• Title/Summary/Keyword: survival Kit

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Comparative Analysis about the Effect of Isolated Phosphatidylcholine and Sodium Deoxycholate for the Viability of Adipocyte (Phosphatidylcholine과 Sodium Deoxycholate가 지방세포 생존에 미치는 영향의 비교 분석)

  • Rha, Eun-Young;Kang, Jo-A;Lee, Jung-Ho;Oh, Deuk-Young;Seo, Je-Won;Moon, Suk-Ho;Ahn, Sang-Tae;Rhie, Jong-Won
    • Archives of Plastic Surgery
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    • 제37권5호
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    • pp.531-534
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    • 2010
  • Purpose: Lipobean$^{(R)}$s, widely used in lipodissolving techniques, contain phosphatidylcholine and sodium deoxycholate as its main substances. They have been approved only as medication for liver disease by the FDA. However, they have been used under various clinical settings without exact knowledge of its action mechanism. The authors designed an in vitro study to analyze the effects of different concentrations of phosphatidylcholine and sodium deoxycholate on adipocytes and other types of cells. Methods: Human adipose-derived stem cell were cultured and induced to differentiate into adipocytes. Fibroblasts extracted from human inferior turbinate tissue, and MC3T3-E1 osteoblast lines were cultured. Phosphatidylcholine solution dissolved with ethanol was applied to the culture medium at differing concentrations (1, 4, 7, 10 mg/mL). The sodium deoxycholate solution dissolved in DMSO applied to the medium at differing concentrations (0.07, 0.1. 0.4. 0.7 mg/mL). Cells were dispersed at a concentration of $5{\times}10^3$ cells/well in 24 well plates, and surviving cells were calculated 1 day after the application using a CCK-8 kit. Results: The number of surviving cells of adipocytes, fibroblasts and osteoblasts decreased as the concentration of sodium deoxycholate increased. However, all types of cells that had been processed in a phosphatidylcholine showed a cell survival rate of over 70% at all concentrations. Conclusion: This study shows that sodium deoxycholate is the more major factor in destroying adipocytes, and it is also toxic to the other cells. Therefore, we conclude that care must be taken when using Lipobean$^{(R)}$s as a method of reducing adipose tissue, for its toxicity may destroy other nontarget cells existing in the subcutaneous tissue layer.

A Study on the Inhibitory Effect of Yeongdamsagantang on Alzheimer in $A{\beta}-oligomer-induced$ Neuro 2A Cell Lines (($A{\beta}-oligomer$로 유도된 Neuro2A 세포주에서 용담사간탕(龍膽瀉肝湯)의 치매 억제 효과)

  • Kim, Hae-Su;Shin, Yoo-Jeong;Park, Jong-Hyuk;Kim, Seung-Mo;Paek, Kyung-Min;Park, Chi-Sang
    • The Journal of Korean Medicine
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    • 제29권2호
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    • pp.151-164
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    • 2008
  • Objective: To investigate the effects of Yeongdamsagantang (YDGT) on apoptosis of neuronal cells that can result in dementia. Method: The water extract of the YDGT was tested in vitro for its beneficial effects on neuronal survival and neuroprotective functions, particularly in connection with $A{\beta}$ oligomer-related dementias. $A{\beta}$ oligomers derived from proteolytic processing of the ${\beta}-amyloid$ precursor protein (APP), including the $amyloid-{\beta}$ peptide $(A{\beta})$, play a critical role in the pathogenesis of Alzheimer's disease. A neuroblastoma cell line stably expressing an $A{\beta}$ oligomerassociated neuronal degeneration was used to investigate if YDGT inhibits formation of $A{\beta}$ oligomer. To measure the ATP generating level in mitochondrial membrane, luciferin/luciferase luminescence kit (Promega) and luminator was used, and to survey the protein's apparition, confocal microscopy was used. Result: $A{\beta}oligomer$ had a profound attenuation in the increase in CT105 expressing neuro2A cells from YDGT. Experimental evidence indicates that YDGT protected against neuronal damage from cells, but its cellular and molecular mechanisms remain unknown. We demonstrated that YDGT inhibited formation of $amyloid-{\beta}$ $(A{\beta})$ oligomers, which were the behavior, and possibly causative, features of AD. The decreased $A{\beta}$ oligomer in the presence of YDGT was observed in the conditioned medium of this $A{\beta}oligomer-secreting$ cell line under in vitro. In the cells, YDGT significantly attenuated mitochondrion-initiated apoptosis. Conclusion: (i) a direct $A{\beta}$ oligomer toxicity and the apoptosis initiated by the mitochondria; and (ii) multiple cellular and molecular neuroprotective mechanisms, including attenuation of apoptosis and direct inhibition of $A{\beta}$ oligomer aggregation, underlie the neuroprotective effects of YDGT.

