• Journal of Food Hygiene and Safety
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• v.33 no.4
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• pp.280-288
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• 2018

In this study, an approach for the analysis of the five cephalopod species (octopus, long-arm octopus, squid, wet-foot octopus, beka squid) consumed in the Republic of Korea is developed. The samples were collected from the Southeast Asian countries Thailand, Indonesia, Vietnam, and China. The SYBR-green-based real-time qPCR method, based on the mitochondrial DNA genome of the five cephalopods was developed and validated. The intergroup variations in the mitochondrial DNA are evident in the bioinformatic analysis of the mitochondrial genomic DNA sequences of the five groups. Some of the highly-conserved and slightly-variated regions are identified in the mitochondrial cytochrome-c-oxidase subunit I (COI) gene, 16s ribosomal RNA (16s rRNA) gene, and 12s ribosomal RNA (12s rRNA) gene of these groups. To specify each five cephalopod groups, specific primer sets were designed from the COI, 16s rRNA and 12s rRNA regions. The specific primer sets amplified the DNA using the SYBR-green-based real-time PCR system and 11 commercially secured animal tissues: Octopus vulgaris, Octopus minor, Todarodes pacificus, Dosidicus gigas, Sepia esculenta, Amphioctopus fangsiao, Amphioctopus aegina, Amphioctopus marginatus, Loliolus beka, Loligo edulis, and Loligo chinensis. The results confirmed by a conveient way to calculate relative amplification levels between different samples in that it directly uses the threshold cycles (Ct)-value range generated by the qPCR system from these samples. This genomic DNA-based molecular technique provides a quick, accurate, and reliable method for the taxonomic classification of the animal tissues using the real-time qPCR.

• Journal of Nutrition and Health
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• v.37 no.9
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• pp.786-793
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• 2004

Women with menopause or rats with ovariectomy is associated with increased body weight, body fat and insulin resistance, which are components of metabolic syndrome. Increased prevalence of metabolic syndrome after menopause might be associated with mitochondrial dysfunction, since mitochondrial oxidative and phosphorylation activity is strongly correlated with insulin sensitivity. Although estradiol replacement prevents the metabolic syndrome, harmful effect of estradiol hampers the casual usage to prevent the metabolic syndrome. It has been reported that genistein has a mild estrogenic activity, decreases fat mass in mice and has an antidiabetic role in diabetic rats. Although insulin resistance is closely related to mitochondrial functions, there has not been yet any study in regard to the effect of dietary genistein on mitochondrial function in the insulin resistant female subjects induced by ovariectomy or similar situation. The present study investigated whether the supplementation of genistein in the high fat diet affected the mitochondrial function of high fat fed ovariectomized rats. Female Sprague Dawley rats (8 weeks old) were assigned to the following groups: sham-operated+ high fat diet (S, n=6); sham-operated + high fat diet with 0.1% genistein (S + G, n=7); ovariectomized + high fat diet (OVX, n=8); ovariectomized + high fat diet with 0.1% genistein (OVX+ G, n=8). Ovariectomy significantly increased body weight compared with S group. Genistein consumption in ovariectomized (OVX + G) rats decreased body weight gain compared with OVX rats. Liver weights were increased by ovariectomy. The hepatic mitochondrial protein density expressed as mg per g liver was lower in the OVX group than in the S group. However, OVX + G group showed the increased mitochondrial protein density similar to the level of S group. When mRNA levels of genes related to mitochondria such as peroxisome proliferator-activated receptor ${\gamma}$ coactivator 1 (PGC-1) and cytochrome c oxidase subunit III (COX III) were measured, there were decreases in the mRNA levels of PGC-1 and COX III in S + G, OVX and OVX + G group. The activity of cytochrome c oxidase was not different between groups. We could observe the decrease in succinate dehydrogenase (SDH) activity per g liver in OVX rats. Genistein supplement increased SDH activity. In conclusion, genistein supplementation to the OVX rats enhanced mitochondrial function by increasing mitochondrial protein density and SDH activity. The improvement in mitochondrial function by genistein can contribute to the improvement in metabolic syndrome.

