• Title/Summary/Keyword: Functional characterization

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Molecular characterization and functionality of rumen-derived extracellular vesicles using a Caenorhabditis elegans animal model

  • Hyejin Choi;Daye Mun;Sangdon Ryu;Min-jin Kwak;Bum-Keun Kim;Dong-Jun Park;Sangnam Oh;Younghoon Kim
    • Journal of Animal Science and Technology
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    • v.65 no.3
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    • pp.652-663
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    • 2023
  • The rumen fluids contain a wide range of bacteria, protozoa, fungi, and viruses. The various ruminal microorganisms in the rumen provide nutrients by fermenting the forage they eat. During metabolic processes, microorganisms present in the rumen release diverse vesicles during the fermentation process. Therefore, in this study, we confirmed the function of rumen extracellular vesicles (EVs) and their interaction with the host. We confirmed the structure of the rumen EVs by transmission electron microscope (TEM) and the size of the particles using nanoparticle tracking analysis (NTA). Rumen EVs range in size from 100 nm to 400 nm and are composed of microvesicles, microparticles, and ectosomes. Using the Caenorhabditis elegans smart animal model, we verified the interaction between the host and rumen EVs. Exposure of C. elegans to rumen EVs did not significantly enhance longevity, whereas exposure to the pathogenic bacteria Escherichia coli O157:H7 and Staphylococcus aureus significantly increased lifespan. Furthermore, transcriptome analysis showed gene expression alterations in C. elegans exposed to rumen EVs, with significant changes in the metabolic pathway, fatty acid degradation, and biosynthesis of cofactors. Our study describes the effect of rumen EV interactions with the host and provides novel insights for discovering biotherapeutic agents in the animal industry.

Candidate Gene Analysis to Rice Bacterial Leaf Blight Resistance of Korean Races of Xoo (Xanthomonas oryzae) in Rice Genetic Resources by GWAS Analysis

  • Myung Chul Lee;Yu-Mi Choi;Myoung-Jae Shin;Hyemyeong Yoon;Sukyeung Lee;Kebede Taye Desta
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2020.08a
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    • pp.49-49
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    • 2020
  • Bacterial leaf blight (BLB), caused by X. oryzae pv. oryzae(Xoo), is one of the most destructive diseases of rice due to its high epidemic potential. Understanding BLB resistance at a genetic level is important to further improve the rice breeding that provides one of the best approaches to control BLB disease. In the present investigation, a total of 10,000 accessions of rice germplasm were tested to resistance degree of four Korean isolated races (K1, K2, K3 and K3a) of Xoo by bioassay and a diverse 268 accessions was selected to the genome-wide association study (GWAS) using high quality 34,724 SNPs to identify the associated with resistance loci. LOC_Os04g53160 of chromosome 4 was significantly associated with K1 race resistant. LOC_Os11g46230 and LOC_Os11g47150 of chromosome 11 were highly associated with K2 and K3 races as 23.7 and 27.4 of -log(P) value, but K3a resistant loci was weakly associated at LOC_Os03g55270 of chromosome 3. The results of the GWAS validate known gene of BLB resistant and identified novel loci of R genes that provide useful targets for further investigation to help the breeding system and identified gene and QTL provide valuable sources for further functional characterization.

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A streamlined pipeline based on HmmUFOtu for microbial community profiling using 16S rRNA amplicon sequencing

  • Hyeonwoo Kim;Jiwon Kim;Ji Won Cho;Kwang-Sung Ahn;Dong-Il Park;Sangsoo Kim
    • Genomics & Informatics
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    • v.21 no.3
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    • pp.40.1-40.11
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    • 2023
  • Microbial community profiling using 16S rRNA amplicon sequencing allows for taxonomic characterization of diverse microorganisms. While amplicon sequence variant (ASV) methods are increasingly favored for their fine-grained resolution of sequence variants, they often discard substantial portions of sequencing reads during quality control, particularly in datasets with large number samples. We present a streamlined pipeline that integrates FastP for read trimming, HmmUFOtu for operational taxonomic units (OTU) clustering, Vsearch for chimera checking, and Kraken2 for taxonomic assignment. To assess the pipeline's performance, we reprocessed two published stool datasets of normal Korean populations: one with 890 and the other with 1,462 independent samples. In the first dataset, HmmUFOtu retained 93.2% of over 104 million read pairs after quality trimming, discarding chimeric or unclassifiable reads, while DADA2, a commonly used ASV method, retained only 44.6% of the reads. Nonetheless, both methods yielded qualitatively similar β-diversity plots. For the second dataset, HmmUFOtu retained 89.2% of read pairs, while DADA2 retained a mere 18.4% of the reads. HmmUFOtu, being a closed-reference clustering method, facilitates merging separately processed datasets, with shared OTUs between the two datasets exhibiting a correlation coefficient of 0.92 in total abundance (log scale). While the first two dimensions of the β-diversity plot exhibited a cohesive mixture of the two datasets, the third dimension revealed the presence of a batch effect. Our comparative evaluation of ASV and OTU methods within this streamlined pipeline provides valuable insights into their performance when processing large-scale microbial 16S rRNA amplicon sequencing data. The strengths of HmmUFOtu and its potential for dataset merging are highlighted.