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Survival, isolation and characterization of Bacillus cereus from Sunshik (선식에서 Bacillus cereus의 분리 및 특성 조사와 열에 대한 사멸률 연구)

  • Cho, Yong-Sun;Jung, Eun-Young;Lee, Myung-Ki;Yang, Cheul-Young;Shin, Dong-Bin
    • Journal of Food Hygiene and Safety
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    • 제23권4호
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    • pp.343-347
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    • 2008
  • Recently, Sunshik has been issued because of easy-cook and well-being food. Sunshik basically was made of the heated cereals. Amount of spore-forming Bacillus cereus was detected and it has been caused some problem of food safety. B. cereus was isolate from 57 out of 161 Sunshik samples resulting in the isolation rate of 35.4%. Quantitative analysis of 57 samples showed that 21 samples were less than 100 CFU/g, 33 samples were between 100 and 1,000 CFU/g and distinctively even 3 (1.9%) samples had over 1,000 CFU/g. Typical morphology of B. cereus isolated from Sunshik was observed on MYP agar and then further characteristics was identified by using VITEK 2 (Biomeriux, France). 53 strains out of 57 strains isolated from Sunshik (about 93.0%) produced diarrheal enterotoxin in brain heart infusion broth which was detected by the Bacillus cereus enterotoxin reversed passive latex agglutination test kit (Oxoid England). The D-values of the B. cereus spores were $75^{\circ}C$ (37.1mim), $80^{\circ}C$ (22.5mim), $85^{\circ}C$ (4.9mim), and $90^{\circ}C$ (3.1mim) respectively. The Z-value was calculated $12.8^{\circ}C$ in Sunshik sample inoculated with B. cereus. Therefore, the management of B. cereus in Sunshik is required for the food-safety.

Quality characteristics of Weissella confusa strain having gluten degradation activity from salted seafood (젓갈로부터 분리된 글루텐 분해능을 가지는 Weissella confusa 균주와 특성)

  • Yoon, Jong Young;Hwang, Kwontack
    • Food Science and Preservation
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    • 제23권6호
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    • pp.883-889
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    • 2016
  • A new lactic acid bacteria with gluten-degrading activity which was isolated from salted sea foods (traditional Korea fermented food), identified as Weissella confusa (99%) by use of API kit and 16S rRNA sequencing, and designated as W. confusa. When the W. confusa cultured for 48 hours at $30^{\circ}C$ in a MRS medium containing 1% gluten, 45% of gluten was founded to be degraded. W. confusa showed 85% of survival rate at pH 3, and 94% tolerance at 0.1% oxgall, which indicates that W. confusa would survive in stomach of human. Experiments on the thermostability was confirmed that it has a stability of 70% in $50^{\circ}C$. W. confusa inhibited the growth of some pathogen, except for S. aureus. Results in this study suggest that using W. confusa for fermentation of grain flour containing gluten would be desirable to prepare the gluten-free foods needed for those who suffer from celia disease and gluten allergy.

INDUCTION OF APOPTOSIS IN ORAL CANCER CELL LINE THROUGH AN RECOMBINANT HCCS-1 ADENOVIRUS (재조합 HCCS-1 아데노바이러스를 이용한 구강암 세포주의 세포사멸 유발)

  • Kim, Chang-Hyen;Lee, Dong-Ju;Lee, ll-Kyu;Kim, Myung-Jin;Kim, Jin-Woo;Pyo, Sung-Woon
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • 제31권4호
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    • pp.306-311
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    • 2005
  • Despite advances in surgery, radiotherapy, and chemotherapy, the survival of patients with oral squamous cell carcinoma has not significantly improved over the past several decades. Gene therapy is currently under investigation and shows us new possibility of cancer curing method. This experiment was undergone to find out the cell growth inhibition effect and evidence of apoptosis by HCCS-1(human cervical cancer suppressor-1), one of the candidates of tumor suppressor gene, transducted to human oral cancer cell line. To determine the efficiency of the adenovirus as a gene delivery vector cell line was transducted with LacZ gene and analysed with X-gal staining. Northern blot was performed to confirm the transfection with HSCC-1 gene and cell viability was assessed by cell cytotoxicity assay using cell count kit(CCK). To show the evidence of apoptosis, DNA fragmentation assay and flow cytometry(FACS) were performed. We had successfully construct the recombinant HSCC-1 adenovirus(Ad5CMV-HCCS-1), and importation efficiency was 20% at 2 MOI(multiplicity of infection), 80% at 20 MOI. Northern blot analysis showed that a single 0.6kb mRNA transcript was expressed in Ad5CMV-HCCS-1 transducted cell lines. As a result of CCK, when comparing to control subjects, transducted group showed 50% growth inhibition. In DNA fragmentation assay, according to increasing of MOI, DNA volume was diminished. In FACS analysis, DNA distribution showed fragmentation. This results imply that HCCS-1gene has growth inhibition effect in human oral cancer cell lines through apoptosis induction.

