• 생약학회지
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• 제49권2호
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• pp.145-154
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• 2018

Aconiti Kusnezoffii Tuber has been traditionally used to treat the symptoms of rheumatoid arthritis and joint pain. The main constituents are diterpenoid alkaloids such as benzoylmesaconine, benzoylaconine, mesaconitine, aconitine, and hypaconitine. In Korea, Aconiti Kusnezoffii Tuber is officially defined as the tubers of Aconitum kusnezoffii Reichb., A. ciliare Decasisne, and A. triphyllum Nakai. On the other hand, only the tuber of A. kusnezoffii is to be used in China. In order to identify the botanical origin of Aconiti Kusnezoffii Tuber circulated in Korea, we analyzed 24 samples of Aconiti Kusnezoffii Tuber obtained from local markets for comparative DNA analysis. The sequence analysis of nrRNA ITS 1 was useful to distinguish Aconitum species and revealed that the roots of A. karakolicum were circulated in Korean markets without discretion. HPLC quantitative analysis showed that aconitine was detected at the highest amount in A. karakolicum. Authentic diterpenoid alkaloids were coinjected for quantification of aconitine-type ingredients. All data were statistically grouped by Principal Component Analysis (PCA). This study suggests that both molecular and chemical analyses should be utilized for the standardization and the quality control for Aconiti Kusnezoffii Tuber.

• Journal of Microbiology and Biotechnology
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• 제20권10호
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• pp.1463-1470
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• 2010

E. coli has long been widely used as a host system for the manufacture of recombinant proteins intended for human therapeutic use. When considering the impurities to be eliminated during the downstream process, residual host cell DNA is a major safety concern. The presence of residual E. coli host cell DNA in the final products is typically determined using a conventional slot blot hybridization assay or total DNA Threshold assay. However, both the former and latter methods are time consuming, expensive, and relatively insensitive. This study thus attempted to develop a more sensitive real-time PCR assay for the specific detection of residual E. coli DNA. This novel method was then compared with the slot blot hybridization assay and total DNA Threshold assay in order to determine its effectiveness and overall capabilities. The novel approach involved the selection of a specific primer pair for amplification of the E. coli 16S rRNA gene in an effort to improve sensitivity, whereas the E. coli host cell DNA quantification took place through the use of SYBR Green I. The detection limit of the real-time PCR assay, under these optimized conditions, was calculated to be 0.042 pg genomic DNA, which was much higher than those of both the slot blot hybridization assay and total DNA Threshold assay, where the detection limits were 2.42 and 3.73 pg genomic DNA, respectively. Hence, the real-time PCR assay can be said to be more reproducible, more accurate, and more precise than either the slot blot hybridization assay or total DNA Threshold assay. The real-time PCR assay may thus be a promising new tool for the quantitative detection and clearance validation of residual E. coli host cell DNA during the manufacturingprocess for recombinant therapeutics.

• International Journal of Industrial Entomology and Biomaterials
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• 제23권2호
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• pp.223-229
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• 2011

Cyclophilins are originally identified as cytosolic binding protein of the immunosuppressive drug cyclosporine A. They have an activity of peptidyl prolyl cis/trans-isomerases (PPIase), which may play important roles in protein folding, trafficking, assembly and cell signaling. In this study, we report the cloning and characterization of a Bombyx mori cyclophilin A (bCypA) cDNA. The full-length cDNA of bCypA consist of 947 nucleotides with a polyadenylation signal sequence AATAAA and contain an open reading frame of 498 nucleotides encoding a polypeptide of 166 amino acids. The deduced amino acid sequence of bCypA shares a central peptidyl prolyl cis/trans-isomerase and a cyclosporin-A-binding domain with other cyclophilin sequences. Relative quantification real-time (RT) PCR analysis shows that mRNA transcripts of bCypA are detected in all the investigated tissues and highest expression level in the skin of 3-day-old 5 instar larva. Also, bCypA had PPIase activity on the proline-containing peptides. Accordingly, we suggest that bCypA is a new member of the cyclophilin A (CyPA) family and will be useful for quality control of bioactivity recombinant proteins with proline-containing peptides.

