• 미생물학회지
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• 제31권2호
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• pp.166-174
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• 1993

Streptomyces coelicolor (Muller) 의 세포에 $100 \mu$M 의 과산화수소를 1시간 동안 처리하여 과산화수소 스트레스중에 생성되는 단백질을 L-[$^{35}S$] methionine 을 이용하여 순간 표지하였다. 단백질을 추출하여 2차원 겔 전기영동으로 분석한 결과 지수 성장기의 세포가 가지는 총세포 단백질 중 약 100개의 단백질의 합성이 과산화수소에 의해 증가되는 것을 관찰하였다. 이들을 Pin(peroxide inducible) 단백질이라고 명명하고 과상화수소 처리 후 발현이 증가되는 시간에 따라 세 그룹으로 나누었다. 약 60 개의 Pin 단백질은 과산화수소 처리후 20분 이내에 발현이 증가하여 1시간동안 지속적으로 다량 합성되었다. 정체 성장기의 세포에서는 62개의 단백질의 합성이 과산화수소에 의해 증가되었으며, 이 중 21 개의 단백질은 지수성장기의 Pin 단백질과 일치하였다. 과산화수소에 대한 저항성이 증가한 세가지 돌연변이체의 단백질을 지수 성장기에서 추출하여 2 차원 겔 전기영동으로 분석한 결과, 각각의 경우 15, 17, 15개의 Pin 단백질을 야생형보다 항상적으로 다량합성하는 것을 관찰하였다. 이 pin 단백질 중 9개 (D74.7 a, E76.0c, E23.3, F50.7, F47.2a, F25.5, G39.6b, G24.0, H39.6a) 는 두가지 돌연변이체 모두에게 증가되었다. 따라서 이 단백질들은 S. coelicolor 가 과산화수소 스트레스에 대응하는데 있어 중요한 역할을 담당한다고 추정된다.

• 생물정신의학
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• 제14권3호
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• pp.177-183
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• 2007

목 적: 성상세포(astrocyte)에서 생산되어 신경세포의 증식과분화에 관여하는 S100B 단백이 정신분열병의 진행과 증상론과 관련이 있다는 연구가 계속 진행되고 있다. 이에 정신분열병 환자와 정상대조군의 혈청 S100B 농도를 비교하고, 정신분열병 환자의 증상 양상과 S100B 농도와의 연관성을 연구하였다. 방 법: DSM-IV-TR 진단 기준에 따라 정신분열병으로 진단받은 환자 중 정신분열병 최초 발병 환자 혹은 최소 6개월 간 약물 치료를 하지 않은 정신분열병 환자 21명과 정상대조군 27명의 혈청 S100B 농도를 비교하였으며, 정신분열병 환자의 PANSS 전체점수와 양성증상점수, 음성 증상점수 등과 S100B의 혈청농도 간의 상관관계에 대해서 알아보았다. 결 과: 정신분열병 환자에서의 S100B 혈청농도($0.074{\pm}0.039$ ng/ml)와 정상대조군에서의 S100B 혈청 농도($0.072{\pm}0.030$ng/ml)에는 통계적인 차이는 없었다(p=0.925). 또한, 음성증상점수와 S100B 혈청농도(${\rho}$=0.410, p=0.065 ; 표 3)의 상관관계 및 양성증상점수와 S100B 혈청농도 (${\rho}$=-0.390, p=0.080 ; 표 3)의 상관관계는 통계적으로 유의하지 않았다. 결 론: 이번 연구에서 S100B 혈청농도와 정신분열병과의 관련성을 뚜렷하게 발견하기 어려웠다. 이러한 S100B의 역할과 연관성을 확인하기 위해서 연구대상의 확대와 CSF에서의 농도 측정, 장기적 추적 검사, 다른 관련 물질과의 연관성에 대한 추가적인 연구가 필요하다.

