• 한국융합학회논문지
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• 제13권1호
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• pp.101-107
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• 2022

Tertomotide는 항암제로 개발된 펩타이드 백신이다. 그러나 동물실험과 임상시험에서 염증성 증상이 완화되는 현상이 발견된바 있다. 이에 tertomotide가 항염물질로 작용하는지 확인하기 위하여 직접적인 항염활성과 그 작용기전을 조사하였다. 이를 위해 LPS 또는 PMA에 의해 활성화된 monocyte에 tertomotide를 처리한 후 염증성 cytokine 생산과 관련된 신호전달과정을 관찰하였다. Monocyte에서 tertomotide는 TNF-α, IL-1β, IL-8 등 염증성 싸이토카인의 생산을 감소시켰고 NF-κB 신호를 감쇄시켰으며 또한 TNF-α에 의한 ERK1/2와 P38 MAPK의 활성화를 저해하였다. 이 결과는 tertomotide 처치에 따른 염증성 질환 완화가 NF-κB/STAT3의 신호의 감쇄와 항염활성에 의한 것이라고 설명할 수 있고 이를 활용하여 신규 항염 약물의 도출이 가능할 것으로 판단된다. 이는 면역학적 활성을 목표로 계산화학적으로 설계된 물질의 생물학적 성질을 활용하여 새로운 약물을 도출하는 융합연구의 예시가 될 것이다.

• BMB Reports
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• 제47권11호
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• pp.649-654
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• 2014

Neutrophils play an important role in the initiation of innate immunity against infection and injury. Although many different types of G-protein coupled receptors are functionally expressed in neutrophils, no reports have demonstrated functional expression of umami taste receptor in these cells. We observed that mouse neutrophils express the umami taste receptor T1R1/T1R3 through RNA sequencing and quantitative RT-PCR analysis. Stimulation of mouse neutrophils with L-alanine or L-serine, which are ligands for the umami taste receptor, elicited not only ERK or p38 MAPK phosphorylation but also chemotactic migration. Moreover, addition of L-alanine or L-serine markedly reduced the production of several cytokines including $TNF-{\alpha}$ induced by lipopoly-saccharide (LPS) through inhibition of $NF-{\kappa}B$ activity or STAT3 phosphorylation in neutrophils. Our findings demonstrate that neutrophils express the umami taste receptor, through which tastants stimulate neutrophils, resulting in chemotactic migration, and attenuation of LPS-induced inflammatory response.

• Molecular & Cellular Toxicology
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• 제2권3호
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• pp.193-201
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• 2006

Cephalexin, one of most widely prescribed cephalosporin, has been reported to cause acute renal failure as a side effect in human and experimental animals. Although numerous animal studies have been reported for the cephalosporin nephrotoxicity, the molecular and cellular nephrotoxic mechanisms of cephalexin are still unknown. This investigation evaluated the time-dependent gene expression profile of kidney in mouse during cephalexin induced nephrotoxicity. C57BL/6 female mice were administered either saline or 1,000 mg/kg cephalexin intraperitoneally. Mice were sacrificed at 3, 6, and 24 hr after administration. Blood biochemical and histopathological results indicated cephalexin induced nephrotoxicity. Microarray experiment carried out using Affymetrix $GeneChip^{(R)}$. There were 198 informative genes that were significantly expressed >5-fold versus control at 3, 6, and 24 hr (p<0.01), of which 156 and 42 were up-and down-regulated, respectively. Major classes of up-regulated genes at 3, 6 hr included those involved in MAPK/Jak-STAT signaling pathway and immune response such as cytokine-cytokine receptor interaction and complement and coagulation cascades. At 24 hr, up-regulated genes were mainly involved in regeneration/repair and immune response; down-regulated genes were generally associated with transporters and intermediary metabolism. Among the up-regulated genes at 24 hr, several potential biomarkers on nephrotoxicity such as Kim-1, Fga, Timp1, and Slc34a2 were clustered in a same category. In addition, Tnfrsf12a and Lcn2 which were consistently up-regulated (>5 fold) were also included as potential biomarkers. These results may provide clues for elucidating the mechanism of cephalexin induced nephrotoxicity and evaluating potential biomarkers to assess nephrotoxicity.

