• Title/Summary/Keyword: Synthetic biology

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Potentiality of Oligodeoxynucleotides as An Inducer for Antifungal Peptide in Two Lepidopteran Insects, Bombyx mori and Galleria mellonella

  • Kim, Iksoo;Lee, Young-Shin;Lee, Kwang-Sik;Cha, So-Young;Kang, Pil-Don;Sohn, Bong-Hee;Lee, In-Hee;Jin, Byung-Rae;Hwang, Jae-Sam
    • International Journal of Industrial Entomology and Biomaterials
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    • v.8 no.1
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    • pp.95-99
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    • 2004
  • Synthetic oligodeoxynucleotides (ODNs) containing unmethylated CpG dinucleotides in particular base contexts are known to induce immunity in vertebrate cells. In insect, however, it was recent to find out that ODNs induces insect immunity as other immune inducer such as lipopolysaccharide. However, the finding was solely based on one lepidopteran insect, Bombyx mori, and the expression of insect immunity was neither dependent on numbers of CpG repeats nor methylation of CpG repeats within ODNs. Instead, foreignness of DNA has been suggested to be a key factor governing induction of antibacterial peptide. In this study, we expanded our previous understanding to the potentiality of ODNs as an immune inducer for antifungal peptide in Galleria mellonella and B. mori. To do this, a defensin-type antifungal peptide gene, reported from G. mellonella was cloned and partially sequenced from G. mellonella and B. mori successfully and utilized as a probe in the Northern blot analysis. We found out that ODNs also work as an immune inducer for antifungal peptide in the fat body and midgut of G. mellonella and B. mori larvae. Also, induction pattern of antifungal peptide was irrelevant to the numbers of CpG repeats within ODNs as previously reported on the induction pattern of antibacterial peptides.

Evaluation of Some Insecticides and Environmental Friendly Agricultural Materials against Winter Cherry Bug, Acanthocoris sordidus (Coreidae, Hemiptera) (꽈리허리노린재(Acanthocoris sordidus, Coreidae, Hemiptera)에 대한 몇 가지 살충제와 친환경농자재의 활성평가)

  • Kang, Chan-Yeong;Ryu, Tae-Hee;Kwon, Hye-Ri;Yu, Yong-Man;Youn, Young-Nam
    • The Korean Journal of Pesticide Science
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    • v.20 no.2
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    • pp.159-164
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    • 2016
  • The winter cherry bug (Acanthocoris sordidus Thunberg, Coreidae, Hemiptera) is a insect pest that have a piercing sucking type mouthpart, and it is injured to solanaceous plants including pepper plant. For the control of the winter cherry bug, 20 organic synthetic insecticides and 7 environmental friendly agricultural materials (EFAMs) were selected with single formulation. Fenitrothion, fenthion and phenthoate were significantly higher mortality to adults and 3rd instar nymphs of winter cherry bug as over 95 and 100 % after 72 hours with treatment. Otherwise, in case of EFAMs, there is no significantly higher effective materials to adults and 3rd instar nymphs of winter cherry bug. Eighty percentage of Sophora flavescens was showed only around 50% mortality with the high variation after 72 hours with treatment against adult of the winter cherry bug.

The Effect of Cyclosporin A on Osteoblast in vitro (Cyclosporin A가 in vitro에서 조골세포에 미치는 영향)

