoris. Their EPS concentrations in glucose and lactose media were higher than other published strains of Lc. lactis subsp. cremoris. Low-fat fermented milk separately prepared with APL15 and APL16 reached pH 4.3 and acidity 0.8% with a viable count of 9 log colony-forming units/mL. The physical properties of both products were superior to the control yogurt, showing significant improvements in syneresis and viscosity (p<0.05). Our low-fat products had appropriate sensory scores in appearance and texture according to sensory evaluation. Although decreasing viable cells of strains during the 21-day storage test, low-fat fermented milk made by APL15 exhibited stable physicochemical properties, including pH value, acidity, syneresis and sufficient viable cells throughout the storage period. Conclusion: This study demonstrated that Lc. lactis subsp. cremoris APL15 isolated from TRFM had good fermentation abilities to produce low-fat fermented milk. These data indicate that EPS-producing lactic acid bacteria have great potential to act as natural food stabilizers for low-fat fermented milk.

Growth, Clonability, and Radiation Resistance of Esophageal Carcinoma-derived Stem-like Cells

  • Li, Jian-Cheng;Liu, Di;Yang, Yan;Wang, Xiao-Ying;Pan, Ding-Long;Qiu, Zi-Dan;Su, Ying;Pan, Jian-Ji
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.8
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    • pp.4891-4896
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    • 2013
  • Objective: To separate/enrich tumor stem-like cells from the human esophageal carcinoma cell line OE-19 by using serum-free suspension culture and to identify their biological characteristics and radiation resistance. Methods: OE-19 cells were cultivated using adherent and suspension culture methods. The tumor stem-like phenotype of CD44 expression was detected using flow cytometry. We examined growth characteristics, cloning capacity in soft agar, and radiation resistance of 2 groups of cells. Results: Suspended cells in serum-free medium formed spheres that were enriched for CD44 expression. CD44 was expressed in 62.5% of suspended cells, but only in 11.7% of adherent cells. The suspended cells had greater capacity for proliferation and colony formation in soft agar than the adherent cells. When the suspended and adherent cells were irradiated at 5 Gy, 10 Gy, or 15 Gy, the proportion of CD44+ suspended cells strongly and weakly positive for CD44 was 77.8%, 66.5%, 57.5%; and 21.7%, 31.6%, 41.4%, respectively. In contrast, the proportion of CD44+ adherent cells strongly positive for CD44 was 18.9%, 14.%, and 9.95%, respectively. When the irradiation dose was increased to 30 Gy, the survival of the suspended and adherent cells was significantly reduced, and viable CD44+ cells were not detected. Conclusion: Suspended cell spheres generated from OE-19 esophageal carcinoma cells in serum-free stem medium are enriched in tumor stem-like cells. CD44 may be a marker for these cells.

Ultraviolet Radiation-Induced Apoptosis is Inversely Correlated with the Expression Level of Poly(ADP-ribose) Polymerase

  • Oh, Kyu Seon;Lee, Dong Wook;Chang, Jeong Hyun;Moon, Yong Suk;Um, Kyung ll
    • Animal cells and systems
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    • v.5 no.1
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    • pp.77-83
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    • 2001
  • The present study was conducted to elucidate whether the expression level of poly(ADP-ribose) polymerase (PARP) is related to the ultraviolet radiation (UV)-induced apoptosis. After treatment of the mammalian cell lines HeLa S3 and Chinese hamster ovary (CHO) with 50 J/m2 UV, induction of apoptosis was determined by several means during 24 h post-incubation. Incidence of apoptosis was much lower in CHO than HeLa S3 cells based on the percentage of apoptotic cells in terms of morphological changes in nucleus or direct counting of viable cells and qualitative or quantitative DNA fragmentation. Interestingly, when the expression level of PARP was measured by western blotting, the amounts of PARP that was retained at each time point inversely correlated with the incidences of apoptosis in these cells. Concomitant with generation of the 85 kDa fragment, 116 kDa PARP disappeared in HeLa S3 within 6 h after UV treatment, whereas a fair amounts of 116 kDa band was still retained in CHO cells at 36 h post-incubation. This inverse relationship was also observed in the adaptive response system, in which cells weve treated with a high dose of UV after pretreatment with a low dose. As expected, typical adaptive responses appeared in CHO cells but not in HeLa cells, showing greater cell viability and lesser DNA fragmentation. During the adaptive response in CHO cells, PARP was expressed at much higher level compared to the single, high dose-treated cells. Interestingly, even though PARP was induced at 6 h post-incubation In both cell types, its expression was more prominent in CHO cells. Thus, our data indicate that the retained level of intact PARP against UV damage inversely correlates with incidence of apoptosis in mammalian cells, and also suggest that a machinery to protect the PARP degradation against UV damage exists in CHO but not in HeLa S3 cells.