• Journal of Korean Neurosurgical Society
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• 제39권4호
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• pp.271-276
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• 2006

Objective : Despite the recent progress that has been made in intracerebral monitoring, it is still difficult to quantify the exact extent of primary brain damage after severe head injury. In this work, we investigate the role of S-100B protein as a serum marker of brain damage after severe head injury. Methods : 21 patients with severe head injury [GCS score <9] were selected for this prospective study. A venous blood sample was taken as soon as possible after head injury and the serum concentration of S-100B protein was measured daily for five consecutive days. The serum level of S-100B protein was compared with the patients' outcome. The outcome was measured twice, at hospital discharge and after 6 months of follow-up using the Glasgow Outcome Scale[GOS]. Results : Those patients who died within two weeks [after head injury] had a significantly higher serum S-100B value than those who survived [median, 9.64ug/L versus 2.91ug/L]. Seven [78%] of the nine patients who died had a maximum S-100B value of 2ug/L or higher, while three [25%] of the twelve surviving patients showed a maximum S-100B protein value of more than 2ug/L [P<005]. Conclusion : These results indicate that S-100B protein appears to be the most reliable index for estimating the extent of brain damage.

• Asian-Australasian Journal of Animal Sciences
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• 제30권10호
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• pp.1478-1485
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• 2017

Objective: The effects of vaccinating 18-day-old chicken embryos with the combination of recombinant Eimeria profilin plus Clostridium perfringens (C. perfringens) NetB proteins mixed in the Montanide IMS adjuvant on the chicken immune response to necrotic enteritis (NE) were investigated using an Eimeria maxima (E. maxima)/C. perfringens co-infection NE disease model that we previously developed. Methods: Eighteen-day-old broiler embryos were injected with $100{\mu}L$ of phosphate-buffered saline, profilin, profilin plus necrotic enteritis B-like (NetB), profilin plus NetB/Montanide adjuvant (IMS 106), and profilin plus Net-B/Montanide adjuvant (IMS 101). After post-hatch birds were challenged with our NE experimental disease model, body weights, intestinal lesions, serum antibody levels to NetB, and proinflammatory cytokine and chemokine mRNA levels in intestinal intraepithelial lymphocytes were measured. Results: Chickens in ovo vaccinated with recombinant profilin plus NetB proteins/IMS106 and recombinant profilin plus NetB proteins/IMS101 showed significantly increased body weight gains and reduced gut damages compared with the profilin-only group, respectively. Greater antibody response to NetB toxin were observed in the profilin plus NetB/IMS 106, and profilin plus NetB/IMS 101 groups compared with the other three vaccine/adjuvant groups. Finally, diminished levels of transcripts encoding for proinflammatory cytokines such as lipopolysaccharide-induced tumor necrosis $factor-{\alpha}$ factor, tumor necrosis factor superfamily 15, and interleukin-8 were observed in the intestinal lymphocytes of chickens in ovo injected with profilin plus NetB toxin in combination with IMS 106, and profilin plus NetB toxin in combination with IMS 101 compared with profilin protein alone bird. Conclusion: These results suggest that the Montanide IMS adjuvants potentiate host immunity to experimentally-induced avian NE when administered in ovo in conjunction with the profilin and NetB proteins, and may reduce disease pathology by attenuating the expression of proinflammatory cytokines and chemokines implicated in disease pathogenesis.

• Asian-Australasian Journal of Animal Sciences
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• 제29권5호
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• pp.731-738
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• 2016

Glucagon-like peptide-2 (GLP-2) is important for intestinal barrier function and regulation of tight junction (TJ) proteins, but the intracellular mechanisms of action remain undefined. The purpose of this research was to determine the protective effect of GLP-2 mediated TJ and transepithelial electrical resistance (TER) in lipopolysaccharide (LPS) stressed IPEC-J2 cells and to test the hypothesis that GLP-2 regulate TJ and TER through the phosphatidylinositol 3-kinase (PI3K)-protein kinase B (Akt)-mammalian target of rapamycin (mTOR) signaling pathway in IPEC-J2 cells. Wortmannin and LY294002 are specific inhibitors of PI3K. The results showed that $100{\mu}g/mL$ LPS stress decreased TER and TJ proteins occludin, claudin-1 and zonula occludens protein 1 (ZO-1) mRNA, proteins expressions (p<0.01) respectively. GLP-2 (100 nmol/L) promote TER and TJ proteins occludin, claudin-1, and zo-1 mRNA, proteins expressions in LPS stressed and normal IPEC-J2 cells (p<0.01) respectively. In normal cells, both wortmannin and LY294002, PI3K inhibitors, prevented the mRNA and protein expressions of Akt and mTOR increase induced by GLP-2 (p<0.01) following with the significant decreasing of occludin, claudin-1, ZO-1 mRNA and proteins expressions and TER (p<0.01). In conclusion, these results indicated that GLP-2 can promote TJ's expression and TER in LPS stressed and normal IPEC-J2 cells and GLP-2 could regulate TJ and TER through the PI3K/Akt/mTOR pathway.