  • Kim, Jae-Woo;Lee, Hyun-Jung;Kang, Jung-Hwa;Ohk, Seung-Ho;Choi, Bong-Kyu;Yoo, Yun-Jung;Cho, Kyoo-Sung;Choi, Seong-Ho
    • Journal of Periodontal and Implant Science
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    • v.30 no.4
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    • pp.747-757
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    • 2000
  • Cyclosporin A(CsA) is an immunosuppressive agent widely used for preventing graft rejecting response in organ transplantation. The basic properties of CsA to osteoblast has not been well known yet. A better understanding of the mechanisms of CsA function on bone could provide valuable information regarding basic properties of bone remodeling, pharmacotherapeutic intervention in metabolic bone disease, and the consequences of immunosuppression in bone physiology. The purpose of this study was to investigate the effect of CsA on osteoblast by evaluating parameters of proliferation, collagen synthetic activity, alkaline phosphatase activity, and ALP mRNA expression in mouse calvarial cell. 1. CsA ($3{\mu}g/m{\ell}$) treated mouse calvarial cell showed statistically significant increase in cell proliferation.(P<0.05) 2. CsA($1,\; 3{\mu}g/m{\ell}$) treated MC3T3 cell line showed statistically significant increase in cell proliferation. 3. The amount of collagen of CsA($3{\mu}g/m{\ell}$) treated mouse calvarial cell was decreased statistically significantly. 4. Alkaline phosphatase activity was increased statistically significantly in CsA treated group($1{\mu}g/m{\ell}$). 5. mRNA expression of ALP was increased in CsA treated group These results suggest that CsA could affect bone remodeling by modulating proliferation & differentiation of osteoblast.

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Inhibitory Effect of a Phosphatidyl Ethanolamine Derivative on LPS-Induced Sepsis

  • Lee, Chunghyun;An, Hyun-Jung;Kim, Jung-In;Lee, Hayyoung;Paik, Sang-Gi
    • Molecules and Cells
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    • v.27 no.2
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    • pp.251-255
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    • 2009
  • Sepsis is the leading cause of death in critically ill patients. Today, around 60% of all cases of sepsis are caused by Gram-negative bacteria. The cell wall component lipopolysaccharide (LPS) is the main initiator of the cascade of cellular reactions in Gram-negative infections. The core receptors for LPS are toll-like receptor 4 (TLR4), MD-2 and CD14. Attempts have been made to antagonize the toxic effect of endotoxin using monoclonal antibodies against CD14 and synthetic lipopolysaccharides but there is as yet no effective treatment for septic syndrome. Here, we describe an inhibitory effect of a phosphatidylethanolamine derivative, PE-DTPA (phosphatidylethanolamine diethylenetriaminepentaacetate) on LPS recognition. PE-DTPA bound strongly to CD14 ($K_d$, $9.52{\times}10^{-8}M$). It dose dependently inhibited LPS-mediated activation of human myeloid cells, mouse macrophage cells and human whole blood as measured by the production of tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) and nitric oxide, whereas other phospho-lipids including phosphatidylserine and phosphatidylethanolamine had little effect. PE-DTPA also inhibited transcription dependent on $NF-{\kappa}B$ activation when it was added together with LPS, and it rescued LPS-primed mice from septic death. These results suggest that PE-DTPA is a potent antagonist of LPS, and that it acts by competing for binding to CD14.

Production of Exopolysaccharides and İndole Acetic Acid (IAA) by Rhizobacteria and Their Potential against Drought Stress in Upland Rice

  • Tetty Marta Linda;Jusinta Aliska;Nita Feronika;Ineiga Melisa;Erwina Juliantari
    • Journal of Microbiology and Biotechnology
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    • v.34 no.6
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    • pp.1239-1248
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    • 2024
  • Peatlands are marginal agricultural lands due to highly acidic soil conditions and poor drainage systems. Drought stress is a big problem in peatlands as it can affect plants through poor root development, so technological innovations are needed to increase the productivity and sustainability of upland rice on peatlands. Rhizobacteria can overcome the effects of drought stress by altering root morphology, regulating stress-responsive genes, and producing exopolysaccharides and indole acetic acid (IAA). This study aimed to determine the ability of rhizobacteria in upland rice to produce exopolysaccharides and IAA, identify potential isolates using molecular markers, and prove the effect of rhizobacteria on viability and vigor index in upland rice. Rhizobacterial isolates were grown on yeast extract mannitol broth (YEMB) medium for exopolysaccharides production testing and Nutrient Broth (NB)+L-tryptophan medium for IAA production testing. The selected isolates identify using sequence 16S rRNA. The variables observed in testing the effect of rhizobacteria were germination ability, vigour index, and growth uniformity. EPS-1 isolate is the best production of exopolysaccharides (41.6 mg/ml) and IAA (60.83 ppm). The isolate EPS-1 was identified as Klebsiella variicola using 16S rRNA sequencing and phylogenetic analysis. The isolate EPS-1 can increase the viability and vigor of upland rice seeds. K. variicola is more adaptive and has several functional properties that can be developed as a potential bioagent or biofertilizer to improve soil nutrition, moisture and enhance plant growth. The use of rhizobacteria can reduce dependence on the use of synthetic materials with sustainable agriculture.

