• 제목/요약/키워드: Co-incubation

검색결과 506건 처리시간 0.023초

영지의 ${\beta}$-glucan성 다당류에 의해 활성화된 흰쥐 간내 Kupffer 세포의 NO, TNF-${\alpha}$ 및 TGF-${\beta}$ 형성 (Nitric Oxide, TNF-${\alpha}$ and TGF-${\beta}$ Formation of Rat Kupffer Cell Activated by the ${\beta}$-Glucan from Ganoderma lucidum)

  • 한만덕;이준우;정훈;김용석;나수정;윤경하
    • 한국미생물·생명공학회지
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    • 제27권1호
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    • pp.28-34
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    • 1999
  • Ganoderan (GAN), an immunomodulating ${\beta}$-glucan from mushroom Ganoderma lucidum, was evaluated for its ability to induce formation of nitric oxide (NO), tumor necrosis factor-${\alpha}$(TNF-${\alpha}$) and transforming growth factor (TGF-${\beta}$) from rat Kupffer cell in vitro. Hepatic macrophages activated by GAN significantly elevated concentration of NO and TNF-${\alpha}$ in cultured medium, but not significantly elevated that of TGF-${\beta}$. GAN-activated Kupffer cells secrete 14.9${\mu}$M (p<0.01) of NO and 2619.5${\rho}$g/ml (p<0.01) of TNF-${\alpha}$after 36hr of incubation at 37$^{\circ}C$. The results revealed that GAN enhanced 4-fold production of NO and 19 fold formation of TNF-${\alpha}$ compared to the control. The proliferation of GAN-activated Kupffer cells was inhibited as compared with its negative control. Comparing the activity among glucans derived from microorganisms, highly branched zymosan, glucomannan from Saccharomyces cerevisiae, significantly increased TNF-${\alpha}$ and NO production. These results indicate that the ${\beta}$-glucan from G. lucidum activates rat Kupffer cell and secretes NO and TNF-${\alpha}$. It also suggest that rat Kupffer cell posses certain receptor for ${\beta}$-anomeric glucan.

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정지환과 시스플라틴의 신경교아세포종에 대한 세포고사 기전연구 (Mechanisms of Apoptosis by Combination with Jeongjihwan and Cisplatin in Human Glioblastoma Cells)

  • 신학수;이선우;이민구;윤종민;이인;신선호;문병순
    • 대한한의학회지
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    • 제26권2호
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    • pp.1-12
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    • 2005
  • Objectives: Malignant gliomas are often treated with cisplatin (cis-diamminedichloroplatinum(II), CDDP) and radiation but results remain unsatisfactory. Since malignant glioma displays moderate resistance to conventional therapy, a new treatment modality is needed to improve the outcome of patients with these tumors. The aim of this study was to investigate the effects of the combined use of Jongjihwan(JJH) and cisplatin(CDDP) on cultured malignant glioma cells, A172. Methodss & Results: The combined use of cisplatin and Jeongjihwan had synergistic effects on Al72 cells during 24 hr-incubation, This treatment resulted in a decrease of cell viability, Which was revealed as apoptosis Characterized by activation of caspase-3 protease as well as cleavage of poly ADP-ribose polymerase (PARP) with change of mitochondria membrane potential transition. The expression of members of the Bcl-2 protein family was modulated during co-treatment with Jeongjihwan and cisplatin. Activation of caspase-3 and mitochondrial alterations were central to co-treatment with Jeongjihwan and cisplatin-induced apoptosis. Conclusions: We conclude that co-treatment with Jeongjihwan and cisplatin-induced activation of the mitochondrial pathway enables cell death. Also, we suggest the combined theory of JJH and cisplatin could be a useful method for glioblastoma.

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Immunopathogenesis of COVID-19 and early immunomodulators

  • Lee, Kyung-Yil;Rhim, Jung-Woo;Kang, Jin-Han
    • Clinical and Experimental Pediatrics
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    • 제63권7호
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    • pp.239-250
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    • 2020
  • The novel coronavirus disease 2019 (COVID-19) is spreading globally. Although its etiologic agent is discovered as severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), there are many unsolved issues in COVID-19 and other infectious diseases. The causes of different clinical phenotypes and incubation periods among individuals, species specificity, and cytokine storm with lymphopenia as well as the mechanism of damage to organ cells are unknown. It has been suggested that in viral pneumonia, virus itself is not a direct cause of acute lung injury; rather, aberrant immune reactions of the host to the insults from viral infection are responsible. According to its epidemiological and clinical characteristics, SARS-CoV-2 may be a virus with low virulence in nature that has adapted to the human species. Current immunological concepts have limited ability to explain such unsolved issues, and a presumed immunopathogenesis of COVID-19 is presented under the protein-homeostasis-system hypothesis. Every disease, including COVID-19, has etiological substances controlled by the host immune system according to size and biochemical properties. Patients with severe pneumonia caused by SARS-CoV-2 show more severe hypercytokinemia with corresponding lymphocytopenia than patients with mild pneumonia; thus, early immunomodulator treatment, including corticosteroids, has been considered. However, current guidelines recommend their use only for patients with advanced pneumonia or acute respiratory distress syndrome. Since the immunopathogenesis of pneumonia may be the same for all patients regardless of age or severity and the critical immune-mediated lung injury may begin in the early stage of the disease, early immunomodulator treatment, including corticosteroids and intravenous immunoglobulin, can help reduce morbidity and possibly mortality rates of older patients with underlying conditions.

