• 한국동물위생학회지
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• 제32권1호
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• pp.61-76
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• 2009

At the present study, it was aimed to explore the molecular genetic characterization of multiple antimicrobial resistant Salmonella spp. isolates from pigs and cattle. A total of 138 Salmonella Typhimurium (S. Typhimurium) isolates were typed with phage, among them, 83.3% of S. Typhimurium tested could divide into a 10 phage types. Definitive type 193 (DT193) (25.4%) and DT195 (24.6%) were exhibited as the dominant types. DT104 and U302 were found from pigs and cattle. On the other hand, S. Enteritidis had 6 phage types, of them, phage type 21 (PT21) and PT11b were the popular types. In the plasmid profiles, 135 of S. Typhimurium isolates were exhibited 1 to 6 plasmid bands which molecular weight ranged from 90 to 2kb. 35 isolates (25.4%) harbored a 90kb plasmid which is thought to be the serotype specific virulence plasmid. Two of twenty five S. Enteritidis had common plasmids at 2 and 1.5kb. With multiplex polymerase chain reaction, virulence genes (invA and spvC) were detected from all Salmonella spp. from 167 of S. Typhimurium, S. Enteritidis and chloramphenicol resistant S. Schwarzengrund, but some drug resistant genes, such as PSE-1, cml/tetR and flo were not determined but other drug resistant genes, for example TEM and int were found. The detection rates of spvC, TEM and int gene was 35.3%, 29.3% and 72.5%, respectively. The TEM gene was highly popular in S. Typhimurium, which was detected from ampicillin and amoxicillin resistant strains as 95.9%. int gene was able to detect from all the isolates identified as multidrug resistsnt (MDR), particularly DT193 was thought as the most prevalent virulence and multidrug resistance isolate. The major plasmid profile and drug resistance pattern of DT193 were 90, 40, 10.5, 6.3, 3.0kb and ACCbDNaPSSuT, respectively. MDR was commonly found in other phage types, particularly DT104, U302 and DT203.

• 한국동물위생학회지
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• 제32권1호
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• pp.49-59
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• 2009

At the present study, it was aimed to explore the states of antimicrobial resistant Salmonella spp. isolates from 3,850 pigs (2,732 ileocecocolic lymphnodes and 1,118 cecal contents) and 1,764 cattle (965 cecal lymphnodes and 799 cecal contents) slaughtered in Busan province from December 2000 to November 2001. Among 5,614 samples, 457 of Salmonella spp. were isolated from pig lymphnodes (13.5%), pig cecal contents (4.4%), cattle lymphnodes (3.5%) and cattle cecal contents (0.5%). Salmonella spp. were showed different isolation ratio, that was 10.8% in summer, 9.0% in autumn, 8.4% in spring and 5.0% in winter. As a result of serotyping, B group (65.4%) were identified as the most common in pigs and cattle, in order of $C_1$ (14.0%), $D_1$ (5.5%), $C_2$ (4.2%), $E_1$ (4.2%) and L (3.5%). 34 serotypes were found, among them, Salmonella Typhimurium (S. Typhimurium) (21.0%) was the most common serotype from pigs and cattle. The major serotypes were in order of S. Derby (15.3%), S. Schwarzengrund (14.7%), S. Typhimurium var Copenhagen (9.2%), S. Mbandaka (5.7%), S. Enteritidis (5.5%) and S. Ruiru (3.5%). The most common serotype was S. Typhimurium in pigs, and S. Ruiru in cattle. S. Ruiru was firstly isolated from pigs and cattle in Korea. In antimicrobial susceptibility test, all the isolates were demonstrated susceptibility to norfloxacin and ofloxacin. But the isolates were showed resistance other antibiotics in order of doxycycline (68.3%), tetracycline (67.8%), penicillin (54.5%) and streptomycin (52.5%). S. Typhimurium were exhibited resistance to ampicillin (34.8%), chloramphenicol (36.2%), streptomycin (94.9%), sulfamethoxazole/trimethoprim (34.8%) and tetracycline (97.8%). There were 53 strains (38.4%) which had multi drug resistant (MDR) isolates, resistant to more than 6 antimicrobial agents. The most common resistance patterns of MDR isolates were ampicillin, chloramphenicol, carbenicillin, doxycycline, nalidixic acid, penicillin, streptomycin, sulfamethoxazole/trimethoprim and tetracycline (ACCbDNaPSSuT).

