• Title/Summary/Keyword: Alternative protein

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Beneficial Effects of Fermented Cricket Powder as a Hair Growth Promoting Agent in a Mice Model (생쥐 모델에서 쌍별 귀뚜라미 발효 분말의 발모 촉진제로서의 유익한 효과)

  • Hwang, Jihye;Hwang, Ui Wook
    • Journal of Life Science
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    • v.32 no.3
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    • pp.196-201
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    • 2022
  • Insects have been proposed as new protein sources for human nutrition, and protein availability is affected by insect characteristics. Fermentation can be used to obtain a variety of insect-based ingredients and products with unique properties, but its effect on protein availability is unknown. Fermented cricket (Gryllus bimaculatus) powder consists mainly of protein, and its oral administration has been reported to improve hair growth in androgenetic alopecia. The purpose of this study was to evaluate the hair-promoting activity of fermented cricket powder in an animal model using male C57BL/6 mice (25-30 g). The abdominal hair of the mouse (2x2.5 cm) was gently removed, and the groups fed as follows: Intact controls (no cricket powder); cricket powder only; and fermented cricket powder only. Food was applied daily for 11 weeks. Observational and physical examinations were performed and the results of the different groups compared. The application of fermented cricket powder significantly (p<0.01) promoted hair growth compared to the intact controls. The C57BL/6 results confirmed increased growth after seven weeks when the proportion of anagen follicles had increased by about 125% and 120% in the control and cricket powder groups, respectively. In conclusion, fermented cricket powder can be seen as a promising alternative alopecia treatment because it promotes hair growth, and, given the powder's composition, trace elements such as amino acids may have contributed to these effects.

Discovering the Anti-cancer Effects of Ligusticum Chuanxiong through Network-based Pharmacology Analysis and Molecular Docking: An Inquiry into Natural Products (네트워크 기반 약리학 분석 및 분자 도킹을 통한 천궁의 항암 효과 예측: 천연물에 대한 탐구)

  • Do Kyung Han;Jee Won Shon;Eui Suk Sung;Youn Sook Kim;Won G. An
    • Journal of Life Science
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    • v.33 no.11
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    • pp.876-886
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    • 2023
  • In some cases of head and neck cancers (HNC), surgical interventions may result in the loss of organs and/or changes to their functions, thereby significantly affecting the patient's quality of life. As a result, the surgical treatment of HNC patients is often limited to specific cases, and alternative treatment modalities, such as chemotherapy, are considered. However, serious adverse effects caused by chemotherapy, such as severe nausea and vomiting, necessitate the need for the development of adjunctive methods to minimize patient suffering. Chuanxiong, Ligusticum chuanxiong (L. chuanxiong), is a natural herb used in Eastern medicine to treat cerebrovascular disorders and headaches. This study aimed to predict the effect and potential of L. chuanxiong as an auxiliary anticancer drug through network-based pharmacology and molecular docking analysis. The study results showed that 40 out of 41 genes of L. chuanxiong shared common targets of HNC and their proteins could be used to target HNC cells to prevent cancer progression. The results of the functional enrichment analysis confirmed that L. chuanxiong is associated with the neuroactive-ligand metabolism and neurotransmitter pathways, indicating its potential medicinal value as an adjuvant in HNC treatment. Lastly, our findings demonstrated that the active ingredient of L. chuanxiong, (Z)-Ligustilide, has the ATP binding site of heat shock protein 90, a protein known to promote the activation of cancer cells. These results suggest that L. chuanxiong is a promising candidate for developing auxiliary anticancer drugs, and further research could potentially lead to the discovery of newer and safer anti-cancer agents.

An Efficient Method for Production of Extracellular Human Tissue Factor in Escherichia coli (인간조직인자 세포외 부분의 효과적인 제조 방법)

  • Yoo, Hwan-Goo;Park, Yang-Jin;Lee, Woo-Yiel
    • Journal of Life Science
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    • v.19 no.5
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    • pp.561-565
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    • 2009
  • Human Tissue factor is an essential enzyme activator that forms a catalytic complex with factor VII/ VIIa, and catalyzes both the extrinsic and intrinsic blood coagulation cascades. The extracellular domain of human tissue factor is responsible for association with the biological partner. The efficient procedures for preparing biologically active human tissue factor are essential for the preclinical and clinical studies with coaguligands. An expression vector in Escherichia coli has been constructed to direct the production of extracellular human tissue factor without a fusion protein or a $His_6$ at the N-terminus. The recombinant human tissue factor was expressed in large amounts as a non-native state in E. coli. The recombinant protein was simply renatured during the DEAE-sephacel chromatographic purification procedure. Our expression and purification system does not require a protease treatment or an additional chromatographic step to remove a fusion contaminant, which provides a very useful alternative to conventional expression systems for the production of human tissue factor.