Quantitation of Plasma Apolipoprotein A-I with a Sandwich Type Enzyme-Linked Immunosorbent Assay Using Monoclonal Antibodies

  • Lee, Min-Gyu;Kang, Jae-Seon;Jeong, Jae-Yeon;Jue, Dae-Myung;Kim, Hack-Joo
    • BMB Reports
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    • 제30권6호
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    • pp.390-396
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    • 1997
  • A sandwich-type enzyme-linked immunosorbent assay (ELISA) for the quantification of human apolipoprotein A-I (apoA-I) was developed using monoclonal antibodies. For this assay, we used three monoclonal antibodies to trap and detect apo A-I. HDAI16 and HDA15 monoclonal antibodies were used for trapping apoA-I and HDAI8 monoclonal antibody was for detecting apoA-I. These three monoclonal antibodies were produced by immunizing mice with high density lipoprotein (HDL) isolated from human plasma. By immunoblot analysis, these three monoclonal antibodies were specific to apoA-I and showed no cross-reactivities with other plasma proteins. The results of competition assays for epitope cross-reactivity test also verified that these monoclonal antibodies identified separate and distinct epitopes on HDL and apoA-I. Affinity constants of monoclonal antibodies were measured by ELISA. Their association constants ranged from $10^7$ to $10^8$ $M^{-1}$. For this assay, pure apoA-I was isolated by affinity chromatography using monoclonal antibodies. In this sandwich assay, the amount of HRP-labeled HDAI8 bound to apoA-I trapped by HDAI16 and HDAI5 was proportional to apoA-I concentration in the range of 0 to 500ng/ml. ApoA-I concentration in plasma was calculated from the linear regression equation of standard curve. The precision and reliability of the assays are reflected in the low intra-and interassay coefficients of variation that averaged 3.25% and 4.30%, respectively. This assay is sensitive, simple, reproducible, convenient in incubation interval, and does not use radioisotope: thus it can be widely applied in clinical laboratories.

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Identification of Lactobacillus ruminus SPM0211 Isolated from Healthy Koreans and Its Antimicrobial Activity against Some Pathogens

  • Yun Ji-Hee;Yim Dong-sool;Kang Jin-Yang;Kang Byung-Yong;Shin Eun-ah;Chung Myung-Jun;Kim Soo-Dong;Baek Dae-Heoun;Kim Kyungjae;Ha Nam-Joo
    • Archives of Pharmacal Research
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    • 제28권6호
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    • pp.660-666
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    • 2005
  • The intestinal microbiota are important to the host with regard to resistance they impart against bacterial infections and their involvement in mediating metabolic functions. Lactic acid producing bacteria such as Lactobacillus play an important physiological role in these matters. The aim of the present study was to isolate Lactobacillus sp. that inhibits enteric pathogens. Initially, 17 isolates from healthy Koreans were collected on Lactobacillus selective medium. Resistance of the isolates to antibiotics including rifampicin, streptomycin, clindamycin and vancomycin was measured. One of the isolate was identified as Lactobacillus ruminus on the basis of bacterial cell morphology, cultural characteristic and biochemical characteristics, 16S rRNA sequence analysis and PCR-RAPD. Antimicrobial activity of the bacterium against Vancomycin Intermediate Resistant Staphylococcus aureus (VISA) and Vancomycin-Resistant Enterococci (VRE) was measured. About $10^4$ cells of VISA or VRE were mixed with 1, 5, and 9 mL of L. ruminus SPM 0211 and the final volume was adjusted to 10 mL with brain heart infusion (BHI) broth. The cell suspension was incubated for 3, 6, 9, and 24 h, serially diluted and then plated on BHI agar plates. As numbers of L. ruminus SPM 0211 were increased, viable cell count of VISA and VRE decreased. The strongest antimicrobial activity of SPM 0211 was observed after 9 h incubation in any mixture, almost completely inhibiting the growth of these two bacteria. The results suggest that the freshly isolated L. ruminus SPM 0211 may be used as a pro-biotic microbe that prevents the colonization of enteric pathogens and can thereby promote good gastrointestinal health.

