• BMB Reports
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• 제54권2호
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• pp.98-105
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• 2021

The clustered regularly interspaced short palindromic repeats (CRISPR) system is a family of DNA sequences originally discovered as a type of acquired immunity in prokaryotes such as bacteria and archaea. In many CRISPR systems, the functional ribonucleoproteins (RNPs) are composed of CRISPR protein and guide RNAs. They selectively bind and cleave specific target DNAs or RNAs, based on sequences complementary to the guide RNA. The specific targeted cleavage of the nucleic acids by CRISPR has been broadly utilized in genome editing methods. In the process of genome editing of eukaryotic cells, CRISPR-mediated DNA double-strand breaks (DSB) at specific genomic loci activate the endogenous DNA repair systems and induce mutations at the target sites with high efficiencies. Two of the major endogenous DNA repair machineries are non-homologous end joining (NHEJ) and homology-directed repair (HDR). In case of DSB, the two repair pathways operate in competition, resulting in several possible outcomes including deletions, insertions, and substitutions. Due to the inherent stochasticity of DSB-based genome editing methods, it was difficult to achieve defined single-base changes without unanticipated random mutation patterns. In order to overcome the heterogeneity in DSB-mediated genome editing, novel methods have been developed to incorporate precise single-base level changes without inducing DSB. The approaches utilized catalytically compromised CRISPR in conjunction with base-modifying enzymes and DNA polymerases, to accomplish highly efficient and precise genome editing of single and multiple bases. In this review, we introduce some of the advances in single-base level CRISPR genome editing methods and their applications.

• 식품과학과 산업
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• 제53권4호
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• pp.410-421
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• 2020

농·식품 분야의 중장기 연구개발 계획 수립과 관련해서, 해외 주요국가의 R&D 정책 및 투자 동향을 파악하여 기초자료로 제공하고자 한다. 해외 주요국의 동향을 살펴본 결과, 선진국을 중심으로 농·식품 분야 R&D에 대한 지속적인 정부 지원과 함께 효율적인 자원관리의 중요성을 강조하고 있다. 미국이나 유럽의 경우 농산업 생산성에 대한 공공투자를 줄이고, 기후변화·식량안보·식품안전·국민건강·자연환경 등의 사회문제 부각에 따라 지속가능성 측면에서 경제와 환경 간의 조화, 인체영양과 건강뿐만 아니라 동식물 건강, 통합적 생태계 접근 등을 강조하고 있다. 특히 식품 분야는 식품안전, 영양과 건강을 중심으로 공공부문이, 식품가공, 제조는 민간부문이 주로 투자하는 편이다. 한편 일본이나 중국의 경우에는 민간투자보다 공공부문에서의 투자가 크게 앞서고 있으며, 정부가 중장기 연구방향을 제시하는 등 연구개발혁신을 주도하는 입장이다. 비록 국가마다 시급한 현안사항과 대응방안이 다르지만, 농·식품 분야 연구개발혁신 및 발전전략의 효율성을 제고하기 위해 공공과 민간의 상호보완적 역할 분담을 바탕으로 긴밀한 협력활동이 이루어지고 있다.

• Biomolecules & Therapeutics
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• 제30권1호
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• pp.48-54
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• 2022

GPR43 (also known as FFAR2), a metabolite-sensing G-protein-coupled receptor stimulated by short-chain fatty acid (SCFA) ligands is involved in innate immunity and metabolism. GPR43 couples with Gα_i/o and Gα_q/11 heterotrimeric proteins and is capable of decreasing cyclic AMP and inducing Ca²⁺ flux. The GPR43 receptor has additionally been shown to bind β-arrestin 2 and inhibit inflammatory pathways, such as NF-κB. However, GPR43 shares the same ligands as GPR41, including acetate, propionate, and butyrate, and determination of its precise functions in association with endogenous ligands, such as SCFAs alone, therefore remains a considerable challenge. In this study, we generated novel synthetic agonists that display allosteric modulatory effects on GPR43 and downregulate NF-κB activity. In particular, the potency of compound 187 was significantly superior to that of pre-existing compounds in vitro. However, in the colitis model in vivo, compound 110 induced more potent attenuation of inflammation. These novel allosteric agonists of GPR43 clearly display anti-inflammatory potential, supporting their clinical utility as therapeutic drugs.

