• 센서학회지
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• 제15권4호
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• pp.237-244
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• 2006

Atomic force microscope (AFM) has become a common tool for the structural and physical studies of biological macromolecules, mainly because it provides the ability to perform experiments with samples in a buffer solution. In this study, structure of proteins and nucleic acids has been studied in their physiological environment that allows native intermolecular complexes to be formed. Cr and Au were deposited on p-Si (100) substrate by thermal evaporation method in sequence with the thickness of $200{\AA}$ and $500{\AA}$, respectively, since Au is adequate for immobilizing biomolecules by forming a self-assembled monolayer (SAM) with semiconductor-based biosensors. The SAM, streptavidin and biotin interacted each other with their specific binding energy and their adsorption was analyzed using the Bio-AFM both in a solution and under air environment. A silicon nitride tip was used as a contact tip of Bio-AFM measurement in a solution and an antimony doped silicon tip as a tapping tip under air environment. Actual morphology could also be obtained by 3-dimensional AFM images. The length and agglomerate size of biomolecules was measured in stages. Furthermore, $R_{a}$ (average of surface roughness) and $R_{ms}$ (mean square of surface roughness) and surface density for the adsorbed surface were also calculated from the AFM image.

• 한국잠사곤충학회지
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• 제50권1호
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• pp.15-19
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• 2012

최근 다양한 생체재료를 이용하여 연골재생과 관련한 많은 연구가 진행되고 있다. 실크단백질은 생체적합성이 뛰어나며, 우수한 기계적 강도를 가지고 있는 천연 고분자 물질로서 최근 생체재료로 사용하기위한 연구가 세계적으로 많이 이루어지고 있다. 본 연구는 실크단백질이 연골재생에 효과가 있는지를 확인하기위하여 수행되었다. 우리는 연골세포를 코뼈로부터 분리하고, 3종류의 배지 (DMEM, DMEM/F12, RPMI)와 서로 다른 농도의 ascorbic acid를 사용하여 최적 배양조건을 확립하였다. 그 결과 우리가 분리한 연골세포는 10% FBS와 $100{\mu}M$ ascorbic acid가 함유된 DMEM배지에서 가장 잘 생장하였다. 연골에 대한 실크의 영향을 관찰하기위해서, 실크 피브로인 용액을 제작하고 이를 멸균한것과 멸균하지 않은 것으로 구분하여 연골세포 배양 시 첨가하여 연골분화에 대한 마커인자인 제2형 콜라겐의 발현량을 측정하였다. 멸균하지 않은 실크 피브로인 첨가시 제2형 콜라겐의 발현량이 2.7배 증가하였으나, 멸균된 실크 피브로인의 첨가는 제2형 콜라겐의 발현량을 오히려 감소시켰다. 또한 실크 피브로인은 제10형 콜라겐의 발현을 증가시키는 것을 확인하였다. 이 효과는 특히 연골세포를 3차원 배양할 때 더 컸다. 본 연구결과를 통하여 우리는 연골을 재생하는데 있어서 실크 단백질을 가능성을 보았으며, 향후 연구에서 연골재생과 실크의 관계를 좀 더 정밀하게 파악하고자 한다.

• Nutritional Sciences
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• 제7권1호
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• pp.23-30
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• 2004

As a central component of a novel protein kinase cascade, the activation of the mitogen-activated protein (MAP) kinase cascade has attracted considerable attention. We sought to determine the effect of exercise and diet on the activation of the extracellular-signal regulated protein kinase (ERK) 1/2 and the p38 MAP kinase pathways in rat soleus muscle. Forty-eight Sprague-Dawley rats were assigned to one of two dietary conditions: high-carbohydrate (CHO) or high-fat (FAT). Animals having each dietary condition were further divided into one of three subgroups: a sedentary control group that did not exercise (NT), a group that performed 8 weeks of treadmill running and was sacrificed 48 h after their final treadmill run (CE), and a group that was sacrificed immediately after their final routine exercise training (AE). A high-fat diet did not have any significant effect on phosphorylated and total forms of ERK 1/2 or p38 MAP kinase. In chronically trained muscle that was taken 48 h after the last training, phosphorylated ERK 1/2 significantly increased only in the FAT but not in the CHO groups. In the case of total ERK 1/2, it increased significantly for both groups. In contrast, both phosphorylated and total forms of p38 MAP kinase decreased markedly compared to sedentary muscle. In muscle that was taken immediately after a last bout of exercise, phosphorylated ERK 1/2 increased in both groups but statistical significance was seen only in the CHO group. Total ERK 1/2 in acutely stimulated muscle increased only in the CHO-AE group even though the degree was much lower than the phosphorylated status. Muscle that was taken immediately after the routine training increased in phosphorylation status of p38 MAP kinase for both dietary conditions. However, statistical significance was seen only in the CHO group owing to a large variation with FAT. In conclusion, a high-fat diet per se did not have any notable effect versus a high-carbohydrate diet on MAP kinase pathways. However, when diet (either CHO or FAT) was combined with exercise and/or training, there was differentiated protein expression in MAP kinase pathways. This indicates MAP kinase pathways have diverse control mechanisms in slow-twitch fibers.