Pseudomonas sp. HJ에 의한 포도당으로부터 Poly(3-Hydroxybutyric-Co-3-Hydroxyvaleric) Acid의 생합성에 대한 보조기질의 영향 (Effect of Cosubstrate on tile Production of Poly(3-Hydroxybutyric-Co-3-Hydroxyvaleric) Acid from Glucose by Pseudomonas sp, HJ)

  • 손홍주;고명선이상준
    • KSBB Journal
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    • 제11권5호
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    • pp.586-592
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    • 1996
  • 본 연구에서는 단일 탄소원인 포도당으로부터 P PHA를 생산하는 콩시균 Pseudomonas sp. HJ를 보다 정확하게 동정하기 위하여 수치분류를 실시하 였으며, 또한 보조기질을 첨가하였을때의 PHB/HV 합성 양상을 검토하였다. Pseudomonas sp. HJ는 각 대조균주들중에서 Pseudomonas picketti와 가장 유 사도가 높은 것으로 나타났다(78.8%). 그러나 수치 분류에 서 통일 균종에 포함될 수 있는 유샤도는 85 % 정도인 것으로 얄려져 있으므로, 본 공시균 P Pseudomonas sp. HJ는 Pseudomonas 속의 다른 종 (species)이거나 새로운 종(species)일 가능성이 높 았다. 보조기 질로 hexadecane과 propionic acid를 각각 0.4%씩 첨가했을 경우, 농도증가에 비례하여 H HV 함량은 점차 증가하여 각각 60.8 mol%, 44.9 mol%까지 축적되었다. PHB/HV내의 HV 함량은 p propionic acid냐 hexadecane의 농도를 달리하여 첨가함으로써 조절할 수 있었다. 보조기질로 propl- onic acid 0.2 %를 대수기 중기 인 배양 18시간에 첨 가하였을때 균체 생육이 가장 우수하였으나, HV monomer의 비율은 대수기 말기인 배양 24시간대에 첨가하였을때 49.6 mol%로 가장 높았다.

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Screening of Cholesterol-lowering Bifidobacterium from Guizhou Xiang Pigs, and Evaluation of Its Tolerance to Oxygen, Acid, and Bile

  • Zhang, Rujiao;He, Laping;Zhang, Ling;Li, Cuiqin;Zhu, Qiujin
    • 한국축산식품학회지
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    • 제36권1호
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    • pp.37-43
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    • 2016
  • Cardiovascular and cerebrovascular diseases seriously harm human health, and Bifidobacterium is the most beneficial probiotic in the gastrointestinal tract of humans. This work aimed to screen cholesterol-lowering Bifidobacterium from Guizhou Xiang Pig and evaluate its tolerance to oxygen, acid, and bile. Twenty-seven aerotolerant strains with similar colony to Bifidobacterium were isolated through incubation at 37℃ in 20% (v/v) CO2-80% (v/v) atmospheric air by using Mupirocin lithium modified MRS agar medium, modified PTYG with added CaCO3, and modified PTYG supplemented with X-gal. Ten strains with cholesterol-lowering rates above 20% (w/w) were used for further screening. The selected strains’ tolerance to acid and bile was then determined. A combination of colony and cell morphology, physiological, and biochemical experiments, as well as 16S rRNA gene-sequence analysis, was performed. Results suggested that BZ25 with excellent characteristics of high cholesterol-removal rate of 36.32% (w/w), as well as tolerance to acid and bile, was identified as Bifidobacterium animalis subsp. lactis. To further evaluate Bifidobacterium BZ25’s growth characteristic and tolerance to oxygen, culture experiments were performed in liquid medium and an agar plate. Findings suggested that BZ25 grew well both in environmental 20% (v/v) CO2-80% (v/v) atmospheric air and in 100% atmospheric air because BZ25 reached an absorbance of 1.185 at 600 nm in 100% atmospheric air. Moreover, BZ25 was aerotolerant and can grow in an agar medium under the environmental condition of 100% atmospheric air. This study can lay a preliminary foundation for the potential industrial applications of BZ25.