Adnexal Masses: Clinical Application of Multiparametric MR Imaging & O-RADS MRI (난소-자궁부속기 종괴: 다중기법 MR 영상의 임상 적용과 O-RADS MRI)

  • So Young Eom;Sung Eun Rha
    • Journal of the Korean Society of Radiology
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    • v.82 no.5
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    • pp.1066-1082
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    • 2021
  • Incidental adnexal masses considered indeterminate for malignancy are commonly observed on ultrasonography. Multiparametric MRI is the imaging modality of choice for the evaluation of sonographically indeterminate adnexal masses. Conventional MRI enables a confident pathologic diagnosis of various benign lesions due to accurate tissue characterization of fat, blood, fibrous tissue, and solid components. Additionally, functional imaging sequences, including perfusion- and diffusion-weighted imaging, improve the diagnostic efficacy of conventional MRI in differentiating benign from malignant adnexal masses. The ovarian-adnexal reporting and data system (O-RADS) MRI was recently designed to provide consistent interpretations in assigning risk of malignancy to ovarian and other adnexal masses, and to provide a management recommendation for each risk category. In this review, we describe the clinical application of multiparametric MRI for the evaluation of adnexal masses and introduce the O-RADS MRI risk stratification system.

Production Process and Physiochemical Characterization of Low-sugar Yuzu Syrup (대체 감미료를 활용한 저당 유자 시럽 제조 및 이화학적 특성 조사)

  • Yunha Bak;Bo-Bae Lee;Ae Eun Im;Jeong-Yong Cho;Seung-Hee Nam
    • The Korean Journal of Food And Nutrition
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    • v.37 no.2
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    • pp.67-79
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    • 2024
  • Yuzu (Citrus junos) undergoes traditional processing to create preserved yuzu with a sugar content of 50%. This study aimed to produce low-sugar yuzu syrup using artificial or natural sweeteners instead of sugar. Among artificial sweeteners, maltitol showed minimal browning and lack of deposition at high temperatures. The addition of xanthan gum (0.2%) and carboxymethyl cellulose (0.2%) resolved the issue of layer separation and viscosity decrease of yuzu syrup. Alternatively, grain syrup, a natural sweetener, improved viscosity and homogeneity without the additives. Yuzu syrups were developed using yuzu juice and preserved yuzu, with maltitol (20~40%) or grain syrup (50~70%) as the sweetener. Yuzu syrups containing 35% maltitol (M35) or 55% grain syrup (G55) had less than 5% and 10% free sugar, respectively. These syrups exhibited taste patterns similar to commercial yuzu syrup in analysis using an electronic tongue. Furthermore, M35 and G55 contained yuzu flavonoids at concentrations of 19.82 mg/g and 24.09 mg/g, respectively. Antioxidant activity (DPPH, ABTS radical scavenging) of M35 and G55 was equivalent to 10.55, 17.59 mg/100 g of Vitamin C and 97.39, 33.92 mg/100 g of Vitamin C, respectively. Consequently, M35 and G55 offer promising alternatives to preserved yuzu, providing low-sugar yuzu syrups enriched with functional ingredients.

Formulation and Quality Evaluation of Chicken Nuggets Supplemented with Beef and Chicken Livers