• Journal of Microbiology and Biotechnology
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• 제19권5호
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• pp.511-519
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• 2009

A chimeric gene encoding enhanced green fluorescent protein (EGFP) and a S-layer protein from Lactobacillus brevis KCTC3102, and/or two copies of the Fe-binding Z-domain, a synthetic analog of the B-domain of protein A, was constructed and expressed in Escherichia coli BL21(DE3). The S-layer fusion proteins produced in a 500-1 fermentor were likely to be stable in the range of pH 5 to 8 and $0^{\circ}C$ to $40^{\circ}C$. Their adhesive property enabled an easy and rapid immobilization of enzymes or antibodies on solid materials such as plastics, glass, sol-gel films, and intestinal epithelial cells. Owing to their affinity towards intestinal cells and immunoglobulin G, the S-layer fusion proteins enabled the adhesion of antibodies to human epithelial cells. In addition, feeding a mixture of the S-layer fusion proteins and antibodies against neonatal calf diarrhea (coronavirus, rotavirus, Escherichia coli, and Salmonella typhimurium) to Hanwoo calves resulted in 100% prevention of neonatal calf diarrhea syndrome (p<0.01), whereas feeding antibodies only resulted in 56% prevention.

• Journal of Periodontal and Implant Science
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• 제46권1호
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• pp.2-9
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• 2016

Purpose: Porphyromonas gingivalis and Tannerella forsythia have been implicated as the major etiologic agents of periodontal disease. These two bacteria are frequently isolated together from the periodontal lesion, and it has been suggested that their interaction may increase each one's virulence potential. The purpose of this study was to identify proteins on the surface of these organisms that are involved in interbacterial binding. Methods: Biotin labeling of surface proteins of P. gingivalis and T. forsythia and liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis was performed to identify surface proteins involved in the coaggregating activity between P. gingivalis and T. forsythia. Results: It was found that three major T. forsythia proteins sized 161, 100, and 62 kDa were involved in binding to P. gingivalis, and P. gingivalis proteins sized 35, 32, and 26 kDa were involved in binding to T. forsythia cells. Conclusions: LC-MS/MS analysis identified one T. forsythia surface protein (TonB-linked outer membrane protein) involved in interbacterial binding to P. gingivalis. However, the nature of other T. forsythia and P. gingivalis surface proteins identified by biotin labeling could not be determined. Further analysis of these proteins will help elucidate the molecular mechanisms that mediate coaggregation between P. gingivalis and T. forsythia.

• 한국미생물·생명공학회지
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• 제16권5호
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• pp.389-392
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• 1988

Escheriachia coli pSL 2-1 clone은 Bacillus sphaericus 1593의 모기살충 유전자를 클로닝한 재조합 DNA이다. 이 클론이 생산하는 살충독소 단백질의 분자량을 SDS-polyacrylamide gel을 이용하여 측정했다. B. sphaericus 1593균이 생산하는 독소결정체를 분리하여 전기영동을 한 결과는 6개의 단백질밴드(43, 58, 64, 100, 113, 130Kd)가 형성되었으나, 독소결정체를 알칼리 pH로 용해하여 전기영동을 하면 2개의 단백질 밴드(43과 64Kd)만이 나타났다. 그러나 대장균 pSL2-1균이 생산하는 독소단백질을 Sephadex G-200으로 정제하여 모기유충에 살충력이 있는 단백질을 전기영동한 결과는 42Kd만이 나타났다. LC50은 2 $\mu\textrm{g}$/$m\ell$이었다. B. sphaericus와 pSL2-1 clone 생산하는 살충단백질은 42Kd 단백질인 것으로 생각된다.

