• BMB Reports
• /
• v.54 no.9
• /
• pp.458-463
• /
• 2021

Cytokines activate inflammatory signals and are major mediators in progressive β-cell damage, which leads to type 1 diabetes mellitus. We recently showed that the cell-permeable Tat-CIAPIN1 fusion protein inhibits neuronal cell death induced by oxidative stress. However, how the Tat-CIAPIN1 protein affects cytokine-induced β-cell damage has not been investigated yet. Thus, we assessed whether the Tat-CIAPIN1 protein can protect RINm5F β-cells against cytokine-induced cytotoxicity. In cytokine-exposed RINm5F β-cells, the transduced Tat-CIAPIN1 protein elevated cell survivals and reduced reactive oxygen species (ROS) and DNA fragmentation levels. The Tat-CIAPIN1 protein reduced mitogen-activated protein kinases (MAPKs) and NF-κB activation levels and elevated Bcl-2 protein, whereas Bax and cleaved Caspase-3 proteins were decreased by this fusion protein. Thus, the protection of RINm5F β-cells by the Tat-CIAPIN1 protein against cytokine-induced cytotoxicity can suggest that the Tat-CIAPIN1 protein might be used as a therapeutic inhibitor against RINm5F β-cell damage.

• Proceedings of the Korean Society of Crop Science Conference
• /
• 2017.06a
• /
• pp.166-166
• /
• 2017

OsTAT encodes a twin-arginine translocator (TAT) pathway signal protein. It contains a TRANS membrane domain and a chloroplast transit peptide. mRNA transcription profiling of OsTAT1 revealed that it is highly overexpressed in the leaves corroborating reports on its role in chloroplast. Moreover, its level of expression is more pronounced during earlier stages (germination, 3-leaf stage, and maximum tillering) of growth in rice. A lower disease progress curve of bacterial blight is evident in transgenic lines compared with the wild type, Dongjin indicating its involvement in immunity to Xoo. Expression pattern following infection of Xoo strain K2 depicts highest levels at 4 and 8 hour post-inoculation which implies crucial induction of resistance during early response. This study initially reports a new overview on the biological functions of plant's TAT pathway. Further molecular and genetic analyses are underway to provide detailed involvement of OsTAT in disease resistance.

• Journal of Families and Better Life
• /
• v.13 no.1
• /
• pp.199-206
• /
• 1995

부모의 사랑은 가정환경을 통하여 자녀의 대인관계나 사회화에 있어서 최초의 모델이 될 수 있고 자녀의 성격형성이나 문제행동에 깊은 영향을 준다. 이 연구는 청년기 자녀의 잠재된 갈등요인 즉 청년들에게 영향을 주는 요인이 무엇인가를 면접조사와 TAT를 통한 방법으로 시행하였다. 면접조사 결과 부모는 거부적 과보호, 무관심으로 청년들의 갈등을 일으키고 형제간의 갈등이 있는 것으로 나타났다. TAT결과는 부모상에서 부모의 모습은 무관심하거나, 간섭이 심하고 강요적이었으며 환경반응은 가족간의 사랑과 이해가 없는 경우로 나타났다. 불안 갈등요인도 형제간의 갈등, 부모에 대한 사랑의 부족, 결핍 등으로 나타났다.

• BMB Reports
• /
• v.39 no.3
• /
• pp.253-262
• /
• 2006

Parkinson's disease (PD) is a common neurodegenerative disorder and is characterized by the progressive loss of dopaminergic neurons in the substantia nigra. Although many studies showed that the aggregation of $\alpha$-synuclein might be involved in the pathogenesis of PD, its protective properties against oxidative stress remain to be elucidated. In this study, human wild type and mutant $\alpha$-synuclein genes were fused with a gene fragment encoding the nine amino acid trans activator of transcription (Tat) protein transduction domain of HIV-l in a bacterial expression vector to produce a genetic in-frame WT Tat-$\alpha$-synuclein (wild type) and mutant Tat-a-synucleins (mutants; A30P and A53T), respectively, and we investigated the protective effects of wild type and mutant Tat-$\alpha$-synucleins in vitro and in vivo. WT Tat-$\alpha$-synuclein rapidly transduced into an astrocyte cells and protected the cells against paraquat induced cell death. However, mutant Tat-$\alpha$-synucleins did not protect at all. In the mice models exposed to the herbicide paraquat, the WT Tat-$\alpha$-synuclein completely protected against dopaminergic neuronal cell death, whereas mutants failed in protecting against oxidative stress. We found that these protective effects were characterized by increasing the expression level of heat shock protein 70 (HSP70) in the neuronal cells and this expression level was dependent on the concentration of transduced WT Tat-$\alpha$-synuclein. These results suggest that transduced Tat-$\alpha$-synuclein might protect cell death from oxidative stress by increasing the expression level of HSP70 in vitro and in vivo and this may be of potential therapeutic benefit in the pathogenesis of PD.

