• Title/Summary/Keyword: viability test

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Effects of Curcuma longa L. on Some Kinds of Cancer Cells (강황이 수종의 암세포에 미치는 영향)

  • Yoon, Joo-Ho;Kim, Jin-Sung;Yoon, Sang-Hyub;Ryu, Bong-Ha
    • The Journal of Internal Korean Medicine
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    • v.27 no.2
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    • pp.429-443
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    • 2006
  • Objectives : The Purpose of this study was to identify anti-tumor effects of Curcuma longa L. on some kinds of cancer cells through molecular biologic methods. Materials & Methods : We used 4 kinds of cancer cell lines such as glioma cells(A172), cervical cancer cells(HeLa), Prostate cancer cells(PC3), lung cancer cells(A549). We injected the boiled extract of Curcuma longa L. $5{\mu}g,\;10{\mu}g$ to culture media(ml) for 24 hours. We measured the cytotoxic effect on 4 kinds of cancer cells through trypan blue exclusion test and the suppressive effect on viability of 4 kinds of cancer cells via MTT assay. We measured the change of mitochondria membrane potential via flow cytometry. The quantitative RT-PCR was used to examine the effect on the revelation of Bcl-2 and Bax which genes are related to apoptosis. We examined the effect on the revelation of Bcl-2 protein and Bax protein by western blot analysis. Results: 1. Extract of Curcuma longa L. showed significant cytotoxic effect on A172, HeLa, PC3 compared to the control group with density dependent manner. 2. Extract of Curcuma longs L. showed significant suppressive effect on viability of A172, HeLa, PC3 compared to the control group with density dependent manner. 3. Curcuma longs L. induced apoptosis by decreasing the membrane potential of mitochondria in A172, HeLa, PC3. 4. In the test about the revelation of genes related to apoptosis, the revelation of Bcl-2 decreased and the revelation of Bax increased in A172. HeLa, PC3 treated with Curcuma longa L. with density dependent manner. 5. In the test about the revelation of protein related to apoptosis, the protein levels of Bcl-2 decreased and the protein levels of Bax increased in A172, HeLa, PC3 treated with Curcuma longa L. Conclusions: This experiment shewed that Curcuma longs L. has anti-tumor effect on glioma, cervical, Prostate cancer cells except on lung cancer. We hope that anti-tumor effects of Curcuma longa L. will be more Practically identified.

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Whey Protein Concentrate, Pullulan, and Trehalose as Thermal Protective Agents for Increasing Viability of Lactobacillus plantarum Starter by Spray Drying

  • Sun, Haiyue;Hua, Xiaoman;Zhang, Minghao;Wang, Yu;Chen, Yiying;Zhang, Jing;Wang, Chao;Wang, Yuhua
    • Food Science of Animal Resources
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    • v.40 no.1
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    • pp.118-131
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    • 2020
  • It is necessary to add protective agents for protecting the probiotic viability in the preparation process of probiotics starter. In this study, we used whey protein concentrate (WPC), pullulan, trehalose, and sodium glutamate as the protective agent and optimized the proportion of protective agent and spray-drying parameters to achieve the best protective effect on Lactobacillus plantarum. Moreover, the viable counts of L. plantarum in starter stored at different temperatures (-20℃, 4℃, and 25℃) for 360 days were determined. According to response surface method (RSM), the optimal proportion of protective agent was 24.6 g/L WPC, 18.8 g/L pullulan, 16.7 g/L trehalose and 39.3 g/L sodium glutamate. The optimum spray-drying parameters were the ratio of bacteria to protective agents 3:1 (v: v), the feed flow rate 240 mL/h, and the inlet air temperature 115℃ through orthogonal test. Based on the above results, the viable counts of L. plantarum was 12.22±0.27 Log CFU/g and the survival rate arrived at 85.12%. The viable counts of L. plantarum stored at -20℃ was more than 1010 CFU/g after 200 days.