INHIBITORY EFFECT OF OCTYL-PHENOL AND BISPHENOL A ON CALCIUM SIGNALING IN CARDIOMYOCYTE DIFFERENTIATION OF MOUSE EMBRYONIC STEM CELLS

  • J.-H. LEE;Y.-M. YOO;E.-M. JUNG;CH. AHN;E.-B. JEUNG
    • The Korean Journal of Physiology and Pharmacology
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    • v.70 no.3
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    • pp.435-442
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    • 2019
  • Endocrine-disrupting chemicals (EDCs) have structures similar to steroid hormones and can interfere with hormone synthesis and normal physiological functions of reproductive organs. For example, sex steroid hormones influence calcium signaling of the cardiac muscle in early embryo development. To confirm the effect of progesterone (P4), octyl-phenol (OP), and bisphenolA(BPA) on early differentiation of mouse embryonic stem cells(mESCs) into cardiomyocytes, mESCs were treated with P4, OP, and BPA two days after attachment and media were replaced every two days. In addition, cells were treated with mifepristone (RU486), a synthetic steroid that has an affinity for progesterone receptor (Pgr), for one day starting on day 11. Beating ratio was decreased with P4, OP, and BPA treatment. The Pgr mRNA level was significantly increased in the P4-, OP- and BPA-treated groups. However, the mRNA level of the calcium channel gene (Trpv2), contraction-related genes (Ryr2, Cam2, and Mylk3) and cardiac development and morphogenesis genes (Rbp4, Ly6e, and Gata4) were significantly decreased in the P4-, OP-, and BPA-treated groups. Interestingly, treatment with RU486 rescued the altered calcium channel gene, contraction-related genes, and cardiac development and morphogenesis genes. P4, OP, and BPA treatments reduced the intracellular calcium level. Taken together, these results indicate that EDCs (OP and BPA) has a structure similar to that of endogenous steroid hormones such as progesterone and estrogen, and OP and BPA act like progesterone to inhibit and disrupt cardiomyocyte differentiation of mESCs.

Induction of caspase-dependent apoptosis in melanoma cells by the synthetic compound (E)-1-(3,4-dihydroxyphenethyl)-3-styrylurea

  • Kim, Ji-Hae;Jang, Young-Oh;Kim, Beom-Tae;Hwang, Ki-Jun;Lee, Jeong-Chae
    • BMB Reports
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    • v.42 no.12
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    • pp.806-811
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    • 2009
  • Recently, various phenolic acid phenethyl ureas (PAPUs) have been synthesized from phenolic acids by Curtius rearrangement for the development of more effective anti-oxidants. In this study, we examined the anti-tumor activity and cellular mechanism of the synthetic compound (E)-1-(3,4-dihydroxyphenethyl)-3-styrylurea (PAPU1) using melanoma B16/F10 and M-3 cells. Results showed that PAPU1 inhibited the cell proliferation and viability, but did not induce cytotoxic effects on primary cultured fibroblasts. PAPU1 induced apoptotic cell death rather than necrosis in melanoma cells, a result clearly proven by the shift of cells into sub-$G_1$ phase of the cell cycle and by the substantial increase in cells positively stained with TUNEL or Annexin V. Collectively, this study revealed that PAPU1 induced apoptosis in a caspase-dependent manner, suggesting a potential role as a cancer chemopreventive agent for melanoma cells.