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Freeze Drying of Lactic Acid Bacteria Fermented Food Prepared from Egg White Powder and Casein Supplemented with Growth Stimulating Agent (생육촉진물질이 첨가된 난백분말과 카제인으로 만든 젖산균 발효식품의 동결건조)

  • Ko, Young-Tae;Lee, Eun-Ju
    • Korean Journal of Food Science and Technology
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    • v.31 no.5
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    • pp.1337-1344
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    • 1999
  • Lactic acid bacteria fermented foods were prepared from egg white powder, casein and growth stimulating agent. pH change and growth of Lactbacillus acidophilus(KCTC 2182) during freeze drying were studied. The effects of freeze drying on sensory evaluation. hardness and volatile aroma compounds in freeze dried sample or reconstituted sample were also studied. Freezing and freeze drying did not affect pH of fermented samples. Number of viable cells in original fermented samples was markedly reduced during freezing or freeze drying. When number of viable cells in original fermented sampler was considered at 100%. survival ratio of viable cells after freezing was $72.0{\sim}82.4%$ and that after freeze dying $10.0{\sim}20.4%$. When sensory properties of original fermented samples were compared with those of freeze dried/reconstituted samples, sensory properties of original samples were generally better than those of freeze dried/reconstitute samples. However, the reconstitution property and the acceptability of freeze dried samples were good. Volatile aroma compounds in original fermented samples were reduced during freeze drying. The reduction degree of volatile aroma compounds varied with sample.

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Freeze Drying of Fermented Milk Prepared from Milk and Fruit Juices (유유와 과즙으로 만든 발효유의 동결건조)

  • Ko, Young-Tae;Oh, Mi-Hwa
    • Korean Journal of Food Science and Technology
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    • v.30 no.6
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    • pp.1448-1455
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    • 1998
  • Fermented milk was prepared from milk or mixture of milk and apple juice/grape juice, and it was freeze dried. pH change and growth of Lactobacillus acidophilus (KCTC 2182) during freeze drying were studied. The effects of freeze drying on sensory evaluation and volatile aroma compounds in freeze dried sample or reconstituted sample were also studied. Freezing and freeze drying did not affect pH of fermented milk. Number of viable cells of L. acidophilus was markedly reduced during freezing or freeze drying. When number of viable cells in original fermented milk was considered as 100%, survival ratio of viable cells after freezing was $64.5{\sim}85.2%$ and that after freeze drying was $10.0{\sim}21.1%$. When sensory properties of original fermented milk prepared from juice-milk (ratio 15:35) were compared with those of freeze dried/reconstituted sample, sensory properties of original sample were better than those of freeze dried/reconstituted sample. Ethanol, diacetyl, butanol and acetoin were detected in all of original samples and freeze dried/reconstituted samples while acetone was detected in samples containing high amount of grape juice. Volatile aroma compounds in original fermented milk were reduced during freeze drying. L. acidophilus produced ethanol, diacetyl and acetoin during fermentation.

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Physicochemical properties of granular formulation using Paenibacillus sp. AC-1 as a microbial fungicide (미생물 살균제 Paenibacillus sp. AC-1 입제의 물리화학적 특성)

  • Oh, Kyeong-Seok;Lee, Young-Kee;Lee, Jae-Kook;Kim, Jin-Hwa
    • The Korean Journal of Pesticide Science
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    • v.9 no.3
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    • pp.262-267
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    • 2005
  • In order to commercialize Paenibacillus sp. AC-1 and to minimize its harmful side effects, four granular formulations were prepared using AC-1 powder, adjuvant, and carrier and then their physicochemical properties of the formulations were investigated. Out of the carriers tested, the best one was talc for the formulation. Viable cells was stabilized during the formulating process. Viable cells in the granules formulated with Paenibacillus sp. AC-1 powder were stabilized at storage temperature range ($4{\sim}50^{\circ}C$) after 12 weeks. The release rate of viable cells from granules into water under a static condition were eluted over 90% in 7 hours and breakdown rates of particle were 100% in 1 day. Among the tested formulations, granular formulation comprising of 20% of Paenibacillus sp. AC-1 powder, 7% of polycarboxylate as surface active agent, 1% of sodium polyacrylate as adjuvant, the rest as carrier showed to be best.