• Title/Summary/Keyword: OGT

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Quantitative Analytical Method for Eighteen Marker Compounds in Ongyeong-tang, a Traditional Herbal Formula, Using High-Performance Liquid Chromatography for Quality Assessment

  • Kim, Jung-Hoon;Shin, Hyeun-Kyoo;Seo, Chang-Seob
    • The Korea Journal of Herbology
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    • v.30 no.4
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    • pp.11-20
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    • 2015
  • Objectives : Ongyeong-tang (OGT) is a traditional herbal formula used to cure gynaecological disorders. OGT consists of 12 herbal medicines containing various bioactive components. Therefore, the development of suitable analytical method for the marker compounds is necessary for the quality control of OGT. Methods : Determination of the 18 marker compounds in OGT preparations was quantitatively performed by high-performance liquid chromatography-photodiode array detection analysis. The marker compounds were separated on a reversed-phase C 18 column and the analytical method was successfully validated, which was applied to compare OGT extracts from laboratory preparation and commercial OGT granules. Results : Limit of detection and limit of quantification values were in the ranges of $0.001-0.016{\mu}g/mL$ and $0.003-0.047{\mu}g/mL$, respectively. Precision was 0.03-3.71 % within a day and 0.03-3.81 % over four consecutive days. Recovery of marker compounds ranged from 90.63-108.26 %, with relative standard deviation (RSD) values < 4.0 %. Reproducibility was < 2.5 % of the RSD value. The 18 marker compounds were stable within 16 h at $10^{\circ}C$, with the RSD value < 3.5 %. Quantitative analysis results showed that the quantities of the 18 marker compounds varied among OGT samples. Pearson coefficient evaluation and principal component analysis demonstrated that an OGT water extract produced by a laboratory method clearly differed from commercial OGT granules. Conclusions : The developed analytical method was simple, precise, and reliable. Therefore, it can be used for the quality assessment of OGT preparations.

Length polymorphism in OGT between Korean native pig, Chinese Meishan, and the Western pig breeds

  • Nam, Yoon Seok;Kim, Doo-Wan;Kim, Myoung-Jik;Cho, Kyu-Ho;Kim, Jong Gug
    • Journal of Animal Science and Technology
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    • v.57 no.3
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    • pp.12.1-12.5
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    • 2015
  • Background: The Korean native pig (KNP) is generally thought to have come from northern China to the Korean peninsula approximately 2000 years ago. KNP pigs were at the brink of extinction in the 1980s, since then efforts have been made to restore the breed by bringing together the remaining stocks in South Korea. As a result, KNP was registered as a breed in 2006. To find additional breed-specific markers that are distinct among pig breeds, variations in O-linked N-acetylglucosamine transferase (OGT) were investigated. OGT is located on chromosome X and catalyzes the post-translational addition of a single O-linked-${\beta}$-N-acetylglucosamine to target proteins. Findings: Length polymorphism in the intron 20 of OGT was identified. The intron 20 of OGT from Duroc, Landrace, and Yorkshire breeds was 281-bp longer than that from either KNP or Chinese Meishan pigs. The difference between the Western pig breeds (BB genotype) and KNP or Meishan pigs (AA genotype) was due to an inserted 276-bp element and the 5-bp ACTTG. Conclusions: The polymorphism in OGT identified in this study may be used as an additional marker for determining the breed of origin among Meishan and the Western pig breeds. The length polymorphism suggests that the locus near OGT is not fixed in KNP. This marker would be relevant in determining the breed of origin in crossbred pigs between KNP pigs with known genotypes and the Western pig breeds with BB genotypes, thus confirming the contribution of the X chromosome from each breed.

Polymorphism in the intron 20 of porcine O-linked N-acetylglucosamine transferase