• Journal of Korean Society of Forest Science
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• v.101 no.3
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• pp.517-525
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• 2012

In this study, a 4-year-old mountain ginseng was mixed and ripened with 4-year-matured persimmon vinegar, and then it was diluted 5 times and orally administerd to rats. Afterwards, by analyzing the protein expression rate which affects both the carbohydrate metabolism and the lipid metabolism, this study examined the anti-obesity effect of the fusion material. The rats were divided into a control group (CON), a persimmon vinegar group (PV) and a mountain ginseng+persimmon vinegar fusion material group (MPV). The weight gain rate was found to be low in PV and MPV, and the concentration of glucose was also low in PV and MPV. However, GLUT-2 was found to be significantly high in these two groups on the contrary. Both the concentration of free fatty acid and CPT-1 protein expression rate were high in PV and MVP, but MVP was higher than PV. Cytochrome C oxidase was found to be higher in MPV than in CON. AMPK, $PPAR-{\gamma}$ and $PGC1-{\alpha}$ were all high in PV and MPV, but MPV was higher than PV. All the results above verified the thermogenesis effect of the fusion material, leading to an increase of energy metabolism, and it was thought that the fusion material could be effectively used for anti-obesity. However, it seems necessary to verify the anti-obesity effect through various further studies.

• Journal of Life Science
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• v.24 no.6
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• pp.626-632
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• 2014

The genetic structure and phylogenetic relationship were investigated in Korean red spotted grouper populations using the nucleotide sequence polymorphisms of the mitochondrial DNA (mtDNA) cytochrome c oxidase subunit I (COI) gene. The COI gene was sequenced showed 99.1-99.8% identity with the EF607565 sequence previously reported. A total of twenty haplotypes were found, and the Korean population showed nineteen haplotypes. Among those, Hap_03 and Hap_08 showed Jeju-do and China-specific COI sequences, respectively. However, Hap_07 had twelve COI sequences from South Korea and records from Hong Kong and Taiwan. Neighbor-joining (NJ) trees constructed from the phylogenetic analyses based on the polymorphisms of the COI haplotypes showed a monophyletic branching pattern within the genus Epinephelus. This indicated that the red spotted grouper populations had evolved from common maternal ancestors. In addition, the Hap_08, which had the COI sequence recorded only from China Sea, was found in the middle of the NJ tree nearby Hap_07 and showed a close relationship with Hap_07. This indicates that Chinese red spotted grouper is also maternally related to other populations in East Asia. Consequently, East Asian red spotted grouper populations are maternally related, as well as sharing the same evolutionary history, and are still affected by the East Asian ocean current (Kuroshio). These findings help to explain the genetic structure and phylogenetic relationship of red spotted grouper and also contribute to research on artificial breeding and industrialization.

• Journal of Food Hygiene and Safety
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• v.34 no.5
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• pp.502-507
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• 2019

Cephalopods are one of the most important fishery resources in the world because of their desirable taste and nutritional value. In south Korea, one of the countries in which a large amount of seafood is consumed, cephalopods (e.g., octopus, squid, and cuttlefish) have an annual consumption rate of over 400,000 metric tons. In this study, octopus and squid products (n=28) sold on the market were monitored by analyzing sequences of DNA barcode markers (cytochrome c oxidase subunit I and 16S ribosomal RNA genes). For species identification, the NCBI BLAST database was screened with the sequences and analyzed as a query. In this BLAST search, twelve squid products showed 99-100% sequence identity to Dosidicus gigas (n=3) and Todarodes pacificus (n=9). In the case of the other 16 products that were declared using octopus as raw materials on the labels, six products were identified as Cistopus taiwanicus (n=1), Amphioctopus marginatus (n=1), Scaeurgus unicirrhus (n=1), and Dosidicus gigas (n=3). Monitoring results indicated that a significant percentage (37.5%) of mislabeling was present in octopus products sold on the South Korean market.

• Korean Journal of Clinical Laboratory Science
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• v.51 no.1
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• pp.64-70
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• 2019

Dirofilaria immitis (D. immitis) is a filarial nematode that causes cardiopulmonary dirofilariasis in dogs. In the late stages of infection, infected dogs show one or more symptoms and advanced heart disorder with perivascular inflammation. To detect D. immitis specifically and efficiently in the early stages of infection, a duplex TaqMan qPCR assay was developed based on previous studies using primers and probes specialized to detect D. immitis cytochrome c oxidase subunit I (COI) and dog glyceraldehyde-3-phosphate dehydrogenase (GAPDH). As positive controls, plasmid DNAs were constructed from D. immitis COI or dog GAPDH and a TA-cloning vector. Simplex and duplex TaqMan qPCR assays were performed using the specific primers, probes, and genomic or plasmid DNA. The duplex reaction developed could detect D. immitis COI and dog GAPDH in the same sample simultaneously after optimization of the primer concentrations. The limit of detection was 25 copies for the simplex and duplex assays, and both showed good linearity, high sensitivity, and excellent PCR efficiency. The duplex assays for pathogen detection reduce the costs, labor, and time compared to simplex reactions. Therefore, the duplex TaqMan qPCR assay developed herein will allow efficient D. immitis detection and quantification from a large number of samples simultaneously.

