• Clinical and Experimental Vaccine Research
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• 제13권3호
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• pp.232-241
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• 2024

Purpose: Brucellosis, a zoonotic infectious disease, is a worldwide health issue affecting animals and humans. No effective human vaccine and the complications caused by the use of animal vaccines are among the factors that have prevented the eradication of the disease worldwide. However, bio-engineering technologies have paved the way for designing new targeted and highly efficacious vaccines. In this regard, the study aimed to evaluate immunity induced by mannosylated niosome containing Brucella recombinant trigger factor/Bp26/Omp31 (rTBO) chimeric protein in a mouse model. Materials and Methods: rTBO as chimeric antigen (Ag) was expressed in Escherichia coli BL21 (DE3) and, after purification, loaded on niosome and mannosylated niosome. The characteristics of the nanoparticles were assessed. The mice were immunized using rTBO, niosome, and mannosylated niosome-rTBO in intranasal and intraperitoneal routes. Serum antibodies (immunoglobulin [Ig]A, IgG, IgG1, and IgG2a) and splenocyte cytokines (interferon-gamma, interleukin [IL]-4, and IL-12) were evaluated in immunized mice. Finally, immunized mice were challenged by B. melitensis and B. abortus. A high antibody level was produced by niosomal antigen (Nio-Ag) and mannosylated noisomal antigen (Nio-Man-Ag) compared to the control after 10, 24, and 38 days of immunization. The IgG2a/IgG1 titer ratio for Nio-Man-Ag was 1.2 and 1.1 in intraperitoneal and intranasal methods and lower than one in free Ag and Nio-Ag. Cytokine production was significantly higher in the immunized animal with Ag-loaded nanoparticles than in the negative control group (p<0.05). Moreover, cytokine and antibody levels were significantly higher in the injection than in the inhalation method (p<0.05). Results: The combination of mannosylated noisome and rTBO chimeric proteins stimulate the cellular and humoral immune response and produce cytokines, playing a role in developing the protective acquired immune response in the Brucella infectious model. Also, the intraperitoneal route resulted in a successful enhancement of cytokines production more than intranasal administration. Conclusion: Designing an effective vaccine candidate against Brucella that selectively induces cellular and humoral immune response can be done by selecting a suitable nanoniosome formulation as an immunoadjuvant and recombinant protein as an immune response-stimulating Ag.

• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2002년도 국제심포지엄
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• pp.97-97
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• 2002

Human eryhropoietin (EPO) is acidic glycoprotein hormone that plays key role in hematopoiesis by facilitating differentiation of erythrocyte and formation of hemoglobin (Hb) and is used for the treatment of anemia. Human EPO is consist of 166 amino acids which is modified by three N-glycosylations (24, 38, 83) and single O-glycosylation (126). N-glycosylation is reported to be related to the cellular secretion and activity of EPO. In this study, we examined effects of mutagenesis in glycosylation site of recombinat hEPO for the cellular secretion during production from cultured CHO cell. We produced rhEpo which was cloned by PCR from human liver cDNA (TaKaRa) in cultured CHO cell. Using supernatant of the culture, ELISA assay and western analysis were performed. To estimate biological activity, 20IU of rhuEpo was subcutaneously injected into four ICR mice. After 8 days, HCT level was increased average 13 per cent, RBC was increased ca. 2${\times}$10$\^$6//${\mu}\ell$. In disease model Rat (anemia c-kit, WSRC-WS/WS), HCT was increased ca. 12%, RBC was increased ca. 1.6${\times}$10$\^$6//${\mu}\ell$. These results suggests that rhEpo we produced has biological activity. To remove glycosylation site by substituting 24, 38, 83, and 126th asparagine (or serine) with glutamic acid, overlapping -extension site-directed mutagenesis was performed. To add novel glycosylation sites, 69, 105th leucine was mutated to asparagine. Mutant EPO construct was transfected into CHO cell. Supernatant of the cell culture was analyzed using ELISA assay with monoclonal anti-EPO antibody (Medac, Germany). Since, several reports for mutagenesis of glycosylation sites showed case-by-case results, we examined both transient expression and stable expression. Addition of novel glycosylation sites resulted no secretion while deletion mutants had little effect except some double deletion mutants (24/83 and 38/83) and triple mutant. We suggest that not single but combination of glycosyl group affect secretion of EPO.

