• Title/Summary/Keyword: Small protein

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Evaluation of circulating IGF-I and IGFBP-3 as biomarkers for tumors in dogs

  • Song, Doo-Won;Ro, Woong-Bin;Sur, Jung-Hyang;Seung, Byung-Joon;Kang, Hyun-Min;Kim, Jong-Won;Park, See-Hyoung;Park, Hee-Myung
    • Journal of Veterinary Science
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    • v.22 no.6
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    • pp.77.1-77.10
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    • 2021
  • Background: Serum-based parameters are considered non-invasive biomarkers for cancer detection. In human studies, insulin-like growth factor-I and II (IGF-I and IGF-II) and insulin-like growth factor binding protein-3 (IGFBP-3) are useful as diagnostic or prognostic markers and potential therapeutic targets. Objectives: This study examined the diagnostic utility of circulating IGF-I, IGF-II, and IGFBP-3 levels in healthy dogs and dogs with tumors. Methods: The serum concentrations of these biomarkers in 86 dogs with tumors were compared with those in 30 healthy dogs using an enzyme-linked immunosorbent assay (ELISA). Results: The ELISA results showed no difference between healthy dogs and dogs with tumors in the serum IGF-II concentrations. On the other hand, there was a significant difference in the circulating IGF-I and IGFBP-3 levels between healthy dogs and dogs with tumors. The concentrations of serum IGF-I (median [interquartile range], 103.4 [59.5-175] ng/mL) in dogs with epithelial tumors were higher than those (58.4 ng/mL [43.5-79.9]) in healthy dogs. Thus, the concentrations of serum IGFBP-3 (43.4 ng/mL [33.2-57.2]) in dogs with malignant mesenchymal tumors were lower than those (60.8 ng/mL [47.6-70.5]) in healthy dogs. Conclusions: The serum IGF-I and IGFBP-3 levels can be used as diagnostic biomarkers in dogs with tumors.

Roles of Carbohydrate-Binding Module (CBM) of an Endo-β-1,4-Glucanase (Cel5L) from Bacillus sp. KD1014 in Thermostability and Small-Substrate Hydrolyzing Activity

  • Lee, Jae Pil;Shin, Eun-Sun;Cho, Min Yeol;Lee, Kyung-Dong;Kim, Hoon
    • Journal of Microbiology and Biotechnology
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    • v.28 no.12
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    • pp.2036-2045
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    • 2018
  • An endo-${\beta}$-1,4-glucanase gene, cel5L, was cloned using the shot-gun method from Bacillus sp.. The gene, which contained a predicted signal peptide, encoded a protein of 496 amino acid residues, and the molecular mass of the mature Cel5L was estimated to be 51.8 kDa. Cel5L contained a catalytic domain of glycoside hydrolase (GH) family 5 and a carbohydrate-binding module family 3 (CBM_3). Chromatography using HiTrap Q and CHT-II resulted in the isolation of two truncated forms corresponding to 50 (Cel5L-p50) and 35 kDa (Cel5L-p35, CBM_3-deleted form). Both enzymes were optimally active at pH 4.5 and $55^{\circ}C$, but had different half-lives of 4.0 and 22.8 min, respectively, at $70^{\circ}C$. The relative activities of Cel5L-p50 and Cel5L-p35 for barley ${\beta}$-glucan were 377.0 and 246.7%, respectively, compared to those for carboxymethyl-cellulose. The affinity and hydrolysis rate of pNPC by Cel5L-p35 were 1.7 and 3.3 times higher, respectively, than those by Cel5L-p50. Additions of each to a commercial enzyme set increased saccharification of pretreated rice straw powder by 17.5 and 21.0%, respectively. These results suggest CBM_3 is significantly contributing to thermostability, and to affinity and substrate specificity for small substrates, and that these two enzymes could be used as additives to enhance enzymatic saccharification.

Small Molecule-Based Strategy Promotes Nucleus Pulposus Specific Differentiation of Adipose-Derived Mesenchymal Stem Cells