• Journal of Korean Medical Science
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• 제33권47호
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• pp.303.1-303.10
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• 2018

Background: Cell division cycle 6 (CDC6) is an essential regulator of DNA replication and plays important roles in the activation and maintenance of the checkpoint mechanisms in the cell cycle. CDC6 has been associated with oncogenic activities in human cancers; however, the clinical significance of CDC6 in prostate cancer (PCa) remains unclear. Therefore, we investigated whether the CDC6 mRNA expression level is a diagnostic and prognostic marker in PCa. Methods: The study subjects included 121 PCa patients and 66 age-matched benign prostatic hyperplasia (BPH) patients. CDC6 expression was evaluated using real-time polymerase chain reaction and immunohistochemical (IH) staining, and then compared according to the clinicopathological characteristics of PCa. Results: CDC6 mRNA expression was significantly higher in PCa tissues than in BPH control tissues (P = 0.005). In addition, CDC6 expression was significantly higher in patients with elevated prostate-specific antigen (PSA) levels (> 20 ng/mL), a high Gleason score, and advanced stage than in those with low PSA levels, a low Gleason score, and earlier stage, respectively. Multivariate logistic regression analysis showed that high expression of CDC6 was significantly associated with advanced stage (${\geq}T3b$) (odds ratio [OR], 3.005; confidence interval [CI], 1.212-7.450; P = 0.018) and metastasis (OR, 4.192; CI, 1.079-16.286; P = 0.038). Intense IH staining for CDC6 was significantly associated with a high Gleason score and advanced tumor stage including lymph node metastasis stage (linear-by-linear association, P = 0.044 and P = 0.003, respectively). Conclusion: CDC6 expression is associated with aggressive clinicopathological characteristics in PCa. CDC6 may be a potential diagnostic and prognostic marker in PCa patients.

• Biomolecules & Therapeutics
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• 제29권3호
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• pp.321-330
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• 2021

Oxidative stress plays a crucial role in the development of neuronal disorders including brain ischemic injury. Thioredoxin 1 (Trx1), a 12 kDa oxidoreductase, has anti-oxidant and anti-apoptotic functions in various cells. It has been highly implicated in brain ischemic injury. However, the protective mechanism of Trx1 against hippocampal neuronal cell death is not identified yet. Using a cell permeable Tat-Trx1 protein, protective mechanism of Trx1 against hydrogen peroxide-induced cell death was examined using HT-22 cells and an ischemic animal model. Transduced Tat-Trx1 markedly inhibited intracellular ROS levels, DNA fragmentation, and cell death in H2O2-treatment HT-22 cells. Tat-Trx1 also significantly inhibited phosphorylation of ASK1 and MAPKs in signaling pathways of HT-22 cells. In addition, Tat-Trx1 regulated expression levels of Akt, NF-κB, and apoptosis related proteins. In an ischemia animal model, Tat-Trx1 markedly protected hippocampal neuronal cell death and reduced astrocytes and microglia activation. These findings indicate that transduced Tat-Trx1 might be a potential therapeutic agent for treating ischemic injury.