• Animal Bioscience
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• 제36권1호
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• pp.19-28
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• 2023

Objective: The objective of this study was to perform genome (genome wide association studies [GWAS]) and chromosome (CWAS) wide association analyses to identify single nucleotide polymorphisms (SNPs) associated with growth traits in registered Simmental and Simbrah cattle. Methods: The phenotypes were deregressed BLUP EBVs for birth weight, weaning weight direct, weaning weight maternal, and yearling weight. The genotyping was performed with the GGP Bovine 150k chip. After the quality control analysis, 105,129 autosomal SNP from 967 animals (473 Simmental and 494 Simbrah) were used to carry out genotype association tests. The two association analyses were performed per breed and using combined information of the two breeds. The SNP associated with growth traits were mapped to their corresponding genes at 100 kb on either side. Results: A difference in magnitude of posterior probabilities was found across breeds between genome and chromosome wide association analyses. A total of 110, 143, and 302 SNP were associated with GWAS and CWAS for growth traits in the Simmental-, Simbrah- and joint -data analyses, respectively. It stands out from the enrichment analysis of the pathways for RNA polymerase (POLR2G, POLR3E) and GABAergic synapse (GABRR1, GABRR3) for Simmental cattle and p53 signaling pathway (BID, SERPINB5) for Simbrah cattle. Conclusion: Only 6,265% of the markers associated with growth traits were found using CWAS and GWAS. The associated markers using the CWAS analysis, which were not associated using the GWAS, represents information that due to the model and priors was not associated with the traits.

• Journal of Microbiology and Biotechnology
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• 제33권12호
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• pp.1657-1670
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• 2023

This study aimed to evaluate the effects of Limosilactobacillus fermentum and Lactiplantibacillus plantarum isolated from human feces coordinating with inulin on the composition of gut microbiota and metabolic profiles in db/db mice. These supplements were administered to db/db mice for 12 weeks. The results showed that the Lactobacillaceae coordinating with inulin group (LI) exhibited lower fasting blood glucose levels than the model control group (MC). Additionally, LI was found to enhance colon tissue and increase the levels of short-chain fatty acids. 16S rRNA sequencing revealed that the abundance of Corynebacterium and Proteus, which were significantly increased in the MC group compared with NC group, were significantly decreased by the treatment of LI that also restored the key genera of the Lachnospiraceae_NK4A136_group, Lachnoclostridium, Ruminococcus_gnavus_group, Desulfovibrio, and Lachnospiraceae_UCG-006. Untargeted metabolomics analysis showed that lotaustralin, 5-hydroxyindoleacetic acid, and 13(S)-HpODE were increased while L-phenylalanine and L-tryptophan were decreased in the MC group compared with the NC group. However, the intervention of LI reversed the levels of these metabolites in the intestine. Correlation analysis revealed that Lachnoclostridium and Ruminococcus_gnavus_group were negatively correlated with 5-hydroxyindoleacetic acid and 13(S)-HpODE, but positively correlated with L-tryptophan. 13(S)-HpODE was involved in the "linoleic acid metabolism". L-tryptophan and 5-hydroxyindoleacetic acid were involved in "tryptophan metabolism" and "serotonergic synapse". These findings suggest that LI may alleviate type 2 diabetes symptoms by modulating the abundance of Ruminococcus_gnavus_group and Lachnoclostridium to regulate the pathways of "linoleic acid metabolism", "serotonergic synapse", and" tryptophan metabolism". Our results provide new insights into prevention and treatment of type 2 diabetes.

• Nutrition Research and Practice
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• 제18권1호
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• pp.46-61
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• 2024

BACKGROUND/OBJECTIVES: An increasing life expectancy in society has burdened healthcare systems substantially because of the rising prevalence of age-related metabolic diseases. This study compared the effects of animal protein hydrolysate (APH) and casein on metabolic diseases using aged mice. MATERIALS/METHODS: Eight-week-old and 50-week-old C57BL/6J mice were used as the non-aged (YC group) and aged controls (NC group), respectively. The aged mice were divided randomly into 3 groups (NC, low-APH [LP], and high-APH [HP] and fed each experimental diet for 12 weeks. In the LP and HP groups, casein in the AIN-93G diet was substituted with 16 kcal% and 24 kcal% APH, respectively. The mice were sacrificed when they were 63-week-old, and plasma and hepatic lipid, white adipose tissue weight, hepatic glucose, lipid, and antioxidant enzyme activities, immunohistochemistry staining, and mRNA expression related to the glucose metabolism on liver and muscle were analyzed. RESULTS: Supplementation of APH in aging mice resulted in a significant decrease in visceral fat (epididymal, perirenal, retroperitoneal, and mesenteric fat) compared to the negative control (NC) group. The intraperitoneal glucose tolerance test and area under the curve analysis revealed insulin resistance in the NC group, which was alleviated by APH supplementation. APH supplementation reduced hepatic gluconeogenesis and increased glucose utilization in the liver and muscle. Furthermore, APH supplementation improved hepatic steatosis by reducing the hepatic fatty acid and phosphatidate phosphatase activity while increasing the hepatic carnitine palmitoyltransferase activity. Furthermore, in the APH supplementation groups, the red blood cell (RBC) thiobarbituric acid reactive substances and hepatic H₂O₂ levels decreased, and the RBC glutathione, hepatic catalase, and glutathione peroxidase activities increased. CONCLUSIONS: APH supplementation reduced visceral fat accumulation and alleviated obesity-related metabolic diseases, including insulin resistance and hepatic steatosis, in aged mice. Therefore, high-quality animal protein APH that reduces the molecular weight and enhances the protein digestibility-corrected amino acid score has potential as a dietary supplement for healthy aging.