  • Hua, Jianming;Shen, Ning;Wang, Jingkai;Tao, Yiqing;Li, Fangcai;Chen, Qixin;Zhou, Xiaopeng
    • Molecules and Cells
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    • v.42 no.9
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    • pp.661-671
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    • 2019
  • Adipose tissue-derived mesenchymal stem cells (ADSCs) are promising for regenerating degenerated intervertebral discs (IVDs), but the low efficiency of nucleus pulposus (NP)-specific differentiation limits their clinical applications. The Sonic hedgehog (Shh) signaling pathway is important in NP-specific differentiation of ADSCs, and Smoothened Agonist (SAG) is a highly specific and effective agonist of Shh signaling. In this study, we proposed a new differentiation strategy with the use of the small molecule SAG. The NP-specific differentiation and extracellular matrix (ECM) synthesis of ADSCs were measured in vitro, and the regenerative effects of SAG pretreated ADSCs in degenerated IVDs were verified in vivo. The results showed that the combination of SAG and transforming growth factor-${\beta}3$ ($TGF-{\beta}3$) is able to increase the ECM synthesis of ADSCs. In addition, the gene and protein expression levels of NP-specific markers were increased by treatment with SAG and $TGF-{\beta}3$. Furthermore, SAG pretreated ADSCs can also improve the disc height, water content, ECM content, and structure of degenerated IVDs in vivo. Our new differentiation scheme has high efficiency in inducing NP-specific differentiation of ADSCs and is promising for stem cell-based treatment of degenerated IVDs.

Effects of diet and roughage quality, and period of the day on diurnal feeding behaviour patterns of sheep and goats under subtropical conditions

  • Moyo, Mehluli;Adebayo, Rasheed Adekunle;Nsahlai, Ignatius Verla
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.5
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    • pp.675-690
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    • 2019
  • Objective: This study investigated the effect of diet and roughage quality (RQ) on dry matter intake, duration and number of daytime and night-time eating bouts, idling sessions and ruminating activities in small ruminants. Methods: In Exp 1 and 2, RQ was improved by urea treatment of veld hay, while diet quality was improved by supplementing with Lucerne hay (Exp 3), sunflower meal and lespedeza (Exp 4), fish meal (Exp 5a), and sunflower meal (Exp 5b). In all experiments goats and sheep were blocked by weight and randomly allocated to experimental diets. Day-time (06:00 to 18:00 h) and night time (18:00 to 06:00 h) feeding behaviour activities were recorded. Results: RQ affected rumination index in Exp 1, but not in Exp 2, 3, and 5. Time spent eating and ruminating was affected by RQ (Exp 1, 3, and 4), period of day (all experiments) and their interaction (Exp 1). Intake rates (g/bout and g/min) were similar across diets. Period of day affected the duration of rumination sessions (Exp 1, 2, and 3); diet or RQ affected the duration of eating bouts (Exp 3) and rumination sessions (Exp 1 and 2). RQ had a significant effect on the duration of eating sessions in Exp 3 only, whilst period of day affected this same behaviour in Exp 2 and 3. Generally, goats and sheep fed on roughage alone ruminate at night and eat more during the day but those fed a roughage and supplemented with Lucerne hay spent more time ruminating than eating. Time spent eating and ruminating had positive correlations to crude protein and feed intake. Intake rates had strong positive correlations to intake. Conclusion: Chewing time, number of eating and ruminating sessions, and duration of eating bouts are physiologically controlled in small ruminants, though chewing time requires isometric scaling during modelling of intake.

Processing and Quality Characteristics of a Cultured Recessive Small-sized Abalone Haliotis discus hannai Extract (양식 참전복(Haliotis discus hannai) 열성패 추출물의 제조 및 품질특성)

  • Cho, Jun-Hyun;Nam, Hyeon-Gyu;Oh, Kwang-Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.51 no.6
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    • pp.640-646
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    • 2018
  • To determine whether there are differences in the food component characteristics of the cultured recessive small-sized abalone Haliotis discus hannai (SA; 30-40 each/kg) and middle-sized abalone (MA; 10-15 each/kg), the proximate compositions, fatty acid and total amino acid compositions of these two species were evaluated. Additionally, extraction methods were performed on the SA to asses the quality characteristics of the resulting. In terms of proximate composition, MA had lower moisture and carbohydrate levels and higher crude protein contents than SA. The total amino acid contents of MA and SA were 15,734.4 and 11,379.1 mg/100 g, respectively, the major amino acids were glutamic acid, aspartic acid, serine, glycine, alanine, leucine, arginine and lysine, and the major fatty acids were 16:0, 18:0, 18:1n-9, 18:1n-7, 20:4n-6, 20:5n-3, and 22:5n-3. The pH levels and total nitrogen and amino nitrogen contents of the hot-water extract (WE) and scrap enzyme hydrolysate (SE) samples from the SA were 6.32 and 6.05, 1.36% and 1.52%, and 342.1 and 403.1 mg/100 g, respectively. The extraction yields and free amino acid contents from SA were 1,317 and 440 mL/kg, and 8,721.1 and 9,070.7 mg/100 g, respectively, and the concentrations of major components were as follows: arginine, glycine, glutamic acid, alanine and lysine. Additionally, the complex extract (WE+SE) was superior to the traditional extract (WE) in terms of extraction yield, amino-nitrogen content, and organoleptic qualities but not odor.