  • Kim, Jong Gug;Nonneman, Dan;Kim, Doo-Wan;Shin, Sangsu;Rohrer, Gary A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.8
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    • pp.1086-1092
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    • 2017
  • Objective: O-linked N-acetylglucosamine (O-GlcNAc) transferase (OGT) catalyzes the addition of O-GlcNAc and GlcNAcylation has extensive crosstalk with phosphorylation to regulate signaling and transcription. Pig OGT is located near the region of chromosome X that affects follicle stimulating hormone level and testes size. The objective of this study was to find the variations of OGT between European and Chinese pigs. Methods: Pigs were tested initially for polymorphism in OGT among European and Chinese pigs by polymerase chain reaction and sequencing at the U.S. Meat Animal Research Center (USMARC). The polymorphism was also determined in an independent population of pigs including European and Chinese Meishan (ME) breeds at the National Institute of Animal Science (NIAS, RDA, Korea). Results: The intron 20 of OGT from European and Chinese pigs was 514 and 233 bp, respectively, in the pigs tested initially. They included 1 White composite (WC) boar and 7 sows ($2Minzu{\times}WC$, $2Duroc\;[DU]{\times}WC$, $2ME{\times}WC$, $1Fengzing{\times}WC$) at USMARC. The 281-bp difference was due to an inserted 276-bp element and GACTT in European pigs. When additional WC and ME boars, the grandparents that were used to generate the $1/2ME{\times}1/2WC$ parents, and the 84 boars of 16 litters from mating of $1/2ME{\times}1/2WC$ parents were analyzed, the breeds of origin of X chromosome quantitative trait locus (QTL) were confirmed. The polymorphism was determined in an independent population of pigs including DU, Landrace, Yorkshire, and ME breeds at NIAS. OGT was placed at position 67 cM on the chromosome X of the USMARC swine linkage map. Conclusion: There was complete concordance with the insertion in European pigs at USMARC and NIAS. This polymorphism could be a useful marker to identify the breed of origin of X chromosome QTL in pigs produced by crossbreeding Chinese and European pigs.

Saponarin content and biosynthesis-related gene expression in young barley (Hordeum vulgare L.) seedlings

  • Lee, HanGyeol;Woo, So-Yeun;Ra, Ji-Eun;Lee, Kwang-Sik;Seo, Woo Duck;Lee, Jeong Hwan
    • Journal of Plant Biotechnology
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    • v.46 no.4
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    • pp.247-254
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    • 2019
  • Flavonoids are widely distributed secondary metabolites in plants that have a variety biological functions, as well as beneficial biological and pharmacological activities. In barley (Hordeum vulgare L.), for example, high levels of saponarin accumulate during primary leaf development. However, the effect of saponarin biosynthetic pathway genes on the accumulation of saponarin in barley is poorly understood. Accordingly, the aim of the present study was to examine the saponarin contents and expression levels of saponarin biosynthetic pathway genes [chalcone synthase (CHS), chalcone isomerase (CHI), and UDP-Glc:isovitexin 7-O-glucosyltransferase (OGT)] during early seedling developmental and under several abiotic stress conditions. Interestingly, the upregulation of HvCHS, HvCHI, and HvOGT during early development was associated with saponarin accumulation during later stages. In addition, exposure to abiotic stress conditions (e.g., light/dark transition, drought, and low or high temperature) significantly affected the expression of HvCHS and HvCHI but failed to affect either HvOGT expression or saponarin accumulation. These findings suggested that the expression of HvOGT, which encodes an enzyme that catalyzes the final step of saponarin biosynthesis, is required for saponarin accumulation. Taken together, the results of the present study provide a basis for metabolic engineering in barley plants, especially in regards to enhancing the contents of useful secondary metabolites, such as saponarin.

An Exploratory Study of the Effect of Ongyeong-tang Administration on Insomnia Disorder in Patients with Neurological Diseases: A Retrospective Chart Review Study

  • Seungwon Kwon;Han-Gyul Lee;Woo-Sang Jung;Sang-Kwan Moon;Ki-Ho Cho
    • The Journal of Internal Korean Medicine
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    • v.43 no.6
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    • pp.1122-1133
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    • 2022
  • Objectives: The aim of this study was to retrospectively evaluate the efficacy and safety of Ongyeong-tang (OGT) in the treatment of insomnia in patients afflicted with neurological diseases, according to the medical records of one medical institution. OGT is a herbal prescription that originates from the "Synopsis of Prescriptions of the Golden Chamber" and has been used for menopausal symptoms, uterine diseases, skin diseases, insomnia, and other neuropsychiatric symptoms. Methods: Medical records of patients who visited Kyung Hee University Korean Medicine Hospital from January 1, 2021 to June 30, 2022, received outpatient or inpatient treatment, and met the inclusion criteria were used in this study. Results: Twenty-four patients satisfied the inclusion criteria and were included. The Pittsburgh Sleep Quality Index-Korean version (PSQI-K) score was used to evaluate the outcomes before and after treatment. After an average of 14.42±9.2 days of OGT extract intake, 20 patients (83.3%) experienced improvement. The remaining four (16.7%) had no change in their symptoms, but none demonstrated worsening of symptoms. The total PSQI-K score showed a statistically significant decrease from 13.08±4.54 to 10.42±4.58 (p<0.001). Sleep quality, sleep duration, sleep disturbance, use of sleeping medications, and daytime dysfunction also showed statistically significant improvements. Conclusions: Administration of OGT extract may be a relatively safe and effective therapeutic option for insomnia in patients with neurological diseases.

