The primary therapeutic approach for Brucella species infections has mainly been based on antibiotic treatment. However, the development of vaccines for brucellosis control remains controversial. Furthermore, there is currently no licensed vaccine available for human brucellosis. This study aims to evaluate the effect of a combination of recombinant protein vaccines against Brucella (B.) abortus infection using a mouse model. Two B. abortus genes, namely dapB and gpm, were cloned and expressed in competent Escherichia (E.) coli DH5α using the pCold-TF vector. Successfully cloned vectors were subjected to PCR amplification using specific primer pairs. The apparent sizes of dapB and gpm were detected at 807 bp and 621 bp, respectively. Besides, the purified recombinant proteins dapB and gpm were detected using SDS-PAGE electrophoresis with correct sizes of 82.86 kDa and 87.61 kDa, respectively. These recombinant proteins were used to immunize mice as a combined subunit vaccine (CSV) to elicit host immunity against B. abortus infection. Mice immunized with CSV exhibited increased proliferation of CD4+ and/or CD8+ T cells at week 7th and 9th before sacrifice, in comparison to the control group. Notably, CSV immunization showed a significant decrease in bacterial burden in the spleen compared to the control group. Altogether, CSV using dapB and gpm induced host adaptive immune response against Brucella infection, suggesting its potential as an effective new subunit vaccine candidate.
Extensive research and testing continue to be conducted for the development of vaccines targeting zoonotic diseases such as brucellosis. In this study, the potential of the DapB as a recombinant protein vaccine to effectively combat Brucella abortus 544 infection in BALB/c mice was evaluated. Western blotting assay results showed that recombinant protein DapB reacted with Brucella-positive serum, indicating its potential immunoreactivity. In vivo results showed that the peripheral blood CD4+ and CD8+ T cell population significantly increased in the DapB-immunized mice group after the first, second and third blood collection, compared to the control group that received PBS. Additionally, at the fourth blood collection, an increase in CD4+ T cell activation was observed in three vaccination groups compared to PBS negative control group. These results indicate the potential of DapB in stimulating cellular immunity. Fourteen days after infection, the bacterial load in the spleen was evaluated. The reduction in bacterial replication in the spleen by both DapB and RB51 highlights their protective efficacy against Brucella infection. These findings contribute to the ongoing efforts in developing effective vaccines against brucellosis and provide valuable insights for further research in this field.
본 연구에서는 2010년 여름에 천수만에서 저층해수를 채집하여 용존산소와 영양염 농도를 측정하였다. 또한 benthic chamber내의 해수시료를 시계열로 채집하는 자동화된 Benthic Lander를 설치하여 해수-퇴적물간 영양염 플럭스를 측정하였다. 오염된 인공호수 유출수가 들어오는 천수만 북쪽에서는 저층수의 용존산소는 2 mg/l로 hypoxia의 존재 가능성이 확인되었다. 반면 남쪽 천수만 입구의 저층 용존산소는 5 mg/l이었다. 영양염은 용존산소와 반대의 분포 경향을 보였고, N/P ratio의 변화는 hypoxia에 의해 발생된 인산염의 탈착과 용출 때문으로 보인다. 만 북쪽 해역의 유기탄소 산화율과 산소소비율은 남쪽 만 입구 해역보다 약 2배 큰 값을 보였고, 영양염 benthic flux는 천수만 북쪽에서 4내지 6배 높았다. 이러한 결과는 해수-퇴적물간 물질 플럭스를 정확히 추정하기 위해서는 hypoxia의 역할에 대한 이해가 중요하다는 점을 시사해준다.
Mung bean (Vigna radiata L.) is an increasingly important human food source, as well as a new functional agent, mainly due to its potent antioxidant activity. This study was conducted to determine antioxidant activity of fractions from mung bean seeds by measuring DPPH radical scavenging activity and to quantify the flavonoids by means of HPLC analysis. Vitexin and isovitexin were present in both ethanol and water extracts in highest amount. Flavonoids, vitexin and isovitexin were quantified from 195 germplasms of mung beans and their concentrations varied by 4.7 fold. Especially, the breeding line KM99004-4B-2 (Suwon28/KM94004), which has grown in Jeollanamdo Agricultural Research and Extension Services, showed the highest amount (15.88 mg/g) of total flavonoids. The vitexin portion was averaged 70.73±1.38%. High positive correlation (r=0.96***) between vitexin and isovitexin contents showed. However, the flavonoid content showed very low correlation with the 24 growth and ecological characteristics. Seed coats of mung beans had the highest flavonoid amount, showing 50~70 times more than cotyledons. Flavonoid contents in the seed, the cotyledon, and the seed coat were decreased as the seed imbibition time increased.