• 한국응용과학기술학회지
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• 제37권4호
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• pp.744-754
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• 2020

본 연구는 파킨슨 질환(Parkinson's disease) 마우스 모델을 대상으로 지구성 운동과 MitoQ 섭취가 뇌의 흑질의 미토콘드리아 기능에 미치는 영향을 확인하는데 목적이 있다. 파킨슨 질환을 유도하기 위해 C57BL/6 수컷 마우스를 대상으로 복강 내 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP) 25mg/kg과 흡수를 돕기 위한 probenecid 250mg/kg을 이용하여 주 2회 5주간 총 10회 투여하였다. 실험 집단은 생리식염수를 투여하는 집단(Normal Conrol (NC), n=10), MPTP 투여집단(MPTP Control (MC), n=10), MPTP 투여 + MitoQ 투여집단(MPTP + MitoQ (MQ), n=10), MPTP 투여 + 운동집단(MPTP + Exercise (ME), n=10), MPTP 투여 + MitoQ 투여 + 운동집단(MPTP + MitoQ + Exercise (MQE), n=10) 총 5 집단으로 구성하였으며, 운동집단은 지구성 운동을 실시하였고 MitoQ집단은 점진적으로 250μmol로 늘리면서 5주간 섭취하였다. 연구결과 Rotarod-test에서 MC 집단에 비해 처치 집단은 운동 기능 저하의 개선을 보였다. 또한 MC 집단에 비해 처치 집단은 tyrosine hydroxylase의 수준의 증가와 알파시누클린(α-synuclein) 단백질 축적을 감소시켰다. 그리고 미토콘드리아 생합성에 주요조절 인자인 PGC-1α와 항산화 효소인 Catalase 발현이 MC 집단에 비해 처치 집단에서 증가해 미토콘드리아 기능을 개선했으며, 세포사멸 조절인자인 Bcl-2의 증가와 Bax의 감소를 통해 세포사멸을 완화했다. 따라서 5주간의 지구성 운동과 MitoQ 섭취는 파킨슨 질환에서 나타나는 병리학적 특징을 완화하고 운동기능을 향상시키는데 효과적인 것으로 나타났다.

• 한국임상수의학회지
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• 제27권3호
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• pp.240-245
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• 2010

대퇴골 머리의 골괴사는 특발성이며 진행성 질환이다. 골괴사증 연구를 위한 다양한 동물모델이 보고되었지만, 현재까지 표준화된 동물모델은 완성되지 않았다. 본 연구에서 랫드에서 세가지 외과적인 방법으로 유발한 골괴사모델을 비교하였다. 20마리의 SD 랫드 (24주령, 숫컷)를 각 5마리씩 대조군과 세개의 실험군으로 구분하였다. 세가지 외과적인 방법으로 골괴사를 유발하였다. 원인대를 자르고, 대퇴골목의 골막을 절개 (S군), 철사를 대퇴골목에 결찰 (W군), W군과 동일한 방법으로 대퇴골목에 철사를 감은 후, 전기소락기의 끝을 철사에 접촉시켜 소락, 그리고 철사를 제거 (B군). 2주후, 랫드는 안락사하고 대퇴골 머리와 목을 채취하였다. H/E 염색, 사프라닌 O염색, TUNEL 염색을 실시하여 대퇴골 머리의 골과 연골에 골괴사 병변을 평가하였다. 모든 외과적인 방법 (S, W, B군)에서 골괴사가 2주간의 단기간에 유발되었다. 연골부분에 대한 유의적인 괴사변화는 B군에서만 관찰되었다. 철사를 통한 소락 후, 철사를 제거하는 변형된 외과적 방법은 다른 방법에 비해 더욱 효과적으로 골괴사 실험모델을 완성하였다.

• 대한수의학회지
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• 제31권2호
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• pp.155-161
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• 1991

The mechanisms of demyelination in Theiler's murine encephalomyelitis virus (TMEV)-induced chronic central nervous system(CNS) disease are still unclear and are probably multifactoral. This study was intended to culture spinal cord cells isolated from TMEV-induced demyelinating mice. By Percoll density centrifugation of enzymatically dissociated tissue, the cells were collected and then cultured on poly-L-lysine-coated plastic coverslips for 2 weeks. Oligodendrocytes, astrocytes and macrophages were identified using cell-type specific markers. Viral antigens were not present in oligodendrocytes and in astrocytes by double immunofluorescence. Affected mouse oligodendrocytes had less capacities of sheet formation and galactocerebroside immunoreactivity than those of control cell 3. These findings support the hypothesis that immune mediated mechanisms play an important role in the process of demyelination in this animal model.