• 한국동물위생학회지
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• 제41권1호
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• pp.57-61
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• 2018

Canine peripheral nerve sheath tumors (PNSTs) are spindle cell tumors that arise from Schwann cells, perineural cells, fibroblasts or all of them. Based on the morphology and biologic behavior, PNSTs are divided into benign PNST (BPNST) and malignant PNST (MPNST) forms. The aim of this study is to diagnose the two cases of neoplastic tissue samples with features of PNSTs by the histopathology and immunohistochemistry. The study was performed using two specimens from small animal clinic. The first case, A was a mass, 3~4 cm in diameter, extruded from vaginal mucosa of 10-year-old spayed female mixed-breed dog. And the second case, B was a subcutaneous mass, 1.5 cm in diameter, which is originated from right hind leg of 9-year-old castrated male mixed-breed dog. Two cases were stained with hematoxylin and eosin (H&E) for histopathological examination. And also immunohistochemistry (IHC) was performed by the avidin-biotin peroxidase complex (ABC) method with antibodies specific for the following proteins: S-100 protein, smooth muscle actin (SMA) and epidermal growth factor receptor (EGFR). In results, Antoni B schwannoma pattern characterized by pleomorphic, round and fusiform polygonal cells was seen in A. In B, Antoni A pattern, densely packed spindle cells arranged in interlacing bundles was seen in addition to Antoni B pattern. In IHC, cytoplasms of neoplastic cells were diffusely labeled for S-100 expression in A and B. For SMA, both A and B show negative expression. And for EGFR, A shows negative expression but B shows partially positive expression in areas of Antoni B schwannoma pattern. The histopathologic features of two cases coupled with the S-100 immunoreactivity led to a diagnosis of PNST. For SMA, both A and B show negative expression. The diagnosis of A will be a BPNST with the negative result and B will be a MPNST with the positive result for EGFR.

• 한국환경농학회지
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• 제25권1호
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• pp.7-13
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• 2006

비닐하우스는 노지보다 UV-B 선량이 적고 외기와 차단되어 노지와 다른 생육환경 하에서 작물이 생육하게 된다. 본 연구는 UV-B 저선량 상태인 비닐하우스 재배 잎들깨에 UV-B를 처리하여 수량과 생리변화를 검토하고자 수행하였다. UV-B는 형광램프를 이용하여 잎들깨 수관으로부터 150, 120, 90 cm에서 처리하여 각각 노지자연량, 노지자연량+50% 증가량, 노지자연량+100% 증가량을 다른 광의 간섭이 없는 밤 9시부터 2시간씩 30일간 처리하였다. UV-B 처리에 의한 잎들깨 수량은 비닐하우스, 노지 자연량 150% 증가량, 노지자연량, 노지자연량+100% 증가량 처리 순이었다. 7월 20일의 자연 UV-B 선량은 노지 13.6 kJ/일, 비닐하우스 4.9 kJ/일로 비닐하우스의 UV-B 선량은 노지보다 64% 감소되는 것으로 측정되었다. UV-B가 균일하게 처리된 잎들깨의 단백질을 추출하여 2차원전기영동으로 분리하고, 이미지분석하여 발현량을 분석한 결과 UV-B에 의해 33개의 단백질 발현이 변화되었으며 이중 10개가 동정되었다. 동정된 단백질의 기능별로 분류하면 광합성과 관련된 것이 40%, 스트레스 및 스트레스 방어와 관련된 것이 60%였다. UV-B에 의해 발현이 감소한 단백질은 광합성과 관련된 ATP synthase CF1 alpha chain이었고 발현이 증가한 단백질은 DNA recombination and repair protein recF, Heat shock protein 21, Catalase, Galactinol synthase, S-adenosyl-L-methionine, Calcium-dependent protein kinase(CDPK)-like 로 주로 스트레스 및 스트레스 방어와 관련된 단백질들로서 UV-B는 잎들깨 세포내 DNA와 광합성기구를 손상시켜 광합성에 저해적으로 작용하여 수량을 감소시킨 것으로 판단되었다.