The effect of photodynamic therapy on Aggregatibacter actinomycetemcomitans attached to surface-modified titanium

  • Cho, Kyungwon;Lee, Si Young;Chang, Beom-Seok;Um, Heung-Sik;Lee, Jae-Kwan
    • Journal of Periodontal and Implant Science
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    • v.45 no.2
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    • pp.38-45
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    • 2015
  • Purpose: The purpose of this study was to evaluate the effect of photodynamic therapy (PDT) using erythrosine and a green light emitting diode (LED) light source on biofilms of Aggregatibacter actinomycetemcomitans attached to resorbable blasted media (RBM) and sandblasted, large-grit, acid-etched (SLA) titanium surfaces in vitro. Methods: RBM and SLA disks were subdivided into four groups, including one control group and three test groups (referred to as E0, E30, E60), in order to evaluate the effect of PDT on each surface. The E0 group was put into $500{\mu}L$ of $20{\mu}M$ erythrosine for 60 seconds without irradiation, the E30 group was put into erythrosine for 60 seconds and was then irradiated with a LED for 30 seconds, and the E60 group was put into erythrosine for 60 seconds and then irradiated with a LED for 60 seconds. After PDT, sonication was performed in order to detach the bacteria, the plates were incubated under anaerobic conditions on brucella blood agar plates for 72 hours at $37^{\circ}C$, and the number of colony-forming units (CFUs) was determined. Results: Significant differences were found between the control group and the E30 and E60 groups (P<0.05). A significantly lower quantity of CFU/mL was found in the E30 and E60 groups on both titanium disk surfaces. In confocal scanning laser microscopy images, increased bacterial death was observed when disks were irradiated for a longer period of time. Conclusions: These findings suggest that PDT using erythrosine and a green LED is effective in reducing the viability of A. actinomycetemcomitans attached to surface-modified titanium in vitro.

Inhibiting Effect of Injinoryung-san-Ga-Samchilgun on Liver Fibrosis in Rats (인진오령산가삼칠근이 흰쥐의 간섬유화 억제에 미치는 영향)

  • Kim, Dong-Woo;Kim, Young-Chul;Ko, Heung
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.22 no.4
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    • pp.863-870
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    • 2008
  • The aim of this study was to investigate that Injinoryung-san-Ga-Samchilgun(IJORS) has an inhibitory effect on the development of liver fibrosis in rats. The influence of IJORS on liver stellate cell viability in rat was measured by the MTT assay, and proliferation was measured by the BrdU assay. The mRNA expression of procollagen type $1{\alpha}2$, ${\alpha}-SMA$, TIMP1, and TIMP2 all of which are associated with liver fibrosis, were analyzed by RT-PCR. The inhibitory effect of IJORS on procollagen production in hepatic stellate cell was examined using by enzyme immuno assay(procollagen Type 1 C-Peptide EIA). And after IJORS was orally administered to experimental rats with thioacetamide(TAA)-induced liver fibrosis for 4 weeks, the body weight, liver function test, complete blood and the change of portal pressure were measured. IJORS prevented hepatic stellate cell viability and proliferation in a dose-dependent manner. IJORS reduced the mRNA expression of procollagen type $1{\alpha}2$, ${\alpha}-SMA$ and TIMP1 and the production of procollagen protein. IJORS inhibited the increase of AST, ALT, WBC and portal pressure in rats administered by TAA. IJORS is considered to prevent liver fibrosis by inhibiting the activation of stellate cell and production of procollagen and prevent the progress of liver fibrosis by inhibiting the inflammation of liver tissue complicated in many liver disease.

The Effect of Topical Application of Heparin with Microneedling on Skin Flap Survival (미세바늘을 이용한 헤파린의 국소 도포가 피부 피판 생존에 미치는 영향)

  • Yang, Eunjung;Kim, Sugwon
    • Archives of Plastic Surgery
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    • v.36 no.3
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    • pp.254-261
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    • 2009
  • Purpose: The objective of this study was to evaluate the heparin effect for a viability of random - pattern dorsal flap in hairless mouse. Methods: A caudally - based random dorsal flap, measuring $1.5{\times}5cm$, was designed and heparin was applied topically after microneeding. Twenty five male hairless mice were randomly divided into control (Group1, n=5); received only microneedling (Group 2, n=5), only heparin(Group3, n=5), microneedling with saline(Group 4, n=5), and microneedling with heparin group(group5,n=5) to the flap during 7 days. The number of the capillaries were compared between the experimental groups and control group with respect to neovascularization after heparin application using imaging analysis program under hematoxylin - eosin stain. The capillary blood flow was measured by laser Doppler flowmetry. After seven days each animal was evaluated for the percentage area of the flap survival. Mann - Whitnety U test and Kruskal - Wallis statistical analysis of survival relationships was performed. Results: It can be observed increased number of the blood vessels in the experimental groups however it was not statistically significant. Blood flow of the haparin with microneedling group maintained higher than other experimental groups. Treated microneeding and heparin mice were significantly better flap viability than in controls (flap survival 67% and $54.4mm^2$ respectively; p<.01). Positive correlation was shown between flap survival rate and laser Doppler flux value only at first day after surgery. Conclusion: Heparin has a beneficial effect on capillary flow and improve peripheral circulatory disturbances in random pattern flaps.

