• Title/Summary/Keyword: protein stabilization

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Tissues Expression, Polymorphisms Identification of FcRn Gene and Its Relationship with Serum Classical Swine Fever Virus Antibody Level in Pigs

  • Liu, Yang;Wang, Chonglong;Liu, Zhengzhu;Xu, Jingen;Fu, Weixuan;Wang, Wenwen;Ding, Xiangdong;Liu, Jianfeng;Zhang, Qin
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.8
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    • pp.1089-1095
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    • 2012
  • Neonatal Fc receptor (FcRn) gene encodes a receptor that binds the Fc region of monomeric immunoglobulin G (IgG) and is responsible for IgG transport and stabilization. In this report, the 8,900 bp porcine FcRn genomic DNA structure was identified and putative FcRn protein included 356 amino acids. Alignment and phylogenetic analysis of the porcine FcRn amino acid sequences with their homologies of other species showed high identity. Tissues expression of FcRn mRNA was detected by real time quantitative polymerase chain reaction (Q-PCR), the results revealed FcRn expressed widely in ten analyzed tissues. One single nucleotide polymorphism (SNP) (HQ026019:g.8526 C>T) in exon6 region of porcine FcRn gene was demonstrated by DNA sequencing analysis. A further analysis of SNP genotypes associated with serum Classical Swine Fever Virus antibody (anti-CSFV) concentration was performed in three pig populations including Large White, Landrace and Songliao Black pig (a Chinese indigenous breed). Our results of statistical analysis showed that the SNP had a highly significant association with the level of anti-CSFV antibody (At d 20; At d 35) in serum (p = 0.008; p = 0.0001). Investigation of expression and polymorphisms of the porcine FcRn gene will help us in further understanding the molecular basis of the antibody regulation pathway in the porcine immune response. All these results indicate that FcRn gene might be regarded as a molecular marker for genetic selection of anti-CSFV antibody level in pig disease resistance breeding programmes.

Effects of Proline and Gelatin on hCTLA4Ig Production in Transgenic Rice Suspension Cells (형질전환 벼 현탁세포를 이용한 hCTLA4Ig 생산에서 proline과 gelatin이 미치는 영향)

  • Song, Mi-Na;Cheon, Su-Hwan;Kwon, Jun-Young;Choi, Sung-Hun;Kim, Dong-Il
    • KSBB Journal
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    • v.24 no.3
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    • pp.246-252
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    • 2009
  • Rice cells were transformed to express human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig) using RAmy3D promoter. hCTLA4Ig was produced and secreted into culture media inducibly when sugar was depleted. The obstacles of this system are the cell death and release of proteases by sugar starvation. These problems resulted in the losses of stability and productivity of hCTLA4Ig. Therefore, the effects of proline as an inhibitor of cell death were investigated. When 4 mM proline was added in sugar-free media, the cell death and release of proteases were reduced. As a consequence, the production of hCTLA4Ig was enhanced. In addition, the effects of protein stabilizers such as gelling agents were studied. It was found that the application of 0.01 g/L gelatin led to an increase in hCTLA4Ig production. This increase might be originated from the stabilization of hCTLA4Ig. In conclusion, the production of hCTLA4Ig could be enhanced by the additions of proline and gelatin in transgenic rice cell cultures.

Betaine-γ-aminobutyric Acid Transporter 1 (BGT-1/mGAT2) Interacts with the PDZ Domain of Munc-18 Interacting Proteins (Mints) (Betaine-γ-aminobutyric acid transporter 1 (BGT-1/mGAT2)과 Munc-18-interacting (Mint) 단백질의 PDZ 결합)

  • Kim, Sang-Jin;Jeong, Young-Joo;Choi, Sun-Hee;Choi, Chun-Yeon;Jun, Hee-Jae;Moon, Il-Soo;Seog, Dae-Hyun;Jang, Won-Hee
    • Journal of Life Science
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    • v.22 no.9
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    • pp.1159-1165
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    • 2012
  • The action of neuronally released ${\gamma}$-aminobutyric acid (GABA) is terminated by uptake into the neurons by GABA transporters (GATs). The mechanism underlying the stabilization and regulation of GAT2 has not yet been elucidated. We used the yeast two-hybrid system to identify proteins that interact with and, thereby, regulate betaine-${\gamma}$-aminobutyric acid transporter 1 (BGT-1/mGAT2). We found an interaction between BGT-1/mGAT2 and Munc-18-interacting proteins (Mints). The "T-H-L" motif at the C-terminal end of BGT-1/mGAT2 was essential for the interaction with Mint2 in the yeast two-hybrid assay. Mint2 bound to the tail region of BGT-1/mGAT2, but not to other GAT members. When co-expressed in HEK-293T cells, Mint2 was co-immunoprecipitated with BGT-1/mGAT2. In addition, we demonstrated the cellular co-localization of BGT-1/mGAT2 and Mint2 in the cells. These results suggest that Mint2 contributes to the regulation of BGT-1/mGAT2.

