• Title/Summary/Keyword: Bt protein

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Antimelanogenic of Artemisia fukudo Makino Extract in Melanoma Cells (큰비쑥 추출물의 멜라닌 생성 억제 효과)

  • Kim, Min-Jin;Kim, Seoyeon;Hyun, Kwang Hee;Kim, Duk Soo;Kim, Seung-Young;Hyun, Chang-Gu
    • KSBB Journal
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    • v.32 no.3
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    • pp.233-237
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    • 2017
  • Melanin is one of the most important factors affecting skin color. Melanogenesis is the bioprocess of melanin production by melanocytes in the skin and hair follicles and is mediated by several enzymes, such as tyrosinase, tyrosinase related protein (TRP)-1, and TRP-2, MITF. In this study, we investigated the effect of Artemisia fukudo Makino extracts on tyrosinase activity and melanin production as natural products of whitening functional cosmetics. Melanin content in murine B16F10 melanoma cells were decreased by Artemisia fukudo Makino extracts in a dose-dependently. In addition, the inhibition of tyrosinase activity of Artemisia fukudo Makino extracts showed to decrease tyrosinase activity as the concentration of ${\alpha}-MSH$ was increased. Furthermore, western blot analysis revealed that Artemisia fukudo Makino extracts significantly downregulated the expression of tyrosinase, TRP-1 which treat of ${\alpha}-MSH-induced$ melanogenesis in murine B16F10 melanoma cells. As a result, Artemisia fukudo Makino extract showed functionalities as an effective whitening agent to inhibit melanin formation.

Insecticidal Toxin and Research Trends of Photorhabdus, Entomopathogenic Bacteria (곤충살충성 세균 Photorhabdus의 Insecticidal Toxin과 연구동향)

  • Jang, Eun-Kyung;Shin, Jae-Ho
    • Microbiology and Biotechnology Letters
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    • v.38 no.2
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    • pp.117-123
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    • 2010
  • BT toxin is produced by a soil bacterium Bacillus thuringiensis and has long been used as a biological insecticide without any competition. Recently, Photorhabdus, a symbiotic bacterium from entomopathogenic nematodes, family Heterorhabditae, has been researched and discussed as alternatives to B. thuringiensis. Photorhabdus, which lives in the gut of entomopathogenic nematodes, is a highly virulent pathogen of a wide range of insect larvae. When an insect is infected by the nematodes, the bacteria are released into the cadaver, and produce a number of insecticidal toxins. The biological role of the different Photorhabdus toxins in the infection process is still unclear. Photorhabdus toxin complex (Tc) is highly secreted gut-active toxin and has been characterized as a potent three-component (A, B and C) insecticidal protein complex. These components are necessary for full oral activity against insect larvae. The Photorhabdus PirAB binary toxins exhibit a potent injectable activity for Galleria mellonella larvae, and have oral toxicity against mosquitoes and caterpillar pest Plutella xylostella. Other toxin, 'makes caterpillars floppy' (Mcf) showed injectable activity on caterpillars. Recombinant Mcf triggers apoptosis in both insect hemocytes and the midgut epithelium and carries a BH3 domain. In this review, the relationship between the Photorhabdus and the nematode is discussed and recent important insecticidal toxins from Photorhabdus are described.

A Freeze-drying Formulation and Target Specificity of Double-stranded RNA-expressing Bacteria to Control Insect Pests (Double-stranded RNA 발현 세균의 동결건조 제형화와 적용 대상 해충 선택성)

