• 제목/요약/키워드: secretion proteins

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Biochemical Analysis of Physiological Stress Induced by High Frequency Sound Treatment in the Beet Armyworm, Spodoptera exigua (고주파 처리에 따른 파밤나방(Spodoptera exigua)의 생리적 스트레스의 생화학적 분석)

  • Kim, Yong-Gyun;Son, Ye-Rim;Seo, Sam-Yeol;Park, Bok-Ri;Park, Jung-A
    • Korean journal of applied entomology
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    • v.51 no.3
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    • pp.255-263
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    • 2012
  • High frequency sounds disrupt physiological processes, such as feeding behavior, development and immune responses of Spodoptera exigua. We analyzed high frequency sounds with respect to biochemical changes in S. exigua. High frequency sound (5,000 Hz, 95 dB) suppressed protein synthesis and secretion of midgut epithelium. It also significantly inhibited a digestive enzyme activity of phospholipase $A_2$. The gene expression of three different heat shock proteins and apolipophorin III was altered, particularly in midgut tissue in response to high frequency sound treatments. High frequency sound treatments significantly increased sugar and lipid levels in hemolymph plasma. These results suggest that high frequency sounds are a physiological stress that induces biochemical changes in S. exigua.

Growth, Protein and Pigment Content of Rice Seedlings under Phosphorus Deprivation Condition

  • Yun, Song-Joong;Park, Myoung-Ryul;Kim, Young-Doo;Kim, Key-Young;Baek, So-Hyeon
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.48 no.2
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    • pp.103-107
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    • 2003
  • Phosphorus (P) is a macronutrient playing important roles in many plant processes. Significant interest has been devoted to search and utilize genotypic variations in P use efficiency in rice but with little effort to understand its physiological and biochemical bases. In this study, we examined responses to P deprivation of some primary and secondary traits in 3-week-old seedlings of the three genotypes, Sobi-byeo (japonica), Dasan-byeo (japonica $\times$ indica) and Palawan (indica). In general, percent weight due to root was increased up to 26%, but amounts of root protein and proteins secreted from roots were decreased by 11 to 19% and 31 to 51 %, respectively, by 3 to 21 days of P deprivation in the three genotypes. Interestingly, however, responses of Palawan to short-term P deprivation were contrasting to those of Dasan-byeo and Sobi-byeo in seedling weight and contents of shoot protein, chlorophyll and anthocyanin. Seedling weight was not decreased, but shoot protein content was decreased in P-deprived seedlings of Palawan. Contents of chlorophyll in leaves and anthocynin in roots were increased in Dasan-byeo and Sobi-byeo, but decreased in Palawan. The results suggest that responses of protein and pigment synthesis to P deficiency are different in modem and traditional varieties and the difference may at least in part be due to the selection for high yield under highly fertilized conditions.

Construction of CpG Motif-enriched DNA Vaccine Plasmids for Enhanced Early Immune Response

  • Park Young Seoub;Hwang Seung Ha;Choi Cha-Yong
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.1
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    • pp.29-33
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    • 2005
  • A DNA vaccine methodology using eukaryote expression vectors to produce immunizing proteins in the vaccinated hosts is a novel approach to the development of vaccine and immuno-therapeutics, and it has achieved considerable success over several infectious diseases and various cancers. To further enhance its efficiency, attempts were made to develop novel plasmid vectors containing multiple immunostimulatory CpG motifs, for rapid and strong immune response. First, a 2.9 kb compact plasmid vector (pVAC), containing CMV promoter, polycloning site, BGH poly(A) terminator, ampicillin resistance gene and pBR322 origin was constructed. A pVAC-hEPO was also constructed, which contained a human erythropoietin gene, for evaluating the transfection efficiency of naked plasmid DNA both in vitro and in vivo. To examine the adjuvant effect of multi-CpG motifs on naked plasmid DNA, 22 and 44 enriched and unmethylated CpG motifs were introduced into pVAC to generate pVAC-ISS1 and pVAC-ISS2, respectively. $100{\mu}g$ of pSecTagB, pVAC, pVAC-ISS1 or pVAC-ISS2 were each injected intramuscularly into the tibilias anterior muscle of Balb/c mice. The level of interleukin-6 induced in the mice injected with pVAC-ISS1 and pVAC-ISS2 were significantly elevated after 12 hours, which were almost 2 and 2.5 times higher than that in the mice injected with pSecTagB, respectively. These results suggest that DNA vaccine plasmids with enriched CpG motifs can induce rapid secretion of interleukin-6 by lymphocytes. In conclusion, these vectors can contribute to the development of adjuvant-free DNA vaccinations against infectious diseases and various cancers.