Induction of Autolysis and Autoplast Formation of Anaerobic Clostridium thermohydrosulfuricum (혐기성 Clostridium thermohydrosulfuricum의 Autolysis 및 Autoplast 형성유도)

  • 김욱한;박동찬;정기택;이용현
    • Korean Journal of Microbiology
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    • v.27 no.4
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    • pp.357-365
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    • 1989
  • Induction conditions for autolysis and autoplast formation of thermophilic Clostridium thermohydrosulfuricum were studied. The cells in the initial exponential growth phase were well autolysed in Tris-HCl buffer or inorganic buffers containing univalents, such as $K^{+}$ and $Na^{+}$ , and chemicals such as cysteine-HCl, sorbitol and glycerol. Meanwhile, autolysis induction was slightly inhibited by divalents, such as $Mg^{2+}, Mn^{2+}, Ca^{2+}, Ni^{2+}$, and strongly by divalents, such as $Fe^{2+}, Cu^{2+}$ and citric acid. The autolysis was stimulated when the cells were grown in the medium containing ampicillin that inhibites cell wall synthesis, meanwhile, it was slightly inhibited by nucleic acids and protein synthesis inhibitors. The optimal pH and temperature for the induction of autolysis were 7.5 and $60^{\circ}C$, respectively. On the other hand, the cells were autoplasted without lysozyme treatment during autolysis due to the stabilization of protoplasmic membrane in the presence of divalents such as $Mg^{2+}, Mn^{2+}, Ca^{2+}, Ni^{2+}$. Autoplast formation was mostly induced at $37^{\circ}C$ in 50mM Tris-HCl buffer (pH 7.5) containing 20 mM $MgCl^{2}$ and 0.3 M glycerol, and in the late exponential growth phase growing cell.

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A Patient Diagnosed with Spinocerebellar Ataxia Type 5 associated with SPTBN2: Case Report (SPTBN2와 연관된 spinocerebellar ataxia type 5를 진단받은 환자)

  • Hur, Min woo;Ko, Ara;Lee, Hyun Joo;Lee, Jin Sung;Kang, Hoon-Chul
    • Journal of the Korean Child Neurology Society
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    • v.25 no.3
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    • pp.200-203
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    • 2017
  • Spinocerebellar ataxias (SCAs) are autosomal dominant neurodegenerative disorders which disrupt the afferent and efferent pathways of the cerebellum that cause cerebellar ataxia. Spectrin beta non-erythrocytic 2 (SPTBN2) gene encodes the ${\beta}-III$ spectrin protein with high expression in Purkinje cells that is involved in excitatory glutamate signaling through stabilization of the glutamate transporter, and its mutation is known to cause spinocerebellar ataxia type 5. Three years and 5 months old boy with delayed development showed leukodystrophy and cerebellar atrophy in brain magnetic resonance imaging (MRI). Diagnostic exome sequencing revealed that the patient has heterozygous mutation in SPTBN2 (p.Glu1251Gln) which is a causative genetic mutation for spinocerebellar ataxia type 5. With the patient's clinical findings, it seems reasonable to conclude that p.Glu1251Gln mutation of SPTBN2 gene caused spinocerebellar ataxia type 5 in this patient.

A Study on the Antioxidant and MMPs Protein Expression Inhibitive Effect of Punica granatum L. Extract and Its Stabilization with Liquid Crystal Emulsion (석류추출물의 항산화와 MMPs 단백질 발현 억제 및 액정 유화물에서의 안정화에 관한 연구)

  • Roh, Jin-Sun;Yeom, Hyeon-Ji;Oh, Min-Jeong;Lee, Jin-Young
    • Journal of Life Science
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    • v.31 no.2
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    • pp.164-174
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    • 2021
  • This study confirmed the potential of Punica granatum L. extract for functional activity verification and cosmetic development. The electron-donating ability of Punica granatum L. extract was shown 60.6% at a 1,000 ㎍/ml concentration. Its ABTS+ radical scavenging ability was shown 93.9% at a 1,000 ㎍/ml concentration. Additionally, the inhibitive effects of elastase and collagenase inhibition effects were measured as 30% and 47.2%, respectively, at a 1,000 ㎍/ml concentration. To determine the effect of Punica granatum L. extract on the proliferation of fibroblasts (CCD-986sk), cell viability was measured using a 3-[4,5-dimethyl-thiazol-2-yl]-2,5-diphenyl-tetrazoliumbromide (MTT) assay. As a result, survival rates of 130% or higher at a 500 ㎍/ml concentration or less were confirmed. According to the results of Western blot with Punica granatum L. extract, the expression inhibition rates of matrix metalloproteinase-1 (MMP-1), matrix metalloproteinase-2 (MMP-2), and matrix metalloproteinase-3 (MMP-3) were decreased by 23.2%, 81.9%, and 69.2%, respectively, at a 100 ㎍/ml concentration. Based on the results above, O/W liquid crystal cream with 0.1% Punica granatum L. extract was prepared. The stabilities were tested at 4, 25, 45, and 50℃. By checking the pH, change over time, and stability by temperature, it was confirmed that all were stable for one month. Thus, Punica granatum L. extract shows potential as a natural material for cosmetics.

