• Parasites, Hosts and Diseases
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• v.42 no.4
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• pp.195-200
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• 2004

We investigated the induction of resistance to Haemaphysalis longicornis infestation in rabbits that had been immunized with recombinant H. longicornis P27/30 protein. The success of immunological control methods is dependent upon the use of potential key antigens as tick vaccine candidates. Previously, we cloned a gene encoding 27 kDa and 30 kDa proteins (P27/30) of H. longicornis, and identified P27/30 as a troponin I-like protein. In this study, rabbits that were immunized with recombinant P27/30 expressed in Escherichia coli showed the statistically significant longer feeding duration for larval and adult ticks (P<0.05), low engorgement rates in larval ticks (64.4%), and an apparent reduction in egg weights, which suggest that H. longicornis P27/30 protein is a potential candidate antigen for a tick vaccine. These results demonstrated that the recombinant P27/30 protein might be a useful vaccine candidate antigen for biological control of H. longicornis.

• Molecules and Cells
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• v.39 no.4
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• pp.299-309
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• 2016

We have previously reported the role of miR-217 in anti-cancer drug-resistance. miRNA array and miRNA hybridization analysis predicted miR-30a-3p as a target of miR-217. miR-30a-3p and miR-217 formed a negative feedback loop and regulated the expression of each other. Ago1 immunoprecipitation and co-localization analysis revealed a possible interaction between miR-30a-3p and miR-217. miR-30a-3p conferred resistance to anti-cancer drugs and enhanced the invasion, migration, angiogenic, tumorigenic, and metastatic potential of cancer cells in CAGE-dependent manner. CAGE increased the expression of miR-30a-3p by binding to the promoter sequences of miR-30a-3p, suggesting a positive feedback loop between CAGE and miR-30a-3p. miR-30a-3p decreased the expression of p53, which showed the binding to the promoter sequences of miR-30a-3p and CAGE in anti-cancer drug-sensitive cancer cells. Luciferase activity assays showed that p53 serves as a target of miR-30a. Thus, the miR-30a-3p-CAGE-p53 feedback loop serves as a target for overcoming resistance to anti-cancer drugs.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 2000.04a
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• pp.193-198
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• 2000

PCBs를 분해하는 bphABC유전자를 plasmid vactor pMFB2에 유전자조작한 Pseudomonas putida AC30(pMFB2)를 PCBs에 오염된 연안해역의 해수와 저니로 만든 microcosm에 도입한 결과, 각각 도입 4일과 7일만에 사멸하였다. 그러나, 도입한 P. putida AC30(pMFB2)는 사멸하였지만, 연안해수와 저니 microcosm에서 plasmid pMFB2가 전이한 토착미생물이 검출되었다. 도입한 P. putida AC30(pMFB2)의 생잔실패의 원인을 분석한 결과 공경 0.2$\mu\textrm{m}$의 filter를 통과하는 물질과 생물이 가장 크게 영향을 미치는 것으로 나타났다. 유전자조작 P. putida AC30(pMFB2)의 도입과 bphABC유전자의 토착미생물로의 전이에 따른 토착미생물군집에 미치는 영향을 개체수 변동으로 조사한 결과, 토착미생물 군집에 미치는 영향은 보이지 않았다. P. putida AC30(pMFB2)의 도입에 의한 PCBs의 생분해성을 분석하였다. 그러나, 도입한 유전자조작 균주가 생잔에 실패함으로써 잔류하고 있는 PCBs의 농도변화는 보이지 않았다.