Studies on Vitamin Mineral Interactions in Relation to Passive Transfer of Immunoglobulins in Buffalo Calves

  • Sikka, P.;Lal, D.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.6
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    • pp.825-830
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    • 2006
  • Status of blood minerals and their absorption by neonate calves as influenced by fat soluble vitamins supplementation in their respective mothers, mineral supplementation in calves themselves has been evaluated. The objective was to know the impact of antioxidant vitamin supplementation to advance pregnant buffaloes, on enhanced acquired immunity during first few hours after birth, in relation to weight gain in buffalo calves. Advance pregnant buffaloes (n = 30) consisting of average body weight of $550{\pm}15$ kg and of 4-6 parity were fed on 25 kg green (green Jawar-Sorghum bicolor), 2-3 kg wheat straw and 3-4 kg concentrate mixture individually per day. Intramuscular injections of vitamin triplex A $D_3$ E consisting of -2,500,000 IU of vit A -Palmitate; 2,500,000 IU of vitamin $D_3$ and 1,000 IU of vit E (dl-alpha tocopherol acetate) were given per dose, a month prior to parturition, twice at 15 days interval to 15 dams. Rest of the 15 pregnant buffaloes served as negative controls. Secretion of immune proteins, immunoglobulin (Ig) enhanced by 80% in colostrum. The blood serum levels of Zn, Cu, Ca, Mg were measured from birth to 90 days in calves. A significant (p<0.05) difference between the blood serum Zn levels of calves born to vitamin supplemented and non-supplemented dams was measured and a positive correlation between blood serum Zn levels and injections of vitamins was identified. Association of Zn and Cu with passive immunity status has been identified in these calves. A significant positive correlation between Zn and Cu was also identified which showed a change under the impact of vitamin supplementation in buffaloes. The study signifies the role of micronutrients supplementation in dams prior to parturition, in calf immunity development. The study indicates significant mineral - vitamins interactions during this process.

Expression of Serratia marcescens Metalloprotease(SMP)Gene in Escherichia coli and Serratia marcescens (대장균과 Serratia marcescens에서 Serratia marcescens Metalloprotease(SMP) 유전자의 발현)

  • Kim, Ki-Seok;Jung, Jae-Yeon;Park, Kun-Sik;Kim, Tae Un;Byun, Si Myung;Shin, Yong Chul
    • Microbiology and Biotechnology Letters
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    • v.23 no.3
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    • pp.288-296
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    • 1995
  • To investigate high-level expression of Serratia marcescens metalloprotease (SMP) in Escherichia coli and S. marcescens, we constructed various recombinant plasmids: pSP2, containing SMP gene and lac promoter; pKSP2, containing SMP gene and tac promoter; pTSP2, containing SMP gene, trc99a promoter, and lacI$^{q}$. The recombinant E. coli (pKSP2) strain expressed SMP to a high-level, about 36% of total cellular proteins but accumulated inactive SMP precursors intracellularly, which indicated that E. coli does not have activation and secretion system for SMP. To overproduce active SMP, we transformed S. marcescens with the recombinant plasmids by a modified CaCl$_{2}$ method. The recombinant S. marcescens ATCC27117 (pSP2) containing lac promoter for SMP transcription produced 530 U/ml of active SMP on LB broth, which is about 5.1 times of the SMP yield, 105 U/ml of a control strain, S. marcescens ATCC27117 (pUC19). However, S. marcescens ATCC27117 (pKSP2) containing tac promoter for SMP transcription did not grow healthy and hardly produced SMP. To overcome a harmful effect of the strong tac promoter, we constructed a regulatory plasmid pTSP2 containing a strong trc99a promoter and its repressor gene lacI$^{q}$. When S. marcescens ATCC27117 (pTSP2) was induced with 1.0 mM IPTG after 9 hr cultivation, 2,200 U/ml of SMP was obtained in LB broth, which is about 21 times of that of a control strain.