• Asian-Australasian Journal of Animal Sciences
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• 제31권12호
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• pp.1963-1973
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• 2018

Objective: This study was conducted to evaluate the effect of dietary energy and lipase supplementation on growth performance, nutrient digestibility, serum profiles, intestinal morphology, small intestinal digestive enzyme activities, biochemical index of intestinal development and noxious gas emission in weaning pigs. Methods: A total of 240 weaning pigs ([Yorkshire${\times}$Landrace]${\times}$Duroc) with an average body weight (BW) of $7.3{\pm}0.12kg$ were used in this 28-d experiment. Weaning pigs were randomly allocated to 4 dietary treatments in a $2{\times}2$ factorial arrangement with 2 levels of energy (net energy = 2,470 kcal/kg for low energy diet and 2,545 kcal/kg for basal diet) and 2 levels of lipase (0 and 1.5 U/g of lipase) according to BW and sex. There were 6 replications (pens) per treatment and 10 pigs per pen (5 barrows and 5 gilts). Results: Weaning pigs fed the low energy diet had lower (p<0.05) gain-to-feed ratio (G:F) throughout the experiment, apparent digestibility of dry matter, nitrogen, ether extract, and gross energy during d 0 to 14, average daily gain during d 15 to 28, lipase activity in duodenum and ileum and protein/DNA in jejunum (p<0.05), respectively. Lipase supplementation had no effect on growth performance but affected apparent nutrient digestibility (p<0.05) on d 14 and enhanced lipase activity in the duodenum and ileum and protease activity in duodenum and jejunum of pigs (p<0.05) fed the low energy diet. Lipase reduced serum low-density lipoprotein cholesterol (LDL-C) and triglyceride (TG), $NH_3$ production (p<0.05) from the feces. Conclusion: The low energy diet decreased G:F throughout the experiment and nutrient digestibility during d 0 to 14 as well as lipase activity in duodenum and ileum. Lipase supplementation increased nutrient digestibility during d 0 to 14 and exerted beneficial effects on lipase activity in duodenum and ileum as well as protease activity in duodenum and jejunum, while reduced serum LDL-C, TG and fecal $NH_3$.

• 현장농수산연구지
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• 제23권1호
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• pp.117-128
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• 2021

한우의 성장단계별 부위 근육발달을 이해하는 것은 도체율 개선에 따른 소득증대와 증체율 증가에 따른 생산효율 향상에 긍정적인 영향을 미친다. 본 연구에서는 한우의 등심과 사태 유래 근육위성세포를 분리 후 세포단위의 발달 및 분화를 비교하여 transcriptome 단위의 작용기전을 제시하였다. 한우의 부위별 근육 유래 근육위성세포의 근섬유의 양은 4일에 가장 높게 나타났고 이후 감소하였다. 한우의 근육위성세포의 발달 단계에 따라 발현되는 총 전사체 유전자의 종류는 사태근육 위성세포에서 높게 나타났다. 등심과 사태 근육 유래 위성세포의 발달단계에 따라 유의적인 차등 유전자 453개를 찾아냈고 이를 이용한 기능유 전체 분석이 필요하다. 등심과 사태유래 근육위성세포를 이용한 동일조건 분화 비교에서 사태유래 근육 위성세포의 분화 시 myosin complex, skeletal muscle contraction, troponin complex, skeletal muscle tissue development 와 같은 근섬유 형성관련 유전자의 발현이 높게 나타나는 것으로 보아 같은 개체의 근육조직에서도 부위별로 차등 발달이 되고 있다는 것을 알 수 있다. 기존 연구에서는 근육의 성장에 대한 이해를 위해 사양과 영양관련 시험이 많이 이루어졌다. 향후 세포단위의 연구들이 많이 이루어져 작용기작에 대한 생물정보 자료를 추가로 적용한다면 한우의 정밀사양을 적용할수 있는 바탕이 마련될 것이다. 또한 근육위성세포의 연구는 추후 동물실험 윤리제도 강화에 따른 비동물 전임상 screening 시험 활용과 대체단백질 산업의 주요 이슈인 배양육 소재 개발 연구와 같이 축산시험연구의 지속적인 확장성에 많은 영향을 미칠 것으로 생각된다.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2001년도 춘계학술발표대회
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• pp.69-69
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• 2001