• IMMUNE NETWORK
• /
• v.6 no.4
• /
• pp.192-198
• /
• 2006

Background: Investigating strategy to enhance efficiency of gene transfer via adenovirus is critical to sustain gene expression in targeted cells or tissues to regulate immune responses. However, the use of adenovirus as a gene delivery method has been limited by the native tropism of the virus. In this study, the critical parameter is to improve the efficient binding of viral particles to the plasma membrane prior to cellular uptake. Methods: Human immunodeficiency virus (HIV-1) trans-acting activator of transcription (TAT), a protein transduction domain, was fused to the ectodomain of the coxsackie-adenovirus receptor (CAR). The CAR-TAT protein was produced from a Drosophila Schneider 2 cells (S2) transfected with CAR-TAT genes. The function of CARTAT was analyzed the efficiency of adenoviral gene transfer by flow cytometry, and then immunizing AdVGFP with CAR-TAT was transduced on dendritic cells (DCs). Results: S2 transfectants secreting CAR-TAT fusion protein has been stable over a period of 6 months and its expression was verified by western blot. Addition of CAR-TAT induced higher transduction efficiency for AdVGFP at every MOI tested. When mice were vaccinated with DC of which adenoviral transduction was mediated by CAR-TAT, the number of IFN-${\gamma}$ secreting T-cells was increased as compared with those DCs transduced without CAR-TAT. Conclusion: Our data provide evidence that CAR-TAT fusion protein enhances adenoviral transduction and immunogenecity of transgenes on DCs and may influence on the development of adenoviral-mediated anti-tumor immunotherapy.

• Korean Journal of Veterinary Service
• /
• v.17 no.2
• /
• pp.95-113
• /
• 1994

To establish an effective diagnostic measure for detection of the antibodies against Actinobacillus pleuropneumoniae, the methods for tube agglutination test (TAT), plate agglutination test (PAT), micro-agglutination test(MAT) and agar-gel immunodiffusion test(ID) were improved and standarized, and the comparative studies were carried out. The results obtained through the experiments were summarized as follows. 1. The rabbit hyperimmune sera to reference serotypes 1 to 6 were cross-tested with TAT, PAT, MAT and ID. In the homologous systems, the range of antibody titers in TAT was 80 to 640, showing the cross-reaction in serotypes 3, 4, 5 and 6. The range of antibody titers in PAT was 4 to 64, showing the cross-reaction in serotypes 3, 4, 5 and 6. In ID, the range of antigen titers was 8 to 32, and cross-reaction was observed in serotype 5. 2. The optimal concentration of antigen in PAT and MAT were 100mg /ml and 1.25mg /ml respectively. The most sensitive reaction in MAT was observed in 52$^{\circ}C$ for 18hrs. 3. In ID, the most promising antigen and the buffer for agar-gel were EDTA-treated antigen and 0.05M tris buffer (pH 7.2), respectively. 4. By the tests for 200 swine sera, it was found that the frequency of positive reaction were 203 in TAT, 240 in PAT and 163 in ID. 5. When compared the titers of TAT with those of MAT for 200 swine sera, MAT showed the higher titer than TAT being increased by relative correlation. Int was found that the titer for positive readings were 20 in TAT and 40 in MAT. 6. when compared the results of ID with those of TAT for 200 swine sera, all sera with TAT titer under 10 were negative in ID. Of the sera with TAT titer 20 and 40, 55.1% nd 91.8% were positive in ID, respectively. All sera with TAT titer above 80 were positive in ID. In comparison of ID and MAT, all sera with MAT titer under 20 were negative in ID. Of the sera with MAT titer 40 and 80, 24.7% and 93.9% were positive in ID, respectively. All sera with MAT titer over 160 showed positive in ID. 7. In conclusion, the established MAT showed high sensitivity but low specificity, wherease ID revealed low sensitivity but high specificity.