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High-Level Secretory Expression of Human Procarboxypeptidase B by Fed-Batch Cultivation of Pichia pastoris and its Partial Characterization

  • Kim, Mi-Jin;Kim, Sang-Hyuk;Lee, Jae-Hyung;Seo, Jin-Ho;Lee, Jong-Hwan;Kim, Jong-Hyun;Kim, Yeon-Hee;Nam, Soo-Wan
    • Journal of Microbiology and Biotechnology
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    • v.18 no.12
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    • pp.1938-1944
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    • 2008
  • The procpb gene encoding human procarboxypeptidase B (proCPB, GeneBank access code AJ224866) was cloned and its Pichia expression plasmid, $pPIC9{\alpha}$/hproCPB (9.2 kb), was constructed, in which procpb was under the control of the AOXl promoter and connected to the downstream of the mating factor ${\alpha}$-1 ($MF{\alpha}1$) signal sequence. The plasmid was linearized by digestion with Sacl, and integrated into the genome of P. pastoris strain GS115. By culturing of Pichia transformant on methanol medium, the human proCPB was successfully expressed and secreted into the culture supernatant. Moreover, Western blot analysis of the extracellular proteins showed proCPB bands clearly at a molecular mass of 45 kDa, confirming the expression of proCPB with its right size. The CPB activity reached about 3.5 U/ml and 12.7 U/ml in the flask and fermentor batch cultures of Pichia transformant, respectively. No CPB enzyme activity was found in the intracellular fraction. When the fed-batch cultivation was performed with methanol and glycerol mixture as a feeding medium, the extracellular CPB activity was increased to 42.0 U/ml, which corresponds to a 3.3-fold higher level of CPB activity than that of batch culture. The $K_m$ and $k_{cat}$ values of recombinant human CPB enzyme for hippuryl-$_L$-Arg as a substrate were estimated to be 0.16 mM and $11.93\;sec^{-1}$, respectively.

Human umbilical cord blood mesenchymal stem cells engineered to overexpress growth factors accelerate outcomes in hair growth

  • Bak, Dong Ho;Choi, Mi Ji;Kim, Soon Re;Lee, Byung Chul;Kim, Jae Min;Jeon, Eun Su;Oh, Wonil;Lim, Ee Seok;Park, Byung Cheol;Kim, Moo Joong;Na, Jungtae;Kim, Beom Joon
    • The Korean Journal of Physiology and Pharmacology
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    • v.22 no.5
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    • pp.555-566
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    • 2018
  • Human umbilical cord blood mesenchymal stem cells (hUCB-MSCs) are used in tissue repair and regeneration; however, the mechanisms involved are not well understood. We investigated the hair growth-promoting effects of hUCB-MSCs treatment to determine whether hUCB-MSCs enhance the promotion of hair growth. Furthermore, we attempted to identify the factors responsible for hair growth. The effects of hUCB-MSCs on hair growth were investigated in vivo, and hUCB-MSCs advanced anagen onset and hair follicle neogeneration. We found that hUCB-MSCs co-culture increased the viability and up-regulated hair induction-related proteins of human dermal papilla cells (hDPCs) in vitro. A growth factor antibody array revealed that secretory factors from hUCB-MSCs are related to hair growth. Insulin-like growth factor binding protein-1 (IGFBP-1) and vascular endothelial growth factor (VEGF) were increased in co-culture medium. Finally, we found that IGFBP-1, through the co-localization of an IGF-1 and IGFBP-1, had positive effects on cell viability; VEGF secretion; expression of alkaline phosphatase (ALP), CD133, and ${\beta}-catenin$; and formation of hDPCs 3D spheroids. Taken together, these data suggest that hUCB-MSCs promote hair growth via a paracrine mechanism.