The Effect of Speed-dependent with Body Weight Supported Treadmill Training on the Ambulation of Stroke (속도-의존적 체중지지 트레드밀 보행이 뇌졸중 환자의 보행에 미치는 영향)

  • Kim, Jwa-Jun;Rho, Min-He;Goo, Bong-Oh;Ahn, So-Youn
    • The Journal of Korean Physical Therapy
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    • v.17 no.3
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    • pp.339-350
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    • 2005
  • This study is the quasi-experimental study on the gait training rehabilitation. The purpose of this study is to prepare the baseline data for most suitable of gait while we were scrutinizing how the walking characters, functional walking ability, gait quality of stroke patients were affected by the gait on BWSTT (Body Weight Supported Treadmill Training) through the change of treadmill velocity and body weight support. To accomplish this purpose, this study used thirty subjects, more than 3 months post stroke, for rehabilitation who were divided between two gait training groups they received the neurophysiological physical therapy. For 6 weeks, 5 times a week for 15 minutes per session, the BWSTT group participated in 30 sessions structured speed-dependent treadmill training with 30% body weight supported, and the ratio of body weight support was gradually decreased as the patients advanced the capability of more self-support. The OGT(Over Ground Training) group received the same quantity of equal sessions like BWSTT. Firstly, we measured the absolute improvement of walking velocity (m/s), capacity(min/m) and cadence(steps/min) among walking characters. Secondly, we measured the functional walking ability such as Functional Ambulatory Category(FAC, score out of 5), Modified Motor Assesment Scale(MMAS, score out of 6) and Gait Quality Chart(score out of 41). Data analysis was performed with using SPSS 10.0 win program. The descriptive analysis was used to obtain average and standard deviation. The independent t-test and the paired t-test were used to compare both the groups about pre and post training test. Treatment effects were established by pre and post assessment. Subjects tolerated the training well without side-effects. Therefore, the results of this study were as follows; 1. There was a more significant difference from the improvement of walking velocity(0.09m/s), endurance(4.53min/m), cadence(4.20steps/min), FAC(0.26score), MMAS(0.33 score) and hip joint and pelvic of gait quality(0.39 score) ever before in the BWSTT group(p<.05). 2. There was a more significant increase from the walking velocity(0.01m/s) in the OGT group(p<.05). 3. There was a more statistical significant increase from comparing the average of walking velocity in both groups ever before(0.42m/s in BWSTT group and 0.31m/s in OGT group)(p<.05). There was a statistical significant difference from the average of cadence in both groups(61.87step/min in BWSTT group and 3.60steps/min in OGT group)(p<.05). As we can see from above, the findings suggest that BWSTT may be more effective than the OGT for improving some gait parameters such as gait velocity and cadency. This conclusion also suggest that BWSTT is more effective for the improvement of gait of stroke patients.

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O-GlcNAcylation of NF-κB Promotes Lung Metastasis of Cervical Cancer Cells via Upregulation of CXCR4 Expression

