• Microbiology and Biotechnology Letters
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• v.49 no.1
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• pp.75-87
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• 2021

Type I Interferons (IFNα) are known for their role as biological anticancer agents owing to their cell-apoptosis inducing properties. Development of an appropriate, cost-effective host expression system is crucial for meeting the increasing demand for proteins. Therefore, this study aims to develop codon-optimized IFNα-2b in L. lactis NZ3900. These cells express extracellular protein using the NICE system and Usp₄₅ signal peptide. To validate the mature form of the expressed protein, the recombinant IFNα-2b was screened in a human colorectal cancer cell line using the cytotoxicity assay. The IFNα-2b was successfully cloned into the pNZ8148 vector, thereby generating recombinant L. lactis pNZ8148-SP_Usp45-IFNα-2b. The computational analysis of codon-optimized IFNα-2b revealed no mutation and amino acid changes; additionally, the codon-optimized IFNα-2b showed 100% similarity with native human IFNα-2b, in the BLAST analysis. The partial size exclusion chromatography (SEC) of extracellular protein yielded a 19 kDa protein, which was further confirmed by its positive binding to anti-IFNα-2b in the western blot analysis. The crude protein and SEC-purified partial fraction showed IC₅₀ values of 33.22 ㎍/ml and 127.2 ㎍/ml, respectively, which indicated better activity than the metabolites of L. lactis NZ3900 (231.8 ㎍/ml). These values were also comparable with those of the regular anticancer drug tamoxifen (105.5 ㎍/ml). These results demonstrated L. lactis as a promising host system that functions by utilizing the pNZ8148 NICE system. Meanwhile, codon-optimized usage of the inserted gene increased the optimal protein expression levels, which could be beneficial for its large-scale production. Taken together, the recombinant L. lactis IFNα-2b is a potential alternative treatment for colorectal cancer. Furthermore, its activity was analyzed in the WiDr cell line, to assess its colorectal anticancer activities in vivo.

• Journal of Life Science
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• v.33 no.1
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• pp.34-42
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• 2023

In a signal transduction network, from the perception of stress signals to stress-responsive gene ex- pression, binding of various transcription factors to cis-acting elements in stress-responsive promoters coordinate the adaptation of plants to abiotic stresses. Among the AP2/ERF transcription factor family genes, group VII ERF genes, such as RAP2.12, RAP2.2, RAP2.3, AtERF73/HRE1, and AtERF71/ HRE2, are known to be involved in the response to hypoxia stress in Arabidopsis. In this study, we dissected the HRE1 promoter to identify hypoxia-responsive region(s). The 1,000 bp upstream promoter region of HRE1 showed increased promoter activity in Arabidopsis protoplasts and transgenic plants under hypoxia conditions. Analysis of the promoter deletion series of HRE1, including 1,000 bp, 800 bp, 600 bp, 400 bp, 200 bp, 100 bp, and 50 bp upstream promoter regions, using firefly luciferase and GUS as reporter genes indicated that the -200 to -100 region of the HRE1 promoter is responsible for the transcriptional activation of HRE1 in response to hypoxia. In addition, we identified two putative hypoxia-responsive cis-acting elements, the ERF-binding site and DOF-binding site, in the -200 to -100 region of the HRE1 promoter, suggesting that the expression of HRE1 might be regulated via the ERF transcription factor(s) and/or DOF transcription factor(s). Collectively, our results suggest that HRE1 contains hypoxia-responsive cis-acting elements in the -200 to -100 region of its promoter.