• 한국해양바이오학회지
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• 제2권3호
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• pp.127-132
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• 2007

The innate immune response which is seen as the initial defense mechanism induced upon foreign invasion has been well documented in higher vertebrates. This has also been observed in fish infected with NNV. However, the fish immune system based on fully established genome project has not been fully elucidated. Therefore, in this review, we hope to correlate NNV infection in fish that has devastated the aquaculture industry, to its host immune system. Further, we discuss the potential preventive measures in overcoming the widespread of this neurodisease.

• International Journal of Internet, Broadcasting and Communication
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• 제12권3호
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• pp.131-138
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• 2020

The cabbage extract of the research does not show cytotoxicity, and thus can be used safely. In an experiment performed on an animal model with liver injury induced by a drug (APAP), it could be seen that the cabbage extract exhibited the effects of protecting liver and improving liver function by effectively reducing AST and ALT which are liver injury markers, indicating that the cabbage extract is effective as a pharmaceutical composition for preventing or treating liver disease. In particular, the cabbage extract was effective in treating inflammation of the liver by reducing the expression of the inflammatory mediators iNOS and COX-2 and the proinflammatory cytokine IL-1β, which are involved in acute inflammatory reactions accompanying liver injury. In the research, an extract of cabbage heat-treated at a temperature of 100 to 150℃ had a better liver function-improving effect or anti-inflammatory effect than an extract of raw cabbage.

• Clinical and Experimental Reproductive Medicine
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• 제43권1호
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• pp.1-8
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• 2016

Granulosa cell tumors (GCTs) are rare sex cord-stromal tumors that have been studied for decades. However, their infrequency has delayed efforts to research their etiology. Recently, mutations in human GCTs have been discovered, which has led to further research aimed at determining the molecular mechanisms underlying the disease. Mouse models have been important tools for studying GCTs, and have provided means to develop and improve diagnostics and therapeutics. Thus far, several genetically modified mouse models, along with one spontaneous mouse model, have been reported. This review summarizes the phenotypes of these mouse models and their applicability in elucidating the mechanisms of granulosa cell tumor development.

• 한국가금학회:학술대회논문집
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• 한국가금학회 2002년도 가을 학술발표논문집
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• pp.46-58
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• 2002

포도당 대사에서 중심적인 역할을 담당하고 있는 호르몬인 인슐린의 작용기전은 이전에 알려진 것보다 매우 다양하고 복잡한 세포내 신호전달 체계에 의해서 수행된다. 지난 10여년간 이러한 세포 신호전달 흐름내 중간물질인 여러 가지 단백질의 역할에 대해서 많은 연구가 진행되어 적지 않은 성과를 거두고 있다. 근래에는 해당 유전자에 대해 분자생물학적인 조작을 가하여 실험동물 체내에서 이러한 중간 단백질이 발현되지 못하도록 한 후, 각 조직에서의 인슐린 신호전달 체계를 더욱 심도 있게 연구하고 있다. 여러 연구방법 가운데서, 특히 조직 특이적으로 특정 유전자를 제거하여 태어난 개체를 이용하고, 또한 이들을 서로 교배시켜 나타나는 현상을 연구하여 비정상적인 포도당대사를 이해하는데 많은 도움을 받고 있다. 동물종에 따른 대사상의 차이점 등으로 인해 적지 않은 한계를 갖고 있지만, 동물 질환모델은 포도당 대사와 같은 생체 내에서 중요한 작용을 깊게 이해하고 더 나아가서는 비정상적인 대사를 밝히는데 핵심적인 역할을 수행하고 있다. 기초분야의 성과를 활용하는 응용학문인 축산분야에서도 이러한 접근방법과 연구 결과는 시사하는 바가 있을 것이다.

• Progress in Superconductivity
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• 제12권2호
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• pp.124-128
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• 2011

We have developed a high-$T_C$ SQUID magnetometer system to measure magnetocardiograms of laboratory rats. White noise of the measurement system was about 50 fT/$Hz^{1/2}$ when measured in a magnetically shielded box. We optimized the measurement position to obtain clear MCG wave from rat's small heart by using grid measurements. With the optimization, the MCG signal was successfully detected with the peak amplitude of about 50 pT. We could observe well defined P-, QRS-, and T-wave from the rat MCG. The results suggest that the developed system has a strong potential to monitor the progress of heart disease model using laboratory rat.