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Development of diagnostic kit(Test-MT) for the microplate latex agglutination test of toxoplasmosis in animal (Latex 응집반응을 이용한 동물의 톡소플라즈마병 진단용 kit 개발에 관한 연구)

  • Suh, Myung-deuk;Joo, Hoo-don;Maass, Daivd
    • Korean Journal of Veterinary Research
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    • v.35 no.3
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    • pp.583-593
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    • 1995
  • The present study was conducted to develop a toxoplasma latex agglutination test antigen(Test-MT) and evaluate the toxoplasma latex agglutination(LA) test using a newly-made "Test-MT kit" by comparing with the Toxo-MT kit(Eiken chemical co, Tokyo). Also, the specifity and sensitivity test were made by comparing with IFA test and IgG-ELISA. Tachyzoite suspensions of Toxoplasma gondii(RH strain) were ultracentrifuged for 30min at $60,000{\times}g(4^{\circ}C)$ and the supernatant was used as a water-lysate antigen. Polystyrene latex particles of $1.0{\mu}m$ in diameter(Polyscience co) were used for the preparation of sensitized latex-antigen supension(Test-MT). The frequency distribution of LA titers in Test-MT showed two peaks at <1:32 and 1:128. The borderline titer for positive test in Test-MT was determined to be 1:64. But the frequency distribution of LA tites in Toxo-MT showed two peaks at <1:16 and 1:64. The positive borderline was determined to be 1:32. Agreement of reactions between Test-MT and Toxo-MT kit by LA test was shown 92.5% in bovine sera and 97.0% in swine sera, respectively. From the results obtained here it was determined that the sensitized latex-antigen, Test-MT kit, for the microtiter agglutination test prepared as same as by the procedure described in the previous paper(Suh and Lee, 1993) was useful as a highly specific, sensitive and stable immunotiteration reagent for serodiagnosis of toxoplasma infection in animal sera.

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Resistance to Toxoplasma gondii Infection in Mice Treated with Silk Protein by Enhanced Immune Responses

  • Moon, Joung-Ho;Pyo, Kyoung-Ho;Jung, Bong-Kwang;Chun, Hyang-Sook;Chai, Jong-Yil;Shin, Eun-Hee
    • Parasites, Hosts and Diseases
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    • v.49 no.3
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    • pp.303-308
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    • 2011
  • This study investigated whether elevated host immune capacity can inhibit T. gondii infection. For this purpose, we used silk protein extracted from Bombyx mori cocoons as a natural supplement to augment immune capacity. After silk protein administration to BALB/c mice for 6 weeks, ratios of T lymphocytes ($CD4^+$ and $CD8^+$ T-cells) and splenocyte proliferative capacities in response to Con A or T. gondii lysate antigen (TLA) were increased. Of various cytokines, which regulate immune systems, Th1 cytokines, such as IFN-${\gamma}$, IL-2, and IL-12, were obviously increased in splenocyte primary cell cultures. Furthermore, the survival of T. gondii (RH strain)-infected mice increased from 2 days to 5 or more days. In a state of immunosuppression induced by methylprednisolone acetate, silk protein-administered mice were resistant to reduction in T-lymphocyte ($CD4^+$ and $CD8^+$ T-cells) numbers and the splenocyte proliferative capacity induced by Con A or TLA with a statistical significance. Taken together, our results suggest that silk protein augments immune capacity in mice and the increased cellular immunity by silk protein administration increases host protection against acute T. gondii infection.

Anti-inflammatory Function of the Sophora japonica Extract Rutin: The Inhibitory Effect of Rutin of Korean Sophora japonica on the Productions of NO and TNF-alpha from Mouse Peritoneal Macrophages (괴화 추출물 루틴의 소염작용: 쥐의 복강대식세포로부터 NO와 TNF-alpha 생산에 있어서 괴화 루틴에 의한 억제효과)