The cell cycle phase in which donor nuclei exist prior to nuclear transfer is an important factor governing developmental rates of reconstituted embryos. It was suggested that quiescent G0 and cycling G1 cells could support normal development of reconstituted embryos. In a quest of optimized donor nuclei treatment prior to nuclear transfer, this study was undertaken to examine the cell cycle characteristics of bovine fetal and adult somatic cells when cultured under a variety of culture treatments and the cell cycle change with the lapse of time after trypsinization. This was archived by measuring the DNA content of cells using flow cytometry, Cultured fetal fibroblast cells, adult skin and muscle cells, and cumulus cells were divided by 3 culture treatments; 1) grown to 60-70% confluency (cycling), 2) serum starved culture, 3) culture to confluency. Trypsinized cells were fixed by 70% ethanol and stained with propidium iodide. For one experiment, trypsinized cells were resuspended in DMEM＋10% FBS and incubated for 1.5, 3 and 6 h with occasional shaking before ethanol fixation. Cell cycle phases were determined by flow cytometry enabling calculation of percentages of G0＋G1, S and G2＋M. The majority of cells were in G0＋Gl stage regardless of origin of cells. Cultures that were serum starved or cultured to confluency contained significantly (P<0.05) higher percentages of cells in G0＋G1 (89.5-95.4%). For every cell lines and culture treatments, percentages of cells in existing in G0＋G1 increased with decreasing of the cell size from large to small. In the serum starved and confluency groups, about 98% of small cells were in G0＋G1 Serum starved culture contained higher percentages of small-sized cells (38.5-66.9%) than cycling and confluent cultures regardless of cell lines (P<0.05). After trypsinization of fetal fibroblast and adult skin cells that were serum starved and cultured to confluency, the percentages of cells in G0＋G1 significantly increased by incubation for 1.5(95.7-99.5%) and 3.0 h (95.9-98.6%). The results suggest that the efficient synchronization of bovine somatic cells in G0＋G1 for nuclear transfer can be established by incubation for a limited time period after trypsinization of serum starved or confluent cells.

• Journal of Ginseng Research
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• 제47권2호
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• pp.337-346
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• 2023

Background: Ginsenoside Rb2, a major active component of Panax ginseng, has various physiological activities, including anticancer and anti-inflammatory effects. However, the mechanisms underlying the rejuvenation effect of Rb2 in human skin cells have not been elucidated. Methods: We performed a senescence-associated β-galactosidase staining assay to confirm cellular senescence in human dermal fibroblasts (HDFs). The regulatory effects of Rb2 on autophagy were evaluated by analyzing the expression of autophagy marker proteins, such as microtubule-associated protein 1A/1B-light chain (LC) 3 and p62, using immunoblotting. Autophagosome and autolysosome formation was monitored using transmission electron microscopy. Autophagic flux was analyzed using tandem-labeled GFP-RFP-LC3, and lysosomal function was assessed with Lysotracker. We performed RNA sequencing to identify potential target genes related to HDF rejuvenation mediated by Rb2. To verify the functions of the target genes, we silenced them using shRNAs. Results: Rb2 decreased β-galactosidase activity and altered the expression of cell cycle regulatory proteins in senescent HDFs. Rb2 markedly induced the conversion of LC3-I to LC3-II and LC3 puncta. Moreover, Rb2 increased lysosomal function and red puncta in tandem-labeled GFP-RFP-LC3, which indicate that Rb2 promoted autophagic flux. RNA sequencing data showed that the expression of DNA damage-regulated autophagy modulator 2 (DRAM2) was induced by Rb2. In autophagy signaling, Rb2 activated the AMPK-ULK1 pathway and inactivated mTOR. DRAM2 knockdown inhibited autophagy and Rb2-restored cellular senescence. Conclusion: Rb2 reverses cellular senescence by activating autophagy via the AMPK-mTOR pathway and induction of DRAM2, suggesting that Rb2 might have potential value as an antiaging agent.