• 한국미생물·생명공학회지
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• 제17권4호
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• pp.392-396
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• 1989

Facultative methylotroph인 Methylobacterium organophilum을 유일한 탄소원으로 메탄올을 0.5% (v/v) 함유한 최소 합성배지를 이용하여 pH 6.8과 3$0^{\circ}C$에서 배양하였다. 세포의 증식은 배지 내의 여러 성분들에 의해 영향을 받았으며 이로부터 최대증식속도를 얻을 수 있는 최적 합성배지 조성을 확립하였다. 배지조성 중 질소원이 결핍되면 세포의 증식이 감소하며 Fe$^{+2}$ 또는 Mn$^{+2}$ 이온의 결핍은 세포의 비증식속도를 감소시켰다. 탄소원인 메탄올은 농도를 증가시킬수록 세포의 비증식속도가 감소하는 메탄올의 기질 저해성을 확인하였으며, 4%(v/v) 이상의 메탄을 농도에서는 세포의 증식이 완전히 저해되었다. 이러한 메탄올의 기질 저해성은 간헐식 유가배양법을 이용하여 극복하고 PHB의 축적을 촉진하는 영양분 제한(nutrient-deficiency) 실험을 수행하였다. 합성배지의 영양분 중 NH$_{4}^{+}$, SO$_{4}^{-2}$, $K^+$ 또는 PO$_{4}^{-3}$ 이온을 결핍시켰을 때 M. organophilum의 PHB 생산이 증가하였으며 칼륨이온을 결핍시켰을때 세포의 건조 중량에 대한 PHB 함량은 최대 58%를 얻었다.

• Asian-Australasian Journal of Animal Sciences
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• 제18권2호
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• pp.165-169
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• 2005

This study was performed to offer effective basic data for selection and improvement of Korean dairy cattle through identifying distributional properties among candidate genes (bovine butyrophilin, signal transducers and activators of transcription 5a, and prolactin hormone). In this study, polymorphisms of candidate genes were identified and the relationships between loci and production traits of each gene were analyzed using frozen semen of Holstein bulls (19 proven and 77 candidates). In butyrophilin (BTN) locus, polymorphisms information contents (PIC) value of BTN2 (0.372) was higher than those of others (BTN1; 0.155, BTN3; 0.254, BTN4; 0.169). As a result of analysis of genotyping STAT5a, using single stranded conformational polymorphism (SSCP) method and microsatellite locus, PIC values were 0.189 and 0.457, respectively. And PIC value of prolactin hormone gene was 0.176. In the relationships between genotypes and production traits, BTN3 was associated with 305-day production traits (p<0.05). PTAs for B allele were such as 110.43, 88.28 and 75.25 in BTN1, 3, 4 and these values were higher than those of A allele, but in the case of BTN2, A allele with 154.19 was higher than that of B allele. The results obtained from using candidate genes may be used as an useful index for the genetic improvement of dairy cattle population in Korea, and further studies are needed.

• Asian Pacific Journal of Cancer Prevention
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• 제16권6호
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• pp.2489-2493
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• 2015

Background: Loss of heterozygosity (LOH) on chromosomal regions is crucial in tumor progression and this study aimed to identify genome-wide LOH in pancreatic cancer. Materials and Methods: Single-nucleotide polymorphism (SNP) profiling data GSE32682 of human pancreatic samples snap-frozen during surgery were downloaded from Gene Expression Omnibus database. Genotype console software was used to perform data processing. Candidate genes with LOH were screened based on the genotype calls, SNP loci of LOH and dbSNP database. Gene annotation was performed to identify the functions of candidate genes using NCBI (the National Center for Biotechnology Information) database, followed by Gene Ontology, INTERPRO, PFAM and SMART annotation and UCSC Genome Browser track to the unannotated genes using DAVID (the Database for Annotation, Visualization and Integration Discovery). Results: The candidate genes with LOH identified in this study were MCU, MICU1 and OIT3 on chromosome 10. MCU was found to encode a calcium transporter and MICU1 could encode an essential regulator of mitochondrial $Ca^{2+}$ uptake. OIT3 possibly correlated with calcium binding revealed by the annotation analyses and was regulated by a large number of transcription factors including STAT, SOX9, CREB, NF-kB, PPARG and p53. Conclusions: Global genomic analysis of SNPs identified MICU1, MCU and OIT3 with LOH on chromosome 10, implying involvement of these genes in progression of pancreatic cancer.

• Asian Pacific Journal of Cancer Prevention
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• 제15권2호
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• pp.937-943
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• 2014

Polymorphisms in miRNA binding sites have been shown to affect miRNA binding to target genes, resulting in differential mRNA and protein expression and susceptibility to common diseases. Our purpose was to predict SNPs (single nucleotide polymorphisms) within miRNA binding sites of inflammatory genes in relation to gastric cancer. A complete list of SNPs in the 3'UTR regions of all inflammatory genes associated with gastric cancer was obtained from Pubmed. miRNA target prediction databases (MirSNP, Targetscan Human 6.2, PolymiRTS 3.0, miRNASNP 2.0, and Patrocles) were used to predict miRNA target sites. There were 99 SNPs with MAF>0.05 within the miRNA binding sites of 41 genes among 72 inflammation-related genes associated with gastric cancer. NF-${\kappa}B$ and JAK-STAT are the two most important signaling pathways. 47 SNPs of 25 genes with 95 miRNAs were predicted. CCL2 and IL1F5 were found to be the shared target genes of hsa-miRNA-624-3p. Bioinformatic methods could identify a set of SNPs within miRNA binding sites of inflammatory genes, and provide data and direction for subsequent functional verification research.

