• 한국축산식품학회지
• /
• 제43권4호
• /
• pp.625-638
• /
• 2023

Among various biological agents, bacteriocins are important candidates to control Listeria monocytogenes which is a foodborne pathogen. In this study, a novel bacteriocin, named agilicin C7, was isolated from Ligilactobacillus agilis C7 showing inhibitory activity against L. monocytogenes. Agilicin C7 biosynthesis gene was characterized by bioinformatics analyses and heterologously expressed in Escherichia coli for further study. The anti-listeria activity of recombinant agilicin C7 (r-agilicin C7) was lost by proteases and α-amylase, suggesting that agilicin C7 is a glycoprotein. r-Agilicin C7 has wide pH and thermal stability and is also stable in various organic solvents. It destroyed L. monocytogenes by damaging the integrity of the cell envelope. These properties of r-agilicin C7 indicate that agilicin C7 is a novel amylase-sensitive anti-listerial Class IId bacteriocin. Physicochemical stability and inhibitory activity against L. monocytogenes of r-agilicin C7 suggest that it can be applied to control L. monocytogenes in the food industry, including dairy and meat products.

• 한국미생물·생명공학회지
• /
• 제33권2호
• /
• pp.117-122
• /
• 2005

Beijerinckia indica를 통한 Heteropolysaccharide-7(PS-7)생산 최적화를 위해 다양한 탄소원과 질소원을 첨가하여 다른 배지 조성들이 PS-7에 미치는 영향을 조사하였다. 높은 C/N ratio에서 PS-7 생산을 배양 40시간까지 효과적으로 증가시킬 수 있었으며 낮은 C/N ratio에선 탄소원으로 첨가한 glucose가 균체의 생산에 사용되었을뿐 PS-7의 생산은 낮았다. 하지만, 높은 C/N ratio에서는 배양액의 점도로 균체의 생산이 정체하는 것으로 보아 본 실험에서는 20g/l glucose와 $NH_{4}NO_3$ 7.5 mM으로 33.3의 C/N ratio가 PS-7 생산에 적절한 비율인 것으로 관찰되었다. 다양한 탄소원을 첨가한 결과, B. indica는 PS-7 생산을 위하여 탄소원으로 hexose 또는 glucose-generating 당을 효율적으로 이용할 수 있었다. Beijerinckia indica를 이용한 PS-7 생산을 위해 첨가한 다양한 탄소원 중 sucrose 농도가 최고의 PS-7(6.96 g/L)을 생산 하였다. 51 배양기를 이용한 배양에서 탄소원 20 g/L glucose, 질소원 7.5 mM $NH_{4}NO_3$이 포함된 MSM 배지내 배양된 B. indica의 세포 농도가 최고 2.5 g/L이였고, 생산된 최고 PS-7농도는 7.5 g/L(35174 cp)으로 나타났다. 질소원이 완전히 소모된 12시간 이후, 7.5 mM의 $NH_{4}NO_3$, glutamine, glutamate을 첨가한 MSM 배지에서의 B. indica 배앙 결과, 질소원이 B. indica 내에서 탄소원 대사를 조절하는 역할을 하는 것을 관찰할 수 있었으며 최종적으로 PS-7의 생산을 조절할 수 있었다.

• 생명과학회지
• /
• 제18권2호
• /
• pp.162-166
• /
• 2008

본 연구는 플라스크 배양을 통해 B. indica L3 균주를 가지고 PS-7을 생산할 수 있는 최적 조건을 조사하기 위해서 수행하였다. RSM을 통한 배지의 최적화 실험에서는 whey lactose, glucose, ammonium nitrate 세 가지의 독립변수의 조성에 따라 배양 72시간에 해당하는 DCW, PS-7, 그리고 점도의 변화에 대한 결과를 관찰하였다. 결정 계수(R square)의 값은 PS-7 생산량, DCW, 점성에서 각각 0.72, 0.64, 0.85로 신뢰성 있는 값을 획득하였다. Whey lactose와 glucose의 조합에 따른 PS-7 생산에 미치는 효과를 살펴보면 whey lactose (2%), glucose (1%)에서 최적의 PS-7 생산을 나타내었다. 결론적으로 glucose에 whey lactose를 기반으로 하는 배지 첨가와 C/N 비율이 PS-7의 생산에 큰 영향을 미침을 알 수 있었다.