  • Lee, Mu-Hong;Jeong, Jae-Hyun;Jeong, Myeong-Soo;Chang, Sung-Ho;Her, Erk
    • Korean Journal of Medicinal Crop Science
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    • v.18 no.2
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    • pp.105-112
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    • 2010
  • Korean Sophora japonica has been found to posses an anti-inflammatory activity. In this study, Korean Sophora japonica extract, rutin, was used to know whether rutin inhibits to produce inflammatory mediators NO and TNF-$\alpha$ from the mouse peritoneal macrophages that were treated an inflammatory agent LPS. The rutin-1 hr pretreated macrophages were incubated with LPS for 0.5~5 hrs, and then collected the supernatant and the cell lysate for measurements of the level of iNOS, NO, TNF-$\alpha$ mRNA, TNF-$\alpha$, and p-NF-${\kappa}B$. Minimal and maximal effective doses of the rutin on them were 1 and $100{\mu}g/ml$, respectively. The maximal effective dose of rutin certainly inhibted the productions of iNOS, NO, TNF-$\alpha$ mRNA, TNF-$\alpha$and p-NF-${\kappa}B$ from the LPS-treated macrophages (p<0.0001). Its $ED_{50}$ for inhibition of TNF-$\alpha$ mRNA and p-NF-${\kappa}B$ was $5{\mu}g/m{\ell}$, and for iNOS, NO, and TNF-$\alpha$ was $10{\mu}g/m{\ell}$. The rutin did not have any cytotoxic effect. As the results, the Sophora japonica rutin could be a good candidate for an anti-inflammatory action.

Comparison of Seropositivity to Trichomonas vaginalis between Men with Prostatic Tumor and Normal Men

  • Kim, Jung-Hyun;Moon, Hong-Sang;Kim, Kyu-Shik;Hwang, Hwan-Sik;Ryu, Jae-Sook;Park, Sung-Yul
    • Parasites, Hosts and Diseases
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    • v.57 no.1
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    • pp.21-25
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    • 2019
  • Trichomoniasis is the most common curable sexually-transmitted infection. Most Trichomonas vaginalis-infected men are asymptomatic and can remain undiagnosed and untreated, and this has been thought to result in chronic persistent prostatic infection. Chronic inflammation is regarded as the major factor in the pathogenesis and progression of benign prostatic hyperplasia (BPH) and prostatic cancer (PCa). The aim of this study is to identify seropositivity to T. vaginalis in men with prostate tumors (BPH or PCa) visited to Hanyang University Hospital. A total of 183 men were enrolled between October 2013 and November 2014. They consisted of 139 with BPH (mean age: $64.0{\pm}0.07$) and 44 with prostate cancer (mean age: $73.3{\pm}0.18$). We carried out ELISA to identify the seropositivity to T. vaginalis. Mixed lysate antigen extracted from 8 strains of T. vaginalis was used in the ELISA. Also 58 male outpatients visited to Health Promotion Center in Hanyang University Hospital were evaluated for comparing group. As a results, seropositivity to T. vaginalis in patients with prostatic diseases was 19.7% (BPH: 18.7%, PCa: 22.7%) and it was significantly higher than the 1.7% of the comparing healthy group (P=0.001). Therefore, prostatic tumor showed higher seropositivity against T. vaginalis than normal men. As far as we know, this is the first report about seroprevalence in prostatic tumor in Korea.

Anti-Endotoxin 9-Meric Peptide with Therapeutic Potential for the Treatment of Endotoxemia

  • Krishnan, Manigandan;Choi, Joonhyeok;Choi, Sungjae;Kim, Yangmee
    • Journal of Microbiology and Biotechnology
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    • v.31 no.1
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    • pp.25-32
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    • 2021
  • Inflammatory reactions activated by lipopolysaccharide (LPS) of gram-negative bacteria can lead to severe septic shock. With the recent emergence of multidrug-resistant gram-negative bacteria and a lack of efficient ways to treat resulting infections, there is a need to develop novel anti-endotoxin agents. Antimicrobial peptides have been noticed as potential therapeutic molecules for bacterial infection and as candidates for new antibiotic drugs. We previously designed the 9-meric antimicrobial peptide Pro9-3 and it showed high antimicrobial activity against gram-negative bacteria. Here, to further examine its potency as an anti-endotoxin agent, we examined the anti-endotoxin activities of Pro9-3 and elucidated its mechanism of action. We performed a dye-leakage experiment and BODIPY-TR cadaverine and limulus amebocyte lysate assays for Pro9-3 as well as its lysine-substituted analogue and their enantiomers. The results confirmed that Pro9-3 targets the bacterial membrane and the arginine residues play key roles in its antimicrobial activity. Pro9-3 showed excellent LPS-neutralizing activity and LPS-binding properties, which were superior to those of other peptides. Saturation transfer difference-nuclear magnetic resonance experiments to explore the interaction between LPS and Pro9-3 revealed that Trp3 and Tlr7 in Pro9-3 are critical for attracting Pro9-3 to the LPS in the gram-negative bacterial membrane. Moreover, the anti-septic effect of Pro9-3 in vivo was investigated using an LPS-induced endotoxemia mouse model, demonstrating its dual activities: antibacterial activity against gram-negative bacteria and immunosuppressive effect preventing LPS-induced endotoxemia. Collectively, these results confirmed the therapeutic potential of Pro9-3 against infection of gram-negative bacteria.