Chemical Synthesis and Determination of Biological Activity of the Epidermal Growth Factor-Like Domain of Mouse Betacellulin

  • Shin, Song-Yub;Kang, Shin-Won;Ha, Jong-Myung
    • BMB Reports
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    • v.28 no.2
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    • pp.87-93
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    • 1995
  • To investigate the biological functions of the EGF-like domain of mouse betacellulin (BTC), mouse BTC(33-80), a 48-residue peptide corresponding to the EGF-like domain, was synthesized by stepwise solidphase methods using a 9-fluorenylmethoxycarbonyl (Fmoc) strategy. The homogeneity of synthetic mouse BTC(33-80) was confirmed by analytical reversed phase (RP)-HPLC, amimo acid analysis, and fast atom bombardment mass spectrometer (FAB-MS). Three disulfide bond pairings of synthetic mouse BTC(33-80) were established by amino acid analysis of cysteine-containing fragments derived from thermolytic digestion. These were consistent with the pairings of EGF and transforming growth factor ($TGF-{\alpha}$). The EGF-Iike domain of mouse BTC showed equipotent activity in both EGF-receptor binding on A-431 epidermoid carcinoma cells, and mitogenesis on NIH-3T3 fibroblast cells, as compared with authentic h-EGF. Results suggest that the EGF-Iike domain of BTC plays a significant role in mitogenic activity with an EGF-receptor mediated system.

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Synthesis and Biological Activities of Aklyl Thiosulfi(o)nates (Alkyl thiosulfi(o)nate 화합물의 합성과 생리활성)

  • Jung, Hyun-Jin;Kyung, Kyu-Hang;Jung, Yi-Sook;Kyung, Suk-Hun
    • Applied Biological Chemistry
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    • v.51 no.3
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    • pp.183-187
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    • 2008
  • Alkyl thiosulfi(o)nates, analogs of allyl-2-propene-1-thiosulfinate isolated from Allium sativum and having antibacterial activity, were chemically synthesized and their biological activities were investigated. Alkyl thiosulfinates were prepared by oxidation of corresponding disulfides with organic peroxy acid, while alkyl thiosulfonates could be obtained by oxidation of the alkyl thiosulfinates using sodium periodate. All synthetic thiosulfi(o)nates showed antibacterial activity against Staphylococcus aureus B33 and antifungal activity against Candida utilis ATCC42416. Further more synthetic alkyl thiosulfonates displayed antioxidant activity and have also prevention effect of platelet aggregation induced by collagen in rat.

Analysis of the Potent Platelet Glycoprotein IIb-IIIa Antagonist from Natural Sources

  • Kang, In-Cheol;Kim, Doo-Sik
    • BMB Reports
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    • v.31 no.5
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    • pp.515-518
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    • 1998
  • Adhesive interaction of the platelet glycoprotien IIb-IIIa (GP IIb-IIIa) with a plasma protein, such as fibrinogen, plays an important role in thrombosis and hemostasis. The specific sequence Arg-Gly-Asp (RGD) is critical for the binding of fibrinogen to platelet. To examine and characterize the GP IIb-IIIa antagonist from natural sources, we have developed a simple enzyme-linked immunosorbant assay (ELISA) system. The GP IIb-IIIa complex was purified to homogeneity from platelet Iysates by the combination of two affinity chromatographic methods using the synthetic RGD peptide (GRGDSPK)-immobilized Sepharose and wheat germ lectin-Sepharose. The synthetic peptide GRGDSP inhibits GP IIb-IIIa binding to immobilized fibrinogen with an $IC_{50}$ of $1.5\;{\mu}M$. Venoms of three different snake species and a Korean scolopendra extract have strong antagonistic activities for the binding of human fibrinogen to the platelet GP IIb-IIIa complex. The $IC_{50}$ values of the snake venom s and scolopendra were in the range of $5.5\;{\mu}g$ to $60\;{\mu}g$. These results provide meaningful information for developing antiplatelet agents.

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