• Journal of Applied Biological Chemistry
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• v.65 no.1
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• pp.57-62
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• 2022

Excessive activation and aggregation of platelets is a major cause of cardiovascular disease. Therefore, inhibition of platelet activation and aggregation is considered an attractive therapeutic target in preventing and treating cardiovascular diseases. In particular, strong platelet activation and aggregation by collagen secreted from the vascular endothelium are characteristic of vascular diseases. Artesunate is a compound extracted from the plant roots of Artemisia or Scopolia species, and has been reported to be effective in anticancer and Alzheimer's disease fields. However, the effect and mechanism of artesunate on collagen-induced platelet activation and aggregation have not been elucidated. In this study, the effect of artesunate on collagen-induced human platelet aggregation was confirmed and the mechanism of action of artesunate was clarified. Artesunate inhibited the phosphorylation of PI3K/Akt and Mitogen-activated protein kinases, which are phosphoproteins that are known to act in the signal transduction process when platelets are activated. In addition, artesunate decreased TXA₂ production and decreased granule secretion in platelets such as ATP and serotonin release. As a result, artesunate strongly inhibited platelet aggregation induced by collagen, a strong aggregation inducer secreted from vascular endothelial cells, with an IC₅₀ of 106.41 µM. These results suggest that artesunate has value as an effective antithrombotic agent for inhibiting the activation and aggregation of human platelets through vascular injury.

• Science of Emotion and Sensibility
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• v.25 no.4
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• pp.45-52
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• 2022

Recently, interest in and demand for sensors that recognize physical activity and their products are increasing. In particular, the development of wearable materials that are flexible, stretchable, and able to detect the user's biological signals is drawing attention. In this study, an experiment was conducted to improve the dip-coating efficiency of a single-walled carbon nanotube dispersion solution after fine holes were made in a hydrophobic material with a micro needle. In this study, dip-coating was performed with a material that was not penetrated, and comparative analysis was performed. The electrical conductivity of the sensor was measured when the sensor was stretched using a strain universal testing machine (Dacell Co. Ltd., Seoul, Korea) and a multimeter (Keysight Technologies, Santa Rosa, CA, USA) was used to measure resistance. It was found that the electrical conductivity of a sensor that was subjected to needling was at least 16 times better than that of a sensor that was not. In addition, the gauge factor was excellent, relative to the initial resistance of the sensor, so good performance as a sensor could be confirmed. Here, the dip-coating efficiency of hydrophobic materials, which have superior physical properties to hydrophilic materials but are not suitable due to their high surface tension, can be adopted to more effectively detect body movements and manufacture sensors with excellent durability and usability.

• Journal of Life Science
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• v.33 no.11
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• pp.956-965
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• 2023

In plants, there are many CCCH zinc finger proteins consisting of three cysteine residues and one histidine residue, which bind to zinc ions with finger configuration. CCCH-type zinc finger proteins are divided into tandem CCCH-type zinc finger (TZF) and non-TZF proteins: TZF proteins contain exactly two tandem CCCH-type zinc finger motifs whereas non-TZF proteins have fewer or greater than two CCCH-type zinc finger motifs. The functions of TZF genes, especially plant-specific RR-TZF genes, have been well studied in several plants, whereas the functional roles of non-TZF genes have not been adequately researched compared to TZF genes. Many non-TZF genes have been identified as being involved in the responses to biotic and abiotic stresses, such as pathogen, high salt, drought, cold, heat, and oxidative stresses. Some non-TZF proteins bind to RNA and are involved in the post-transcriptional regulation of stress-responsive genes in the cytoplasm. In addition, other non-TZF proteins act as transcriptional activators or repressors that regulate the expression of stress-responsive genes in the nucleus. Despite these studies, stress signal transduction and upstream and downstream genes of non-TZF genes have not been sufficiently researched, suggesting that additional studies of the functions of non-TZF genes' functions in plants' stress responses are needed. In this review, we describe non-TZF genes involved in biotic abiotic stress responses in plants and their molecular functions.