Ginsenoside F1 Modulates Cellular Responses of Skin Melanoma Cells

  • Yoo, Dae-Sung;Rho, Ho-Sik;Lee, Yong-Gyu;Yeom, Myung-Hun;Kim, Duck-Hee;Lee, Sang-Jin;Hong, Sung-Youl;Lee, Jae-Hwi;Cho, Jae-Youl
    • Journal of Ginseng Research
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    • 제35권1호
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    • pp.86-91
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    • 2011
  • Ginsenoside (G)-F1 is an enzymatic metabolite generated from G-Rg1. Although this metabolite has been reported to suppress platelet aggregation and to reduce gap junction-mediated intercellular communication, the modulatory activity of G-F1 on the functional role of skin-derived cells has not yet been elucidated. In this study, we evaluated the regulatory role of G-F1 on the cellular responses of B16 melanoma cells. G-F1 strongly suppressed the proliferation of B16 cells up to 60% at 200 ${\mu}g/mL$, while only diminishing the viability of HEK293 cells up to 30%. Furthermore, G-F1 remarkably induced morphological change and clustering of B16 melanoma cells. The melanin production of B16 cells was also significantly blocked by G-F1 up to 70%. Interestingly, intracellular signaling events involved in cell proliferation, migration, and morphological change were up-regulated at 1 h incubation but down-regulated at 12 h. Therefore, our results suggest that G-F1 can be applied as a novel anti-skin cancer drug with anti-proliferative and anti-migration features.

2015년 국내 중동호흡기증후군 유행 양상 (Middle East Respiratory Syndrome Outbreak in Korea, 2015)

  • 최은화
    • Pediatric Infection and Vaccine
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    • 제22권3호
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    • pp.131-135
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    • 2015
  • Since April 2012, more than 1,600 laboratory-confirmed human infections with Middle East Respiratory Syndrome Coronavirus (MERS-CoV) have been reported, occurring primarily in countries in the Arabian Peninsula; the majority in Saudi Arabia. The MERS outbreak in Korea, which began in May 2015 through the importation of a single case who had recently traveled to Bahrain, the United Arab Emirates, Saudi Arabia, and Qatar. As of November 28th, 186 secondary and tertiary cases had been reported; 38 deaths, mainly associated with underlying chronic illnesses, were reported. One case was exported to China and has been recorded as the first MERS case in China. Thirty-seven confirmed cases were associated with the index case, who was hospitalized from May 15 to May 17. Emergency room at one of the nation's largest hospitals had been affected by hospital-to-hospital and intra-hospital transmissions of MERS-CoV, resulting in an outbreak of 90 infected patients. The vast majority of 186 confirmed cases are linked to a single transmission chain associated with health facilities. The median age of patients is 55 years, with a range of 16 to 87 years. The majority (61%) of patients are men. Twenty-five (14%) of the cases involve healthcare workers. The overall median incubation period was six days, but it was four days for secondary cases and six days for tertiary cases. There has been no evidence of airborne transmission and sustained human-to-human transmission in communities. Intensified public health measures, including contact tracing, quarantine and isolation of all contacts and suspected cases, and infection prevention and control have brought the MERS-CoV under control in Korea. Since 4 July no new cases have been reported.

Effects of Hydroxychloroquine Co-administered with Chemotherapeutic Agents on Malignant Glioma Cell Lines : in vitro Study

  • Park, Yong-Sook;Choi, Jae-Young;Chang, Jong-Hee;Park, Yong-Gou;Chang, Jin-Woo
    • Journal of Korean Neurosurgical Society
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    • 제38권1호
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    • pp.47-53
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    • 2005
  • Objective : Anti-malaria drugs may modulate tumor resistance to chemotherapeutic agents, but it has not been proven effective in the treatment of malignant gliomas. The aim of this study was to determine whether adequate pre-clinical data on co-administration of chemotherapeutic agents with anti-malaria drugs on malignant cell lines could be obtained that would warrant its further potential consideration for use in a clinical trial for malignant gliomas. Methods : Two malignant glioma cell lines [U87MG, T98G] were treated with chemotherapeutic agents alone or with anti-malaria drugs. Cells were incubated with drugs for 4 days. Following the 4-day incubation, drug sensitivity assays were performed using 3-[4,5-dimethyl-2-thiazol-2-yl] 2,5-diphenyltetrazolium bromide [MTT] assay following optimization of experimental conditions for each cell lines and cell viability was calculated. Results : In all of four chemotherapeutic agents[doxorubicin. vincrisitne, nimustine, and cisplatin], the cell viability was found to be markedly decreased when hydroxychloroquine was co-administered on both U87MG and T98G cell lines. The two way analysis of variance[ANOVA] yielded a statistically significant two-sided p-value of 0.0033[doxorubicin], 0.0005[vincrisitne], 0.0007[nimustine], and 0.0003[cisplatin] on U87MG cell lines and 0.0006[doxorubicin], 0.0421[vincrisitne], 0.0317[nimustine], and 0.0001[cisplatin] on T98G cell lines, respectively. However, treatment with chloroquine and primaquine did not induce a decrease in cell viability on both U87MG and T98G cell lines. Conclusion : Our data support further consideration of the use of hydroxychloroquine prior to systemic chemotherapy to maximize its tumoricidal effect for patients with malignant gliomas.