  • Ali, Akhtar;Kim, Sung Hwan;Kim, Min Jun;Choi, Mee Young;Kang, Sang Soo;Cho, Gyeong Jae;Kim, Yoon Sook;Choi, Jun-Young;Choi, Wan Sung
    • Molecules and Cells
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    • v.40 no.7
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    • pp.476-484
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    • 2017
  • C-X-C chemokine receptor 4 (CXCR4) stimulates cancer metastasis. NF-${\kappa}B$ regulates CXCR4 expression in cancer cells, and O-GlcNAc modification of NF-${\kappa}B$ promotes its transcriptional activity. Here, we determined whether CXCR4 expression is affected by O-GlcNAcylation of NF-${\kappa}B$ in lung metastasis of cervical cancer. We found elevated levels of O-linked-N-actylglucosamine transferase (OGT) and O-GlcNAcylation in cervical cancer cells compared to those in non-malignant epithelial cells and detected increased expression of NF-${\kappa}B$ p65 (p65) and CXCR4 in cervical cancer cells. Knockdown of OGT inhibited the O-GlcNAcylation of p65 and decreased CXCR4 expression levels in HeLa cells. Thiamet G treatment increased O-GlcNAcylated p65, which subsequently enhanced CXCR4 expression levels. Inhibition of O-GlcNAcylation by 6-Diazo-5-oxo-L-norleucine (DON) treatment decreased p65 activation, eventually inhibiting CXCR4 expression in HeLa cells. Lung tissues from mice engrafted with OGT-knockdown HeLa cells (shOGT) exhibited lower expression of Ki-67 and HPV E6 and E7 oncogenes compared to lung tissues from mice engrafted with control HeLa cells (shCTL). In addition, lung tissues from mice engrafted with shOGT cells exhibited lower p65 and CXCR4 immunoreactivity compared to tissues from mice engrafted with shCTL cells. Taken together, our data suggest that p65 O-GlcNAcylation promotes lung metastasis of cervical cancer cells by activating CXCR4 expression.

Production of O-GlcNAc Modified Recombinant Proteins in Escherichia coli

  • LIM, KI HONG;CHANG HOON HA;HYO IHL CHANG
    • Journal of Microbiology and Biotechnology
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    • v.12 no.2
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    • pp.306-311
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    • 2002
  • O-linked N-acetylglucosamine (O-GlcNAc) is an abundant posttranslationally modified compound in eukaryotic cells. Human O-GlcNAc transferase (OGT) was produced as a maltose binding protein (MBP) fusion protein, which showed significant catalytic activity to modify recombinant Sp1, transcription factor. To facilitate the production of O-GlcNAc modified proteins, instead of using the tedious in vitro glycosylation reaction or expression in eukaryotic cells, a MBP-fusion OGT expression vector (pACYC184-MBPOGT) was constructed using pACYC184 plasmid, which could coexist with general prokaryotic expression vectors containing ColE1 origin. By cotransforming pACYC184-MBPOGT and pGEX-2T vectors into Escherichia coli BL21, intracellular O- GlcNAcylated proteins could be obtained by a simple purification procedure. It is expected that this may be a useful tool for production of O-GlcNAc modified proteins.

Analysis of Human O-GlcNAcase Gene and the Expression of the Recombinant Gene. (사람의 O-linked N-acetyl-$\beta$-D-glucosaminidase 유전자의 분석과 재조합 발현)

  • 강대욱;서현효
    • Korean Journal of Microbiology
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    • v.40 no.2
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    • pp.87-93
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    • 2004
  • Dynamic modification of cytoplasmic and nuclear proteins by O-linked N-acetylglucosamine (O-GlcNAc) on Ser and Thr residues is ubiquitous in higher eukaryotes. And this modification may serve as a signaling mod-ification analogous to protein phosphorylation. Addition and cleavage of O-GlcNAc are catalyzed by O-linked GlcNAc transferase (OGT) and O-linked N-acety1glucosaminidase (O-GlcNAcase), respectively. Two types of human O-GlcNAcase gene were cloned and expressed as three fusion proteins in Escherichia coli. O-GlcNA-case activity showed in the order of thioredoxin fusion> $6{\times}His$ tag> GST fusion. O-GlcNAcase had enzy-matic activity against only ${\rho}$NP-GlcNAc of seven tested substrate analogs. Blast search revealed that O-GlcNAcase has two conserved domains, amino terminal hyaluronidase-like domain and carboxy terminal N-acetyltransferase domain. Extensive deletion studies were done to define catalytically important domains. The deletions of hyaluronidase-like domain and N-acetyltransferase domain abolished enzyme activity. But, N-ter-minal 55 amino acid deletion and C-terminal truncation showed lower activity. Based on deletion analysis, we suggest that hyaluronidase-like domain is essential for enzyme activity and carboxy terminal N-acetyltrans-ferase domain may be modulatory function.