• Title/Summary/Keyword: Protein-protein interactions

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Characterization of Natural Compounds as Inhibitors of NS1 Endonuclease from Canine Parvovirus Type 2

  • So-Hyung Kwak;Hayeong Kim;Hyeli Yun;Juho Lim;Dong-Hyun Kang;Doman Kim
    • Journal of Microbiology and Biotechnology
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    • v.33 no.6
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    • pp.788-796
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    • 2023
  • Canine parvovirus type 2 (CPV-2) has high morbidity and mortality rates in canines. Nonstructural protein 1 (NS1) of CPV-2 has endonuclease activity, initiates viral DNA replication, and is highly conserved. Thus, it is a promising target for antiviral inhibitor development. We overexpressed a 41.9 kDa active recombinant endonuclease in Escherichia coli and designed a nicking assay using carboxyfluorescein and quencher-linked ssDNA as substrates. The optimal temperature and pH of the endonuclease were 37℃ and pH 7, respectively. Curcumin, bisdemethoxycurcumin, demethoxycurcumin, linoleic acid, tannic acid, and α-tocopherol inhibited CPV-2 NS1 endonuclease with IC50 values of 0.29 to 8.03 µM. The extracted turmeric, yerba mate, and sesame cake suppressed CPV-2 NS1 endonuclease with IC50 values of 1.48, 7.09, and 52.67 ㎍/ml, respectively. The binding affinity between curcumin, the strongest inhibitor, and CPV-2 NS1 endonuclease by molecular docking was -6.4 kcal/mol. Curcumin inhibited CPV-2 NS1 endonuclease via numerous hydrophobic interactions and two hydrogen bonds with Lys97 and Pro111 in the allosteric site. These results suggest that adding curcuminoids, linoleic acid, tannic acid, α-tocopherol, extracted turmeric, sesame cake, and yerba to the diet could prevent CPV-2 infection.

Involvement of α1B-adrenoceptors and Rho kinase in contractions of rat aorta and mouse spleen

  • Hadeel A. Alsufyani;James R. Docherty
    • The Korean Journal of Physiology and Pharmacology
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    • v.27 no.4
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    • pp.325-331
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    • 2023
  • α1-adrenoceptors link via the G-protein Gq/G11 to both Ca2+ entry and release from stores, but may also activate Rho kinase, which causes calcium sensitization. This study aimed to identify the subtype(s) of α1-adrenoceptor involved in Rho kinase-mediated responses in both rat aorta and mouse spleen, tissues in which contractions involve multiple subtypes of α1-adrenoceptor. Tissues were contracted with cumulative concentrations of noradrenaline (NA) in 0.5 log unit increments, before and in the presence of an antagonist or vehicle. Contractions produced by NA in rat aorta are entirely α1-adrenoceptor mediated as they are competitively blocked by prazosin. The α1A-adrenoceptor antagonist RS100329 had low potency in rat aorta. The α1D-adrenoceptor antagonist BMY7378 antagonized contractions in rat aorta in a biphasic manner: low concentrations blocking α1D-adrenoceptors and high concentrations blocking α1B-adrenoceptors. The Rho kinase inhibitor fasudil (10 µM) significantly reduced aortic contractions in terms of maximum response, suggesting inhibition of α1B-adrenoceptor mediated responses. In the mouse spleen, a tissue in which all 3 subtypes of α1-adrenoceptor are involved in contractions to NA, fasudil (3 µM) significantly reduced both early and late components to the NA contraction, the early component involving α1B- and α1D-adrenoceptors, and the late component involving α1B- and α1A-adrenoceptors. This suggests that fasudil inhibits α1B-adrenoceptor mediated responses. It is concluded that α1D- and α1B-adrenoceptors interact in rat aorta and α1D-, α1A- and α1B-adrenoceptors interact in the mouse spleen to produce contractions and these interactions suggest that one of the receptors preferentially activates Rho kinase, most likely the α1B-adrenoceptor.