• Korean Journal of Ecology and Environment
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• v.54 no.3
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• pp.240-246
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• 2021

The Chironomidae is a benthic macroinvertebrate commonly found in freshwater ecosystems, along with Ephemeroptera and Trichoptera, which can be used for environmental health assessments. There are approximately 15,000 species of Chironomidae worldwide, but there are limited studies on species identification of domestic Chironomidae larvae. In the present study, we carried out species classification of the Chironomidae larvae that found in Jeju's tap water purification plants using morphological characteristics and genetic identification based on cytochrome c oxidase subunit I (COI) gene of the mitochondrial DNA. Body shape, mentum, antenna, mandible in the head capsule, and claws were observed in the larvae for morphological classification. Analysis of 17 larvae collected from faucets and fire hydrants of domestic tap water purification plants revealed the presence of two species, including 14 Orthocladius tamarutilus and 3 Paratrichocladius tammaater. These results will aid the use of the criteria information about species classification of the Chironomidae for water quality management in water purification plants and diversity monitoring of freshwater environments.

• Journal of Food Hygiene and Safety
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• v.35 no.6
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• pp.574-580
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• 2020

In this study we sought to determine the food fraud by discriminating species of commercial seafood product such as Larimichthys polyactis, Larimichthys crocea, Pennahia argentatus, and Miichthys miiuy, which are difficult to morphologically discriminate. After amplifying the mitochondrial cytochrome c oxidase subunit I gene of the reference fish, the DNA sequences of the amplified PCR products were analyzed. As a result, a 655 bp sequence for species identification was selected for use as DNA barcodes. To confirm the DNA data and primer set, the DNA barcode sequence of each fish was compared to that in that in the NCBI. All of the DNA barcode data were matched with the gene sequence of each fish in the NCBI. A total of 32 processed seafood products (8 L. polyactis, 12 L. crocea, 3 Pennahia argentatus, and 9 Miichthys miiuy) were investigated. Homology of 97% or more in DNA sequences was judged as the same species. As a result of the monitoring, there were no discovered cases of forgery or alteration. However, the use of a raw material name having no matching standard name in the Korea Food Code may cause consumer confusion. Therefore, it is suggested that the standard name or scientific name be co-labeled with the raw material name on seafood products to prevent consumer confusion.

• Journal of Food Hygiene and Safety
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• v.36 no.4
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• pp.331-341
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• 2021

DNA barcode sequences of commercial seafood products, which are difficult to morphologically discriminate, were analyzed to determine cases of food fraud. The gene sequences were analyzed by amplifying the COX I (cytochrome C oxidase subunit I) gene region of mitochondrial DNA, which is mainly used for species identification. The DNA barcode sequences were compared with the gene sequence of each fish registered in the US National Center for Biotechnology. A total of 46 processed seafood products (12 Pagrus majo, 4 Oplegnathus fasciatus, 7 Dentex tumifrons, 2 Acanthopagrus schlegelii, 7 Oreochromis niloticus, 6 Branchiostegus japonicus, 8 Branchiostegus albus) were investigated. Having DNA sequence identity of more than 97% was judged as the same species. As a result of this study, no cases of forgery and alteration were detected. However, some disparities in the commercial names used in local markets and the standard names given in the Korea Food Code were found, which may cause confusion for consumers. It is therefore suggested that the standard name or scientific name be displayed on seafood product labels.

• Journal of Food Hygiene and Safety
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• v.38 no.6
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• pp.496-507
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• 2023

This study investigated species identification and labeling compliance of 48 shrimp products sold in the Korean online markets. Species identification was conducted using the standard DNA barcoding method, using the cytochrome c oxidase subunit I gene. The obtained sequences were compared with those deposited in the NCBI GenBank and BOLD Systems databases. Additionally, phylogenetic analysis was performed to further verify the identified shrimp species. Consequently, 16 shrimp species were identified, including Penaeus vannamei, Pandalus borealis, Palaemon gravieri, Leptochela gracilis, Penaeus monodon, Pleoticus muelleri, Metapenaeopsis dalei, Euphausia pacifica, Lebbeus groenlandicus, Trachypenaeus curvirostris, Argis lar, Metanephrops thomsoni, Metapenaeopsis barbata, Alpheus japonicus, Penaeus chinensis, and Mierspenaeopsis hardwickii. The most prevalent species was Penaeus vannamei, found in 45.8% of the analyzed products. A significant mislabeling rate of 72.9% was found; however, upon excluding generic names such as shrimp, the mislabeling rate reduced to 10.4%. The mislabeling rate was higher in highly-processed products (89.3%) compared with that in minimally-processed products (50%). No correlation was found between the country of origin and mislabeling rate. The results of this study provide crucial data for future monitoring of shrimp products and improving the labeling of shrimp species in Korea.