  • Kim, Eunseong;Kim, Yonggyun
    • Korean journal of applied entomology
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    • v.55 no.2
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    • pp.81-89
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    • 2016
  • Double-stranded RNA (dsRNA) has been applied to control insect pests by its suppressive activity against specific target genes. Integrin is a heterodimer (${\alpha}$ and ${\beta}$) transmembrane protein and plays a critical role in cell-to-cell or cell-to-extracellular matrix interactions in eukaryotes. Suppression of ${\beta}$ subunit integrin gene expression by its specific dsRNA (= dsINT) induces significant mortality against target insects. Furthermore, a recombinant bacterium expressing dsINT is potent to kill target insects. However, it is necessary to develop a formulation technique of the dsRNA-expressing bacteria to apply the bacterial insecticide against field populations. This study formulated the recombinant bacteria by freeze-drying and tested its control efficacy against target insects. The formulation maintained significant insecticidal activity against last instar larvae of Spodoptera exigua. While a commercial Bacillus thuringiensis (Bt) insecticide exhibited only about 60% insecticidal activity against S. exigua last instar, an addition of the dsINT-expressing bacterial formulation significantly enhanced the Bt insecticidal activity. The dsINT-expressing bacterial formulation exhibited relative selectivity to target insects depending on sequence similarity. These results indicate that a freeze-dried form of dsRNA-expressing bacteria keeps its insecticidal activity.

Effect of Various Fat and Oil Types on Weight Gain and Liver Metabolism in Fasting or Normal Diet Rats (불포화도가 다른 유지가 단식과 정상식이 흰쥐의 체중 및 간대사에 미치는 영향)

  • 서화중;박현희
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.22 no.1
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    • pp.19-26
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    • 1993
  • In this study rats in fasting or fed protein free restricted diet including only fat showed much lowered level of serum cholesterol and triglyceride accompanied by utmost weight loss and high level of blood urea nitrogen indicated the tissue degradation, especially in liver with signs of damage or necrosis of hepatic parenchymal cell leading to elevated glutamic pyruvate transaminase value and to death. Rats fed only perilla oil in starvation or as fat source in normal diet dropped down the level of serum cholesterol and triglyceride compared to beef tallow fed rat. But with evidence of glutamic pyruvate transaminase values which was significantly elevated long term ingestion of perilla oil is likely to cause the lesion or any damage of hepatic function.

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Protein Composition and Biological Activities of Bombus ignitus Venom (호박벌 일벌독의 성분 분석 및 생리활성 탐색)

  • Han, Sang-Mi;Lee, Kwang-Gill;Yeo, Joo-Hong;Kweon, Hae-Yong;Woo, Soon-Ok;Yoon, Hyung-Joo;Kim, Me-Ae;Kim, Won-Tae;Baek, Ha-Ju
    • Korean journal of applied entomology
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    • v.46 no.2
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    • pp.229-234
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    • 2007
  • Pure Bombus ignitus venom samples were submitted to two-dimensional gel electrophoresis. A total of 64 excised spots were analyzed by mass spectrometry. Three main proteins resulted in the identification have not been described in other bee venoms before. Dose-dependence against human carcinoma (Hep3B, BT-20, A549 and AGS) were observed from 1ng/ml to 100ng/ml. Expecially, the treatment of 100ng/ml B. ignitus venoms showed the highest cytotoxicity with 55% against hepatocellular carcinoma (Hep3B). The B. ignitus venoms showed strong antimicrobial activities against Enterococcus faecium and Shigella sonnei, and practically antimicrobial activity against the other microorganisms tested. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of E. faecium and S. sonnei, were 0.256ug/ml, respectively.

Coat Color Patterns and Genotypes of Extension and Agouti in Hanwoo and Jeju Black Cattle (제주흑우와 한우에서 Extension, Agouti 유전자형과 모색 출현 양상)