• Journal of Ginseng Research
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• 제46권1호
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• pp.39-53
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• 2022

Ginsenoside Rb₁ (Rb₁), one of the most important ingredients in Panax ginseng Meyer, has been confirmed to have favorable activities, including reducing antioxidative stress, inhibiting inflammation, regulating cell autophagy and apoptosis, affecting sugar and lipid metabolism, and regulating various cytokines. This study reviewed the recent progress on the pharmacological effects and mechanisms of Rb₁ against cardiovascular and nervous system diseases, diabetes, and their complications, especially those related to neurodegenerative diseases, myocardial ischemia, hypoxia injury, and traumatic brain injury. This review retrieved articles from PubMed and Web of Science that were published from 2015 to 2020. The molecular targets or pathways of the effects of Rb₁ on these diseases are referring to HMGB1, GLUT4, 11β-HSD1, ERK, Akt, Notch, NF-κB, MAPK, PPAR-γ, TGF-β1/Smad pathway, PI3K/mTOR pathway, Nrf2/HO-1 pathway, Nrf2/ARE pathway, and MAPK/NF-κB pathway. The potential effects of Rb₁ and its possible mechanisms against diseases were further predicted via Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway and disease ontology semantic and enrichment (DOSE) analyses with the reported targets. This study provides insights into the therapeutic effects of Rb₁ and its mechanisms against diseases, which is expected to help in promoting the drug development of Rb₁ and its clinical applications.

• 한국해양바이오학회지
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• 제14권1호
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• pp.9-24
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• 2022

Low-molecular weight peptides derived from fish collagen exhibit several bioactivities, including antioxidant, antiwrinkle, antimicrobial, antidiabetic, and antihypertension effects. These peptides are also involved in triglyceride suppression and memory improvement. This study aimed to investigate the optimal processing condition for preparing low-molecular weight peptides from flounder skin, and the properties of the hydrolysate. The optimal processing conditions for peptic hydrolysis were as follows: a ratio of pepsin to dried skin powder of 2% (w/w), pH of 2.0, and a temperature of 50℃. Peptic hydrolysate contains several low-molecular weight peptides below 300 Da. Gly-Pro-Hyp(GPHyp) peptide, a process control index, was detected only in peptic hydrolysate on matrix-assisted laser desorption/ionization-time-of-flight(MALDI-TOF) spectrum. 2,2'-azinobis-(3-3-ethylbenzothiazolline-6- sulfonic acid(ABTS) radical scavenging activity of the peptic hydrolysate was comparable to that of 1 mM ascorbic acid, which was used as a positive control at pH 5.5, whereas collagenase inhibition was five times higher with the peptic hydrolysate than with 1 mM ascorbic acid at pH 7.5. However, the tyrosinase inhibition ability of the peptic hydrolysate was lower than that of arbutin, which was used as a positive control. The antibacterial effect of the peptic hydrolysate against Propionibacterium acne was not observed. These results suggest that the peptic hydrolysate derived from a flounder skin is a promising antiwrinkle agent that can be used in various food and cosmetic products to prevent wrinkles caused by ultraviolet radiations.

• 한국환경과학회지
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• 제31권11호
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• pp.993-997
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• 2022

This study was conducted to evaluate the structure and composition (i.e., pH, moisture, total-N, pathogens, and volatile fatty acids) of broiler and duck manure treated with larvae of three insect larvae, namely, Tenebrio molitor, Protaetia brevitarsis seulensis, and Ptecticus tenebrifer. Hatched Tenebrio molitor (n=300), Protaetia brevitarsis seulensis (n=60), and Ptecticus tenebrifer (n=300) were used in this study; specially, the larvae were divided into six treatments with three replicates. The treatments were as follows: T1: 110 g broiler manure + Tenebrio molitor larvae (n=50), T2: 110 g duck manure + Tenebrio molitor larvae (n=50), T3: 125 g broiler manure + Protaetia brevitarsis seulensis larvae (n=10), T4: 125 g duck manure + Protaetia brevitarsis seulensis larvae (n=10), T5: 105 g broiler manure + Ptecticus tenebrifer larvae (n=50), and T6: 105 g duck manure + Ptecticus tenebrifer larvae (n=50). For all the larval treatments, the following results were observed: The moisture content of the duck manure treat with three insect larvae was higher than that of the broiler manure (p<0.05), whereas broiler manure had a higher pH (p<0.05). In addition, the total nitrogen content of broiler manure was higher than that of duck manure (p<0.05). However, the insect larvae did not significantly affect pathogens (E.coli and Salmonella) and the volatile fatty acids (p>0.05). In conclusion, the use of the three insect larvae to create organic nitrogen compost using poultry manure is feasible.