• Biomolecules & Therapeutics
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• v.32 no.5
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• pp.546-555
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• 2024

Aromadendrin is a phenolic compound with various biological effects such as anti-inflammatory properties. However, its protective effects against acute lung injury (ALI) remain unclear. Therefore, this study aimed to explore the ameliorative effects of aromadendrin in an experimental model of lipopolysaccharide (LPS)-induced ALI. In vitro analysis revealed a notable increase in the levels of cytokine/chemokine formation, nuclear factor kappa B (NF-κB) activation, and myeloid differentiation primary response 88 (MyD88)/toll-like receptor (TLR4) expression in LPS-stimulated BEAS-2B lung epithelial cell lines that was ameliorated by aromadendrin pretreatment. In LPS-induced ALI mice, the remarkable upregulation of immune cells and IL-1β/IL-6/TNF-α levels in the bronchoalveolar lavage fluid and inducible nitric oxide synthase/cyclooxygenase-2/CD68 expression in lung was decreased by the oral administration of aromadendrin. Histological analysis revealed the presence of cells in the lungs of ALI mice, which was alleviated by aromadendrin. In addition, aromadendrin ameliorated lung edema. This in vivo effect of aromadendrin was accompanied by its inhibitory effect on LPS-induced NF-κB activation, MyD88/TLR4 expression, and signal transducer and activator of transcription 3 activation. Furthermore, aromadendrin increased the expression of heme oxygenase-1/ NAD(P)H quinone dehydrogenase 1 in the lungs of ALI mice. In summary, the in vitro and in vivo studies demonstrated that aromadendrin ameliorated endotoxin-induced pulmonary inflammation by suppressing cytokine formation and NF-κB activation, suggesting that aromadendrin could be a useful adjuvant in the treatment of ALI.

• Journal of Biomedical Engineering Research
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• v.23 no.1
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• pp.49-60
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• 2002

Receive dynamic focusing with an array transducer can provide near optimum resolution only in the vicinity of transmit focal depth. A customary method to increase the depth of field is to combine several beams with different focal depths, with an accompanying decrease in the frame rate. In this Paper. we Present a simultaneous multiple transmit focusing method in which chirp signals focused at different depths are transmitted at the same time. These chirp signals are mutually orthogonal in a sense that the autocorrelation function of each signal has a narrow mainlobe width and low sidelobe levels. and the crossorelation function of any Pair of the signals has values smaller than the sidelobe levels of each autocorrelation function. This means that each chirp signal can be separated from the combined received signals and compressed into a short pulse. which is then individually focused on a separate receive beamformer. Next. the individually focused beams are combined to form a frame of image. Theoretically, any two chirp signals defined over two nonoverlapped frequency bands are mutually orthogonal In the present work. however, a tractional overlap of adjacent frequency bands is permitted to design more chirp signals within a given transducer bandwidth. The elevation of the rosscorrelation values due to the frequency overlap could be reduced by alternating the direction of frequency sweep of the adjacent chirp signals We also observe that the Proposed method provides better images when the low frequency chirp is focused at a near Point and the high frequency chirp at a far point along the depth. better lateral resolution is obtained at the far field with reasonable SNR due to the SNR gain in Pulse compression Imaging .

• Radiation Oncology Journal
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• v.12 no.3
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• pp.271-284
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• 1994