• Asian Pacific Journal of Cancer Prevention
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• 제17권11호
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• pp.5031-5035
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• 2016

Docetaxel, recognized as a stabilizing microtubule agent, is frequently administrated as a first line treatment for prostate cancers. Due to high side effects of monotherapy, however, combinations with novel adjuvants have emerged as an alternative strategy in cancer therapy protocols. Here, we investigated the combined effects of stattic and docetaxel on the DU145 prostate cancer cell line. Cytotoxicity was evaluated by MTT assay. To understand molecular mechanisms of stattic action, apoptotic related genes including Bcl-2, Mcl-1, Survivin and Bax were evaluated by real-time RT-PCR. Alteration in the expression of pro-apoptotic Bax and anti-apoptotic Bcl-2 genes and Bax/Bcl-2 ratio were investigated via the $2^{{\Delta}{\Delta}CT}$ method. The $IC_{50}$ values for docetaxel and stattic were $3.7{\pm}0.9nM$ and $4.6{\pm}0.8{\mu}M$, respectively. Evaluation of key gene expression levels revealed a noticeable decrease in antiapoptotic Bcl-2 and Mcl-1 along with an increase in pro-apoptotic Bax mRNA levels (p<0.05). Our results suggest that combination of a STAT3 inhibitor with doctaxel can be considered as a potent strategy for induction of apoptosis via increasing Bax mRNA expression.

• Molecules and Cells
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• 제42권12호
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• pp.906-918
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• 2019

MicroRNA-223-3p (miR-223-3p) is one of the potential microRNAs that have been shown to alleviate inflammatory responses in pre-clinical investigations and is highly encased in exosomes derived from bone mesenchymal stem cells (MSC-exosomes). MSC-exosomes are able to function as carriers to deliver microRNAs into cells. Autoimmune hepatitis is one of the challenging liver diseases with no effective treatment other than steroid hormones. Here, we examined whether MSC-exosomes can transfer miR-223-3p to treat autoimmune hepatitis in an experimental model. We found that MSC-exosomes were successfully incorporated with miR-223-3p and delivered miR-223-3p into macrophages. Moreover, there was no toxic effect of exosomes on the macrophages. Furthermore, treatments of either exosomes or exosomes with miR-223-3p successfully attenuated inflammatory responses in the liver of autoimmune hepatitis and inflammatory cytokine release in both the liver and macrophages. The mechanism may be related to the regulation of miR-223-3p level and STAT3 expression in the liver and macrophages. These results suggest that MSC-exosomes can be used to deliver miR-223-3p for the treatment of autoimmune hepatitis.

• Asian-Australasian Journal of Animal Sciences
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• 제30권5호
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• pp.666-671
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• 2017

Objective: Aim of the current comparative study was to evaluate production outputs, reproduction efficiency and functional traits in dual-purpose Fleckvieh and Braunvieh cows, reared under temperate European conditions. Methods: A data-set from 414 Fleckvieh and 42 Braunvieh cows and 799 lactations was analysed. ID tag number, milk yield per milking session, number of steps/interval and milk conductivity were recorded and collected daily using AfiMilk 3.076 A-DU software (Afimilk Ltd., Kibbutz, Israel). Production and milk quality data were taken from the results of the official performance recordings and the reproductive outputs of cows were recorded by the research stations veterinarians. Comparisons between the two genotypes were carried out using the one way analysis of variance protocol, with categorical factor being considered the breed of cows. All the statistical inferences were carried out using Statistica software (StatSoft Inc., Tulsa, OK, USA). Results: Fleckvieh cows significantly outperformed ($p{\leq}0.05$) the Braunvieh herd, with average milk yields of $5,252.1{\pm}35.79kg$ and $4,897.6{\pm}128.94kg$, respectively. Age at first calving was significantly ($p{\leq}0.01$) influenced by the breed, with Fleckvieh heifers being more precocious ($32.8{\pm}0.29mo$) compared to those of Braunvieh breed ($35.7{\pm}0.84mo$). Reproduction efficiency as defined by the number of inseminations per gestation, calving interval, dystocia, days dry and days open, was not influenced by genotype (p>0.05). Incidences of sub-clinical mastitis, clinical mastitis, lameness and abortions were not influenced by the breed factor (p>0.05). Stay-ability of cows was significantly ($p{\leq}0.001$) influenced by genotype, with Braunvieh cows having an average age at culling of $117.88{\pm}11.78$ months compared to $90.88{\pm}2.89$ months in Fleckvieh. Conclusion: Overall, results have shown that genotype significantly influenced milk yield, age at first calving and longevity.