• Journal of Gastric Cancer
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• 제4권3호
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• pp.169-175
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• 2004

Purpose: The tumor suppressor gene p53 has been shown to be a factor in the carcinogenesis or progression of gastric cancer. The mutant p53 has been reported to cause a higher risk of lymph-node metastasis. Futhermore, mutation of the p53 has been linked to a poor prognosis for gastric cancer. The heat shock protein-27 (HSP27), a stress protein, has also been reported to be a poor prognostic factor in ovarian and breast cancers. However, in gastric-cancer patients, controversies exist as to its influence on the prognosis. In the present study, we used an immunohistochemical stain to observe the effects of p53 and HSP27 on the clinicopathological factors and on the prognosis for gastric-cancer patients. Materials and Methods: To evaluate the significance of p53 and HSP27 in gastric cancer patients, we analyzed 212 cases of gastric cancer (stage I.IV). Tissue samples of 212 patients were stained immunohistochemically for the mutant p53 protein and for HSP27. The correlations between protein expression and the clinicopathological factors were investigated. Results: The overall expression rates for p53 and HSP27 were $36.9\%\;and\;27.8\%$, respectively. p53 and HSP27 were correlated to each other because the HSP27 expression rate was higher in the p53-positive group (P=0.046). Statistically, the p53 and the HSP27 expression rates were significantly increased in the case of tumor invasiveness, lymphatic metastasis and vessel involvement. Therefore, they play a role in cancer progression. The 5-year survival rates of the p53-positive and the p53-negative groups were $62.8\%\;and\;60.1\%$, respectively (P=0.793) while the 5-year survival rates for the HSP27-positive and HSP27-negative groups were $54.2\%\;and\;63.1\%$, respectively (P=0.090). Conclusion: p53 and HSP27 were correlated to each other in our immunohistochemical study of gastric carcinomas and they were not independent prognostic factors in gastric- cancer patients. However, further studies are needed to determine their prognostic values for gastric-cancer patients.

• Clinical and Experimental Reproductive Medicine
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• 제29권4호
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• pp.323-335
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• 2002

Object: The present study was accomplished to obtain a gene expression profile of the luminal epithelium during embryo apposition in comparison of implantation (1M) and interimplantation (INTER) sites. Material and Method: The mouse uterine luminal epithelium from IM and INTER sites were sampled on day 4.5 (Day of vaginal plug = day 0.5) by Laser Captured Microdissection (LCM). RNA was extracted from LCM captured epithelium, amplified, labeled and hybridized to microarrays. Results from microarray hybridization were analyzed by Significance Analysis of Microarrays (SAM) method. Differential expression of some genes was confirmed by LCM followed by RT-PCR. Results: Comparison of IM and INTER sites by SAM identified 73 genes most highly ranked at IM, while 13 genes at the INTER sites, within the estimated false discovery rate (FDR) of 0.163. Among 73 genes at IM, 20 were EST/unknown function, and the remain 53 were categorized to the structural, cell cycle, gene/protein expression, immune reaction, invasion, metabolism, oxidative stress, and signal transduction. Of the 24 structural genes, 14 were related especially to extracellular matrix and tissue remodeling. Meanwhile, among 13 genes up-regulated at INTER, 8 genes were EST/unknown function, and the rest 5 were related to metabolism, signal transduction, and gene/protein expression. Among these 58 (53+5) genes with known functions, 13 genes (22.4%) were related with $Ca^{2+}$ for their function. Conclusions: Results of the present study suggest that 1) active tissue remodeling is occurring at the IM sites during embryo apposition, 2) the INTER sites are relatively quiescent than IM sites, and 3) the $Ca^{2+}$ may be a crucial for apposition. Search for human homologue of those genes expressed in the mouse luminal epithelium during apposition will help to understand the implantation process and/or implantation failure in humans.

