• Title/Summary/Keyword: freeze-dried cells

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Imaging of Tumor Proliferation Using Iodine-131-Iodomethyluridine (Iodine-131-Iodomethyluridine을 이용한 종양세포증식의 영상화에 관한 실험적 연구)

  • Min Kyung-Yoon;Kim, Chang-Guhn;Kim, Hyun-Jeong;Lim, Hyung-Guhn;Rho, Ji-Young;Juhng Seon-Kwan;Won Jong-Jin;Yang, David J.
    • The Korean Journal of Nuclear Medicine
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    • v.30 no.3
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    • pp.344-350
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    • 1996
  • Purpose : Noninvasive imaging of tumor cell proliferation could be helpful in the evaluation of tumor growth potential and could provide an early assessment of treatment response. Radiolabeled thymidine, uridine and adenosine have been used to evaluate tumor cell proliferation. These nucleoside analogs are incorporated into DNA during proliferation. Iodine-131-Iodomethyluridine, an analog of Iodine-131-Iododeoxyuridine, is also involved in DNA/RNA synthesis. The purpose of this study was to develop Iodine-131-Iodomethylurdine and image tumor proliferation using Iodine-131-Iodomethyluridine. Materials and Methods : Radiosynthesis of Iodine-131-5-Iodo-2'-O-methyluridine (Iodine-131-Iodomethyluridine) was prepared from 10 mg of 2'-O-methyluridine(Sigma chemical Co., St. Louis, Missouri) and 2.1 mCi(SP. 10Ci/mg) of Iodine-131-labeled sodium iodide in $100{\mu}l$ of water using iodogen reaction. Female Fischer 344 rats were inoculated in the thigh area with breast tumor cells(13765 NF, $10^5$ cells/rat S.C.). After 14 days, the Iodine-131-Iodomethyluridine $10{\mu}Ci$ was injected to three groups of rats(3/group). The percent of injected dose per gram of tissue weight was determined at 0.5-hours, 2-hours, 4-hours, and 24-hours respectively. Tumor bearing rats after receiving Iodine-131-Iodomethyluridine($50{\mu}Ci$ IV) were euthanized at 2 hours after injection. Autoradiography was done using freeze-dried $50{\mu}m$ coronal section. After injection of Iodine-131- Iodomethyluridine ($10{\mu}Ci$/rat, IV) in three breast tumor-bearing rats, planar scintigraphy was taken at 45 minutes, 90 minutes and 24 hours. Results : Iodine-131-Iodomethyluridine was conveniently synthesized using iodogen reaction. The biodistribution showed fast blood clearance and the tumor-to-tissue uptake ratios showed that optimal imaging time was at 2 hours postinjection. Autoradiogram and planar scintigram indicated that tumor could be well visualized. Conclusion : The findings suggest that Iodine-131-Iodomethyluridine, a new radio-iodinated nucleoside, has potential use for evaluation of active regions of tumor growth.

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Insulin Sensitizing and Insulin-like Effects of Water Extracts from Kalopanax pictus NAKAI in 3T3-L1 Adipocyte (엄나무 추출물이 3T3-L1 지방세포에서 인슐린 민감성과 인슐린 유사성 작용에 미치는 영향)

  • Kim, Ho-Kyoung;Park, Sun-Min;Ko, Byoung-Seob
    • Applied Biological Chemistry
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    • v.45 no.1
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    • pp.42-46
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    • 2002
  • Effects of the water extract from Kalopanax pictus NAKAI on insulin-like action and glucose uptake in 3T3-L1 cells were investigated. The bark of K. pictus NAKAI was treated with hot water and the extract was freeze-dried. Total extract of K. pictus NAKAI was fractionated into 6 fractions with increasing gradients from 0 to 100% MeOH on Amberlite XDA-4. Treatment of 3T3-L1 fibroblasts with 1 ${\mu}g/ml$ and 10 ${\mu}g/ml$ of K. pictus NAKAI total extracts significantly increased the differentiation of the cells. When co-treated with inducers such a dexamethasone, 1-methyl-3-isobutylxanthine and insulin, the differentiation was increased at 1 ${\mu}g/ml$ of total extract, but not at 10 ${\mu}g/ml$. In 3T3-L1 adipocytes, glucose uptake was increased by 3.3 times with addition of 0.3 and 3 ${\mu}g/ml$ of Fr. 1 (0-10% MeOH) and Fr. 3 (30% MeOH) at 3 ng/ml insulin. In conclusion, K. pictus NAKAI contains such compounds that play a role of insulin-like action and insulin sensitizer.