• Parasites, Hosts and Diseases
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• v.34 no.2
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• pp.135-142
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• 1996

Antigenic domain of jai or surface protein (p30) of Toxoplosmc Sondii was analyzed after polymerase chain reaction (PCR) of its gene fragments. Hydrophilic or hydrophobic moiety of amino acid sequences were expressed as glutathione S-transferase (G57) fusion proteins. Fragments of p30 gene were as follows: 737, total p30 open reading frame (ORF) ; S28, total ORF excluding N-terminal signal sequence and C-terminal hydrophobic sequence; Al9, N-terminal 2/3 parts of A28; A19, N-terminal 2/3 of S28; P9, C-terminal 2/3 part of S28; Z9. middle 1/3 of S28; and 29, C-terminal 1/3 of S28. respectively. Primer of each fragment was synthesized to include clamp sequence of EcoR I restriction site. PCR amplified DNA was inserted info GST (26 kDa) expression vector, PGEX-47-1 to transform into Escheri,hia coei (.JM105 strain). G57 fusion proteins were expressed with IPTG induction as 63. 54, 45, 45, 35, 36. and 35 kDa proteins measured by SDS-PAGE. Each fusion protein was confirmed with G57 detection kit. Western blot analysis with the serum of a toxoplasmosis patient revealed antigenicity in proteins expressed by T37. S28, and Al9 but not those by Pl8. X9, Y10, and Z9. Antigenicity of p30 seems to be located either in N-terminal 115 part in the presence of middle 1/3 part or in the oligopeptides between margins of the first and second 1/3 parts.

• BMB Reports
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• v.55 no.9
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• pp.447-452
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• 2022

Neurogenic differentiation 1 (NeuroD1) is an essential transcription factor for neuronal differentiation, maturation, and survival, and is associated with inflammation in lipopolysaccharide (LPS)-induced glial cells; however, the concrete mechanisms are still ambiguous. Therefore, we investigated whether NeuroD1-targeting miRNAs affect inflammation and neuronal apoptosis, as well as the underlying mechanism. First, we confirmed that miR-30a-5p and miR-153-3p, which target NeuroD1, reduced NeuroD1 expression in microglia and astrocytes. In LPS-induced microglia, miR-30a-5p and miR-153-3p suppressed pro-inflammatory cytokines, reactive oxygen species, the phosphorylation of c-Jun N-terminal kinase, extracellular-signal-regulated kinase (ERK), and p38, and the expression of cyclooxygenase and inducible nitric oxide synthase (iNOS) via the NF-κB pathway. Moreover, miR-30a-5p and miR-153-3p inhibited the expression of NOD-like receptor pyrin domain containing 3 (NLRP3) inflammasomes, NLRP3, cleaved caspase-1, and IL-1β, which are involved in the innate immune response. In LPS-induced astrocytes, miR-30a-5p and miR-153-3p reduced ERK phosphorylation and iNOS expression via the STAT-3 pathway. Notably, miR-30a-5p exerted greater anti-inflammatory effects than miR-153-3p. Together, these results indicate that miR-30a-5p and miR-153-3p inhibit MAPK/NF-κB pathway in microglia as well as ERK/STAT-3 pathway in astrocytes to reduce LPS-induced neuronal apoptosis. This study highlights the importance of NeuroD1 in microglia and astrocytes neuroinflammation and suggests that it can be regulated by miR-30a-5p and miR-153-3p.

• Korean Journal of Clinical Laboratory Science
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• v.54 no.3
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• pp.201-208
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• 2022

Neuroinflammation is defined as a neurological inflammation within the brain and the spinal cord. In neuroinflammation, microglia are the tissue-resident macrophages of the central nervous system, which act as the first line of defense against harmful pathogens. Dexmedetomidine (Dex) has an anti-inflammatory effect in many neurological conditions. Additionally, the microRNA-30a-5p (miR-30a-5p) mimic has been proven to be effective in macrophages in inflammatory conditions. This study aimed to investigate the synergistic anti-inflammatory effects of both miR-30a-5p and Dex in lipopolysaccharide (LPS)-induced BV2 cells. This study showed that miR-30a-5p and Dex decreased nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) translocation in LPS-induced BV2 cells. MiR-30a-5p and Dex alleviated tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6), LPS-induced phosphorylation c-Jun N-terminal kinases (JNK), extracellular signal-regulated kinase (ERK) and p38. Also, the expression of the NOD-like receptor pyrin domain containing 3 inflammasome (NLRP3), cleaved caspase-1, and ASC was inhibited. Furthermore, LPS-stimulated nitric oxide (NO) production, inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2) expression were attenuated by Dex and miR-30a-5p. Our results indicate that a combination of Dex and miR-30a-5p, attenuates NF-κB activation, the mitogen-activated protein kinase (MAPK) signaling pathway, and inflammatory mediators involved in LPS-induced inflammation and inhibits the activation of the NLRP3 inflammasome in LPS-activated BV2 cells.