Effects of Duchesnea Indica of Colorectal Adenocarcinoma Cells (사매가 대장암 세포에 미치는 영향)

  • Lee, Do-Hyoung;Kim, Jin-Sung;Yoon, Sang-Hyub;Ryu, Ki-Won;Ryu, Bong-Ha
    • The Journal of Internal Korean Medicine
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    • v.26 no.2
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    • pp.310-319
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    • 2005
  • Objectives: The aim is to identify any anti-tumor effects of Duchesnea indica(Andr.) Focke on colon cancer cells. Materials & Methods: Colo201 human adenocarcinoma cells were obtained from American Type Culture Collection. The boiled extract of Duchesnea indica(Andr.) Focke was added (10 and 20 microliters) to cultures and observed at 0, 6, and 12 hours, and at 12-hour intervals thereafter. Morphological changes in colon cancer cells were observed through an inverted microscope, Destruction of colon cancer cells was measured through Trypan blue exclusion testing. Suppression of the viability of colon cancer cells were measured via MTT assay. Anti-cancer mechanisms in the cell cycle of colon cancer cells were analysed via flow cytometry. Results: After introduction of Duchesnea indica(Andr.) Focke to cultures several changes were seen. Significant atrophy of the nucleus and cytoplasm of colon cancer cells was observed, indicating cell injury. Destruction of colon cancer cells was observed in direct proportion to dosage and duration. Suppression of viability of colon cancer cells for each test group was greater than that of the control group increasingly over time(36h, 48h, 60h, 72h), which was statistical significant (p<0.05). Cell numbers of the mitosis phase of the colon cancer cell cycle reduced. Conclusions: Statistcally significant anti-tumor effects of Duchesnea indica(Andr.) Focke were observed in this in vitro experiment. Results support a role for Duchesnea indica(Andr.) Focke in treatment of colon cancer. though it will required progressive research to develop a practical treatment.

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Effects of Storage Time on Bacteria Concentration and Sperm Parameters in Boar Semen (돼지 액상정액 보존 일수에 따른 정액내 세균과 정자 기능의 변화)

  • Chung, Ki-Hwa;Kim, In-Cheol
    • Reproductive and Developmental Biology
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    • v.36 no.3
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    • pp.163-166
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    • 2012
  • This study was conducted to determine the relationship between elapsed time after semen preservation on the changes of bacteria and semen quality. Semen was diluted with BTS(Beltsville Thawing Solution) extender without antibiotic for 7 days and sperm parameter and fertility were measured. Sperm motility was measured by CASA and total bacteria number was counted after 22~24 hr incubation from counting agar plate in which sperm dilute to $10{\sim}10^6$ in 0.9% saline solution and inoculate to agar. Acrosomal integrity was measured by Chlortetracycline (CTC) staining. CTC patterns were uniform fluorescence over the whole head (pattern F), characteristic of incapacitated acrosome-intact spermatozoa; fluorescence-free band in the post-acrosomal region (pattern B), characteristic of capacitated acrosome-intact spermatozoa; and almost no fluorescence over the whole head except for a thin band in the equatorial segment (pattern AR), characteristic of acrosome reacted spermatozoa. Total number of bacteria was significantly increased (p<0.0001) 3 days after preservation. Sperm motility, viability, and morphological abnormality on elapsed time after preservation were lower from 5 ($77.24{\pm}6.47$, p<0.001) and 7 days ($77.24{\pm}6.47$, p<0.001) after preservation compared to 1 ($15.71{\pm}7.18$) and 3 days($18.39{\pm}7.22$) after preservation, respectively. Sperm viability was significantly lower ($53.25{\pm}35.03$, p<0.0001) at 7 days after preservation. Morphological abnormality of sperm was lower (p<0.001) at 1 ($15.71{\pm}7.18$) and 3 ($18.39{\pm}7.22$) days compared to 5 ($21.84{\pm}7.91$) and 7 ($22.59{\pm}9.93$) days after preservation. Acrosomal integrity and capacitation rate (pattern F) were significantly lower (p<0.001) from 5 days after preservation. Based on the data we obtained from this study suggested that semen preserved more than 5 days without antibiotic would not recommend use for artificial insemination.

