• Interdisciplinary Bio Central
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• 제1권2호
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• pp.8.1-8.6
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• 2009

Introduction: It is a challenge to design a protein score function which stabilizes the native structures of many proteins simultaneously. The coarse-grained description of proteins to construct the pairwise-contact score function usually ignores the backbone directionality of protein structures. We propose a new two-body score function which stabilizes all native states of 1,006 proteins simultaneously. This two-body score function differs from the usual pairwise-contact functions in that it considers two adjacent amino acids at two ends of each peptide bond with the backbone directionality from the N-terminal to the C-terminal. The score is a corresponding propensity for a directional alignment of two adjacent amino acids with their local environments. Results and Discussion: We show that the construction of a directional adjacency-score function was achieved using 1,006 training proteins with the sequence homology less than 30%, which include all representatives of different protein classes. After parameterizing the local environments of amino acids into 9 categories depending on three secondary structures and three kinds of hydrophobicity of amino acids, the 32,400 adjacency-scores of amino acids could be determined by the perceptron learning and the protein threading. These could stabilize simultaneously all native folds of 1,006 training proteins. When these parameters are tested on the new distinct 382 proteins with the sequence homology less than 90%, 371 (97.1%) proteins could recognize their native folds. We also showed using these parameters that the retro sequence of the SH3 domain, the B domain of Staphylococcal protein A, and the B1 domain of Streptococcal protein G could not be stabilized to fold, which agrees with the experimental evidence.

• Nuclear Medicine and Molecular Imaging
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• 제41권3호
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• pp.226-233
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• 2007

목적: 광학과 핵의학 및 자기공명 분자영상 기술은 생체내에서 리포터 유전자의 발현을 비침습적으로 평가할 수 있다. 한가지 이상의 유전자 발현을 영상화 할 수 있는 복합분자영상은 유전자의 발현과 유전자 치료 후 효능의 평가를 다양한 방법으로 반복하여 평가할 수 있다는 장점이 있다. 본 연구에서는 핵의학 영상이 가능한 NIS와 광학 영상이 가능한 EGFP 두가지 유전자를 동시에 발현하는 HepG2-Retro-PNRGW (PGKp-NIS-RSVp-EGFP-WPRE) plasmid를 이용한 간암 세포주(HepG2-NE)를 구축하고, NIS와 EGFP 리포터 유전자의 기능 발현을 체내에서 광학영상과 핵의학 영상으로 확인하고자 하였다. 재료 및 방법: pcDNA-NIS로 부터 NIS 유전자를 분리하여 pRetro-PN vector를 만든 후, pLNRGW (LTR-NeoR-RSV-EGFP-WPRE)로부터 RSV-EGFP-WPRE 조각을 분리하여 최종적으로 NIS와 EGFP 유전자가 동시에 발현할 수 있는 pRetro-PNRGW vector를 구축하였다. 구축된 vector를 이용하여 Retro-PNRGW retrovirus를 생산하였으며, 이를 HepG2 세포에 감염시켜 HepG2-NE 세포주를 만들었다. 이 세포주의 NIS 유전자의 발현은 역전사효소 중합효소 연쇄반응으로 mRNA 발현을 확인하였고, EGFP 유전자의 발현은 형광현미경을 통하여 EGFP 단백질이 발현하는 녹색형광을 관찰함으로써 확인하였다. 이중 리포터 유전자 중 NIS 유전자의 기능은 세포에서 방사능 옥소의 섭취량과 유출량의 측정을 통해서 확인하였다. 이렇게 만들어진 세포를 누드마우스에 이식하여 형광 영상, I-123을 이용한 감마카메라 영상과 I-124를 이용한 소동물용 PET 영상을 획득하였다. 결과: NIS와 EGFP의 이중 리포터 유전자를 가지고 있는 HepG2 세포주가 성공적으로 만들어졌다. 세포의 약 50% 정도가 형광 현미경 아래에서 관찰되었다. NIS 유전자의 발현은 역전사효소 중합효소 연쇄반응 실험을 통해서 확인하였고, NIS가 발현된 세포의 방사능옥소 섭취량은 대조군에 비하여 약 9배 정도 높게 나타났다. 방사능옥소 유출량 실험에서는 약 9분에 반 정도의 옥소가 유출되는 것이 확인되었다. 구축된 세포주를 이식한 후 획득한 형광 영상, 감마카메라과 소동물용 PET 영상에서는 반대쪽의 대조군 세포를 이식한 것에 비하여 뚜렷한 형광신호가 보였고, 더 높은 방사능옥소 섭취가 확인되었다. 결론: NIS와 EGFP의 이중 리포터 유전자를 가지는 간암 세포주가 성공적으로 구축되었고, 소동물에서 두 유전자를 각각 치료용 리포터 유전자와 영상 리포터 유전자로의 사용이 가능할 것이라고 생각된다.