• Journal of Chest Surgery
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• v.40 no.2 s.271
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• pp.114-121
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• 2007

Background: Apoptosis plays a crucial role in carcinogenesis, as well as in development and tissue homeostasis. Terminal deoxyribonucleotidyl transferase mediated neck end labelling (TUNEL) and in situ nick end labelling (ISEL) have been used to investigate the apoptosis in tissues. Since the introduction of the M30 monoclonal antibody to overcome drawbacks of TUNEL and ISEL, the apoptosis in various tumors, with the exception of pulmonary carcinomas, has been studied. In this study, attempts were made to examine the correlation of apoptosis in non-small cell carcinomas, using both M30 and the expression of p53 protein, with the clinicopathological factors. Material and Method: Forty five patients with surgically resected non-small cell carcinomas were included. Immunohistochemical staining with M30 and p53 monoclonal antibody were peformed, and their expressions compared with the clinicopathological features. The overall survival time and recurrence-free survival time were calculated, and the factors influencing the survival time analyzed using a univariate analysis. The effects of the expression stati of M30 and p53 on the risks of cancer related to both death and recurrence were evaluated using a multivariate analysis. Result: The p53 positive group had many more M30 positive cells than the p53 negative group (p53 positive group; $61.7{\pm}26.8$ cells vs. p53 negative group; $45.6{\pm}29.6$ cells, p=0.005) and significantly more p53 positive patients showing at least 10 positive cells (apoptotic index, $Al{\ge}1$) on M30 staining (p53 positive group; 52.4% (11/21) vs. p53 negative group 16,7% (4/24), p=0.025). In the univariate analysis, the survival times in relation to smoking (pack-year), performance status (PS) and Al showed significant differences. The multivariate analysis demonstrated the relative risk (R.R) of cancer death increased almost 7.5-fold (R.R 7.482; 95% Cl $1.886{\sim}29.678$; p=0.004) and the risk of recurrence almost 3,8-fold (R.R 3.795; 95% Cl: $1.184{\sim}12.158$; p=0.025) in the high Al (${\ge}1$) compared to the low Al (<1) group. There was no prognostic effect of p53 expression on the survival time or risk of cancer death and recurrence. Conclusion: In non-small cell lung carcinomas, M30 immunohistochemistry was an excellent method for analyzing apoptosis; the high apoptotic index could be an adverse prognostic predictive factor.

• Korean Journal of Animal Reproduction
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• v.24 no.1
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• pp.115-120
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• 2000

We have investigated the effect of short-term immobilization stress on the serum concentrations of cortisol and DHEAS in BALB/c male mice. Serum cortisol and DHEAS concentrations were measured by radioimmunoassay(RIA). We found there were significantly increased in the cortisol levels in 30 min-stressed group(Ⅰ-30N) compared with control(C) group (p＜0.01), and also increased with significance in 120 min-stressed group(I-120N) compared with C group(p＜0.01). Cortisol concentrations were significantly increased in both 30 min-stressed group(Ⅰ-30T), and 120 min-stressed group(Ⅰ-120T) compared with C group(p＜0.01). The sustained increase of cortisol levels were observed in both SG treated and SG non-treated group. Serum cortisol levels were lower in SG treated group than SG non-treated group with significance(p＜0.01). By contrast, DHEAS levels were slightly decreased without significance in Ⅰ-30N, but significantly decreased in Ⅰ-120N compared with C group(p＜0.01). There were slightly decreased in the DHEAS levels in Ⅰ-30T, but significantly decreased in Ⅰ-120T compared with C group(p＜0.01). However, SG treatment did not induce any significant changes of DHEAS levels in both 30 min and 120 min-stressed group. Though short-term immobilization stress, the continuous decline of DHEAS levels were observed. Therefore, these results show that short-term immobilization stress affects the serum concentrations of cortisol and DHEAS in mice.

