• Asian-Australasian Journal of Animal Sciences
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• v.33 no.12
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• pp.2021-2030
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• 2020

Objective: Quantitative polymerase chain reaction (qPCR) has been extensively used in the field of mesenchymal stem cell (MSC) research to elucidate their characteristics and clinical potential by normalization of target genes against reference genes (RGs), which are believed to be stably expressed irrespective of various experimental conditions. However, the expression of RGs is also variable depending on the experimental conditions, which may lead to false or contradictory conclusions upon normalization. Due to the current lack of information for a clear list of stable RGs in bovine MSCs, we conducted this study to identify suitable RGs in bovine MSCs. Methods: The cycle threshold values of ten traditionally used RGs (18S ribosomal RNA [18S], beta-2-microglobulin [B2M], H2A histone family, member Z [H2A], peptidylprolyl isomerase A [PPIA], ribosomal protein 4 [RPL4], succinate dehydrogenase complex, subunit A [SDHA], beta actin [ACTB], glyceraldehyde-3-phosphate dehydrogenase [GAPDH], TATA box binding protein [TBP], and hypoxanthine phosphoribosyltrasnfrase1 [HPRT1]) in bovine bone marrow-derived MSCs (bBMMSCs) were validated for their stabilities using three types of RG evaluation algorithms (geNorm, Normfinder, and Bestkeeper). The effect of validated RGs was then verified by normalization of lineage-specific genes (fatty acid binding protein 4 [FABP4] and osteonectin [ON]) expressions during differentiations of bBMMSCs or POU class 5 homeobox 1 (OCT4) expression between bBMMSCs and dermal skins. Results: Based on the results obtained for the three most stable RGs from geNorm (TBP, RPL4, and H2A), Normfinder (TBP, RPL4, and SDHA), and Bestkeeper (TBP, RPL4, and SDHA), it was comprehensively determined that TBP and RPL4 were the most stable RGs in bBMMSCs. However, traditional RGs were suggested to be the least stable (18S) or moderately stable (GAPDH and ACTB) in bBMMSCs. Normalization of FABP4 or ON against TBP, RPL4, and 18S presented significant differences during differentiation of bBMMSCs. However, although significantly low expression of OCT4 was detected in dermal skins compared to that in bBMMSCs when TBP and RPL4 were used in normalization, normalization against 18S exhibited no significance. Conclusion: This study proposes that TBP and RPL4 were suitable as stable RGs for qPCR study in bovine MSCs.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.33 no.4
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• pp.325-330
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• 2000

Alginate obtained from brown algae had various physicochemical and rheological properties and could used as a dietary fiber, However, alginate has not been widely applied to the food industry, since it had high viscosity, high gelling effect conjugated with some mineral, and low solubility. To improve functionality of alginate, partially develymerized alginates, which was water-soluble dietary fiber were obtained by hydrolysis of alginate from the sea tangle, Laminaria japonicus, heated at $121^{\circ}C$. Effects of depolymerization of alginate on the changes of viscosity and average molecular weight, block composition ratio of mannuronate to guluronate (M/G ratio), chemical properties using $PT-IR, ^1H-NMR, and ^(13)C-NMR$ spectrum were investigated. The average molecular weight and viscosity of the alginate were rapidly decreased with the thermal decomposition, and estimated to be 1,307,415 dalton and 284,000 cps, before heating, 728,106 and 3,940.29 cps after 30 min heating, 102,635 and 22.22 cps after 2.5 hrs heating, 51,205 and 12.05 cps after 3 hrs, and 10,049 and 4.28 cps after 6.5 hrs, respectively. The M/G ratio was increased with the heating time, while MM-block did not show any changes and GG-block diminished. The results of $FT-IR, ^1H-NMR and ^(13)C-NMR$ spectrum suggested that changes of molecular structure did not occur by the thermal decomposition.

• Journal of Nutrition and Health
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• v.52 no.6
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• pp.529-539
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• 2019