• Reproductive and Developmental Biology
• /
• 제39권4호
• /
• pp.105-115
• /
• 2015

Toll-like receptor 7 (TLR7) is critical for the triggering of innate immune response by recognizing the conserved molecular patterns of single-stranded RNA (ssRNA) viruses and mediated antigenic adaptive immunity. To understand how TLR7 distinguish pathogen-derived molecular patterns from the host self, it is essential to be able to identify TLR7 receptor interaction interfaces, such as active sites or R848-agonist binding sites. The functional interfaces of TLR7 can serve as targets for structure-based drug design in studying the TLR7 receptor's structure-function relationship. In contrast to mammalian TLR7, chicken TLR7 (chTLR7) is unknown for its important biological function. Therefore, it has been targeted to mediate contrasting evolutionary patterns of positive selection into non-synonymous SNPs across eleven species using TLR7 conservation patterns (evolutionary conserved and class-specific trace residues), where protein sequence differences to the TLR7 receptors of interest record mutation that have passed positive section across the species. In this study, we characterized the Lys609 residue on chTLR7-ECD homodimer interfaces to reflect the current tendency of evolving positive selection to be transfer into a stabilization direction of the R848-agonist/chTLR7-ECDs complex under the phylogenetically variable position across species and we suggest a potential indicator for contrasting evolutionary patterns of both the species TLR-ECDs.

• The Korean Journal of Physiology and Pharmacology
• /
• 제17권1호
• /
• pp.65-71
• /
• 2013

The transient receptor potential melastatin type 7 (TRPM7) channel is a widely expressed non-selective cation channel with fusion to the C-terminal alpha kinase domain and regarded as a key regulator of whole body $Mg^{2+}$ homeostasis in mammals. However, the roles of TRPM7 during osteoclastogenesis in RAW264.7 cells and bone marrow-derived monocyte/macrophage precursor cells (BMMs) are not clear. In the present study, we investigate the roles of TRPM7 in osteoclastogenesis using methods of small interfering RNA (siRNA), RT-PCR, patch-clamp, and calcium imaging. RANKL (receptor activator of NF-${\kappa}B$ ligand) stimulation did not affect the TRPM7 expression and TRPM7-mediated current was activated in HEK293, RAW264.7, and BMM cells by the regulation of $Mg^{2+}$. Knock-down of TRPM7 by siTRPM7 reduced intracellular $Ca^{2+}$ concentration ($[Ca^{2+}]_i$) increases by 0 mM $[Mg^{2+}]_e$ in HEK293 cells and inhibited the generation of RANKL-induced $Ca^{2+}$ oscillations in RAW264.7 cells. Finally, knock-down of TRPM7 suppressed RANKL-mediated osteoclastogenesis such as activation and translocation of NFATc1, formation of multinucleated cells, and the bone resorptive activity, sequentially. These results suggest that TRPM7 plays an essential role in the RANKL-induced $[Ca^{2+}]_i$ oscillations that triggers the late stages of osteoclastogenesis.

• Asian Pacific Journal of Cancer Prevention
• /
• 제14권1호
• /
• pp.387-392
• /
• 2013

Objective: The aim of the present study was to explore mechanisms by which let-7c suppresses NSCLC cell proliferation. Methods: The expression level of let-7c was quantified by qRT-PCR. A549 and H1299 cells were transfected with let-7c mimics to restore the expression of let-7c. The effects of let-7c were then assessed by cell proliferation, colony formation and cell cycle assay. Mouse experiments were used to confirm the effect of let-7c on tumorigenicity in vivo. Luciferase reporter assays and Western blotting were performed to identify target genes for let-7c. Results: HOXA1 was identified as a novel target of let-7c. MTS, colony formation and flow cytometry assays demonstrated that forced expression of let-7c inhibited NSCLC cell proliferation by inducing G1 arrest in vitro, consistent with inhibitory effects induced by knockdown of HOXA1. Mouse experiments demonstrated that let-7c expression suppressed tumorigenesis. Furthermore, we found that let-7c could regulate the expression of HOXA1 downstream effectors CCND1, CDC25A and CDK2. Conclusions: Collectively, these results demonstrate let-7c inhibits NSCLC cell proliferation and tumorigenesis by partial direct targeting of the HOXA1 pathway, which suggests that restoration of let-7c expression may thus offer a potential therapeutic intervention strategy for NSCLC.

• 마이크로전자및패키징학회지
• /
• 제14권4호
• /
• pp.21-25
• /
• 2007

본 연구에서는 시차주사열량법, 젖음성 시험기, 비커스 경도계와 인장 시험기를 이용하여 Sn-0.7wt%Cu-Xwt%Re(X=$0.01{\sim}1.0$) 솔더의 특성에 관하여 평가하였다. 희토류 금속의 함량이 증가함에 따라 솔더의 용융 온도가 약간 상승하는 경향을 나타내었으나 $0.01{\sim}1.0%$의 희토류 금속이 첨가된 범위에서는 $233.9{\sim}234.7^{\circ}C$의 작은 용융 온도 범위를 갖는 것을 확인할 수 있었다. 솔더의 젖음성 시험 결과로부터 Sn-0.7Cu-0.1Re 솔더의 젖음성이 다른 솔더보다 우수한 것을 확인할 수 있었으며, Sn-0.7Cu-0.1Re 솔더의 젖음성이 Sn-0.7wt%Cu-0.01wt%P 솔더보다 우수한 것을 확인할 수 있었다. 또한 희토류 금속의 함량이 증가할수록 솔더의 경도와 인장 강도가 증가하는 경향을 확인할 수 있었다.