A retrospective study on the efficacy of Roxadustat in peritoneal dialysis patients with erythropoietin hyporesponsiveness

  • Jie Liu;Shuang Li;Fan Yang;Tianyu Li;Rui Li;Yousuf Waheed;Chen Meng;Shulin Li;Kun Liu;Yanshan Tong;Haisheng Xu;Chuankuo Tian;Xinglei Zhou
    • The Korean journal of internal medicine
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    • v.39 no.3
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    • pp.488-500
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    • 2024
  • Background/Aims: Roxadustat, an oral medication for treating renal anemia, is a hypoxia-inducible factor prolyl hydroxylase inhibitor used for regulating iron metabolism and promoting erythropoiesis. To investigate the efficacy and safety of roxadustat in patients undergoing peritoneal dialysis (PD) with erythropoietin hyporesponsiveness. Methods: Single-center, retrospective study, 81 PD patients (with erythropoietin hyporesponsiveness) were divided into the roxadustat group (n = 61) and erythropoiesis-stimulating agents (ESAs) group (n = 20). Hemoglobin (Hb), total cholesterol, intact parathyroid hormone (iPTH), brain natriuretic peptide (BNP), related indicators of cardiac function and high-sensitivity C-reactive protein (hs-CRP) were collected. Additionally, adverse events were also recorded. The follow-up period was 16 weeks. Results: The two groups exhibited similar baseline demographic and clinical characteristics. At baseline, the roxadustat group had a mean Hb level of 89.8 ± 18.9 g/L, while the ESAs group had a mean Hb level of 95.2 ± 16.0 g/L. By week 16, the Hb levels had increased to 118 ± 19.8 g/L (p < 0.05) in the roxadustat group and 101 ± 19.3 g/L (p > 0.05) in the ESAs group. The efficacy of roxadustat in improving anemia was not influenced by baseline levels of hs-CRP and iPTH. Cholesterol was decreased in the roxadustat group without statin use. An increase in left ventricular ejection fraction and stabilization of BNP were observed in the roxadustat group. Conclusions: For PD patients with erythropoietin hyporesponsiveness, roxadustat can significantly improve renal anemia. The efficacy of roxadustat in improving renal anemia was not affected by baseline levels of hs-CRP0 and iPTH.

Urban aquaculture of catfish, Silurus asotus, using biofloc and aquaponics systems (바이오플락과 아쿠아포닉스를 이용한 도심형 양식시스템에서의 메기양식)

  • Kim, Seok Ryel;Jang, Jin Woo;Kim, Bum Ju;Jang, In Kwon;Lim, Hyun Jeong;Kim, Su Kyoung
    • Korean Journal of Environmental Biology
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    • v.37 no.4
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    • pp.545-553
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    • 2019
  • This study was conducted to determine whether the water in which nitrate accumulated during long-term fish culture in an aquaponics system without water exchange could be removed and reused as catfish-culturing water. The catfish (Silurus asotus) were cultured in the urban aquaculture system using BFT (Biofloc Technology) aquaculture and an aquaponics system (two rearing tanks, 3 tons each) without exchanging the rearing water. After 151 days (from March to August) of rearing, 2.8 g of fry had grown to an average weight of 171.3 g (total weight, 56.53 kg) and 235.5 g (total weight 71.1 kg), respectively. The overall survival rate was 65% in the urban aquaculture system. However, the survival rate was 77.7% before separation into the two tanks. The survival rates after the separation were 92.9% and 78.0%. In the early biofloc watermaking process, there was a high mortality rate. After water stabilization, the mortality rate decreased and some mortality occurred during the period when the total amount of suspended solids (TSS) increased. The results of monthly blood analysis of the catfish showed that the AST concentration was significantly higher in April. Blood ALT levels and triglycerides showed no difference in the rearing period and the glucose, cholesterol, and total protein levels were significantly higher in July. There was no difference in the other periods. The plants produced by the aquaponics system using catfish-rearing water were lettuce, basil, chard, and red chicory. These showed smooth growth and a total of 148.85 kg of plants were harvested in five months. It was possible to remove nitric acid from the aquaponics system and reuse it as catfish-rearing water. Maintaining proper plant quantity according to the capacity of the catfish showed that the combination of agricultural and aquatic products was possible.