Multifunctional Probiotic and Functional Properties of Lactiplantibacillus plantarum LRCC5314, Isolated from Kimchi

  • Yoon, Seokmin;Cho, Hyeokjun;Nam, Yohan;Park, Miri;Lim, Ahyoung;Kim, Jong-Hwa;Park, Jaewoong;Kim, Wonyong
    • Journal of Microbiology and Biotechnology
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    • v.32 no.1
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    • pp.72-80
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    • 2022
  • In this study, the survival capacity (acid and bile salt tolerance, and adhesion to gut epithelial cells) and probiotic properties (enzyme activity-inhibition and anti-inflammatory activities, inhibition of adipogenesis, and stress hormone level reduction) of Lactiplantibacillus plantarum LRCC5314, isolated from kimchi (Korean traditional fermented cabbage), were investigated. LRCC5314 exhibited very stable survival at ph 2.0 and in 0.2% bile acid with 89.9% adhesion to Caco-2 intestinal epithelial cells after treatment for 2 h. LRCC5314 also inhibited the activities of α-amylase and α-glucosidase, which are involved in elevating postprandial blood glucose levels, by approximately 72.9% and 51.2%, respectively. Treatment of lipopolysaccharide (LPS)-stimulated RAW 264.7 cells with the LRCC5314 lysate decreased the levels of the inflammatory factors nitric oxide, tumor necrosis factor (TNF-α), interleukin (IL)-1β, and interferon-γ by 88.5%, 49.3%, 97.2%, and 99.8%, respectively, relative to those of the cells treated with LPS alone. LRCC5314 also inhibited adipogenesis in differentiating preadipocytes (3T3-L1 cells), showing a 14.7% decrease in lipid droplet levels and a 74.0% decrease in triglyceride levels, as well as distinct reductions in the mRNA expression levels of adiponectin, FAS, PPAR/γ, C/EBPα, TNF-α, and IL-6. Moreover, LRCC5314 reduced the level of cortisol, a hormone with important effect on stress, by approximately 35.6% in H295R cells. L. plantarum LRCC5314 is identified as a new probiotic with excellent in vitro multifunctional properties. Subsequent in vivo studies may further demonstrate its potential as a functional food or pharmabiotic.

Bioaerosol Exposure and in vitro Activation of Toll-like Receptors in a Norwegian Waste Sorting Plant

  • Eriksen, Elke;Graff, Pal;Pedersen, Ine;Straumfors, Anne;Afanou, Anani K.
    • Safety and Health at Work
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    • v.13 no.1
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    • pp.9-16
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    • 2022
  • Background: The global shift toward greener societies demands new technologies and work operations in the waste-management sector. However, progressive industrial methods do not necessarily consider workers' health. This study characterized workers' exposure to bioaerosols and investigated the bioaerosols' potential to engage the immune system in vitro. Methods: Full shift personal aerosol sampling was conducted over three consecutive days. Dust load was analyzed by gravimetry, fungal and actinobacterial spores were analyzed by scanning electron microscopy, and endotoxin by limulus amebocyte lysate (LAL) assay. In vitro exposure of HEK cells to airborne dust samples was used to investigate the potential of inducing an inflammatory reaction. Results: The total dust exposure level exceeded the recommended occupational exposure limit (OEL) of 5.0 mg/m3 in 3 out of 15 samples. The inhalable endotoxin level exceeded the recommended exposure level by a 7-fold, whereas the fungal spore level exceeded the recommended exposure level by an 11-fold. Actinobacterial spores were identified in 8 out of 14 samples. In vitro experiments revealed significant TLR2 activation in 9 out of 14 samples vs. significant TLR4 activation in all samples. Conclusion: The present study showed that the dust samples contained potentially health-impairing endotoxin, fungi, and actinobacterial levels. Furthermore, the sampled dust contained microbial components capable of inducing TLR activation and thus have the potential to evoke an inflammatory response in exposed individuals.