Whitening Effect of Banana Leaf Extract (바나나잎 추출물의 미백 개선 효과)

  • Hwang, Hyung Seo;Yoo, Dae Sung;Shim, Joong Hyun
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.42 no.1
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    • pp.37-43
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    • 2016
  • This research was carried out to identify the whitening effect of Banana leaf extract. B16F10 cells were used to measure cell viability, mRNA expression, and tyrosinase activity inhibition assay from B16F10 cell. We also carried out clinical test of the cream product containing banana leaf extract. In this study, we elucidated the effects of banana leaf extract on TRP1 / TRP2 / Tyr mRNA expression and tyrosinase activity inhibition. Quantitative real-time PCR showed that banana leaf extract decreased mRNA level of TRP1, TRP2 and Tyr gene and tyrosinase activity inhibition assay also revealed that banana leaf extract 65% decreased melanin production in B16F10 cell. Banana leaf extract cream can whiten the skin darkness induced by ultraviolet. Therefore, we successfully identified the whitening effect of banana leaf extract, and this finding suggested the banana leaf extract is a considerable potent cosmetic ingredient for skin whitening. Based on this, we anticipated further researches about banana leaf extract for mechanism to develop not only cosmetics but healthcare food or medicines.

Viability Assay of Seaweeds Responding to Mountain Fire-Related Pollutants (산불 관련 해양환경오염원들에 대한 해조류의 활력 측정)

  • KANG Se-Eun;JIN Long-Guo;CHOI Jae-Suk;CHO Ji-Young;SHIN Hyun-Woung;HONG Yong-Ki
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.36 no.3
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    • pp.225-229
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    • 2003
  • Plant ash and soil drainage, derived by frequent mountain fires during winter, might cause biological contamination to seaweeds at seashore and river mouse area. To thalli of Ulva pertusa, maximum non-lethal concentration(MNLC), lethal concentration 50 $(LC_{50})$ and minimum lethal concentration (MLC) of pine needle ash were shown as 60, 350 and 550 mg/mL, respectively. The yellow loess and granite sand did not damage at concentrations of 20 and 200 mg/mL, respectively To thalli of Porphyra yezoensis, the MNLC, LC5O, MLC of pine needle ash were shown as 0.08, 0.4 and 1.0 mg/mL, respectively. Effects of yellow loess and granite sand were approximately 1/2 and 1/10 of the ash. To thalli of Undaria pinnatifida, the pine needle ash, yellow loess and granite sand did not damage at the concentration range of 20 to 40 mg/mL. Change of pigments $(chlorophyll\;\alpha,\;lutein,\;\beta-carotene,\;phycoerthrin)$ was also determined at the MNLC, $LC-{50}$ and MLC of pine needle ash. Among three seaweeds tested, P. yezoensis produced the most 2.7-fold of lutein and 2.3-fold of $\beta-carotene$ at $LC-{50}$ of the ash. Thus the P. yezoensis, appeared as a sensitive indicator, could be used as one of test organisms for determination of the biological effect of pollutants contaminated in marine environment.

Cytotoxicity of temporary cements on bovine dental pulp-derived cells (bDPCs) using real-time cell analysis

  • Malkoc, Meral Arslan;Demir, Necla;Sengun, Abdulkadir;Bozkurt, Serife Buket;Hakki, Sema Sezgin
    • The Journal of Advanced Prosthodontics
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    • v.7 no.1
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    • pp.21-26
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    • 2015
  • PURPOSE. To evaluate the cytotoxicity of temporary luting cements on bovine dental pulp-derived cells (bDPCs). MATERIALS AND METHODS. Four different temporary cements were tested: Rely X Temp E (3M ESPE), Ultratemp (Ultradent), GC Fuji Temp (GC), and Rely X Temp NE (3M ESPE). The materials were prepared as discs and incubated in Dulbecco's modified eagle's culture medium (DMEM) for 72 hours according to ISO 10993-5. A real-time cell analyzer was used to determine cell vitality. After seeding $200{\mu}L$ of the cell suspensions into the wells of a 96-well plate, the bDPCs were cured with bioactive components released by the test materials and observed every 15 minutes for 98 hours. One-way ANOVA and Tukey-Kramer tests were used to analyze the results of the proliferation experiments. RESULTS. All tested temporary cements showed significant decreases in the bDPCs index. Rely X Temp E, GC Fuji Temp, and Rely X Temp NE were severely toxic at both time points (24 and 72 hours) (P<.001). When the cells were exposed to media by Ultratemp, the cell viability was similar to that of the control at 24 hours (P>.05); however, the cell viability was significantly reduced at 72 hours (P<.001). Light and scanning electron microscopy examination confirmed these results. CONCLUSION. The cytotoxic effects of temporary cements on pulpal tissue should be evaluated when choosing cement for luting provisional restorations.