THE EFFECT OF BIOPLANT $HTR^{(R)}$(HARD TISSUE REPLACEMENT) ON OSTEOGENESIS OF THE MANDIBULAR BONE DEFECTS IN RABBITS (하악골 결손부의 골형성에 미치는 Bioplant $HTR^{(R)}$의 영향)

  • Choi, Do-Hyun;Ryu, Sun-Youl;Kim, Ok-Joon;Huh, Won-Shil
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.23 no.1
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    • pp.87-94
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    • 2001
  • The present study was performed to investigate the effect of $HTR^{(R)}$ (Hard Tissue Replacement) on osteogenesis in the mandibular bone defects. Eight adult male white rabbits weighing 2.5 to 3.0kg were used. Four bone defects (8mm in diameter and 4mm in depth) were made at the both mandibular body. In the control group, the right mesial bone defect was filled with blood clot and spontaneously healed. In the DFDB group, the right distal bone defect was filled with xenogenic demineralized freeze-dried bone. In the $HTR^{(R)}$ group, the left mesial bone defect was filled with $HTR^{(R)}$. In the $HTR^{(R)}-membrane$ group, the left distal bone defect was filled with $HTR^{(R)}$ and covered with BioMesh membrane. The rabbits were sacrified at 2,4,6 and 9 weeks after the operation and microscopic examination was performed. Results obtained were as follows: In the control and DFDB groups, inflammatory cells and the fibrous connective tissue existed and the bone growth was slower than $HTR^{(R)}$ group by 6 week, and there was intervention of the soft tissue at 9 week. In the $HTR^{(R)}$ group, bone trabeculi extended between the $HTR^{(R)}$ particles without intervention of inflammatory cells and the connective tissue at 4 and 6 weeks. In addition, extensive osseous ingrowth into the $HTR^{(R)}$ particles was observed at 9 week. Bone formation was more active in the $HTR^{(R)}$ group than the control and DFDB groups. There was not obvious difference in the bone healing rate between the $HTR^{(R)}$ and the $HTR^{(R)}-membrane$ group. These results suggest that the $HTR^{(R)}$ promotes osteogenesis in the bone defects and the $HTR^{(R)}$ group has no difference in comparison with the $HTR^{(R)}-BioMesh^{(R)}$ membrane group in bone healing.

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Inhibitory effect of Aralia elata ethanol extract against skin damage in UVB-exposed human keratinocytes and human dermal fibroblasts (두릅순 에탄올 추출물의 인간유래 피부각질형성세포와 피부섬유아세포에서의 자외선에 의한 광노화 억제효과)

  • Yang, Jiwon;Kwak, Chungshil
    • Journal of Nutrition and Health
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    • v.49 no.6
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    • pp.429-436
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    • 2016
  • Purpose: Solar ultraviolet (UV) radiation causes inflammation and matrix metalloproteinase (MMP) overexpression and extracellular matrix depletion, leading to skin photoaging such as wrinkle formation, dryness, and sagging. Activation of MMP is influenced by various molecules such as reactive oxygen species (ROS), proinflammatory cytokines, and transient receptor potential vanilloid type (TRPV)-1, which are increased in UV-irradiated skin cells. Aralia elata (AE) ethanolic extract was reported to inhibit ROS generation caused by UVB-irradiation in keratinocytes. In this study, we investigated the photoprotective effect of AE ethanolic extract on UVB-irradiated human keratinocytes (HaCaT) and human dermal fibroblasts (HDF). Methods: AE was freeze-dried, extracted in 70% ethanol, and concentrated. Skin cells were treated with AE extract for 24 h and then exposed to UVB ($55mJ/cm^2$). After 48 h of incubation, proinflammatory cytokines, MMP-1, type-1 procollagen, and TRPV-1 levels were measured by ELISA or Western blotting. Results: Treatment with AE extract ($100{\mu}g/mL$) significantly inhibited UVB-induced IL-6, IL-8, and $PGE_2$ production in HaCaT by 25.6%, 5.3%, and 70.2%, respectively, and also inhibited elevation of MMP-1 and TRPV-1 caused by UVB irradiation by 20.0% and 41.9%, respectively (p < 0.05). In HDF, AE extract treatment significantly inhibited both elevation of MMP-1 and reduction of type-1 procollagen caused by UVB irradiation (p < 0.05). In addition, type-1 procollagen was elevated by AE extract treatment in normal HDFs (p < 0.05). Conclusion: AE 70% ethanol extract has photoprotective ability via reduction of proinflammatory mediators, TRPV-1 and MMP-1 production, and elevation of collagen synthesis. Our findings suggest that AE extract might be a good natural material to protect against UVB-induced premature skin aging.