• 생명과학회지
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• 제18권7호
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• pp.952-957
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• 2008

본 실험에서는 THP-1 세포의 PMA에 의하여 유도되는 초기 세포부착에 관한 메카니즘을 이해하기 위하여 다양한 요인(혈청, 신규 단백질의 합성, 세포 골격 저해제, 단백질 인신화 저해제)들의 효과를 조사하였다. 또한 본 실험에서는 이들 세포부착의 정도를 일반적으로 세포증식 분석에 사용되고 있는 SRB염색법을 도입하여 세포부착 분석에 간편한 방법의 조건을 확립하였다. PMA에 의한 초기 세포부착에는 배양액중의 혈청의 유무는 영향이 없었으나, 신규 단백질의 합성이 요구되는 것을 확인하였다. 또한 이들 초기 세포부착에 PMA처리에 의한 PKC의 활성화는 필수적이나, 그 하류 활성화 인자로 잘 알려진 MAP-kinase (erk1/2)의 인산화는 필요치 않음을 알 수 있었다. 흥미롭게도 액틴 중합 저해제인 cytochalasin D의 PMA와 공 처리는 오히려 세포부착을 PMA 단독 처리시 보다 증가시켰다. 또한 본 실험에서 사용된 SRB 염색법을 통한 세포부착 분석법은 최근 암 등 다양한 질환의 신약 표적 분자로 주목을 받고 있는 PKC 저해제의 초기 세포 기반 분석에 매우 유용하리라고 생각된다.

• 한국잠사곤충학회지
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• 제52권1호
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• pp.45-51
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• 2014

면역 유도된 천잠(Antheraea yamamai) 유충에서 cecropin-A 유전자를 분리하였고 이 유전자를 Ay-CecA로 명명하였다. 전체 Ay-CecA cDNA 크기는 419 bp로 64개의 아미노산 잔기를 인코딩하는 195 bp ORF로 구성되어 있다. 천잠 CecA 유전자는 22개 잔기의 signal peptide, 4개 잔기의 propeptide 및 항균활성을 갖는 37개 잔기로 구성된 성숙 단백질(mature protein) 영역으로 구성되고 예상 분자량은 4046.81 Da으로 산출되었다. 천잠 CecA의 아미노산 서열은 다른 나비목 곤충에서 분리된 cecropin와 매우 높은 상동성(62 ~ 78%)을 나타냈다. Ay-CecA 유전자의 C말단에 기존에 보고된 곤충의 cecropin에서와 동일하게 C말단 아미드화를 위한 glycine 잔기가 존재하고 있다. 이 펩타이드의 항균활성을 검정하기 위해서 대장균 발현 시스템을 이용하여 활성이 있는 재조합 Ay-CecA 발현에 성공하였다. 발현 기주인 대장균에 대한 재조합 CecA 독성 중화를 위해서 불용성 단백질인 ketosteroid isomerase(KSI) 유전자를 CecA 유전자와 융합하였다. 융합 CecA-KSI 단백질은 대부분 불용성 단백질로 발현되었다. 발현된 융합단백질은 Ni-NTA immobilized metal affinity chromatography(IMAC)에 의해서 정제하였으며 CNBr 반응을 통하여 재조합 CecA를 절단하여 용출하였다. 최종적으로 양이온 교환 chromatography 과정을 통하여 CecA를 순수 정제하였다. 정제된 재조합 Ay-CecA는 그람음성균인 E. cori ML 35, Klebsiella pneumonia 및 Pseudomonas aeruginosa에 대해 매우 높은 항균활성을 나타냈었다. 따라서 본 연구 결과, 높은 항균활성 지닌 CecA는 천잠의 면역 반응에서 중요한 역할을 담당할 것으로 사료된다.