The Daily Dose and Uses of Pinellia Tuber, Vinegar and Eggshell of Goju-tang in Treatise on Cold Damage Diseases (《상한론(傷寒論)》고주탕(苦酒湯)의 반하(半夏)와 고주(苦酒), 계자각(鷄子殼) 용량과 용도)

  • Ku, Tae-Hun;Kim, In-Rak
    • The Korea Journal of Herbology
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    • v.36 no.3
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    • pp.9-14
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    • 2021
  • Objectives : The purpose of this study was to clarify the daily dose and uses of Goju-tang in Treatise on Cold Damage Diseases. Methods : We compared major editions of Treatise on Cold Damage Diseases with the original text of Supplement to the Essential Prescriptions Worth a Thousand Gold and chose the most appropriate text. If none of the existing texts seemed reasonable, We compared texts item by item. When there is nothing adequate even item by item, it was newly modified. These results were compared with currently distributed Pinellia Tuber and standards of eggs. Results : Goju-tang consisted of Goju(vinegar), Pinellia Tuber and an eggshell. Generally, a daily dose was extracted at once and taken 3 times a day, but in Goju-tang, 4 pieces of Pinellia Tuber and one-third Sheong of Goju(solvent) were used, which were one third of average volume. Pinellia Tuber was used cut in small pieces. And it was boiled gradually by low heat. Goju was mainly to reduce the pungency of Pinellia Tuber by cutting raphides of calcium oxalate. The Pot preparing medicines was eggshell which was chosen by its small size, because the volume of Pinellia Tuber and Goju was one-third of average volume. Since an egg shell is consisted of protein, it didn't react with vinegar(Goju). Conclusions : The 4 pieces of Pinellia Tuber and one-third Sheong of Goju(Vinegar) in the Goju-tang were one-third of average dose, and the eggshell was Pot preparing medicines.

Selective cytotoxicity of a novel mitochondrial complex I inhibitor, YK-135, against EMT-subtype gastric cancer cell lines due to impaired glycolytic capacity

  • Yeojin, Sung;Seungbin, Cha;Sang Bum, Kim;Hakhyun, Kim;Seonghwi, Choi;Sejin, Oh;Minseo, Kim;Yunji, Lee;Gino, Kwon;Jooyoung, Lee;Joo-Youn, Lee;Gyoonhee, Han;Hyun Seok, Kim
    • BMB Reports
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    • v.55 no.12
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    • pp.645-650
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    • 2022
  • Epithelial-to-mesenchymal transition (EMT)-subtype gastric cancers have the worst prognosis due to their higher recurrence rate, higher probability of developing metastases and higher chemo-resistance compared to those of other molecular subtypes. Pharmacologically actionable somatic mutations are rarely found in EMT-subtype gastric cancers, limiting the utility of targeted therapies. Here, we conducted a high-throughput chemical screen using 37 gastric cancer cell lines and 48,467 synthetic small-molecule compounds. We identified YK-135, a small-molecule compound that showed higher cytotoxicity toward EMT-subtype gastric cancer cell lines than toward non-EMT-subtype gastric cancer cell lines. YK-135 exerts its cytotoxic effects by inhibiting mitochondrial complex I activity and inducing AMP-activated protein kinase (AMPK)-mediated apoptosis. We found that the lower glycolytic capacity of the EMT-subtype gastric cancer cells confers synthetic lethality to the inhibition of mitochondrial complex I, possibly by failing to maintain energy homeostasis. Other well-known mitochondrial complex I inhibitors (e.g., rotenone and phenformin) mimic the efficacy of YK-135, supporting our results. These findings highlight mitochondrial complex I inhibitors as promising therapeutic agents for EMT-subtype gastric cancers and YK-135 as a novel chemical scaffold for further drug development.

Incidence of bovine tuberculosis and Brucellosis in Hanwoo (Korean native cattle) and dairy cow in Gyeonsangnam-do province from 2020 to 2022 (2020~2022년간 경상남도의 한우와 젖소의 소 결핵병 및 브루 셀라병의 발생률)

  • Cheol-Ho Kim;Yoo-Jeong Choi;Shin-Ho Lee;Dong-Hyun Han;Yoon-Ho Roh;Dong-Bin Lee;Jae-Hyeon Cho;Chung-Hui Kim
    • Korean Journal of Veterinary Service
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    • v.46 no.1
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    • pp.67-74
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    • 2023
  • In the present study, the incidence of bovine tubeculosis (bTB) and brucellosis in Hanwoo (Korean native cattle) and dairy cow in Gyeongsangnam-do was investigated for three years from 2020 to 2022. The incidence bTB tested by tuberculin skin test with purified protein derivative (PPD) and gamma interferon (γ-INF) test with a commercial enzyme-linked immunosorbent assay. From 2020 to 2022, the incidence of bTB showed a decreasing trend in Hanwoo, while an increasing trend in dairy cow. In the case of Brucellosis, the positive rates for Hanwooe gradually increased. However, no brucellosis was found in dairy cow from 2020 to 2022. The increase in the incidence of these diseases is presumed to be related the small scale and poor sanitation facilities of livestock farms in Gyeongsangnam-do, and easy access of wild animals. Therefore, in order to suppress the incidence of the diseases, it is necessary to the farm scale from small to large and to strengthen sanitary facilities on farms.

