• Title/Summary/Keyword: Functional characterization

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Functional Characterization of Drosophila melanogaster CYP6A8 Fatty Acid Hydroxylase

  • Sang-A Lee;Vitchan Kim;Byoungyun Choi;Hyein Lee;Young-Jin Chun;Kyoung Sang Cho;Donghak Kim
    • Biomolecules & Therapeutics
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    • v.31 no.1
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    • pp.82-88
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    • 2023
  • Genomic analysis indicated that the genome of Drosophila melanogaster contains more than 80 cytochrome P450 genes. To date, the enzymatic activity of these P450s has not been extensively studied. Here, the biochemical properties of CYP6A8 were characterized. CYP6A8 was cloned into the pCW vector, and its recombinant enzyme was expressed in Escherichia coli and purified using Ni2+-nitrilotriacetate affinity chromatography. Its expression level was approximately 130 nmol per liter of culture. Purified CYP6A8 exhibited a low-spin state in the absolute spectra of the ferric forms. Binding titration analysis indicated that lauric acid and capric acid produced type I spectral changes, with Kd values 28 ± 4 and 144 ± 20 µM, respectively. Ultra-performance liquid chromatography-mass spectrometry analysis showed that the oxidation reaction of lauric acid produced (ω-1)-hydroxylated lauric acid as a major product and ω-hydroxy-lauric acid as a minor product. Steady-state kinetic analysis of lauric acid hydroxylation yielded a kcat value of 0.038 ± 0.002 min-1 and a Km value of 10 ± 2 µM. In addition, capric acid hydroxylation of CYP6A8 yielded kinetic parameters with a kcat value of 0.135 ± 0.007 min-1 and a Km value of 21 ± 4 µM. Because of the importance of various lipids as carbon sources, the metabolic analysis of fatty acids using CYP6A8 in this study can provide an understanding of the biochemical roles of P450 enzymes in many insects, including Drosophila melanogaster.

X-ray/gamma radiation shielding properties of Aluminium-Bariume-Zinc Oxide nanoparticles synthesized via low temperature solution combustion method

  • K.V. Sathish;K.N. Sridhar;L. Seenappa;H.C. Manjunatha;Y.S. Vidya;B. Chinnappa Reddy;S. Manjunatha;A.N. Santhosh;R. Munirathnam;Alfred Cecil Raj;P.S. Damodara Gupta;B.M. Sankarshan
    • Nuclear Engineering and Technology
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    • v.55 no.5
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    • pp.1519-1526
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    • 2023
  • For the first time Aluminium-BariumeZinc oxide nanocomposite (ZABONC) was synthesized by solution combustion method where calcination was carried out at low temperatures (600℃) to study the electromagnetic (EM) (X/γ) radiation shielding properties. Further for characterization purpose standard techniques like PXRD, SEM, UV-VISIBLE, FTIR were used to find phase purity, functional groups, surface morphology, and to do structural analysis and energy band gap determination. The PXRD pattern shows (hkl) planes corresponding to spinel cubic phase of ZnAl2O4, cubic Ba(NO3)2, α and γ phase of Al2O3 which clearly confirms the formation of complex nano composite. From SEM histogram mean size of nano particles was calculated and is in the order of 17 nm. Wood and Tauc's relation direct energy band gap calculation gives energy gap of 2.9 eV. In addition, EM (X/γ) shielding properties were measured and compared with the theoretical ones using standard procedures (NaI (Tl) detector and multi channel analyzer MCA). For energy above 356 keV the measured shielding parameters agree well with the theory, while below this value slight deviation is observed, due to the influence of atomic/crystallite size of the ZABONC. Hence synthesized ZABONC can be used as a shielding material in EM (X/γ) radiation shielding.

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.

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.

Biological and Molecular Characterization of a Korean Isolate of Clover Yellow Vein Virus Infecting Canavalia ensiformis

  • Bong-Geun Oh;Ho-Jong Ju;Jong-Sang Chung;Ju-Yeon Yoon
    • Research in Plant Disease
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    • v.30 no.2
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    • pp.157-164
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    • 2024
  • Jack bean (Canavalia ensiformis) is one of healthy products for fermented or functional food in Korea and is widely distributed and cultivated worldwide. During August 2022, Jack bean plants showing symptoms of yellow flecks, chlorosis, necrotic spots and mosaic were observed in Jangheung-gun, South Korea. By transmission electron microscopy, flexuous filamentous virus particles of approximately 750×13 nm in size were observed in the symptomatic leaf samples. The infection of a Korean isolate of clover yellow vein virus (ClYVV-Ce-JH) was confirmed using double antibody sandwich enzyme-linked sorbent assay, reverse transcription polymerase chain reaction and high-throughput sequencing. The complete genome sequence of ClYVV-Ce-JH consists of 9,549 nucleotides (nt) excluding the poly (A) tail and encodes 3,072 amino acids (aa), with an AUG start and UAG stop codon, containing one open reading frame that is typical of a potyvirus polyprotein. The polyprotein of ClYVV-Ce-JH was divided into ten proteins and each protein's cleavage sites were determined. The coat protein (CP) and polyprotein of ClYVV-Ce-JH were compared at the nt and aa levels with those of the previously reported 14 ClYVV isolates. ClYVV-Ce-JH shared 92.62% to 99.63% and 93.39% to 98.05% at the CP and polyprotein homology. To our knowledge, this is the first report of a Korean isolate of ClYVV from Jack bean plants and the complete genome sequence of a ClYVV Jack bean isolate in the world.

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.

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.

Novel Qnr Families as Conserved and Intrinsic Quinolone Resistance Determinants in Aeromonas spp.

  • Sang-Gyu Kim;Bo-Eun Kim;Jung Hun Lee;Dae-Wi Kim
    • Journal of Microbiology and Biotechnology
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    • v.34 no.6
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    • pp.1276-1286
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    • 2024
  • The environment has been identified as an origin, reservoir, and transmission route of antibiotic resistance genes (ARGs). Among diverse environments, freshwater environments have been recognized as pivotal in the transmission of ARGs between opportunistic pathogens and autochthonous bacteria such as Aeromonas spp. In this study, five environmental strains of Aeromonas spp. exhibiting multidrug resistance (MDR) were selected for whole-genome sequencing to ascertain their taxonomic assignment at the species-level and to delineate their ARG repertoires. Analyses of their genomes revealed the presence of one protein almost identical to AhQnr (A. hydrophila Qnr protein) and four novel proteins similar to AhQnr. To scrutinize the classification and taxonomic distribution of these proteins, all Aeromonas genomes deposited in the NCBI RefSeq genome database (1,222 genomes) were investigated. This revealed that these Aeromonas Qnr (AQnr) proteins are conserved intrinsic resistance determinants of the genus, exhibiting species-specific diversity. Additionally, structure prediction and analysis of contribution to quinolone resistance by AQnr proteins of the isolates, confirmed their functionality as quinolone resistance determinants. Given the origin of mobile qnr genes from aquatic bacteria and the crucial role of Aeromonas spp. in ARG dissemination in aquatic environments, a thorough understanding and strict surveillance of AQnr families prior to the clinical emergence are imperative. In this study, using comparative genome analyses and functional characterization of AQnr proteins in the genus Aeromonas, novel Aeromonas ARGs requiring surveillance has suggested.