Effects of Bojungikgi-tang on Trimellitic Anhydride-induced Contact Hypersensitivity in BALB/c Mice (보중익기탕(補中益氣湯)이 접촉성 과민반응에 미치는 영향(影響))

  • Park, Yang-Gu;Jung, Myung;Bok, Young-Ok;Lee, Jin-Ho;Kang, Suk-Hoon;Lim, Kyu-Sang;Yun, Young-Gab
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.29 no.1
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    • pp.16-32
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    • 2016
  • Objective : The water extract of Bojungikgi-tang (BTE) composed with Ginseng Radix, Astragali Radix, Angelicae gigantis Radix, Atractylodes Rhizoma alba, Aurantii nobilis Pericarpium, Glycyrrhizae Radix, Cimicifuga Rhizoma, and Bupleuri Radix and it has been traditionally used for chronic diseases or weakness after illness in oriental countries. However, little is known about the effects of aqueous extract of BTE on trimellitic anhydride (TMA)-induced contact hypersensitivity (CHS). Therefore, the aim of this study was to investigate the effects of BTE on TMA-induced CHS in BALB/c mice.Methods : In this study, the extract of BTE was prepared by extracting with distilled water at 100℃ for 2.5 h. The extract was freeze-dried following filtration through 0.45 μm filter. Mice were orally administrated with or without BTE extract of different doses (25-200 ㎎/㎏/day) for 28 days. During the challenge period, mice were externally applied with different doses of BTE extract one time per day 30 min before TMA treatment. I examined the effects of BTE on the serum levels of IgE, IgG1, IgG2a, and IgG2b, nitric oxide (NO) and prostaglandin E2 (PGE2) production in isolated peritoneal macrophages, Th2 cytokine production in isolated spleen cells, ear swelling responses, and the leukocyte infiltration induced by TMA.Results : The orally and externally administration of BTE dose-dependently reduced the serum levels of hapten-specific immunoglobulin E (IgE), IgG1 titer, and PGE2 production as well as ear swelling responses and leukocyte infiltration in TMA-induced BALB/c mice. The levels of NO and PGE2 production from peritoneal macrophages stimulated with TMA-BSA were markedly suppressed by pretreatment with BTE in a concentration dependent manner. Furthermore, the levels of Th2 cytokines (IL-4, IL-5, IL-13) production from spleen cells stimulated with ConA were markedly suppressed by BTE treatment in a concentration dependent manner.Conclusions : These results suggest that BTE treatment suppresses chronic contact hypersensitivity, and it can be assumed that the suppression of ear swelling, serum IgE, NO and PGE2 levels, leukocyte infiltration, and Th2 cytokines in an animal model. Therefore, these properties may contribute to the strong anti-CHS response effect of BTE.

Gagam-Gongjin-dan Extract Attenuates Immune Responses to Ovalbumin in Balb/c Mice (Balb/c 마우스에서 Ovalbumin 면역반응에 대한 가감공진단(加減拱辰丹) 추출물의 억제효과)

  • Kim, Hong-Jun;Hwang, Sung-Yeoun;Mok, Ji-Ye;Hwang, Byung-Soon;Jeong, Seung-Il;Jang, Seon-Il
    • The Korea Journal of Herbology
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    • v.24 no.4
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    • pp.127-135
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    • 2009
  • Objectives : Gagam-Gongjin-dan (GGD) composited with Cervi parvum Cornu, Corni Fructus, Angelica gigantis Radix, Lycii Fructus, Dioscoreae Rhizoma, Citri Pericarpium, Gastrodiae Rihzoma, Agastachis Herba, Cassiae cortkex, Scutellariae Radix, Schisandrae Fructus has been traditionally used for chronic diseases or weakness after illness in oriental countries. However, little is known about the effects of methanol extract of GGD on immune responses to ovalbumin (OVA) plus alum. Therefore, the purpose of this study was to investigate the effects of GGD on immune responses to ovalbumin plus alum in Balb/c mice. Methods : In this study, the extract of GGD was prepared by extracting with methanol for 7 days. The extract was freeze-dried following filtration through vacuum distillation system. Mice were orally administrated with or without GGD extract of different doses (50-200 mg/kg/day) for 30 days. We examined the effects of GGD extract on the serum levels of total IgE, OVA-specific IgE, IgG1, IgG2a, and CTACK/CCL27 production and CCR10 expression in lymph node cells and body weight change and foot pad swelling responses in ovalbumin treated Balb/c. Results : The oral administration of GGD dose-dependently reduced the serum levels of total IgE, OVA-specific immunoglobulin (IgE, IgG1 and IgG2b) and CTACK/CCL27 production in ovalbumin treated BALB/c mice. The levels of CCR10 expression from lymph node cells of OVA treated mice were markedly suppressed by treatment with GGD in a concentration dependent manner. Furthermore, foot pad swelling responses were also markedly suppressed by GGD. However, body weight were significantly increased dose dependently by GGD treatment. Conclusions : These results suggest that GGD treatment suppresses immune responses to ovalbumin, and these properties may contribute to allergic disease care.