  • Han, Sang-Hyun;Cho, In-Cheol;Kim, Jae-Hwan;Ko, Moon-Suck;Kim, Young-Hoon;Kim, Eun-Young;Park, Se-Pill;Lee, Sung-Soo
    • Journal of Life Science
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    • v.21 no.4
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    • pp.494-501
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    • 2011
  • To understand the relationship between coat color inheritance patterns and genotypes of Extension (E) and Agouti (A) loci in cattle, the genotypes for melanocortin-1 receptor (MC1R) and agouti signaling protein (ASIP) were analyzed in Hanwoo, Jeju black cattle (JBC), and their crossbred progeny. Three MC1R alleles ($E^D$, $E^+$, and e) were found in the black-colored JBC population. JBC had no recessive homozygotes (e/e), but this genotype was predominant in the Hanwoo breed. However, MC1R $E^+$/e Hanwoo did not produce a black coat color as they appeared either as brown or solid red. For ASIP, three genotypes (A/A, A/$A^{Br}$, and $A^{Br}/A^{Br}$) were determined by insertion/deletion of an L1-BT element in Hanwoo. The ASIP $A^{Br}$ allele was rarely observed, and no ASIP $A^{Br}/A^{Br}$ homozygotes were detected in the JBC population. Cattle carrying ASIP $A^{Br}$ did not show any agouti-like brindle pigmentation patterns in either breed or their progeny. The coat colors of the crossbred progeny were discriminated by two colors, yellowish-brown versus dark-brown or black, and their coat colors were directly related to the genotypes of the Extension locus, yellowish-brown (e/e) and dark-brown or black ($E^+$/e), but not to the Agouti locus. ASIP genotypes probably did not affect coat color development in the Hanwoo or crossbred progeny. Our results suggest that the ASIP genotypes do not play key roles in coat color variation, but the MC1R genotypes do direct the phenotypes of Hanwoo, JBC, and their progeny.

Safety Assessment of Foods Produced Using Recombinant DNA Techniques

  • Toyoda, Masatake
    • Toxicological Research
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    • v.17
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    • pp.167-171
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    • 2001
  • The introduction of genetically modified crops has raised concerns regarding safety issues over the insertion of foreign genes into plant genomes using recombinant DNA technology. Since 1991 in Japan, 29 foods and 6 food additives have been evaluated, based on the "Guideline for Safety Assessment", before these foods were marketed. The MHW, however, decided that safety assessment of such foods and food additives should be legally imposed. because soon such foods and food additives are expected to circulate globally and a new system for assessing safety of such foods and food additives at a pre-market stage is necessary, in order to avoid the distribution of any genetically modified foods that have had no safety assessment. The MHW published relevant announcements to amend existing regulations on 1 May 2000. "Standards for safety assessment of seed plant" is established based on a concept of substantial equivalence, and applicable to the products which are regarded as equivalent to the existing products used as foods and food additives. The characterization of the food products entails consideration of the molecular characterization. phenotypic and compositional characteristics, key nutrients and toxicants, and toxicity and allergenicity of the introduced proteins, and if there are indications of unintended effects of the modification, whether further safety testing (animal studies etc.) is needed should be considered. Safety and wholesomeness studies with whole foods should be care fully designed in order to avoid nutritional imbalances causing artifacts and uninterpretable results as was the case of Dr. Pusztaiis report. A case study of genetically modified soybeans (glyphosate-tolerant soybeans) on the immune system of rats and mice is shown. Chemical compositions were also compared with those of the non-GM soybeans. The studies failed to detect any differences in immuno-toxic activity.muno-toxic activity.

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Cancer/Testis OIP5 and TAF7L Genes are Up-Regulated in Breast Cancer

  • Mobasheri, Maryam Beigom;Shirkoohi, Reza;Modarressi, Mohammad Hossein
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.11
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    • pp.4623-4628
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    • 2015
  • Breast cancer still remains as the most frequent cancer with second mortality rate in women worldwide. There are no validated biomarkers for detection of the disease in early stages with effective power in diagnosis and therapeutic approaches. Cancer/testis antigens are recently promising tumor antigens and suitable candidates for targeted therapies and generating cancer vaccines. We conducted the present study to analyze transcript changes of two cancer/testis antigens, OIP5 and TAF7L, in breast tumors and cell lines in comparison with normal breast tissues by quantitative real time RT-PCR for the first time. Significant over-expression of OIP5 was observed in breast tumors and three out of six cell lines including MDA-MB-468, T47D and SKBR3. Not significant expression of TAF7L was evident in breast tumors but significant increase was noted in three out of six cell lines including MDA-MB-231, BT474 and T47D. OIP5 has ssignificant role in chromatin organization and cell cycle control during cell cycle exit and normal chromosome segregation during mitosis and TAF7L is a component of the transcription factor IID, which is involved in transcription initiation of most protein coding genes. TAF7Lis located at X chromosome and belongs to the CT-X gene family of cancer/testis antigens which contains about 50% of CT antigens, including those which have been used in cancer immunotherapy.