Approximate Top-k Labeled Subgraph Matching Scheme Based on Word Embedding (워드 임베딩 기반 근사 Top-k 레이블 서브그래프 매칭 기법)

  • Choi, Do-Jin;Oh, Young-Ho;Bok, Kyoung-Soo;Yoo, Jae-Soo
    • The Journal of the Korea Contents Association
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    • v.22 no.8
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    • pp.33-43
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    • 2022
  • Labeled graphs are used to represent entities, their relationships, and their structures in real data such as knowledge graphs and protein interactions. With the rapid development of IT and the explosive increase in data, there has been a need for a subgraph matching technology to provide information that the user is interested in. In this paper, we propose an approximate Top-k labeled subgraph matching scheme that considers the semantic similarity of labels and the difference in graph structure. The proposed scheme utilizes a learning model using FastText in order to consider the semantic similarity of a label. In addition, the label similarity graph(LSG) is used for approximate subgraph matching by calculating similarity values between labels in advance. Through the LSG, we can resolve the limitations of the existing schemes that subgraph expansion is possible only if the labels match exactly. It supports structural similarity for a query graph by performing searches up to 2-hop. Based on the similarity value, we provide k subgraph matching results. We conduct various performance evaluations in order to show the superiority of the proposed scheme.

Effects of Pre- and Post-workout Energy Bar Supplementation on Blood Lactate and Fitness in Young Adults with CrossFit Training: A randomized crossover study

  • Byung-Gul Lim;Xinxing Li;Wook Song
    • Journal of The Korean Society of Integrative Medicine
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    • v.11 no.4
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    • pp.61-71
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    • 2023
  • Purpose : Energy bars are increasingly popular among active individuals, yet their timing and nutrient combinations' impact on exercise adaptation remains unclear. This study aims to address this knowledge gap by investigating whether the combination of pre- and post-workout supplementation can synergistically enhance fitness and alleviate fatigue in trained CrossFit participants. Investigate if combining pre- and post-workout supplements can enhance fitness and blood lactate management in trained CrossFit participants, potentially improving exercise performance for this group. Methods : In a randomized crossover study, 20 trained CrossFit individuals (11 males, 9 females) completed thrice-weekly 60-minute CrossFit sessions for 3 weeks, with a one-week washout period. Participants were randomly assigned to either a chocolate bar group (CH, 45 g, 225 kcal) or an energy bar group (ES, 48 g, 238 kcal, with protein, caffeine, taurine, and BCAAs). For one week, participants consumed two bars of their assigned supplement five minutes before and after workouts. After a washout period, they switched supplements. Blood lactate levels and a visual analog scale (VAS) were assessed before, immediately after, and 30 minutes post-workout. Fitness tests (hand grip, broad jump, sit-ups) were conducted at baseline and 30 minutes post-final workout. Data were analyzed using two-way repeated measures ANOVA (p<.05), 95 % confidence intervals, and magnitude inferences. Results : Hand grip strength (t=-5.60, p=.000), broad jump (t=-3.43, p=.003) and sit up (t=-3.94, p=.001) were significantly increased in the ES group. Compared to CH group, there was a significant time and group interactions for blood lactate level (F=5.51, p=.008) and VAS(F=31.67, p=.000) in the ES group. Conclusion : Pre- and post-workout energy bar supplementation may have a beneficial effect on blood lactate clearance and fitness in trained CrossFit individual. The combination of proprietary supplements taken may provide benefits for removing the blood lactate during high-intensity functional exercise.

A comprehensive longitudinal study of gut microbiota dynamic changes in laying hens at four growth stages prior to egg production

  • Seojin Choi;Eun Bae Kim
    • Animal Bioscience
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    • v.36 no.11
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    • pp.1727-1737
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    • 2023
  • Objective: The poultry industry is a primary source of animal protein worldwide. The gut microbiota of poultry birds, such as chickens and ducks, is critical in maintaining their health, growth, and productivity. This study aimed to identify longitudinal changes in the gut microbiota of laying hens from birth to the pre-laying stage. Methods: From a total of 80 Hy-Line Brown laying hens, birds were selected based on weight at equal intervals to collect feces (n = 20 per growth) and ileal contents (n = 10 per growth) for each growth stage (days 10, 21, 58, and 101). The V4 regions of the 16S rRNA gene were amplified after extracting DNA from feces and ileal contents. Amplicon sequencing was performed using Illumina, followed by analysis. Results: Microbial diversity increased with growth stages, regardless of sampling sites. Microbial community analysis indicated that Firmicutes, Proteobacteria, and Bacteroidetes were the dominant phyla in the feces and ileal. The abundance of Lactobacillus was highest on day 10, and that of Escherichia-shigella was higher on day 21 than those at the other stages at the genus level (for the feces and ileal contents; p<0.05). Furthermore, Turicibacter was the most abundant genus after changing feed (for the feces and ileal contents; p<0.05). The fecal Ruminococcus torques and ileal Lysinibacillus were negatively correlated with the body weights of chickens (p<0.05). Conclusion: The gut microbiota of laying hens changes during the four growth stages, and interactions between microbiota and feed may be present. Our findings provide valuable data for understanding the gut microbiota of laying hens at various growth stages and future applied studies.