Physicochemical Characteristics and Biological Activities of Rice and Amaranth Fermented by Bacillus subtilis KMKW4 (Bacillus subtilis KMKW4를 이용한 백미 및 아마란스 발효물의 이화학적 특성 및 생리활성)

  • Yang, Su-Jin;Lee, Rea-Hyun;Hong, Joo-Heon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.4
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    • pp.540-548
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    • 2015
  • This study examined the cultural characteristics and biological activities of fermented rice and amaranth with Bacillus subtilis KMKW4. These samples were made with various amounts of rice and amaranth [100:0 (R100), 90:10 (R90), 80:20 (R80), and 70:30 (R70)]. B. subtilis KMKW4 was used as starter for the fermentation, and its cultures at the lated logarithmic growth were inoculated for final concentration of 2% (v/v). Number of viable cells of fermented R80 (7.67 log CFU/mL) was greater than those of R100, R90, and R70 (7.48 log CFU/mL, 7.38 log CFU/mL, and 7.09 CFU/mL, respectively) during the fermentation period (120 h). Amylase activities of fermented R80 and R100 were 57.77 U/mL and 19.91 U/mL, respectively. Furthermore, amylase activities of fermented freeze-dried powders of R100 and R80 were 24.31 U/g and 9.12 U/g, respectively. Free sugar contents of R100 and R80 increased after fermentation, and that of R80 (5,454.15 mg/100 g) significantly increased compared to that of R100 (4,274.85 mg/100 g). The free amino acid content of R80 was higher than that of R100. DPPH and superoxide radical scavenging activities of 5 mg/mL of fermented freeze-dried powder (R80) were 44.21% and 89.76%, respectively. ACE inhibition rates and ${\alpha}$-glucosidase inhibitory activities were significantly higher in R80 than R100. This study suggested that fermentation of R80 might be a new potential source of antioxidant, anti-diabetic, and anti-hypertensive agents applicable to grain enzyme-containing foods.

Cytotoxic Effect of X-irradiation of Mouse Tumor Cells in the Presence of Korean Ginseng Extract (마우스 종양세포의 세포독성에 미치는 인삼 추출액과 방사선조사의 병용 효과)

  • Kwon Hyoung-Cheol;Kim Jin-Ki;Kim Jung-Soo;Choi Dong-Seong
    • Radiation Oncology Journal
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    • v.18 no.3
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    • pp.200-204
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    • 2000
  • Purpose : We already reported the results that aqueous extract of Korean ginseng roots showed a marked cytotoxicity. In this study, we investigated whether combined ginseng product with X-irradiation increase the cytotoxicity of tumor cells than X-irradiation or not. Materials and Methods : Fifty gram of Korean ginseng powder mixed with 1 L of distilled water was extracted with reflux flask under condition of $100^{\circ}C$ for 5 hrs. This aquaous ginseng extract was filtered, centrifuged and then was freezed under condition of $-90^{\circ}C$ for 16-18 hrs. The freezing extract was dried with freeze drier, and then diluted. X-irradiation was given to tumor cells by 6 MeV linear accelerator. The cytotoxicity of ginseng in vitro was evaluated from its ability to reduce the clonogenecity of fibrosarcoma (FSa II) cells. In X-irradiation alone group, each 2, 4, 6 and 8 Gy was given to tumor cells. In X-irradiation with ginseng group, 0.2 mg/mL of ginseng extract was exposed to tumor cells for 1 hour before X-irradiation. Results : The yield for 50 g of ginseng extract which was treated with freezing drier was 3.13 g($6.3\%$). Cytotoxicity In vitro was measured as survival fraction which was judged from the curve, at ginseng concentration of 0.001, 0.01, 0.1 and 1 mg/mL were $0.89\pm0.04$, $0.86\pm0.06$, $0.73\pm0.01$ and $0.09\pm0.02$, respectively. Survival fraction at X-irradiation alone of 2, 4, 6 and 8 Gy were $0.81\pm0.07$, $0.42\pm0.08$, $0.15\pm0.02$, $0.03\pm0.01$, respectively. But, suwival fraction in combined group of X-irradiation and ginseng (0.2mg/ml) at each same radiation dose were $0.28\pm0.01$, $0.18\pm0.03$, $0.08\pm0.02$, $0.006\pm0.002$, respectively (p<0.05). Conclusion : The yield for ginseng extract which was treated with freezing drier was $6.3\%$. Cytotoxicty of Fsa 11 in combined ginseng with X-irradiation group was increased than that of X-irradition alone group, and its enhancing effect seemed to be added.