Inhibition of p90RSK activation sensitizes triple-negative breast cancer cells to cisplatin by inhibiting proliferation, migration and EMT

  • Jin, Yujin;Huynh, Diem Thi Ngoc;Kang, Keon Wook;Myung, Chang-Seon;Heo, Kyung-Sun
    • BMB Reports
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    • v.52 no.12
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    • pp.706-711
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    • 2019
  • Cisplatin (Cis-DDP) is one of the most widely used anti-cancer drugs. It is applicable to many types of cancer, including lung, bladder, and breast cancer. However, its use is now limited because of drug resistance. p90 ribosomal S6 kinase (p90RSK) is one of the downstream effectors in the extracellular signal-regulated protein kinases 1 and 2 (ERK1/2) pathway and high expression of p90RSK is observed in human breast cancer tissues. Therefore, we investigated the role of p90RSK in the Cis-DDP resistance-related signaling pathway and epithelial-mesenchymal transition (EMT) in breast cancer cells. First, we discovered that MDA-MB-231 cells exhibited more Cis-DDP resistance than other breast cancer cells, including MCF-7 and BT549 cells. Cis-DDP increased p90RSK activation, whereas the inactivation of p90RSK using a small interfering RNA (siRNA) or dominant-negative kinase mutant plasmid overexpression significantly reduced Cis-DDP-induced cell proliferation and migration via the inhibition of matrix metallopeptidase (MMP)2 and MMP9 in MDA-MB-231 cells. In addition, p90RSK activation was involved in EMT via the upregulation of mRNA expression, including that of Snail, Twist, ZEB1, N-cadherin, and vimentin. We also investigated NF-κB, the upstream regulator of EMT markers, and discovered that Cis-DDP treatment led to NF-κB translocation in the nucleus as well as its promoter activity. Our results suggest that targeting p90RSK would be a good strategy to increase Cis-DDP sensitivity in triple-negative breast cancers.

Production and Characterization of a Recombinant Antibody Neutralizing Botulinum Neurotoxin A (보툴리눔 신경독소 A를 중화하는 재조합 항체의 제조와 특성 분석)

  • Park, Hong-Gyu;Choi, Mieyoung
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.18 no.1
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    • pp.295-301
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    • 2017
  • Botulinum neurotoxin (BoNT/A) is a neurotoxin that selectively attacks the peripheral cholinergic nerve endings. It is produced by Gram -positive, endospore-forming strict anaerobic bacteria, Clostridium botulinum. Since BoNT/A could be a biothreat agent, as well as a contaminator of food and water supplies, the development of sensitive assays for toxin detection and potent antitoxin for the treatment of intoxication is necessary. In this study, for the purpose of producing monoclonal antibodies (mAbs) that are capable of neutralizing Botulinum neurotoxin type A (BoNT/A), scFv (single-chain variable domain fragment) libraries from the rabbit antisera against BoNT/A was fused to a human IgG. The resulting recombinant scFvIgG antibody protein was expressed in stable cell lines and was purified using a protein A affinity chromatography. The efficacy of scFvIgG mAb was confirmed by ELISA and was evaluated for the neutralization of BoNT/A in vivo. Such an in vivo toxin neutralization assay was performed using mice. Although scFvIgG antibody proteins (10 ug) failed to fully protect the mice challenged with BoNT/A (100,000 $LD_{50}$), it significantly prolonged the survival time. These results suggest that scFvIgG mAb may be capable of neutralizing BoNT/A single-chain variable domain fragment.