Expression of Thymosin β4 in Ameloblasts during Mouse Tooth Development

  • Choi, Baik-Dong;Lee, Seung-Yeon;Nho, Tae-Hee;Jeong, Soon-Jeong;Lim, Do-Seon;Bae, Chun-Sik;Jeong, Moon-Jin
    • Applied Microscopy
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    • v.46 no.1
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    • pp.58-66
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    • 2016
  • Thymosin ${\beta}4$ ($T{\beta}4$) has been recently reported to play a role in dentinogenesis by regulating the expression of dentin matrix proteins. Based on previous studies, it is hypothesized that $T{\beta}4$ is associated with the formation of the enamel matrix and thus plays an important role in ameloblast. However, there is no report on the function of $T{\beta}4$ during tooth development so far. Therefore, in this study, we aimed to investigate the expression of $T{\beta}4$ and its function in ameloblasts during mouse tooth development. $T{\beta}4$ was expressed strongly in the tooth bud at the bud stage and in the dental lamina and oral epithelium at the cap stage. In advanced bell stage at postnatal day 4, large elongated ameloblasts were observed and the expression of the $T{\beta}4$ protein was the highest, with the enamel being was thicker than that in the early bell stage. The length of ameloblasts increased from the presecretory to the secretory stage and decreased from the maturation to the protective stage. These results suggest that $T{\beta}4$ participates not only in the proliferation of oral epithelial cells during the early stage of tooth development but also regulates enamel protein secretion in ameloblasts and enamel mineralization.

Health Improvement and Treatment Effect of Probiotics (프로바이오틱스의 건강증진과 질병치료 효능)

  • Baek, Young-Mi;Kim, Kyung-Soon;Lee, Yeon-Weol;Cho, Chong-Kwan;Yoo, Hwa-Seung
    • Journal of Haehwa Medicine
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    • v.19 no.1
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    • pp.25-34
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    • 2010
  • Background : The human being and the bacteria have accomplished a balance and have coexisted each other during long time. Probiotics have evolved with the human host to exist and the result they have operated profitably to human host. So it requires that the functions of probiotics are expounded in view of Traditional Korean Medicine. Aim : Suggest the functions of probiotics in view of Traditional Korean Medicine. Method : The author's research has been performed to review the related references. Results : Probiotics assist the absorption of the lactose, proteins and minerals and product several kinds of vitamins, organic acids. Probiotics suppresses the growth of noxious bacteria and the production of harmful substances or gases. They absorbed and discharge the bile acid, and thus help us maintain the optimal level of blood cholesterol concentration. They can reinforce the immune response of the mucous membrane and control the hypersensitivity immune reaction such as asthma, atopy on the other hand. Probiotics have right functions as above and so can be applied widely in treatment of various disease and symptom. Conclusion : Considering the functions of probiotics in view of Traditional Korean Medicine, they participate in our spleen-earth-system (digestion and synthesis) and liver-wood-system (regulation of digestion, metabolism, internal secretion etc.), assist the function of lung-metal system(respiration and regulation of water metabolism) and regulate wi-chi (reinforce/control immune system). Consequently, hereafter there would be a necessity of control a circumstance in treatment of various diseases under these categories that probiotics should be able to do their right functions inside the human body.

Expression of Sara2 Human Gene in Erythroid Progenitors

  • Jardim, Denis Leonardo Fontes;Cunha, Anderson Ferreira Da;Duarte, Adriana Da Silva Santos;Santos, Camila Oresco Dos;Saad, Sara Terezinha Olalla;Costa, Fernando Ferreira
    • BMB Reports
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    • v.38 no.3
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    • pp.328-333
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    • 2005
  • A human homologue of Sar1, named Sara2, was shown to be preferentially expressed during erythropoiesis in a culture stimulated by EPO. Previous studies, in yeast, have shown that secretion-associated and Ras-related protein (Sar1p) plays an essential role in protein transport from the endoplasmic reticulum to the Golgi apparatus. Here, we report the molecular analysis of Sara2 in erythroid cell culture. A 1250 bp long cDNA, encoding a 198 amino-acid protein very similar to Sar1 proteins from other organisms, was obtained. Furthermore, we also report a functional study of Sara2 with Real-time quantitative PCR analysis, demonstrating that expression of Sara2 mRNA increases during the initial stages of erythroid differentiation with EPO and that a two-fold increase in expression occurs following the addition of hydroxyurea (HU). In K562 cells, Sara2 mRNA was observed to have a constant expression and the addition of HU also up-regulated the expression in these cells. Our results suggest that Sara2 is an important gene in processes involving proliferation and differentiation and could be valuable for understanding the vesicular transport system during erythropoiesis.