• 한국유기농업학회지
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• 제24권4호
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• pp.833-847
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• 2016

가뭄은 농작물의 생산성을 저해하는 주요 원인 중 하나이며, 잎을 먹는 쌈채류의 경우 물 부족 스트레스에 더 치명적일 수 있다. 본 연구는 케일 유묘에 대한 식물생육촉진 균주(PGPB)의 가뭄 내성 효과를 알아보기 위해 수행되었다. 쌈채류의 토양 및 근권 토양으로부 터 형태학적으로 구분되는 146개의 콜로니를 분리하고 온실 생물검정을 통해 케일 생육촉진이 우수한 균주 SB19를 최종 선발하였다. SB19 균주는 케일 재배 토양으로부터 분리하였으며 16S rRNA 유전자 염기서열 분석 결과 Bacillus sp.로 확인되었다. Bacillus sp. SB19 균주를 처리한 케일에 7일 간 수분 부족 스트레스를 유도하고 7일째에 가뭄 피해 조사 후 모든 처리구에 1회 관수하였다. 이후 다시 7일 간 수분 부족 스트레스를 주어 14일째에 케일의 내건성 증진 여부를 조사하였다. 가뭄 조건 7일째에 $10^6$와 $10^7cell\;mL^{-1}$ 농도의 SB19 균주를 처리한 케일에서 무처리와 비교하여 가뭄 스트레스 경감 효과를 보였다. 7일째에 모든 처리구에 관수 후 다시 가뭄 스트레스를 주었을 때에도 $10^6$와 $10^7cell\;mL^{-1}$ 농도의 SB19 균주 처리구에서 무처리구와 비교하여 가뭄 피해 경감 효과가 있었으며, 7일째와 14일째 모두에서 $10^7cell\;mL^{-1}$ 농도의 SB19 균주 처리구에서 가뭄 피해의 완화 정도가 가장 효과적인 것으로 나타났다. $10^6cell\;mL^{-1}$ SB19 균주 처리구에서는 물 부족으로 인한 잎의 노화가 $10^7cell\;mL^{-1}$ 농도 처리구에 비해 빠르게 발생하였다. 본 연구 결과를 바탕으로 유용 미생물과 식물의 상호작용이 식물의 물 이용률을 증진시키는 중요한 역할을 하고 약한 가뭄 조건에서 쌈채류의 품질을 향상시킬 수 있는 방안이 될 수 있을 것이라고 예측한다. 즉, 미생물학적인 환경 스트레스 극복 방법으로서의 가치를 뒷받침하는 것이라 할 수 있다.

• Reproductive and Developmental Biology
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• 제35권1호
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• pp.123-129
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• 2011

The pig has been considered to serve as an appropriate model of human disease. Therefore, establishment of porcine embryonic stem cell lines is important. The purpose of the present study was to further work in this direction. We produced porcine parthenogenetic embryos, and separately aggregated two of each of two-cell ($2{\times}2$), four-cell ($2{\times}4$), and eight-cell ($2{\times}8$) embryos derived by parthenogenesis. After culture for 4 days, the developmental ability of the aggregates and total blastocyst cell numbers were evaluated. The percentage of blastocysts was significantly higher in both $2{\times}4$- and $2{\times}8$-aggregated embryos ($58.3{\pm}1.9%$ and $37.2{\pm}2.8%$, respectively) than in the control or $2{\times}2$-aggregated embryos ($23.6{\pm}1.1%$ and $12.5{\pm}2.4%$, respectively). Total blastocyst cell numbers were increased in the $2{\times}4$- and $2{\times}8$-aggregated embryos (by $44{\pm}3.0%$ and $45{\pm}3.3%$, respectively) compared with those of control or $2{\times}2$-aggregated embryos ($30.5{\pm}2.1%$ and $30.7{\pm}2.6%$, respectively; p<0.05). The levels of mRNA encoding Oct-4 were higher in both the $2{\times}4$- and $2{\times}8$-aggregated embryos than in the control. When blastocysts derived from $2{\times}4$- aggregated embryos or intact normal embryos were cultured on mouse embryonic fibroblast feeder cells to obtain porcine stem cells, blastocysts from aggregated embryos formed colonies that were better in shape compared with those derived from intact blastocysts. Together, the data show that aggregation of porcine embryos not only improves blastocyst quality but also serves as an efficient procedure by which porcine embryonic stem cells can become established.