• Journal of Ginseng Research
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• 제47권5호
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• pp.627-637
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• 2023

Background: Damage to the healthy intestinal epithelial layer and regulation of the intestinal immune system, closely interrelated, are considered pivotal parts of the curative treatment for inflammatory bowel disease (IBD). Plant-based diets and phytochemicals can support the immune microenvironment in the intestinal epithelial barrier for a balanced immune system by improving the intestinal microecological balance and may have therapeutic potential in colitis. However, there have been only a few reports on the therapeutic potential of plant-derived exosome-like nanoparticles (PENs) and the underlying mechanism in colitis. This study aimed to assess the therapeutic effect of PENs from Panax ginseng, ginseng-derived exosome-like nanoparticles (GENs), in a mouse model of IBD, with a focus on the intestinal immune microenvironment. Method: To evaluate the anti-inflammatory effect of GENs on acute colitis, we treated GENs in Caco2 and lipopolysaccharide (LPS) -induced RAW 264.7 macrophages and analyzed the gene expression of proinflammatory cytokines and anti-inflammatory cytokines such as TNF-α, IL-6, and IL-10 by real-time PCR (RT-PCR). Furthermore, we further examined bacterial DNA from feces and determined the alteration of gut microbiota composition in DSS-induced colitis mice after administration of GENs through 16S rRNA gene sequencing analysis. Result: GENs with low toxicity showed a long-lasting intestinal retention effect for 48 h, which could lead to effective suppression of pro-inflammatory cytokines such as TNF-α and IL-6 production through inhibition of NF-κB in DSS-induced colitis. As a result, it showed longer colon length and suppressed thickening of the colon wall in the mice treated with GENs. Due to the amelioration of the progression of DSS-induced colitis with GENs treatment, the prolonged survival rate was observed for 17 days compared to 9 days in the PBS-treated group. In the gut microbiota analysis, the ratio of Firmicutes/Bacteroidota was decreased, which means GENs have therapeutic effectiveness against IBD. Ingesting GENs would be expected to slow colitis progression, strengthen the gut microbiota, and maintain gut homeostasis by preventing bacterial dysbiosis. Conclusion: GENs have a therapeutic effect on colitis through modulation of the intestinal microbiota and immune microenvironment. GENs not only ameliorate the inflammation in the damaged intestine by downregulating pro-inflammatory cytokines but also help balance the microbiota on the intestinal barrier and thereby improve the digestive system.

• Animal Bioscience
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• 제37권3호
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• pp.522-535
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• 2024

Objective: Transition period is considered from 3 weeks prepartum to 3 weeks postpartum, characterized with dramatic events (endocrine, metabolic, and physiological) leading to occurrence of production diseases (negative energy balance/ketosis, milk fever etc). The objectives of our study were to analyze the periodic concentration of serum beta-hydroxy butyric acid (BHBA), glucose and oxidative markers along with identification, and validation of the putative markers of negative energy balance in buffaloes using in-silico and quantitative real time-polymerase chain reaction (qRT-PCR) assay. Methods: Out of 20 potential markers of ketosis identified by in-silico analysis, two were selected and analyzed by qRT-PCR technique (upregulated; acetyl serotonin o-methyl transferase like and down regulated; guanylate cyclase activator 1B). Additional two sets of genes (carnitine palmotyl transferase A; upregulated and Insulin growth factor; downregulated) that have a role of hepatic fatty acid oxidation to maintain energy demands via gluconeogenesis were also validated. Extracted cDNA (complementary deoxyribonucleic acid) from the blood of the buffaloes were used for validation of selected genes via qRTPCR. Concentrations of BHBA, glucose and oxidative stress markers were identified with their respective optimized protocols. Results: The analysis of qRT-PCR gave similar trends as shown by in-silico analysis throughout the transition period. Significant changes (p<0.05) in the levels of BHBA, glucose and oxidative stress markers throughout this period were observed. This study provides validation from in-silico and qRT-PCR assays for potential markers to be used for earliest diagnosis of negative energy balance in buffaloes. Conclusion: Apart from conventional diagnostic methods, this study improves the understanding of putative biomarkers at the molecular level which helps to unfold their role in normal immune function, fat synthesis/metabolism and oxidative stress pathways. Therefore, provides an opportunity to discover more accurate and sensitive diagnostic aids.