• 한국응용곤충학회지
• /
• 제35권3호
• /
• pp.238-242
• /
• 1996

몇가지 협력제의 7-ethoxy precocene II와 협력작용을 통한 활성증가와 작용기작을 구명하기 위하여 mikweed bug, Oncopeltus fasciatus 2령충에 항유약호르몬유사물과 협력제가 처리된 petri dish에 접종하여 생리활성 정도를 측정하였다. 얻어진 결과는 다음과 같다. 7-Ehtoxy precocene II의 반수활성농도는 $1.18\mu$g/$\textrm{cm}^2$이었다. 7-Ethoxy precocene II와 다른 산화저해제인 RO20-9747의 협력비가 14.05로 가장 높았고 그들의 반수활성농도는 $0.084\mu$$g/\textrm{cm}^2$이었다. 7-Ethoxy precocene II와 다른 협력제 1종, 2종, 4종을 조합을 달리하여 처리하였을 때 뚜렷한 협력작용 증가효과를 가져오지 못했다. 이상의 결과에서 다른 협력제들은 곤충의 방어기작을 알라타체를 포함하여 비선택적으로 저해하는 것으로 생각되었으나 RO20-9747은 7-ethoxy precocene II가 알라타체로 이행하는데 까지 곤충의 방어적 산화기작인 monooxygenase 활성을 선택적으로 저해함으로써 7-ethoxy precocene II가 알라타체로 이행량이 증가하여 활성증가를 가져온 것으로 생각되었다.

• Journal of Korean Neurosurgical Society
• /
• 제63권6호
• /
• pp.707-716
• /
• 2020

Objective : The function of B7H3, a member of the B7 family of proteins, in neuroblastoma (NB) remains poorly characterized. Here we examine the expression pattern of B7H3 in clinical NB specimens and characterize the phenotype of B7H3 knock-down in NB cell line. Methods : Immunohistochemical (IHC) staining was carried out to assess the expression of B7H3 in clinical NB specimens. Survival association was analyzed using five Gene Expression Omnibus (GEO) datasets (GSE85047, GSE45480, GSE62564, GSE16476, GSE49710). Clonogenic survival and flow cytometry were performed after B7H3 knockdown to assess the cellular proliferation and cell survival in vitro. Impact of B7H3 silencing on NB growth was examined in vivo using the SH-SY5Y xenograft model. Results : On IHC staining, B7H3 was widely expressed in clinical NB specimens. Analysis of the transcriptional profiles of five GEO datasets clinically annotated NB specimens revealed that decreased B7H3 expression was associated with improved overall survival. B7H3 knockdown suppressed the proliferation of the SH-SY5Y NB model in vitro and in vivo. Cell cycle analysis revealed that B7H3 silencing induced G1/S arrest. This arrest was associated with the suppression of E2F1 expression and induction of Rb expression. Conclusion : Our results demonstrate that B7H3 expression correlate with clinical survival in NB patients. Preliminary studies suggest that B7H3 may mediate the G1/S transition.

• 약학회지
• /
• 제37권2호
• /
• pp.136-142
• /
• 1993

To a suspension of 1-cyclopropyl-6,8-difluoro-1,4-dihydro-7-{3,7-diazabicyclo[3.3.0]oct-1(5)-en-3-yl}-4-oxo-3-quinoline carboxylic acid(C1) in sodium hydroxide solution and water is added dropwise with stirring carbon disulfide. [6R-[6$\alpha$, 7$\beta$(Z)]]-7-[[[2-Amino-4-thiazoly)methoxyimino]-acetyl]amino]-3-[[[[7-( 3-carboxy-1-cyclopropyl-6,8-difluoro-1,4-dihydro-4-oxo-7-guinolonyl)-3,7-diazabicyclo[3.3.0]oct-1(5)-en-3-yl]thioxomethyl]thio]methyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-en-2-carboxylic acid (DACD) was synthesized from 1-cyclopropyl-6,8-difluoro-1,4-dihydro-7-[7-(mercapto) thioxomethyl-[3,7-dia zabicyclo[3.3.0]oct-1(5)-en-3-yl}]-4-oxo-3-quinoline carboxylic acid disodium salt(C2) and cefotaxime. The invitro activity of novel dual-action cephalosporin, DACD, was compared with the in vitro activities of CENO(cefotaxime 3'-norfloxacin dithiocarbamate), cefotaxime, and norfloxacin against a variety of bacterial species. In vitro activity of DACD was superior to that of norfloxacin against Streptococcus pyogenes. Against Gram-positive and Gram-negative bacteria, its activity was almost equal to that of CENO.