• Annals of Pediatric Endocrinology and Metabolism
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• 제23권4호
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• pp.204-209
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• 2018

Purpose: Growth hormone transduction defect (GHTD) is characterized by severe short stature, impaired STAT3 (signal transducer and activator of transcription-3) phosphorylation and overexpression of the cytokine inducible SH2 containing protein (CIS) and p21/CIP1/WAF1. To investigate the role of p21/CIP1/WAF1 in the negative regulation of the growth hormone (GH)/GH receptor and Epidermal Growth Factor (EGF)/EGF Receptor pathways in GHTD. Methods: Fibroblast cultures were developed from gingival biopsies of 1 GHTD patient and 1 control. The protein expression and the cellular localization of p21/CIP1/WAF1 was studied by Western immunoblotting and immunofluorescence, respectively: at the basal state and after induction with $200-{\mu}g/L$ human GH (hGH) (GH200), either with or without siRNA CIS (siCIS); at the basal state and after inductions with $200-{\mu}g/L$ hGH (GH200), $1,000-{\mu}g/L$ hGH (GH1000) or 50-ng/mL EGF. Results: After GH200/siCIS, the protein expression and nuclear localization of p21 were reduced in the patient. After successful induction of GH signaling (control, GH200; patient, GH1000), the protein expression and nuclear localization of p21 were reduced. After induction with EGF, p21 translocated to the cytoplasm in the control, whereas in the GHTD patient it remained located in the nucleus. Conclusion: In the GHTD fibroblasts, when CIS is reduced, either after siCIS or after a higher dose of hGH (GH1000), p21's antiproliferative effect (nuclear localization) is also reduced and GH signaling is activated. There also appears to be a positive relationship between the 2 inhibitors of GH signaling, CIS and p21. Finally, in GHTD, p21 seems to participate in the regulation of both the GH and EGF/EGFR pathways, depending upon its cellular location.

• 미생물학회지
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• 제50권3호
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• pp.201-209
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• 2014

본 연구에서는 고추재배 지역의 오염지 토양으로부터 탄저병원균인 Colletotrichum gloeosporioides에 대하여 항진균 활성이 우수한 균주를 분리하였다. 분리 균주의 생화학적 특성을 조사한 결과 내생포자를 형성하는 Gram-positive이며, 세포크기는 $2.5{\sim}3.0{\times}1.5{\mu}m$으로 짧은 간균으로 siderophore, cellulase와 IAA (Indole-3-acetic acid)을 생산하였다. 16S rRNA 유전자염기서열 분석 결과 Bacillus sp.와 99%의 상동성을 나타내어 Bacillus sp. CS-52로 명명하였다. Bacillus sp. CS-52의 항진균 활성 물질을 생산하기 위한 배양 최적 조건은 0.5% glucose, 0.7% $K_2HPO_4$, 0.2% $K_2HPO_4$, 0.3% $NH_4NO_3$, 0.01% $MnSO_4{\cdot}7H_2O$, 0.15% yeast extract, pH 7과 $30^{\circ}C$로 조사되었으며, 최적화된 배양조건에서 36시간에 최대 성장을 보이며, 고추 탄저병원균에 대하여 60시간 배양조건에서 가장 높은 13.3 mm의 항진균 활성을 보였다. 포자발아억제력은 48시간에 가장 높은 억제력이 보였고, siderophore는 최종배양시간 72시간까지 생성됨을 확인하였다. 식물생장조절물질 IAA와 활성효소인 cellulase의 경우 배양시간 24시간에 최대 생성됨을 확인하였으며, C. gloeosporioides에 대한 실내에서의 항진균활성 검증결과 화학농약 보다 더 높은 70%의 방제가를 나타내었다. 향후 Bacillus sp. CS-52 균주와 배양액을 이용하여 생물학적 친환경방제제로의 제품화가 가능할 것으로 사료된다.