Overexpression of cholinergic receptor nicotinic gamma subunit inhibits proliferation and differentiation of bovine preadipocytes

  • Jiawei, Du;Hui, Zhao;Guibing, Song;Yuan, Pang;Lei, Jiang;Linsen, Zan;Hongbao, Wang
    • Animal Bioscience
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    • 제36권2호
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    • pp.200-208
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    • 2023
  • Objective: Muscle acetylcholine receptors have five alpha subunits (α, β, δ, ε, or γ), and cholinergic receptor nicotinic gamma subunit (CHRNG) is the γ subunit. It may also play an essential role in biological processes, including cell differentiation, growth, and survival, while the role of CHRNG has not been studied in the literature. Therefore, the purpose of this study is to clarify the effect of CHRNG on the proliferation and differentiation of bovine preadipocytes. Methods: We constructed a CHRNG overexpression adenovirus vector and successfully overexpressed it on bovine preadipocytes. The effects of CHRNG on bovine preadipocyte proliferation were detected by Edu assay, cell counting Kit-8 (CCK-8), real-time fluorescence quantitative polymerase chain reaction (RT-qPCR), Western blot and other techniques. We also performed oil red O, RT-qPCR, Western blot to explore its effect on the differentiation of preadipocytes. Results: The results of Edu proliferation experiments showed that the number of EDU-positive cells in the overexpression group was significantly less. CCK-8 experiments found that the optical density values of the cells in the overexpression group were lower than those of the control group, the mRNA levels of proliferating cell nuclear antigen (PCNA), cyclin A2 (CCNA2), cyclin B1 (CCNB1), cyclin D2 (CCND2) decreased significantly after CHRNG gene overexpression, the mRNA levels of cyclin dependent kinase inhibitor 1A (CDKN1A) increased significantly, and the protein levels of PCNA, CCNB1, CCND2 decreased significantly. Overexpression of CHRNG inhibited the differentiation of bovine preadipocytes. The results of oil red O and triglyceride determination showed that the size and speed of lipid droplets accumulation in the overexpression group were significantly lower. The mRNA and protein levels of peroxisome proliferator activated receptor gamma (PPAR class="checkNonKBPoint">γ), CCAAT enhancer binding protein alpha (CEBPα), fatty acid binding protein 4 (FABP4), fatty acid synthase (FASN) decreased significantly. Conclusion: Overexpression of CHRNG in bovine preadipocytes inhibits the proliferation and differentiation of bovine preadipocytes.

Notoginseng leaf triterpenes ameliorates mitochondrial oxidative injury via the NAMPT-SIRT1/2/3 signaling pathways in cerebral ischemic model rats

  • Weijie, Xie;Ting, Zhu;Ping, Zhou;Huibo, Xu;Xiangbao, Meng;Tao, Ding;Fengwei, Nan;Guibo, Sun;Xiaobo, Sun
    • Journal of Ginseng Research
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    • 제47권2호
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    • pp.199-209
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    • 2023
  • Background: Due to the interrupted blood supply in cerebral ischemic stroke (CIS), ischemic and hypoxia results in neuronal depolarization, insufficient NAD+, excessive levels of ROS, mitochondrial damages, and energy metabolism disorders, which triggers the ischemic cascades. Currently, improvement of mitochondrial functions and energy metabolism is as a vital therapeutic target and clinical strategy. Hence, it is greatly crucial to look for neuroprotective natural agents with mitochondria protection actions and explore the mediated targets for treating CIS. In the previous study, notoginseng leaf triterpenes (PNGL) from Panax notoginseng stems and leaves was demonstrated to have neuroprotective effects against cerebral ischemia/reperfusion injury. However, the potential mechanisms have been not completely elaborate. Methods: The model of middle cerebral artery occlusion and reperfusion (MCAO/R) was adopted to verify the neuroprotective effects and potential pharmacology mechanisms of PNGL in vivo. Antioxidant markers were evaluated by kit detection. Mitochondrial function was evaluated by ATP content measurement, ATPase, NAD and NADH kits. And the transmission electron microscopy (TEM) and pathological staining (H&E and Nissl) were used to detect cerebral morphological changes and mitochondrial structural damages. Western blotting, ELISA and immunofluorescence assay were utilized to explore the mitochondrial protection effects and its related mechanisms in vivo. Results: In vivo, treatment with PNGL markedly reduced excessive oxidative stress, inhibited mitochondrial injury, alleviated energy metabolism dysfunction, decreased neuronal loss and apoptosis, and thus notedly raised neuronal survival under ischemia and hypoxia. Meanwhile, PNGL significantly increased the expression of nicotinamide phosphoribosyltransferase (NAMPT) in the ischemic regions, and regulated its related downstream SIRT1/2/3-MnSOD/PGC-1α pathways. Conclusion: The study finds that the mitochondrial protective effects of PNGL are associated with the NAMPT-SIRT1/2/3-MnSOD/PGC-1α signal pathways. PNGL, as a novel candidate drug, has great application prospects for preventing and treating ischemic stroke.