Identification of Selective STAT1 Inhibitors by Computational Approach

  • Veena Jaganivasan;Dona Samuel Karen;Bavya Chandrasekhar
    • Journal of Integrative Natural Science
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    • v.16 no.3
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    • pp.81-95
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    • 2023
  • Colorectal cancer is one of the most common types of cancer worldwide, ranking third after lung and breast cancer in terms of global prevalence. With an expected 1.93 million new cases and 935,000 deaths in 2020, it is more prevalent in males than in women. Evidence has shown that during the later stages of colon cancer, STAT1 promotes tumor progression by promoting cell survival and resistance to chemotherapy. Recent studies have shown that inhibiting STAT1 pathway leads to a reduction in tumor cell proliferation and growth, and can also promote apoptosis in colon cancer cells. One of the recent approaches in the field of drug discovery is drug repurposing. In drug repurposing approach we have virtually screened FDA database against STAT1 protein and their interactions have been studied through Molecular docking. Cross docking was performed with the top 10 compounds to be more specific with STAT1 comparing the affinity with STAT2, STAT3, STAT4, STAT5a, STAT5b and STAT6. The drugs that showed higher affinity were subjected to Conceptual - Density functional theory. Besides, the Molecular dynamic simulation was also carried out for the selected leads. We also validated in-vitro against colon cancer cell lines. The results showed mainly Acetyldigitoxin has shown better binding to the target. From this study, we can predict that the drug Acetyldigitoxin has shown noticeable inhibitory efficiency against STAT1, which in turn can also lead to the reduction of tumor cell growth in colon cancer.

Network pharmacology analysis of Jakyakgamchotang with corydalis tuber for anti-inflammation (작약감초탕 가 현호색의 항염증 기전에 대한 네트워크 약리학적 분석)

  • Young-Sik Kim;Hongjun Kim;Han-bin Park;Seungho Lee
    • Herbal Formula Science
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    • v.32 no.1
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    • pp.39-49
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    • 2024
  • Objectives : The purpose of this study was to investigate the molecular targets and pathways of anti-inflammatory effects of Jakyakgamchotang with corydalis tuber (JC) using network pharmacology. Methods : The compounds in constituent herbal medicines of JC were searched in TCM systems pharmacology (TCMSP). Target gene informations of the components were collected using chemical-target interactions database provided by Pubchem. Afterwards, network analysis between compounds and inflammation-related target genes was performed using cytoscape. Go enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were performed on inflammation-related targets using DAVID database. Results : 70 active compounds related to inflammation were identified, and 295 target genes related to the anti-inflammatory activity of the compound of JC were identified. In the Go biological process DB and KEGG pathway DB, "inflammatory response", "cellular response to lipopolysaccharide", "positive regulation of interleukin-6 production", and "positive regulation of protein kinase B. signaling", "positive regulation of ERK1 and ERK2 cascade", "positive regulation of I-kappaB kinase/NF-kappaB signaling", "negative regulation of apoptotic process", and "PI3K-Akt signaling pathway" were found to be mechanisms related to the anti-inflammatory effects related to the target genes of JC. The main compounds predicted to be involved in the anti-inflammatory effect of JC were quercetin, licochalcone B, (+)-catechin, kaempferol, and emodin. Conclusions : This study provides the molecular targets and potential pathways of JC on inflammation. It can be used as a basic data for using JC for various inflammatory disease in traditional korean medicine clinic.

Generation of ints14 Knockout Zebrafish using CRISPR/Cas9 for the Study of Development and Disease Mechanisms