Effects of Fermented Rapeseed Meal on Growth Performance and Serum Parameters in Ducks

  • Fazhi, Xu;Lvmu, Li;Jiaping, Xu;Kun, Qian;Zhide, Zhang;Zhangyi, Liang
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.5
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    • pp.678-684
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    • 2011
  • A trial was performed to study the effects of feeding a diet containing solid-state fermentation rapeseed meal (FRSM) replaced soybean meal (SBM) on growth performance and serum biochemistry parameters of ducks and then to determine the appropriate proportion of soybean meal replacement. The 75% rapeseed meal and 25% blood meal were mixed and inoculated with the Lactobacillus plantarum and Bacillus subtilis. Over the 21-day fermentation, isothiocyanates were reduced from 72.7 to 14.1 mmol/kg. A total of 1,280 fifteen-day-old Cherry Valley ducks were randomly allocated into 4 dietary treatments, 4 replicate groups of 80 ducks each for a 30-day feeding trial. In four treatment groups, fermentation rapeseed meal replaced soybean meal at 0, 33, 67 or 100%, respectively. Results showed that feed intake of ducks fed 100% FRSM was greater (p<0.05) than SBM and partial FRSM in both the finishing period (31-45 d) and entire feeding period (15-45 d). Daily gain increased gradually in the three treatment groups with augmenting FRSM over in the whole study period. In the growing period (15-30 d), compared with the SBM group, phosphorus and calcium content in serum from the FRSM group was improved (p<0.05). Total protein concentration was lower in ducks fed 100% FRSM than SBM and 33% FRSM (p<0.05). Concentrations of IgM were dramatically higher for animals fed 100% FRSM than in the SBM, 33% FRSM and 67% FRSM groups. In the finishing trail stage (31-45 d), only serum IgG content in 100% FRSM group was improved (p<0.05). Therefore, rapeseed meal fermented with Lactobacillus plantarum and Bacillus subtilis is a promising alternative protein source and fermented rapeseed meal can completely replace soybean meal in duck diet and potentially reduce the cost of duck production.

The Role of Nuclear Receptor Subfamily 1 Group H Member 4 (NR1H4) in Colon Cancer Cell Survival through the Regulation of c-Myc Stability

  • Lee, Yun Jeong;Lee, Eun-Young;Choi, Bo Hee;Jang, Hyonchol;Myung, Jae-Kyung;You, Hye Jin
    • Molecules and Cells
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    • v.43 no.5
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    • pp.459-468
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    • 2020
  • Nuclear receptor subfamily group H member 4 (NR1H4), also known as farnesoid X receptor, has been implicated in several cellular processes in the liver and intestine. Preclinical and clinical studies have suggested a role of NR1H4 in colon cancer development; however, how NR1H4 regulates colon cancer cell growth and survival remains unclear. We generated NR1H4 knockout (KO) colon cancer cells using clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein-9 nuclease (CAS9) technology and explored the effects of NR1H4 KO in colon cancer cell proliferation, survival, and apoptosis. Interestingly, NR1H4 KO cells showed impaired cell proliferation, reduced colony formation, and increased apoptotic cell death compared to control colon cancer cells. We identified MYC as an important mediator of the signaling pathway alterations induced by NR1H4 KO. NR1H4 silencing in colon cancer cells resulted in reduced MYC protein levels, while NR1H4 activation using an NR1H4 ligand, chenodeoxycholic acid, resulted in time- and dose-dependent MYC induction. Moreover, NR1H4 KO enhanced the anti-cancer effects of doxorubicin and cisplatin, supporting the role of MYC in the enhanced apoptosis observed in NR1H4 KO cells. Taken together, our findings suggest that modulating NR1H4 activity in colon cancer cells might be a promising alternative approach to treat cancer using MYC-targeting agents.