• 웰빙융합연구
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• 제6권2호
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• pp.39-46
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• 2023

Purpose: The small-scale environmental impact assessment system in Korea was introduced and implemented in August 2000, but it has a problem that it cannot guarantee implementation due to the large proportion of qualitative reduction measures for each evaluation item. Therefore, when preparing a small-scale environmental impact assessment, research was conducted on how to improve the existing simple listing-type reduction measures and qualitative evaluation standards to quantitative reduction measures and evaluation standards reflecting regional characteristics. Research design, data and methodology: The small-scale environmental impact assessment system in Korea was introduced and implemented in August 2000, but it has a problem that it cannot guarantee implementation due to the large proportion of qualitative reduction measures for each evaluation item. Therefore, when preparing a small-scale environmental impact assessment, research was conducted on how to improve the existing simple listing-type reduction measures and qualitative evaluation standards to quantitative reduction measures and evaluation standards reflecting regional characteristics. Results: As a result of the analysis of qualitative and quantitative factors, the arithmetic sum of the qualitative factors of the total six projects is 160, accounting for 80% of the total number of reduction measures, and the quantitative factors are 40, accounting for 20%. Among them, the number of qualitative reduction measures reached 97.4% for animal and plant items, and more than 90% for air quality, noise and vibration, and eco-friendly resource circulation items. Conclusions: Therefore, it is necessary to avoid establishing qualitative reduction measures and set quantitative measures as the basis, but to specify the specifications, size, and installation location related to the reduction measures, and to calculate the numerical reduction efficiency.

• 한국미생물·생명공학회지
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• 제51권3호
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• pp.268-279
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• 2023

The pharmaceutical industry in Bangladesh produces a diverse range of antibiotics for human and animal use, however, waste disposal management is inadequate. This results in substantial quantities of antibiotics being discharged into water bodies, which provide suitable environment for the growth of antibiotic-resistant bacteria, capable of spreading resistance genes. This study intended for exploring the bacterial antibiotic resistance profile in adjoining aquatic environmental sources of pharmaceutical manufacturing facilities in Bangladesh. Seven surface water samples were collected from the vicinity of two pharmaceutical industries located in the Savar area and 51 Escherichia coli isolates were identified using both phenotypic and genotypic methods. Antibiotic susceptibility tests revealed the highest percentage of resistance against ampicillin, azithromycin, and nalidixic acid (100%) and the lowest resistance against meropenem (1.96%) out of sixteen different antibiotics tested. 100% of the study E. coli isolates were observed with Multidrug resistance phenotypes, with the Multiple Antibiotic Resistance (MAR) value ranging from 0.6-1.0. Furthermore, 69% of the isolates were Extended Spectrum Beta-Lactamases (ESBL) positive as per the Double Disk Diffusion Synergy Test (DDST). ESBL resistance genes bla_TEM, bla_CTX-M-13, bla_CTX-M-15, and bla_SHV were detected in 70.6% (n = 36), 60.8% (n = 32), 54.9% (n = 28), and 1.96% (n = 1) of the isolates, respectively, by Polymerase Chain Reaction (PCR). Additionally, 15.68% (n = 8) of the isolates were positive for E. coli specific virulence genes in PCR. These findings suggest that pharmaceutical wastewater, if not properly treated, could be a formidable source of antibiotic resistance spread in the surrounding aquatic environment. Therefore, continued surveillance for drug resistance among bacterial populations around drug manufacturing facilities in Bangladesh is necessary, along with proper waste disposal management.

• 한국환경과학회지
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• 제33권2호
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• pp.187-191
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• 2024

This study aimed to investigate the effect of treating environmental purification insect larvae to pig manure on crude ash contents and ammonia production. The experiment set up consisted go two groups: 1 kg of each 3^rd instar TM (Tenebrio molitor) and 3^rd instar PBS (Protaetia brevitarsis seulensis) larvae in Experiment 1 or 3^rd and 4^th instar of HI (Hermetia illucens L.) larvae in Experiment 2 were treated with 5 kg of pig manure. In Experiment 1, the crude ash content was higher in TM larvae-treated pig manure at days 0 and 5 (p>0.05), but was similar to that in PBS larvae-treated pig manure over (p>0.05). Ammonia production was observed at day 0 of TM and PBS larvae-treated pig manure (p<0.05), but did not occur thereafter. For Experiment 2, there was significant difference in crude ash content of 3^th and 4^th instar HI larvae-treated pig manure on day 15. Additionally, ammonia production was found in 3^th and 4^th instar HI larvae-treated pig manure at days 0 and 5, but did not continue over time. In conclusion, treating TM, PBS and HI to pig manure changed the crude ash contents and reduced ammonia through the ability to decompose pig manure. Thus, environmental impact can be minimized using environmental purification insect larvae.

• 한국발생생물학회지:발생과생식
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• 제27권4호
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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.