Association of FOSB exon 4 unmethylation with poor prognosis in patients with late-stage non-small cell lung cancer

  • Dong Sun Kim;Won Kee Lee;Jae Yong Park
    • Oncology Reports
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    • v.43 no.2
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    • pp.655-661
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    • 2020
  • Alterations in DNA methylation have a central role in the development and outcome of most human malignancies. Non-small cell lung cancer (NSCLC), the most common lung cancer, leads to the largest number of cancer-related deaths worldwide. FBJ murine osteosarcoma viral oncogene homolog B (FOSB) is a key component of the activator protein-1 transcription factor and regulates gene networks associated within oncogenic transformation. The role of FOSB in the development of NSCLC is still elusive. Therefore, the methylation status of the FOSB gene was investigated in NSCLC and its clinical significance in NSCLC progression was evaluated. The methylation status of the promoter and exon 4 regions of the FOSB gene were analyzed in 176 NSCLC specimens by bisulfite pyrosequencing and the association between FOSB methylation status and patient survival was investigated. Compared to adjacent non-malignant tissues, FOSB promoter exhibited exclusive unmethylation in all malignant tissues and the exon 4 region was found unmethylated in 18 (10.2% of the total) tumor samples. Exon 4 unmethylation was associated with downregulation of its mRNA and tended to occur in patients with lymph node metastasis. Univariate and multivariate analyses revealed that exon 4 unmethylation was significantly associated with unfavorable overall survival in patients with stage II-IIIA NSCLC (log-rank P=0.05, adjusted hazard ratio=2.43, 95% confidence interval=1.04-5.68, P=0.04). FOSB was identified as a novel gene with tumor-specific gene body unmethylation in NSCLC and a novel predictive biomarker for NSCLC prognosis. Moreover, the present results indicated that FOSB may have a tumor suppressor function in the progression of NSCLC.

Association of Genetic Variations with Pemetrexed-Induced Cytotoxicity in Non-Small Cell Lung Cancer Cells (비소세포폐암 세포주에서 pemetrexed의 세포독성과 유전학적 다형성과의 상관성 조사)

  • Yoon, Seong-Ae;Choi, Jung-Ran;Kim, Jeong-Oh;Shin, Jung-Young;Zhang, XiangHua;Kang, Jin-Hyoung
    • Journal of Life Science
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    • v.20 no.1
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    • pp.103-112
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    • 2010
  • Pemetrexed has demonstrated clinical activity in non-small cell lung cancer (NSCLC) as well as other solid tumors. It transports into the cells via reduced folate carrier (RFC) and is polyglutamated by folypolyglutamate synthetase (FPGS). Pemetrexed directly inhibits several folate-dependent enzymes such as thymidylate synthase (TS), dihydrofolate reductase (DHFR), and glycinamide ribonucleotide formyltransferase (GARFT). We investigated the effects of genetic variations and the expression of RFC, FPGS, TS and DHFR enzymes on drug sensitivity to pemetrexed in NSCLC cells. Polymorphisms in RFC, FPGS, and DHFR were genotyped in four NSCLC cells - A549, PC14, HCC-1588, and H226. Real-time RT-PCR and Western blot was performed to evaluate mRNA transcripts and protein of these genes. The cytotoxicity of pemetrexed was measured by SRB assay. In PC14 and H226 cells, increased mRNA expressions of RFC and FPGS were associated with higher cytotoxicity to pemetrexed. 2R/2R genotype of TS and its increased mRNA expression were associated with drug resistance to pemetrexed in A549 cells, whereas 3R/3R genotype in TS with decreased mRNA expression was associated with higher sensitivity in H226 cells. After pemetrexed treatment, an inverse change of DHFR mRNA and protein expression was found. The strongest linkage disequilibrium (LD) was discovered between-1726C>T and -1188A>C SNP of DHFR gene. Our findings suggest the cytotoxic effect of pemetrexed may be associated with genetic polymorphisms and the expression level of genes involved in pemetrexed metabolisms in NSCLC cells.