Discovery of LDD-1075 as a potent FLT3 inhibitor

  • Kyoung Bin Yoon;Hyo Jeong Lee;Hye Jin Chung;Jungeun Lee;Jungil Choi;Jeong Doo Heo;Yong‑Chul Kim;Sun‑Young Han
    • Oncology Letters
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    • v.17 no.5
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    • pp.4735-4741
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    • 2019
  • Fms-like tyrosine kinase 3 (FLT3) is a valuable pharmacological target in the treatment of acute myeloid leukemia (AML). LDD-1075 and LDD-1076 are indirubin derivatives, and LDD-1075 is the ester form of LDD-1076. LDD-1076 exhibited a potent in vitro FLT3 kinase activity inhibition with an IC50 of 7.89 nM, whereas, LDD-1075 demonstrated a relatively weak activity against FLT3 (IC50 of 3.19 µM). In contrast with the results of the FLT3 kinase activity inhibition assay, the LDD-1076 did not affect the growth of the MV4-11 cell line, which harbors the constitutively activated form of the FLT3 mutation. Notably, LDD-1075 exhibited a strong cytotoxic effect against the MV4-11 cells. When LDD-1075 was incubated with the MV4-11 cell lysate, the formation of LDD-1076 was observed. Treatment with LDD-1075 inhibited the FLT3 phosphorylation along with the phosphorylation of the signal transducer and activator of transcription 5 protein, which is a downstream signal transducer of FLT3. Treatment with LDD-1075 induced apoptosis and cell cycle arrest at the G1 phase. The present study demonstrated that the LDD-1076 formed by the bioconversion of LDD-1075 is a potent FLT3 inhibitor with anti-leukemic activity.

Virus-like particles expressing microneme-associated antigen of Plasmodium berghei confer better protection than those expressing apical membrane antigen 1

  • Min-Ju Kim;Ki Back Chu;Keon-Woong Yoon;Hae-Ji Kang;Dong-Hun Lee;Eun-Kyung Moon;Fu-Shi Quan
    • Parasites, Hosts and Diseases
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    • v.62 no.2
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    • pp.193-204
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    • 2024
  • Malaria is a global disease affecting a large portion of the world's population. Although vaccines have recently become available, their efficacies are suboptimal. We generated virus-like particles (VLPs) that expressed either apical membrane antigen 1 (AMA1) or microneme-associated antigen (MIC) of Plasmodium berghei and compared their efficacy in BALB/c mice. We found that immune sera acquired from AMA1 VLP- or MIC VLP-immunized mice specifically interacted with the antigen of choice and the whole P. berghei lysate antigen, indicating that the antibodies were highly parasite-specific. Both VLP vaccines significantly enhanced germinal center B cell frequencies in the inguinal lymph nodes of mice compared with the control, but only the mice that received MIC VLPs showed significantly enhanced CD4+ T cell responses in the blood following P. berghei challenge infection. AMA1 and MIC VLPs significantly suppressed TNF-α and interleukin-10 production but had a negligible effect on interferon-γ. Both VLPs prevented excessive parasitemia buildup in immunized mice, although parasite burden reduction induced by MIC VLPs was slightly more effective than that induced by AMA1. Both VLPs were equally effective at preventing body weight loss. Our findings demonstrated that the MIC VLP was an effective inducer of protection against murine experimental malaria and should be the focus of further development.

Detection of Pathogenic Yersinia Enterocolitica in Drinking Water and Vegetables by Mutiplex-PCR (Multiplex-PCR에 의한 먹는샘물 및 야채류로부터의 병원성 Yersinia enterocolitica의 신속검출)

  • 이택수;박부길;오덕환
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.32 no.1
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    • pp.35-41
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    • 2003
  • The study was conducted to develope a rapid method for the detection of Yersinia enterocolitica in spring water and vegetables via multiplex polymerase chain reaction (PCR) technique using ail, yst, uirF and subgenus-specific Y16S primers. Specificity and sensitivity of multiplex PCR and application of best primers for the detection of Y. enterocolitica from spring water and vegetables were investigeted. Y. enterocolitica ATCC 27729 strains gave 356 bP and 200 bp (Y16S) and 134 bp (yst) bands. but Y. enterocolitica ATCC 9610 and ATCC 23715 strains gave 200 bp and 134 bp bands.In the meanwhile, non-pathogenic Yersinia species, such as Y. frederikseni, Y. inter-media, Y. kristenseni and Y. pseudotuberculosis gave only single 200 bp band, and other bacteria including Escherichia coli O157:H7 ATCC 25392, Shigella dysenteri. Staphylococcu aureus ATCC 25923 and Listeria mo-nocytogenes ATCC 19111 did not show any bands. Among primers, yst and Y16S primer showed the best sensitivity. Seven CFU/mL Y. enterocolitica cells could be detected with yst and Y16S primers and the sensitivity was significantly improved by the further 2nd PCR after 38 cycles of first PCR amplication. Spring water, cabbage and mushroom were inoculated with Y. enterocolitica to determine the sensitivity of multiplex-PCR for the rapid detection of Y. enterocolitica. Multiplex-PCR assay could detect 7 or 70 cells in spring water and vegetables using whole cell lysate with repeating PCR amplication.