  • Liaqat Mehmood;Syeda Afnan Mujahid;Sawera Asghar;Hafiz Ubaid ur Rahman;Nauman Khalid
    • Food Science of Animal Resources
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    • v.44 no.3
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    • pp.620-634
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    • 2024
  • This study explores the potential of utilizing meat byproducts, specifically chicken and beef liver, to enhance the nutritional value of processed foods like chicken nuggets. Proximate analysis was conducted on the livers, including moisture, ash, fat, and protein content, and degradation potential was observed. Antioxidant potential was analyzed through 2,2-diphenyl-1-picrylhydrazyl (DPPH). The total phenolic content (TPC), oxidative stability through peroxide value (POV), and free fatty acid (FFA) were performed to evaluate quality changes during seven-day storage. The radical scavenging activity showed that beef liver has excellent antioxidant capacity (61.55%- and 195.89- mM gallic acid equivalent for DPPH and TPC, respectively) compared to chicken liver and significantly increased the antioxidant potential of nuggets by 5%-10%. POV and FFA values increased with increased storage days for the liver and its incorporation in nuggets. However, the values remained under the 10 meq/kg threshold. Incorporating the livers into chicken nuggets led to a significant (p=0.000) improvement in nutritional content, particularly a 1.5%-2% increase in protein, with a similar increase in mineral content. Texture and sensory evaluations indicated favorable consumer acceptability for liver-enriched nuggets. Overall, this research shows the value of adding liver as a functional ingredient to enhance the nutritional profile of processed foods.

Characterization and Functional Analysis of Obox4 during Oocyte Maturation by RNA Interference (생쥐의 난소와 난자에서의 Obox4의 동정과 RNAi를 이용한 기능연구)

  • Lee, Hyun-Seo;Lee, Kyung-Ah
    • Clinical and Experimental Reproductive Medicine
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    • v.34 no.4
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    • pp.293-303
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    • 2007
  • Objective: Previously, we identified differentially expressed genes between GV and MII stage mouse oocytes using ACP technology. When we study one of GV selective genes, Obox family, we found Obox4 mRNA expression in ovaries that has been reported as expressed exclusively in testis. Therefore, this study was conducted for characterization and functional analysis for Obox4. Methods: Expression of Obox4 mRNA was examined in gonads and oocytes by RT-PCR. To determine the role of Obox4 in oocyte maturation, Obox4 dsRNA was microinjected into the cytoplasm of GV oocytes followed by 16 h of incubation in the plain medium or by 24 h of incubation in the medium containing IBMX. After RNAi, phenotypes and maturation rates were observed, change in mRNA expression was evaluated, and chromosomal status was confirmed by orcein staining. Results: Obox4 has minimal expression in the ovary compared to that of the other family members. When oocytes were cultured for 16 h in M16 medium after RNAi, maturation rate was not changed significantly, compared with that of non-injected or buffer-injected control oocytes. Surprisingly, however, when oocytes were cultured for 24 h in M16 containing IBMX, in which oocytes were supposed to arrest at GV stage, Obox4 RNAi oocytes were advanced to MI and MII. Spindle structure was disappeared and the chromosomes were condensed in the oocytes after Obox4 RNAi. Conclusions: This is the first report on the expression of Obox4 in the ovary and oocytes. Results of the study suggest that Obox4 plays a crucial role in spindle formation and chromosome segregation during meiosis in oocytes. In addition, Obox4 may play an important role in cAMP-dependent signal cascades of GV-arrest in mouse oocytes.

Chemical and Spectroscopic Characterization of Peat Moss and Its Different Humic Fractions (Humin, Humic Acid and Fulvic Acid) (피트모스에서 추출한 휴믹물질(휴믹산, 풀빅산, 휴민)의 화학적 및 분광학적 물질특성 규명)

  • Lee Chang-Hoon;Shin Hyun-Sang;Kang Ki-Hoon
    • Journal of Soil and Groundwater Environment
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    • v.9 no.4
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    • pp.42-51
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    • 2004
  • Peat humin(p-Humin), humic acid(p-HA) and fulvic acid(p-FA) were isolated from Canadian Sphagnum peat moss by dissolution in 0.1M NaOH followed by acid precipitation. After purification cycles, they are characterized for their elemental compositions and, acid/base properties. Functionalities and carbon structures of the humic fractions were also characterized using FT-IR and solid state $^{13}C$-NMR spectroscopy. Those results are compared with one another and with soil humic substances from literatures. Main purpose of this study was to present a chemical and spectroscopic characterization data of humic substance from peat moss needed to evaluate its environmental applicability. The relative proportions of the p-Humin, p-HA and p-FA in the peat moss was $76\%,\;18\%,\;and\;3\%$, respectively, based on the total organic matter content ($957{\pm}32\;g/kg$). Elemental composition of p-Humin were found to be $C_{1.00}H_{1.52}O_{0.79}N_{0.01}$ and had higher H/C and (N+O)/C ratio compared to those of p-HA($C_{1.00}H_{1.09}O_{0.51}N_{0.02}$) and p-FA($C_{1.00}H_{1.08}O_{0.65}N_{0.01}$). Based on the analysis of pH titration data, there are two different types of acidic functional groups in the peat moss and its humic fractions and their proton exchange capacities(PEC, meq/g) were in the order p-FA(4.91) >p-HA(4.09) >p-Humin(2.38). IR spectroscopic results showed that the functionalities of the peat moss humic molecules are similar to those of soil humic substances, and carboxylic acid(-COOH) is main function group providing metal binding sites for Cd(II) sorption. Spectral features obtained from $^{13}C$-NMR indicated that peat moss humic molecules have rather lower degree of humification, and that important structural differences exist between p-Humin and soluble humic fractions(p-HA and p-FA).