The Role of Survival Motor Neuron Protein associated with Function of Spinal Motor Neuron (척수 운동신경원의 기능과 관련된 생존운동신경원 단백질의 역할)

  • Song, Ju-Young;Kown, Young-Shil;Nam, Ki-Won;Song, Ju-Min;Kim, Dong-Hyun;Kim, Suk-Bum;Moon, Dong-Chul;Choi, Ji-Ho;Kim, Jin-Sang
    • The Journal of Korean Physical Therapy
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    • v.13 no.2
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    • pp.433-444
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    • 2001
  • This review highlights the ontogenesis and the differentiation of motor neuron in spinal cord, and introduce the survival motor neuron(SMN) which is associated with growth and survival of motor neurons. The differentiation of floor plate cells and motor neurons in the vertebrate neural tube appears to be induced by signals from the notochord. This signal is Sonic hedgehog(Shh). The early development of motor neurons involves the inductive action of Shh. The SMN gene is essential for embryonic viability. SMN mRNA is also expressed in virtually all cell types in spinal cord, including large motor neurons. The SMN protein is involved in RNA processing and during early embryonic development is necessary fer cell survival. Two SMN genes are present in 5q 13 in humans: the telomeric gene(SMNt), which is the SMA-determining gene, and the centromeric analog gene(SMNc). The majority of transcripts from the SMNt gene are full length but, major transcripts of the SMNc gene have a high degrees of alternative splicing and tend to have little or no exon 7. The SMN is involved in the RNA processing(the biogenesis of snRNPs and pre-mRNA splicing), the anti-apoptotic effects, and regulating gene expression.

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Expression of the S glycoprotein of transmissible gastroenteritis virus (TGEV) in transgenic potato and its immunogenicity in mice

  • Ahn, Dong-Joo;Youm, Jung Won;Kim, Suk Weon;Yoon, Won Kee;Kim, Hyoung Chin;Hur, Tai-Young;Joung, Young Hee;Jeon, Jae-Heung;Kim, Hyun Soon
    • Korean Journal of Veterinary Research
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    • v.53 no.4
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    • pp.217-224
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    • 2013
  • Transgenic plants have been tested as an alternative host for the production and delivery of experimental oral vaccines. Here, we developed transgenic potatoes that express the major antigenic sites A and D of the glycoprotein S from transmissible gastroenteritis coronavirus (TGEV-$S_{0.7}$) under three expression vector systems. The DNA integration and mRNA expression level of the TGEV-$S_{0.7}$ gene were confirmed in transgenic plants by PCR and northern blot analysis. Antigen protein expression in transgenic potato was determined by western blot analysis. Enzyme-linked immunosorbent assay results revealed that based on a dilution series of Escherichia coli-derived antigen, the transgenic line P-2 had TGEV-$S_{0.7}$ protein at levels that were 0.015% of total soluble proteins. We then examined the immunogenicity of potato-derived TGEV-$S_{0.7}$ antigen in mice. Compared with the wild-type potato treated group and synthetic antigen treated group, mice treated with the potato-derived antigen showed significantly higher levels of immunoglobulin (Ig) G and IgA responses.

Novel Preparation and Characterization of the α4-loop-α5 Membrane-perturbing Peptide from the Bacillus thuringiensis Cry4Ba δ-endotoxin

  • Leetachewa, Somphob;Katzenmeier, Gerd;Angsuthanasombat, Chanan
    • BMB Reports
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    • v.39 no.3
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    • pp.270-277
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    • 2006
  • Helices 4 and 5 of the Bacillus thuringiensis Cry4Ba $\delta$-endotoxin have been shown to be important determinants for mosquito-larvicidal activity, likely being involved in membrane-pore formation. In this study, the Cry4Ba mutant protein containing an additional engineered tryptic cleavage site was used to produce the $\alpha4$-$\alpha5$ hairpin peptide by an efficient alternative strategy. Upon solubilization of toxin inclusions expressed in Escherichia coli and subsequent digestion with trypsin, the 130-kDa mutant protoxin was processed to protease-resistant fragments of ca. 47, 10 and 7 kDa. The 7-kDa fragment was identified as the $\alpha4$-loop-$\alpha5$ hairpin via N-terminal sequencing and mass spectrometry, and was successfully purified by size-exclusion FPLC and reversed-phase HPLC. Using circular dichroism spectroscopy, the 7-kDa peptide was found to exist predominantly as an $\alpha$-helical structure. Membrane perturbation studies by using fluorimetric calcein-release assays revealed that the 7-kDa helical hairpin is highly active against unilamellar liposomes compared with the 65-kDa activated full-length toxin. These results directly support the role of the $\alpha4$-loop-$\alpha5$ hairpin in membrane perturbation and pore formation of the full-length Cry4Ba toxin.

