Subunit vaccines are being developed as a potential therapy for preventing microbial pathogen infection. In this study, the immunogenicity of recombinant Brucella (B.) abortus Fe/Mn superoxide dismutase (rFe/Mn SOD) protein as a subunit vaccine against B. abortus was investigated in BALB/c mice model. Brucella Fe/Mn SOD gene was cloned into a pcold-TF DNA vector. The bacterial recombinant protein was expressed using the Escherichia coli DH5α strain with a size of 82.50 kDa. The western blotting assay showed that rFe/Mn SOD reacted with Brucella-positive serum, indicating the potential immunoreactivity of this recombinant protein. After the second and third vaccinations, the peripheral CD4+ T cell population was increased significantly in the rFe/Mn SOD-immunized mice group compared to the PBS control group. Moreover, immunization of this recombinant protein increased the CD4+ T cell population from the first vaccination to the third vaccination. Meanwhile, the CD8+ T cells were slightly enhanced after the second vaccination compared to the first vaccination and compared to control groups. Fourteen days after the bacterial infection, the splenomegaly and the number of bacteria in the spleen were evaluated. The result showed that both rFe/Mn SOD and positive control RB51 decreased the bacterial replication in the spleen and the splenomegaly compared to control groups. Altogether, these results suggested that rFe/Mn SOD could induce host immunity against B. abortus infection.

Morphology of cumulus-oocyte-complexes (COCs) at germinal vesicle (GV) stage as one of the evaluation criteria for oocyte maturation quality after in vitro maturation (IVM) plays important roles on the meiotic maturation, fertilization and early embryonic development in pigs. When cumulus cells of COCs are insufficient, which is induced the low oocyte maturation rate by the increasing of reactive oxygen species (ROS) in porcine oocyte during IVM. The ROS are known to generate including superoxide and hydrogen peroxide from electron transport system of mitochondria during oocyte maturation in pigs. To regulate the ROS production, the cumulus cells is secreted the various antioxidant enzymes during IVM of porcine oocyte. Our previous study showed that Mito-TEMPO, superoxide specific scavenger, improves the embryonic developmental competence and blastocyst formation rate by regulating of mitochondria functions in pigs. However, the effects of Mito-TEMPO as a superoxide scavenger to help the anti-oxidant functions from cumulus cells of COCs on meiotic maturation during porcine oocyte IVM has not been reported. Here, we categorized experimental groups into two groups (Grade 1: G1; high cumulus cells and Grade 2: G2; low cumulus cells) by using hemocytometer. The meiotic maturation rate from G2 was significantly (p < 0.05) decreased (G1: 79.9 ± 3.8% vs G2: 57.5 ± 4.6%) compared to G1. To investigate the production of mitochondria derived superoxide, we used the mitochondrial superoxide dye, Mito-SOX. Red fluorescence of Mito-SOX detected superoxide was significantly (p < 0.05) increased in COCs of G2 compared with G1. And, we examined expression levels of genes associated with mitochondrial antioxidant such as SOD1, SOD2 and PRDX3 using a RT-PCR in porcine COCs at 44 h of IVM. The mRNA levels of three antioxidant enzymes expression in COCs from G2 were significantly (p < 0.05) lower than COCs of G1. In addition, we investigated the anti-oxidative effects of Mito-TEMPO on meiotic maturation of porcine oocyte from G1 and G2. Meiotic maturation and mRNA levels of antioxidant enzymes were significantly (p < 0.05) recovered in G2 by Mito-TEMPO (0.1 μM, MT) treatment (G2: 68.4 ± 3.2% vs G2 + MT: 73.9 ± 1.4%). Therefore, our results suggest that reduction of mitochondria derived superoxide by Mito-TEMPO may improves the meiotic maturation in IVM of porcine oocyte.

In general, the shape of cumulus-oocyte-complexes (COCs) at germinal vesicle (GV) stage is important roles on meiotic maturation of porcine oocyte during in vitro maturation (IVM). Then, mitochondria produce reactive oxygen species (ROS) such as superoxide from electron transport system during oocyte maturation. ROS levels on oocytes are regulated by various antioxidant enzymes in cumulus cells (CCs). However, the effect of mitochondria derived superoxide production from CCs during IVM of porcine oocyte has not been reported. Firstly, we divided groups according to large number of CCs (Grade 1: G1) and small number of CCs (Grade 2: G2). Then, we counted cumulus cells of G1 and G2 oocyte by using haemocytometer. The oocyte maturation rate was significant decreased (p < 0.05) in G2 oocytes than that of G1 oocytes. We measured mitochondria derived superoxide in G1 and G2 COCs by using Mito-SOX staining. Mitochondrial superoxide was higher in G2 COCs than G1 COCs. Then, the mRNA expression levels of antioxidant enzymes (SOD1, SOD2 and PRDX3) in G2 COCs were decreased compared to G1 COCs. To reduce mitochondria derived superoxide, we used Mito-TEMPO as mitochondrial superoxide scavenger. Oocyte maturation rates in both G1 and G2 groups treated with Mito-TEMPO were increased than that of non-treated groups. Mitochondrial superoxide was lower in G1 and G2 groups treated with Mito-TEMPO than that of non-treatment groups. The mRNA expression levels of antioxidant enzymes in G1 and G2 COCs treated with Mito-TEMPO were increased compared to non-treated groups. Based on these findings, we suggest that reduction of mitochondria derived superoxide by Mito-TEMPO assists maturation competence in porcine oocytes.