• 한국수산과학회지
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• 제47권5호
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• pp.495-500
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• 2014

We examined chemical changes in oysters Crassostrea gigas and packing water that were sold after storage at 5, 10, and $20^{\circ}C$. The pH of oysters stored at $5^{\circ}C$ dropped to 5.81 after 10 days of storage, while that of oysters at $10^{\circ}C$ and $20^{\circ}C$ dropped to 5.37 after 8 days and to 5.04 after 4 days, respectively. The glycogen content of oysters stored at $5^{\circ}C$ decreased from 718.89 to 421.85 mg/100g during storage, while that of oysters at $10^{\circ}C$ decreased to 351.49 mg/100 g after 4 days. The turbidity and soluble protein in packing water increased slightly. The viable cell count of oysters did not exceed 6 log CFU/g after 10 days of storage at $5^{\circ}C$, but that of oysters at $10^{\circ}C$ did so after 8 days. Additionally, the viable cell count of packing water was lower than that of oysters. We performed a principal component analysis, where the first principal component (55.03%-57.24%) and second principal component (42.76%-44.97%) described most variation. The first principal component included the pH of oysters and packing water, and the glycogen content of oysters. A Pearson correlation between the first two principal components had a higher R value than that between other components. Freshness was evaluated using the pH of oysters and packing water, and glycogen. We found that soluble protein content was significantly associated with a lower pH and glycogen content.

• 한국균학회지
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• 제26권4호통권87호
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• pp.551-561
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• 1998

S.B, P.S, C.P를 각각 처리한 배지에 배양된 진균류(Aspergillus phoenicis, Rhizopus acidus, Candida albicans)에서 원형질막을 분리하였다. 원형질막 단백질의 함량과 특성을 대조구와 비교하였다. A. phoenicis의 생장은 S.B 처리구에서 평균 64.0%가 감소하였다. R. acidus의 생장은 P.S 처리구에서 평균 69.0%가 감소하였다. 또한 C. albicans의 생장도 S.B 처리구에서 평균 59.5%의 감소를 나타내었다. 여러 진균세포 원형질막에 함유된 단백질의 함량은 S.B 처리구에서 평균 41.0%, 41.7%, 59.5%가 각각 억제되었다. A. phoenicis의 원형질막 단백질의 변화 양상은 처리구에서는 배양 1일째, 2일째는 대조구와 비슷한 경향을 나타내었다. 그러나 배양 1일째 $116\;KD{\sim}97\;KD$ 밴드는 거의 사라졌고 $45\;KD{\sim}29\;KD$ 밴드는 희미하게 관찰되었다. R. acidus는 S.B 처리구에서 배양 중간기부터 $116\;KD{\sim}97\;KD$ 밴드가 소실되었으며, P.S 처리구와 C.P 처리구는 배양초에 소실되기 시작하여 배양 36시간에는 완전히 소실되었다. C. albicans에서는 $116\;KD{\sim}97\;KD$ 밴드가 대조구와 비교하여 배양초에 소실되기 시작하였고 $66\;KD{\sim}45\;KD$밴드는 배양 96시간에 희미하게 나타났다. 특히 C.P 처리구에서는 배양 96시간에 완전히 소실되었다.

• 한국수산과학회지
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• 제47권6호
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• pp.824-831
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• 2014

This study was conducted to investigate the effects of fishmeal replacement with acid-concentrated soybean meal (ACSBM) on growth performance, blood biochemistry, and ingredient digestibility in juvenile olive flounder Paralichthys olivaceus. Six experimental diets were formulated to replace fishmeal protein with ACSBM at 0%, 20%, 30%, 40%, 50%, and 60% (designated ACSBM0, ACSBM20, ACSBM30, ACSBM40, ACSBM50, and ACSBM60, respectively). Triplicate groups of fish (initial fish mean weight: $14.3{\pm}0.03g$) were fed the experimental diets to apparent satiation (twice daily at 08:00 and 18:00 h). After a 12-week feeding trial, a total of 180 healthy fish were randomly distributed into three Guelph system tanks at a density of 60 fish/tank (initial fish mean weight : $50.6{\pm}2.4g$) to test the apparent digestibility coefficients of the ingredients (ACSBM, fishmeal, and soybean meal). Although negative effects were observed with ACSBM40, ACSBM50 and ACSBM60 after 12 weeks of feeding, up to 20% of the fishmeal protein could be successfully replaced with ACSBM without significant growth depression. Hemoglobin and hematocrit values of fish fed the ACSBM50 and ACSBM60 diets were significantly lower than those of fish fed the ACSBM0 diet. Glucose values of fish fed the ACSBM60 diet were significantly higher than those of fish fed the ACSBM0 and ACSBM20 diets. Digestibility of protein in ACSBM and soybean meal was 85.9% and 82.5%, respectively. Results indicated that at least 20% of fishmeal protein can be replaced by ACSBM in diets of juvenile olive flounder without supplementation of limiting amino acids.