• Fisheries and Aquatic Sciences
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• 제17권1호
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• pp.47-57
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• 2014

Thai Jasmine rice (Oryza sativa, long grain Indica var.) is popular in southeastern Asia and China due to its non-glutinous, fluffy texture and fragrant smell. However it has a high starch digestibility, which leads to an increased glycemic index (GI). Therefore it may require modified cooking methods for diabetes patients. The objectives of this study were to optimize the ratio of Thai Jasmine rice, sea tangle, and olive oil (CLTR) based on consumers' acceptance. The GI of plain cooked Thai Jasmine rice (CLR) was measured as a control. Sensory evaluation and response surface methodology were used to determine the optimal ratio. Texture analysis and nutritional evaluation were also performed on the optimal recipe of cooked Jasmine rice with sea tangle. A multiple regression equation was developed in quadratic canonical polynomial models. We used 26 trained Chinese panelists in their forties to rate color, flavor, adhesiveness, and glossiness, which we determined were highly correlated with overall acceptability. The optimal CLTR formula was 34.8% rice, 2.8% sea tangle, 61.9% water, and 0.5% olive oil. Compared to CLR, CLTR had a lower hardness, but a higher springiness and cohesiveness. However, CLR and CLTR had the same adhesiveness and chewiness. The addition of sea tangle and olive oil delayed retro-gradation of starch in CLTR and increased total dietary fiber, and protein and ash contents. The degree of gelatinization, and in vitro protein and starch digestibility of CLTR were lower than those of CLR. Based on Wolver' method, the GI of CLTR (52.9, incremental area under the glycemic-response curve, ignoring the area below fasting, as used for calculating the GI [Inc]) was lower compared with that of CLR (70.94, Inc), which indicates that CLTR is effective in decreasing and stabilizing blood glucose level, owing to its lower degree of gelatinization and starch digestibility. Our results show that CLTR can contribute to the development of a healthier meal for families and the fast food industry.

• BMB Reports
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• 제39권6호
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• pp.677-685
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• 2006

Proteins that are unfolded or misfolded in the endoplasmic reticulum (ER) must be targeted for refolding or degradation to maintain the homeostasis of the ER. Derlin-1 was reportedly implicated in the retro-translocation of misfolded proteins from the ER to the cytosol for degradation. In this report, we showed that Derlin-1 was down-regulated in the endothelial cells derived from human hepatic cavernous hemangioma (CHEC) compared with other tested cells. Electron microscopy analysis showed that ER was aberrantly enlarged in CHEC cells, but not in other tested cells. When overexpressed, Derlin-1 induced the dilated ER to return normal size. This ER dynamic was associated with the activation of unfolded protein response (UPR). In CHEC cells where Derlin-1 was down-regulated, increased expression of the immunoglobulin heavy chain-binding protein (Bip) and UPR-specific splicing of X-box DNA-binding protein 1 (XBP1) mRNA were detected, as compared with that in other tested cells, indicating that UPR was activated. After Derlin-1 overexpression, the extent of UPR activation diminished, as evidenced by decreased expression of Bip, reduced amount of the spliced form of XBP1 ($XBP1_S$), and elevated expression of the unspliced form of XBP1 ($XBP1_U$). Taken together, these findings provide another example of a single protein being able to affect ER dynamic in mammalian cells, and an insight into the possible molecular mechanism(s).