Bee venom is a complex mixture of toxic components that induces immediate local inflammatory and allergic responses. However, the presence and role of superoxide dismutase (SOD) in bee venom have not been previously investigated. Here, we provide the first demonstration that bee venom contains Cu,Zn SOD (SOD3), a novel extracellular component that promotes local inflammation. Bee venom SOD3 was shown to be an oxidant, rather than an antioxidant, that induces the inflammation-signaling molecule H2O2 in vivo. H2O2 plays a pathological role by triggering an immediate local inflammatory response. Furthermore, bee venom SOD3 significantly induced the activation of proinflammatory mediators (TNF-α and COX-2) and cytokines (IL-1β and IL-6) via the overproduction of H2O2 in mice. Our data demonstrate that bee venom SOD3 induced H2O2, which drives an immediate local inflammatory response, indicating a novel mechanism underlying bee venom-induced local inflammation.

Genistein is a product of naturally occurring isoflavones at relatively high levels in soybeans. The harmful effects of ethanol are attributed to the induction of biological processes which lead to an increase in the generation of reactive oxygen species in fetuses. In this study, we investigated the effects of genistein ( and /ml) on gene expressions of the representative cellular antioxidative enzymes in ethanol (1 /ml)-treated mouse fetuses during the critical period (embryonic days 8.5~10.5) of organogenesis using a semi-quantitative RT-PCR analysis. The mRNA levels of cytosolic glutathione peroxidase (GPx), phospholipid hydroperoxide GPx, cytosolic CU,Zn-superoxide dismutase (SOD), and mitochondrial SOD were significantly decreased in ethanol-treated fetuses. However, the mRNA levels of ethanol plus genistein-treated fetuses were significantly higher than those of ethanol alone fetuses. These results indicate that genistein can up-regulate the expressions of GPx and SOD mRNAs reduced by the ethanol treatment in fetuses.

NAC는 GSH의 전구물질로, thiol기를 포함하는 항산화제 중 하나로 잘 알려져 있으며, 방사선 조사 시 발생하는 생체 내 영향을 감소시켜 생체 손상의 방호 및 회복에 도움을 주는 방사선 방어제로 이용된다. S. cerevisiae에서 항산화제 NAC를 전처리 함에 따라 이온화 방사선 조사에 따른 효모의 세포사멸 방어효과 및 superoxide dismutase (SOD), catalase, glutathione peroxidase (GPx)와 같은

참돌꽃 유식물에서 줄기와 뿌리의 부위별로 나타나는 superoxide dismutase(SOD)의 활성과 환경 스트레스 또는 항산화분자 처리에 따른 SOD활성도를 조사하여 다음과 같은 결론을 얻었다. SOD의 활성은 줄기보다는 뿌리에서 높았고 각 기부보다는 선단부에 이를수록 높았으며 이러한 현상은 유식물 생육의 초기보다 후기에 더 상승된 결과를 나타내고 있다. 또한 환경 스트레스에 대한 SOD의 활성도는 유식을 초기생육 상태에서는 NaCl또는 cadmi

We have generated transgenic mice that expressed mouse extracellular superoxide dismutase (EC-SOD) in their skin. In particular, the expression plasmid DNA containing human keratin K14 promoter was used to direct the keratinocyte-specific transcription of the transgene. To compare intron-dependent and intron-independent gene expression, we constructed two vectors. The vector B, which contains the rabbit -globin intron 2, was not effective for mouse EC-SOD overexpression. The EC-SOD transcript was detected in the skin, as determined by Northern blot analysis. Furthermore, EC-SOD protein was detected in the skin tissue, as demonstrated by Western blot analysis. To evaluate the expression levels of EC-SOD in various tissues, we purified EC-SOD from the skin, lungs, brain, kidneys, livers, and spleen of transgenic mice and measured its activities. EC-SOD activities in the transgenic mice skin were approximately 7 fold higher than in wild-type mice. These results suggest that the mouse overexpressing vector not only induces keratinocyte-specific expression of EC-SOD, but also expresses successfully functional EC-SOD. Thus, these transgenic mice appeared to be useful for the expression of the EC-SOD gene and subsequent analysis of various skin changes, such as erythema, inflamation, photoaging, and skin tumors.

본 연구는 OPS 기법에 의한 돼지 수정란의 동결-융해 시 수정란의 발달 단계와 superoxide dismutase (SOD)가 수정란의 생존능력에 미치는 영향을 검토하였다. 돼지 체외수정 배반포는 OPS방법에 의해 동결 후 융해하여 0~10unitsml의 SOD 존재 하에 48시간 체외배양하였다. 동결-융해 후 형태학적으로 정상적인 수정란의 비율은 초기, 중기 및 확장배반포간에 유의적인 차이는 인정되지 않았다(30.8~38.6%). 그러나 발육단계가 높을수록 형태적으로 정상인 수정란의 비율이 높은 경향을 나타냈다. 수정란의 융해 후 48시간 추가 배양했을 때, 발육이 진행된 수정란은 후기배반포기에 동결한 수정란이 38.7%로 유의적으로 높았으며(P<0.05), 1 unit/ml의 SOD를 첨가한 경우 비교적 높은 생존율을 나타내었다. 본 연구의 결과로부터, 수정란의 OPS방법에 의한 동결-융해 후 생존성의 향상을 위해서는 후기배반포기 단계에 동결하는 것이 유리하며, SOD의 첨가는 수정란의 손상을 어느 정도 방지할 수 있을 것으로 사료된다.