Analysis of Prevalence and Survival pattern of Staphylococcus aureus from Dried Seasoned Fishes (조미건어포의 Staphylococcus aureus 오염도 및 생존패턴 분석)

  • Cho, Joon-Il;Lee, Soon-Ho;Choi, Jun-Hyuk;Choi, Eun-Jung;Hwang, In-Gyun
    • Journal of Food Hygiene and Safety
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    • 제26권4호
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    • pp.366-369
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    • 2011
  • In this study, the contamination levels of total aerobic bacteria, E. coli, total coliform and S. aureus of seasoned dried fishes (SDF) in Korea were investigated. A total of 81 SDF samples were purchased randomly from 28 stores. Contamination range of total aerobic bacteria, total coliform and S. aureus were 150~1,700,000, 10~31,000 and 10~220 CFU/g, respectively. E. coli was detected in only one samples in the qualitative test. We have analyzed quantitatively Staphylococcal enterotoxins (SE-type A, C and D) produced by S. aureus contaminated in SDF using a TECRA kit and standard curve. The curve equation was Y = 0.1499 * X + 0.1183 and maximum amount of SEs in SDF was 0.71 ng/ml. Reduction speed of S. aureus in SDF stored at $37^{\circ}C$ was the highest among the samples stored for 8 days at different temperature of 7, 18 and $37^{\circ}C$. On the basis of the results, SDF in Korea can be contaminated by a variety of pathogenic bacteria. Therefore, precautionary measures are necessary for consumer protection, including the improvement of sanitary conditions in the processing plants in Korea.

In vivo Radioprotective Effects of Basic Fibroblast Growth Factor in C3H Mice (Basic Fibroblast Growth Factor (bFGF)의 방사선보호작용에 대한 실험적 연구)

  • Kim, Yeon-Shil;Yoon, Sei-Chul
    • Radiation Oncology Journal
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    • 제20권3호
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    • pp.253-263
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    • 2002
  • Purpose : In order to understand in vivo radiation damage modifying of bFGF on jejunal mucosa, bone marrow and the effect of bFGF on the growth of transplanted mouse sarcoma 180 tumor in mice. Materials and Methods : Mice were treated with $6\;{\mu}g$ of bFGF at 24 hours and 4 hours before exposing to 600 cGy, 800 cGy and 1,000 cGy total body irradiation (TBI), and then exposed to 3,000 cGy local radiation therapy on the tumor bearing thigh. Survival and tumor growth curve were plotted in radiation alone group and combined group of bFGF and irradiation (RT). Histologic examination was performed in another experimental group. Experimental groups consisted of normal control, tumor control, RT (radiation therapy) alone, $6\;{\mu}g$ bFGF alone, combined group of $3\;{\mu}g$ bFGF and irradiation (RT), combined group of $6\;{\mu}g$ bFGF and irradiation (RT). Histologic examination was peformed with H-E staining in marrow, jejunal mucosa, lung and sarcoma 180 bearing tumor. Radiation induced apoptosis was determined in each group with the DNA terminal transferase nick-end labeling method ($ApopTag^{\circledR}$ S7100-kit, Intergen Co.) Results : The results were as follows 1) $6\;{\mu}g$ bFGF given before TBI significantly improved the survival of lethally irradiated mice. bFGF would protect against lethal bone marrow syndrome. 2) $6\;{\mu}g$ bFGF treated group showed a significant higher crypt depth and microvilli length than RT alone group (p<0.05). 3) The bone marrow of bFGF treated group showed less hypocellularity than radiation alone group on day 7 and 14 after TBI (p<0.05), and this protective effect was more evident in $6\;{\mu}g$ bFGF treated group than that of $3\;{\mu}g$ bFGF treated group. 4) bFGF protected against early radiation induced apoptosis in intestinal crypt cell but might have had no antiapoptotic effect in bone marrow stem cell and pulmonary endothelial cells. 5) There was no significant differences in tumor growth rate between tumor control and bFGF alone groups (p>0.05). 6) There were no significant differences in histopathologic findings of lung and mouse sarcoma 180 tumor between radiation alone group and bFGF treated group. Conclusions : Our results suggest that bFGF protects small bowel and bone marrow from acute radiation damage without promoting the inoculated tumor growth in C3H mice. Improved recovery of early responding normal tissue and reduced number of radiation induced apoptosis may be possible mechanism of radioprotective effect of bFGF.