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Efficacy of 20 Select Herbs Against Common Cold (감기 치료 빈용 처방 구성 약물의 효능 연구)

  • Kim, Dong-hyun;Ahn, Hyun-jin;Chiang, Shuen-cheng;Lee, Beom-joon;Jung, Sung-ki;Jung, Hee-jae
    • The Journal of Internal Korean Medicine
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    • v.36 no.3
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    • pp.236-251
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    • 2015
  • Objectives: To investigate the anti-inflammatory, analgesic, anti-pyretic and anti-histamine activities of 20 herbal medicines to test their efficacy in common cold treatment. Methods: For all experiments, the herbal medicines were extracted with 80% ethanol and freeze-dried. To determine the anti-oxidative properties, we tested DPPH-free radical-scavenging activity and xanthine oxidase inhibitory activity. To determine anti-inflammatory and analgesic potential, we investigated acetic acid-induced vascular permeability and writhing test in ICR mice. For anti-pyretic activities, an LPS-induced pyrexia study was conducted in rabbits. To evaluate the anti-histamine activity, we examined compound 48/80-induced systemic anaphylaxis in ICR mice and the release of β-hexosaminidase on rat basophilic leukemia (RBL-2H3) cells. Results: Ephedrae herba, Forsythiae fructus, Cinnamomi ramulus, and Cimicifugae rhizome showed potent free-radical scavenging activities. Gentianae macrophyllae radix inhibited acetic acid-induced vascular permeability. Schizonepetae spica and Cimicifugae rhizome inhibited acetic acid. Cinnamomi ramulus and Angelicae decursivae radix inhibited LPS-induced pyrexia. Angeliace dahuricae radix and Asari radix inhibited compound 48/80. Scutellariae radix, Cinnamomi ramulus, Ephedrae herba, and Zingiberis rhizoma crudus potently inhibited the release of β-hexosaminidase. Conclusions: We examined the anti-inflammatory, analgesic, anti-pyretic and anti-histamine activities of 20 herbal medicines;We examined the anti-inflammatory, analgesic, anti-pyretic and anti-histamine activities of 20 herbal medicines Codonopsis pilosulae radix, Zingiberis rhizoma crudus, and Cinnamomi ramulus showed novel efficacy. These results suggest that some of herbal medicines may be very effective in treating common cold.

A High Viscosity of Curdlan at Alkaline pH Increases Segregational Resistance of Concrete (염기성 pH에서의 고점도 커들란에 의한 콘크리트의 재료분리 억제 효과 증진)

  • 이인영;김선원;이중헌;김미경;조인성;박영훈
    • KSBB Journal
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    • v.14 no.1
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    • pp.114-118
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    • 1999
  • In order to use a polysaccharide, curdlan, as a concrete admixture, we first developed a pilot-scale fermentation process for the mass production of curdlan. We also examined the rheological properties of curdlan, and tested how well the curdlan obtained in this work increased the segregational resistance of the cement slurry. Fermentation was performed in a 300-liter fermenter equipped with 3 disk-turbine impellers. Since curdlan production is stimulated under nitrogen-limiting conditions, the culture pH was shifted from the optimal pH for cell growth (pH 7.0) to the optimal pH for curdlan production (pH 5.5) at the onset of ammonium exhaustion. We obtained a curdlan production of 65 g/L in 120 hr batch cultivation of Agrobacterium species. The insoluble curdlan at the final stage of fermentation was readily harvested by centrifugation together with the cells. The freeze-dried sample contained 78% (w/w) of curdlan. The solubility and viscosity of the curdlan increased with the increase of the solution pH, which enhances the viscosity of concrete since the pH of concrete is extremely high (pH 13.0). Test results of the curdlan as a concrete admixture with cement slurry demonstrated that it prohibits the leakage of water. In conclusion, this work certifies and enlarges curdlan's industrial potential as a concrete admixture.

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