• Pediatric Infection and Vaccine
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• 제24권3호
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• pp.141-145
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• 2017

목적: 농뇨가 동반된 가와사키병(Kawasaki disease [KD]) 환아와 농뇨 동반이 없는 KD 환아의 임상 및 검사실 지표들을 비교하여 농뇨 동반 KD의 임상적 특징을 파악하고자 하였다. 방법: 2015년 1월부터 2016년 12월까지 KD로 입원한 140명(남아 86명, 여아 54명) 환아의 의무기록을 후향적으로 조사하였다. 결과: 48명의 KD 환아(34.3%)에서 농뇨가 관찰되었다. 농뇨 KD군은 비농뇨 KD군보다 높은 C-반응단백(C-reactive protein [CRP]) 값을 보였고 간 효소치가 증가한 환아의 비율이 높았다. 두 군 간의면역글로불린(intravenous immunoglobulin) 치료 저항성과 관상동맥병변(coronary artery lesion) 발생률에 대한 차이점은 없었다. 농뇨 KD군 중 6명(12.5%)은 요로감염(urinary tract infection)을 감별하기 위해 신장 영상검사를 받았고, 농뇨 KD군의 2/3(66.7%)는 KD 표준치료에 추가로 항생제 치료를 받았다. 결론: 농뇨가 동반된 KD 환아들은 농뇨 동반이 없는 KD 환아들보다 더 높은 CRP 값과 더 많은 경우의 간 효소치 증가를 보였다. 이는 농뇨가 동반된 KD 환아가 농뇨 동반이 없는 KD 환아보다 더 심한 전신염증반응을 보인다는 점을 시사한다.

• 대한바이러스학회지
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• 제27권1호
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• pp.39-47
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• 1997

A large number of viruses belonging to Genus Hantavirus in Family Bunyaviridae are etiologic agents for hemorrhagic fever with renal syndrome (HFRS), or hantavirus pulmonary syndrome (HPS). Hantaan (HTN), Seoul (SED), Belgrade (BEL), Puumala (PUU) serotype viruses are well known causative agents for HFRS in Eurasian continent. Among those viruses Hantaan and Seoul serotypes are well known to cause HFRS in Korea, but there are some sporadic incidence by other than Hantaan or Seoul viruses. Recently we have developed the combined Hantaan-Puumala virus vaccine to prevent world-wide occuring HFRS. This combined vaccine is formalin inactivated, suckling mouse and suckling hamster brain extracts for Hantaan and Puumala viruses, respectively. Protein contents of this purified candidate vaccine is $27\;{\mu}g/ml$, which contains 1,024 ELISA antigen units to each virus, but content of myelin basic protein which is causing experimental allergic encephalomyelitis is less than 0.1 ng/ml. Thirty hamsters were given twice at one month interval intra-muscularly and bled on 30 days after each vaccination from retro-orbital sinus vein. Antibody titers were tested against 5 major serotype viruses, Hantaan, Seoul, Belgrade, Puumala and Sin Nombre viruses by IFA and PRNT. The mean IF antibody titers on 30 days after primary shot were 78.4, 68.8, 68.8, 37.9, and 15.6; mean neutralizing antibody titers were 65.4, 12, 6.1, 65.6 and 0.5 against Hantaan, Seoul, Belgrade, Puumala and Sin Nombre viruses, respectively. The mean IF antibody titers on 30 days after booster shot were 686.9, 567.5, 550.4, 516.3, and 430.9; and neutralizing antibody titers were 710.8, 41.9, 24.3, 409.9, and 1.6 against Hantaan, Seoul, Belgrade, Puumala and Sin Nombre viruses, respectively.