Isolation of Agarivorans sp. JS-1 and Characterization of Its β-Agarase (한천분해세균 Agarivorans sp. JS-1의 분리 및 β-아가라제의 특성 규명)

  • Jin Sun Kim;Dong-Geun Lee;Go-Wun Yeo;Min-Joo Park;Sang-Hyeon Lee
    • Journal of Life Science
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    • v.33 no.4
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    • pp.357-362
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    • 2023
  • This report looks at an agar-degrading marine bacterium and characterization of its agarase. Agar-degrading marine bacterium JS-1 was isolated with Marine agar 2216 media from seawater from the seashore of Sojuk-do, Changwon in Gyeongnam Province, Korea. The agar-degrading bacterium was named as Agarivorans sp. JS-1 by phylogenetic analysis based on 16S rRNA gene sequencing. The extracellular agarase was prepared from the culture media of Agarivorans sp. JS-1 and used for characterization. Relative activities at 20℃, 30℃, 35℃, 40℃, 45℃, 50℃, 55℃, and 60℃ were 70%, 74%, 78%, 83%, 87%, 100%, 74%, and 66%, respectively. Relative activities at pH 5, 6, 7, and 8 were 91%, 100%, 90%, and 89%, respectively. Its extracellular agarase showed maximum activity (207 units/l) at pH 6.0 and 50℃ in 20 mM Tris-HCl buffer. The residual activity after heat treatment at 20℃, 30℃, and 50℃ for 30 minutes was 90%, 70%, and 50% or more, respectively. After a 2-hour heat treatment at 20℃, 30℃, 35℃, 40℃, and 45℃, the residual activity was 80%, 68%, 65%, 63%, and 57%, respectively. At 50℃ and above, after heat treatment for 30 minutes, the residual activity was below 60%. Thin layer chromatography analysis suggested that Agarivorans sp. JS-1 produces extracellular β-agarases as they hydrolyze agarose to produce neoagarooligosaccharides such as neoagarohexaose (20.6%), neoagarotetraose (58.5%), and neoagarobiose (20.9%). Agarivorans sp. JS-1 and its thermotolerant β-agarase would be useful in the production of neoagarooligosaccharides, showing functional activity such as inhibition of bacterial growth and delay of starch degradation.

Characterization of Humic and Fulvic Acids Extracted from Soils in Different Depth: Proton Exchange Capacity, Elemental Composition and 13C NMR Spectrum (깊이별 토양 휴믹산과 풀빅산의 특성 분석: 양성자교환용량, 원소성분비, 13C NMR 스펙트럼)

  • Shin, Hyun-Sang;Lee, Chang-Hoon;Rhee, Dong-Sock;Chung, Kun-Ho;Lee, Chang-Woo
    • Analytical Science and Technology
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    • v.16 no.4
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    • pp.283-291
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    • 2003
  • Humic and fulvic acids present in soils of different depth were extracted and their acidic functional groups and structural characteristics were analyzed and compared. The purpose of this study was to present a basic data needed to evaluate the effect of humic substances on depth distribution and migrational behaviour of radioactive elements deposited on soil. Acidic functional groups of the humic and fulvic acids were analyzed by pH titration method, and their proton exchange capacity (PEC, $mq\;g^{-1}$) and average $pK_a$ values were obtained. Structural characteristics of the humic and fulvic acids were analyzed using their CPMAS $^{13}C$ NMR spectra and elemental composition data. pH titration data showed that fulvic acids have higher acidic functional group contents ranging from 5.5 to $7.0meq\;g^{-1}$ compared with that of humic acids ($3.8{\sim}4.8meq\;g^{-1}$). From depth profiles, it has been found that PEC values of humic acids in deeper soil (> 8 cm) were higher than those at the surface soils. Elemental compositions (H/C ratio) and spectral features ($C_{arom}/C_{aliph}$ ratio) obtained from CPMAS $^{13}C$ NMR spectra showed that the aromatic character in humic acids was a relatively higher than that of fulvic acids, while lower in carboxyl carbon content. The aromatic character and carboxyl carbon contents of humic acids tend to increase as soil depth increased, but those of fulvic acid showed little differences by the soil depth range.