Molecular and Cellular Analyses of NCP, a Nuclear and Centrosomal Protein in Mouse Gametes and Early Embryos

  • Oh, Hwa-Soon;Youn, Hong-Hee;Lee, Kwang-Hee;Son, Chae-Ick;Lee, Sang-Ho
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.97-97
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    • 2003
  • For many animals the centrosome consists of a pair of centrioles and surrounding pericentriolar materials (PCMs). PCMs have been known to play roles during cell division. It is known that centrioles are necessary to assemble centrosomal components. However, many types of oocytes undergo meiosis without centrioles. It is known that in nonmurine mammalian species, the sperm introduces an intact proximal centriole unlike sea urchin where two centrioles are introduced. In case of mouse sperm, the presence of centrosome is not clear In this study, a monoclonal antibody was developed to investigate centrosome during mouse germ cell and early embryo development. Results of immunostaining and Western blotting in CHO cells suggest that the monoclonal antibody recognizes a nuclear and centrosomal protein, thus called NCP. The NCP monoclonal antibody was used to screen a cDNA expression library prepared from 12.5 mouse brain to isolate NCP gene. Nucleotide size of NCP gene obtained from immunoscreening was about 5.5kb. It is determined that the NCP may be closely related with pericentriolar material -1 gene (Pcm-1) from the result of sequencing analysis. The molecular weight, 66kDa, calculated by known DNA sequence in database is consistent with that of detected from Western blotting using CHO cell lysates. Therefore, it is assumed that NCP may be alternative splicing form of Pcm-1 of which molecular weight is 228kDa. In mouse oocytes, NCP was distributed in nucleus as in CHO cells. It was shown that the NCP was localized around neck region, probably the centrosome in mouse neck region. Interestingly, dramatic change in distribution of NCP was also shown in male germ cell development. Finally, we observed the cellular distribution of NCP during early embryo development. NCP was detected in nucleus as well as centrosome foci. It is suggested that the centrioles reassembly we occurring in blastocysts and then affects the distribution of NCP.

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Modulating the Properties of Metal-Sensing Whole-Cell Bioreporters by Interfering with Escherichia coli Metal Homeostasis

  • Yoon, Youngdae;Kang, Yerin;Lee, Woonwoo;Oh, Ki-Cheol;Jang, Geupil;Kim, Bong-Gyu
    • Journal of Microbiology and Biotechnology
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    • v.28 no.2
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    • pp.323-329
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    • 2018
  • In Escherichia coli, the transcription of genes related to metal homeostasis is activated by the presence of target metals. The promoter regions of those genes can be fused with reporter genes to generate whole-cell bioreporters (WCBs); these organisms sense the presence of target metals through reporter gene expression. However, the limited number of available promoters for sensing domains restricts the number of WCB targets. In this study, we have demonstrated an alternative method to generate novel WCBs, based on the notion that since the sensing mechanisms of WCBs are related to metal transportation systems, their properties can be modulated by disrupting metal homeostasis. Mutant E. coli strains were generated by deleting the znt-operon genes zntA, which encodes a zinc-export protein, and zntR, which encodes a znt-operon regulatory protein, to investigate the effects on the metal-sensing properties of WCBs. Deletion of zntA increased the sensitivity but abolished the selectivity of cadmium-sensing WCBs, whereas arsenic-sensing WCBs gained sensitivity toward cadmium. When zntR was deleted, cadmium-sensing WCBs lost the ability to detect cadmium, and this was recovered by introducing exogenous zntR. In addition, the metal-binding site of ZntR was genetically engineered to modulate metal selectivity. This study provides a valuable platform for the development of novel E. coli-based WCBs.