• Restorative Dentistry and Endodontics
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• v.30 no.6
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• pp.435-441
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• 2005

The purpose of this study was to evaluate the effect of increasing application time of single bottle adhesives (SBA) to microtensile bond strength (MTBS) of dried dentin. To expose the superficial dentin surfaces, human molars were sectioned perpendicular to the long axis of tooth. $32\%$ phosphoric acid gels were applied for 15s and rinsed. The teeth were randomly assigned to 3 groups ; S group (Single Bond), O group (One-Step), P group (Prime & Bond NT). Each group was divided to 3 subgroups (W: dentin wipe with wet gauge and light cured immediately, D, dentin dried for 30s and light cured immediately, 30: dentin dried for 30s and light cured after applying SBA for 30s). Composite resin was built up on the dentin surface and sectioned to obtain 20 specimens with $1mm^2$ cross sectional area and the MTBS was measured. For Single Bond, the mean MTBS of S-W and S-30 group were higher than that of S-D group statistically (P<0.05). For One-Step, the mean MTBS of O-D group was statistically lower than that of O-W group (P<0.05). For Prime & Bond NT, the mean MTBS of P-30 group was statistically lower than that of P-D group (P<0.05).

• Journal of Oral Medicine and Pain
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• v.30 no.1
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• pp.1-14
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• 2005

The maintenance of good oral health in adults is often hindered by oral malodor and periodontal diseases which are known to be commonly caused by some species of Gram-negative anaerobic bacteria, with low sensitivity to common synthetic antibiotics or antibacterial chemical agents. Therefore the development of a nonharmful natural antibacterial oral rinsing remedy against the causative bacteria is thought to be very important. The purpose of this study is to obtain the basic data for development of a nonharmful natural antibacterial oral rinsing remedy using colloidal silver. The author applied colloidal silver solution with concentration of 10, 30, 50, 80 ppm to some strains in species of Prevotella intermedia, Porphyromonas gingivalis, Fusobaterium nucleatum, and evaluated the effects of colloidal silver on the growth of experimental bacterial strains in aspects of minimal inhibitory concentration (MIC), minimal bactericidal concentration (MBC) and growth pattern after incubation for 24, 48, 72 hours. The obtained results were as follows: MIC of colloidal silver solution against experimental strains was 30 ppm in P. intermedia, 10 or 30 ppm in P. gingivalis, and 30, 50, or 80 ppm in F. nucleatum. And MBC of colloidal silver solution against experimental strains was 30 ppm in P. intermedia, 30 or 50 ppm in P. gingivalis, 30 or 80 ppm in F. nucleatum. Therefore it was concluded that colloidal silver exhibited bacteriostatic or/and bacteriocidal effects against some experimental strain. And the inhibition of growth of experimental strains were markedly or considerably exhibited under 30 ppm$\sim$50 ppm of colloidal silver solution for 48 hours$\sim$72 hours in P. intermedia, 10 ppm$\sim$30 ppm for 24 hours$\sim$48 hours in P. gingivalis, 30 ppm for 24 hours in F. nucleatum. These results indicate that the colloidal silver inhibited effectively the growth of some species of Gram-negative anaerobic bacteria by exhibition of bacteriostatic or/and bacteriocidal effects, and can be used as a possible major ingredient of the nonharmful natural antibacterial oral rinsing remedy to oral malodor and periodontal diseases.