Purpose : Phospholipase C(PLC) isozymes play significant roles in transmembrane signal transduction. PLC-${\gamma}1$ acts as the intracellular effector in signal transduction for cellular proliferation and differentiation. Ras oncoprotein is also involved in cell growth. We determined the biological significance of PLC and ras oncoprotein in regeneration following radiation and the effect of different modes of administration of 5-FU. Materials and Methods : To determine the effect of the administration mode of 5-FU on the regeneration of intestinal mucosa of rats following radiation, we compared the expression of PLC and ras oncoprotein in six groups. Group I had no treatment. Group II received radiation(8 Gy) only. Group III received radiation(8 Gy) and 5-FU(150mg/kg) continuous intravenous (iv) infusion for 12 hours. Group IV received radiation(8 Gy) and 5-FU(750mg/kg) iv bolus injection. Group V received only 5-FU(150mg/kg) continuous iv infusion for 12 hours, Group VI received only 5-FU (150mg/kg) iv bolus injection. Through immunoblotting and immunohistochemistry, we examined the expression of PLC and ras oncoprotein in rat jejunum at 96 hours after radiation or 5-FU administration and at 120 hours after radiation and 5-FU adminstration. We also investigated the histological findings using hematoxylin and eosin stain. Results : In the immunohistochemistry study, PLC-${\gamma}1$ expression was the highest in group III followed by groups II and VI in that order and was weakly positive in groups V and VI. PLC-${\gamma}1$ was hardly detected in the control group. The expression of ras oncoprotein was the same as the PLC-${\gamma}1$ expression for all groups. These results were confirmed by the histological findings regarding the mucosal regeneration. In the immunoblotting analysis, PLC-${\gamma}1$ expression was the highest in group III followed by group IV and II in that order. This difference between the immunoblotting and immunohistochemistry study was due to the high expression of PLC-${\gamma}1$ on the damaged surface epithelium rather than to its expression in the regeneration region as observed in the immunohistochemistry study for group IV. The expression of PLC-${\delta}1$ was positive only in group V and VI, which received both radiation and 5-FU, and the expression of PLC-${\beta}1$ was negligible for all groups. Conclusion : These results suggest that PLC-${\gamma}1$ mediated signal transduetion and ras oncoprotein may have a significant role in mucosal regeneration after radiation, and that continuous iv infusion of 5-FU may induce active regeneration in intestinal mucosa following radiation. In addition, the expression of PLC-${\delta}1$ in combined group of radiation and 5-FU implies that PLC-${\delta}1$ may be involved in signal transduction mediated by concerted action between radiation and 5-FU.

• International Journal of Industrial Entomology and Biomaterials
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• v.12 no.2
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• pp.57-61
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• 2006

Suppressor of cytokine signaling (SOCS) is known to be as a negative feedback regulator in Janus kinase signal transducer and activator of transcription signaling. Highly conserved SOCS box domain was cloned from a Korean malaria vector, Anopheles sinensis. Sequence analysis indicates that it has identity to Anopheles gambiae (96%), Aedes aegypti (94%), Drosophila melanogaster (78%), Mus musculus (72%) and Homo sapiens (72%), respectively. Tissue specificity RT-PCR demonstrated that the expression level of AsSOCS transcript was high at abdomen, midgut, and ovary, whereas developmental expression patterns showed that the level of AsSOCS was high at egg, early pupae, and adult female. On the other hand, RT-PCR analysis after bacterial challenge showed that SOCS mRNA was strongly induced in larvae. In addition, it was also induced by various immune elicitors such as lipoteicoic acid, CpG-DNA, and laminarin. It seems that AsSOCS, repressor of JAK-STAT pathway, is highly conserved in mosquito, and may play an important role in mosquito innate immune response.

• Journal of the Korean Electrochemical Society
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• v.11 no.3
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• pp.176-183
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• 2008

Redox complexes to transport electrodes from bioreactors to electrodes are very important part in electrochemical biosensor industry. A novel osmium redox complexes were synthesized by the coordinating pyridine group having different functional group at 4-position with osmium metal. Newly synthesized osmium complexes are described as ${[Os(dme-bpy)}_2{(ap-im)Cl]}^{+/2+}$, ${[Os(dme-bpy)}_2{(ap-im)Cl]}^{+/2+}$, ${[Os(dmo-bpy)}_2{(ap-im)Cl]}^{+/2+}$, ${[Os(dcl-bpy)}_2{(ap-im)Cl]}^{+/2+}$. We have been studied the electrochemical characteristics of these osmium complex with electrochemical techniques such as cyclic voltammetry and chronoamperommetry. Osmium redox complexes were immobilized on the screen printed carbon electrode(SPE) with deposited gold nanoparticles. The electrical signal converts the osmium redox films into an electrocatalyst for glucose oxidation. Each catalytic currents were related with the potentials of osmium complexes.