  • Ji Hye Jung;Sanghoon Jeon;Heabin Kim;Seung-Hyun Jung
    • Development and Reproduction
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    • v.27 no.4
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    • pp.205-211
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    • 2023
  • INTS14/VWA9, a component of the integrator complex subunits, plays a pivotal role in regulating the fate of numerous nascent RNAs transcribed by RNA polymerase II, particularly in the biogenesis of small nuclear RNAs and enhancer RNAs. Despite its significance, a comprehensive mutation model for developmental research has been lacking. To address this gap, we aimed to investigate the expression patterns of INTS14 during zebrafish embryonic development. We generated ints14 mutant strains using the CRISPR/Cas9 system. We validated the gRNA activity by co-injecting Cas9 protein and a single guide RNA into fertilized zebrafish eggs, subsequently confirming the presence of a 6- or 9-bp deletion in the ints14 gene. In addition, we examined the two mutant alleles through PCR analysis, T7E1 assay, TA-cloning, and sequencing. For the first time, we used the CRISPR/Cas9 system to create a model in which some sequences of the ints14 gene were removed. This breakthrough opens new avenues for in-depth exploration of the role of ints14 in animal diseases. The mutant strains generated in this study can provide a valuable resource for further investigations into the specific consequences of ints14 gene deletion during zebrafish development. This research establishes a foundation for future studies exploring the molecular mechanisms underlying the functions of ints14, its interactions with other genes or proteins, and its broader implications for biological processes.

Effect of Pig Feces and Pig Waste Mixture Compositions on Bio-oil Production by Pyrolysis Process (돈분과 돈슬러리의 성분이 열분해공정에 의한 바이오오일 생산효율에 미치는 영향)

  • Zhu, Kun;Choi, Hong Lim;Shin, Jongdu;Paek, E
    • Journal of the Korea Organic Resources Recycling Association
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    • v.17 no.4
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    • pp.29-35
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    • 2009
  • Pyrolysis is recently used as one of alternative methods of animal waste treatment. In this study bio-oil was produced at $550^{\circ}C$ in an auger reactor through pyrolysis process. Two pig waste mixtures were used, pig feces mixed with rice husks and pig feces mixed with sawdust. The main compositions of hemicellulose, lignin, cellulose, protein, and fat were analyzed chemically. Based on the main composition results obtained, the contents of holocellulose (the sum of hemicellulose and cellulose) and lignin had a significant positive effect on bio-oil production, and there was a significant negative effect of ash content on bio-oil yield. The interactions between the different feedstocks were evaluated, and it was concluded that the interaction between pig feces and rice husks was minimal, whereas the interaction between pig feces and sawdust was significant.

Dietary Tea Catechin Inclusion Changes Plasma Biochemical Parameters, Hormone Concentrations and Glutathione Redox Status in Goats

  • Zhong, Rongzhen;Xiao, Wenjun;Ren, Guopu;Zhou, Daowei;Tan, Chuanyan;Tan, Zhiliang;Han, Xuefeng;Tang, Shaoxun;Zhou, Chuanshe;Wang, Min
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.12
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    • pp.1681-1689
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    • 2011
  • The beneficial effects of tea catechins (TCs) are related not only to their antioxidant potential but also to the improvement of animal meat quality. In this study, we assessed the effects of dietary TC supplementation on plasma biochemical parameters, hormone responses, and glutathione redox status in goats. Forty Liuyang goats were randomly divided into four equal groups (10 animals/group) that were assigned to four experimental diets with TC supplementation at 4 levels (0, 2,000, 3,000 or 4,000 mg TC/kg DM feed). After a 60-day feeding trial, all goats were slaughtered and sampled. Dietary TC treatment had no significant effect on blood biochemical parameters, however, low-density lipoprotein cholesterol (p<0.001), triglyceride (p<0.01), plasma urea nitrogen (p<0.01), and glucose (p<0.001) decreased and total protein (p<0.01) and albumin (p<0.05) increased with the feeding time extension, and day 20 was the turning point for most of changes. Interactions were found in glutathione (p<0.001) and the ratio of reduced and oxidized glutathione (p<0.05) in whole blood between treatment and feeding time. Oxidized glutathione in blood was reduced (p<0.05) by 2,000 mg TC/kg feed supplementation, and a similar result was observed in longissimus dorsi muscle. Though plasma glutathione peroxidase (p<0.01) and glutathione reductase (p<0.05) activities were affected by treatment and feeding time interactions, and glutathione S-transferases activity increased with feeding day extension, no changed values appeared in longissimus dorsi muscle. In conclusion, dietary TC supplementation affected the concentrations of some blood metabolites and accelerated GSH depletion in the blood of goats. In terms of less high-density lipoprotein cholesterol, the highest insulin and IGF-I concentrations, the highest ratio of reduced and oxidized glutathione in plasma, the dosage of 2,000 mg TC/kg feed might be desirable for growing goats to prevent glutathione depletion and keep normal physiological metabolism.