• BMB Reports
• /
• v.51 no.7
• /
• pp.362-367
• /
• 2018

A major feature of type 1 diabetes mellitus (T1DM) is hyperglycemia and dysfunction of pancreatic ${\beta}$-cells. In a previous study, we have shown that Tat-DJ-1 protein inhibits pancreatic RINm5F ${\beta}$-cell death caused by oxidative stress. In this study, we examined effects of Tat-DJ-1 protein on streptozotocin (STZ)-induced diabetic mice. Wild type (WT) Tat-DJ-1 protein transduced into pancreas where it markedly inhibited pancreatic ${\beta}$-cell destruction and regulated levels of serum parameters including insulin, alkaline phosphatase (ALP), and free fatty acid (FFA) secretion. In addition, transduced WT Tat-DJ-1 protein significantly inhibited the activation of $NF-{\kappa}B$ and MAPK (ERK and p38) expression as well as expression of COX-2 and iNOS in STZ exposed pancreas. In contrast, treatment with C106A mutant Tat-DJ-1 protein showed no protective effects. Collectively, our results indicate that WT Tat-DJ-1 protein can significantly ameliorate pancreatic tissues in STZ-induced diabetes in mice.

• IMMUNE NETWORK
• /
• v.11 no.4
• /
• pp.216-222
• /
• 2011

Background: The ligand for CD137 (CD137L; also called 4-1BBL) is mainly expressed on activated APCs such as dendritic cells, B cells and macrophages. Even though CD137L functions as a trigger of the CD137 signaling pathway for T cell activation and expansion, engagement of CD137L can deliver a signal leading to the production of proinflammatory cytokines in macrophages. Methods: We generated cell-permeable TAT-CD137L cytoplasmic domain fusion protein (TAT-CD137Lct) and examined its ability to initiate the CD137L reverse signaling pathway. Results: Treatment of TAT-CD137Lct induced the production of high levels of IL-6 and TNF-${\alpha}$ mRNAs and proteins in peritoneal macrophages. TAT-CD137Lct increased phosphorylation of Erk, p38 MAPK and Jnk, and activated transcription factors C/EBP and CREB. However, TAT-CD137Lct did not visibly affect the degradation of the inhibitor of NF-${\kappa}B$ ($IkB{\alpha}$). We further demonstrated that JNK activation was required for TAT-CD137Lct-induced production of TNF-${\alpha}$, while activation of Erk and p38 MAPK were involved in IL-6 and TNF-${\alpha}$ production. Conclusion: Our results suggest that TATCD137Lct is an effective activator for the CD137L reverse signaling pathway.

• Journal of Radiopharmaceuticals and Molecular Probes
• /
• v.5 no.2
• /
• pp.135-144
• /
• 2019

Targeted alpha therapy (TAT) based on metallic radionuclides has attracted a lot of attention lately due to its impressive therapeutic efficacy displayed in couple of clinical studies for cancer. Representative metallic radionuclides emitting alpha-particle include 225Ac, 213Bi, and 227Th, and there have been variety of TAT formulations based on different targeting moiety and chelating agents. In this review, we introduce strategies to label metallic radioisotopes with biomolecules and look at some of recent preclinical and clinical results of TAT for cancer.

• BMB Reports
• /
• v.39 no.1
• /
• pp.76-83
• /
• 2006

Pyridoxine-5-P oxidase catalyses the terminal step in the biosynthesis of pyridoxal-S-P, the biologically active form of vitamin $B_6$ Which acts as an essential cofactor. Here, a human brain pyridoxine-5-P oxidase gene was fused with a gene fragment encoding the HIV-1 Tat protein transduction domain (RKKRRQRRR) in a bacterial expression vector to produce a genetic in-frame Tat-pyridoxine-5-P oxidase fusion protein. Expressed and purified Tat-pyridoxine-5-P oxidase fusion protein transduced efficiently into PC12 cells in a time- and dose-dependent manner when added exogenously to culture media. Once inside the cells, the transduced Tat-pyridoxine-5-P oxidase protein showed catalytic activity and was stable for 48 h. Moreover, the formation of pyridoxal-5-P was increased by adding exogenous Tat-pyridoxine-5-P oxidase to media pre-treated with the vitamin $B_6$ precursor pyridoxine. In addition, the intracellular concentration of pyridoxal-S-P was markedly increased when Tat-pyridoxal kinase was transduced together with Tat-pyridoxine-5-P oxidase into cells. These results suggest that the transduction of Tat-pyridoxine-5-P oxidase fusion protein presents a means of regulating the level of pyridoxal-5-P and of replenishing this enzyme in various neurological disorders related to vitamin $B_6$.