Purpose: Sprouts of evening primrose (Oenothera laciniata, OL) were reported to have high contents of flavonoids and potent antioxidant activity. This study examined the antioxidant and antiobesity activities of OL sprouts to determine if they could be a natural health-beneficial resource preventing obesity and oxidative stress. Methods: OL sprouts were extracted with 50% ethanol, evaporated, and lyophilized (OLE). The in vitro antioxidant activity of OLE was examined using four different tests. The antiobesity activity and in vivo antioxidant activity from OLE consumption were examined using high fat diet-induced obese (DIO) C57BL/6 mice. Results: The IC₅₀ for the 2,2-diphenyl-1-picryl-hydrazyl (DPPH) radical scavenging and superoxide dismutase (SOD)-like activities of OLE were 26.2 ㎍/mL and 327.6 ㎍/mL, respectively. OLE exhibited the ferric reducing antioxidant power (FRAP) activity of 56.7 ㎍ ascorbic acid eq./mL at 100 ㎍/mL, and an increased glutathione level by 65.1% at 200 ㎍/mL compared to the control in the hUC-MSC stem cells. In an animal study, oral treatment with 50 mg or 100 mg of OLE/kg body weight for 14 weeks reduced the body weight gain, visceral fat content, fat cell size, blood leptin, and triglyceride levels, as well as the atherogenic index compared to the high fat diet control group (HFC) (p < 0.05). The blood malondialdehyde (MDA) level and the catalase and SOD-1 activities in adipose tissue were reduced significantly by the OLE treatment compared to HFC as well (p < 0.05). In epididymal adipose tissue, the OLE treatment reduced the mRNA expression of leptin, PPAR-γ and FAS significantly (p < 0.05) compared to HFC while it increased adiponectin expression (p < 0.05). Conclusion: OLE consumption has potent antioxidant and antiobesity activities via the suppression of oxidative stress and lipogenesis in DIO mice. Therefore, OLE could be a good candidate as a natural resource to develop functional food products that prevent obesity and oxidative stress.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.46 no.6
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• pp.733-738
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• 2013

This study investigated the inhibitory effect of high hydrostatic pressure (HHP) treatments on histamine production in mackerel Scomber japonicus. Changes in viable cell counts, histamine contents, pH and VBN of mackerel fillet (stored at $4^{\circ}C$ for 25 days) were examined under HPP (200, 300, and 400 MPa). HPP treatments reduced viable cell counts by 2-3 log cycles during storage. Viable cells of mackerels treated with 400 MPa did not appear for 5 days. Histamine production was nearly suppressed by 300 and 400 MPa HPP treatments after 25 days. Furthermore, mackerels treated with HPP showed significantly lower VBN values compared with the control. Additionally, pH values were not affected by the treatments during storage periods. These results suggest that HPP treatment decreased histamine contents in mackerel muscles. Based on our results, HPP treatment may reduce scombroid fish poisoning by decreasing histamine production in mackerel during $4^{\circ}C$ storage.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.33 no.5
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• pp.388-392
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• 2000

To improve functionality and characteristics of alginate from the sea tangle, Laminaria japonicus, partially depolymerized alginates (HAG-10, average molecular weight 10,000; HAG-50, average molecular weight 50,000; HAG-100, average molecular weight 100,000) were obtained with hydrolysis of alginate by heating at $12^{\circ}C$. Effects of the depolymerization on physicochemical properties were investigated in the antimutagenicity and binding capacity of cholesterol, glucose and cadmium. In the Ames mutagenicity test using Salmonella typhimurium TA 100, HAG-10, HAG-50, HAG-100 and intact alginate reduced effectively the mutagenicities induced by aflatoxin $B_1 (AFB_1)$ and N-methyl-N'-nitro-N-nitrosoguanidine(MNNG) and HAG-10 showed the strongest antimutagenicity among the tested samples. The binding capacity of cholesterol, glucose and cadmium at different pH in vitro depended highly on molecular weight of alginate, and the changes in binding capacity at different pH was not different.

• Microbiology and Biotechnology Letters
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• v.42 no.1
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• pp.93-98
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• 2014

This study was conducted to investigate the inhibitory effect of Ecklonia cava (EC) and Eisenia bicyclis (EB) ethanol extract on histamine production in mackerel. Changes in viable cell counts, histamine contents, pH and VBN of mackerel fillet treated with ethanol extracts during 25 days at 4 were measured. Treatments of EC and EB ethanol extract had reduced growth of viable cells by 2 log cycles during storage. Production of histamine was decreased by EC and EB extracts (115 and 96 ppm) when compared to the control at 5 days (384 ppm). The pH of mackerels treated with EC and EB extracts were no different, while the pH of the control increased during storage. Furthermore, the VBN of mackerels treated with EC and EB extracts were significantly decreased when compared to the control. In conclusion, EC and EB extract may reduce scombroid fish poisoning by decreasing histamine production in mackerel during refrigerated storage.

• Journal of Veterinary Clinics
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• v.26 no.6
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• pp.528-533
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• 2009

In the current study, the mesenchymal stem cells (MSCs) isolated and propagated from the human umbilical cord blood (UCB) were tested for their capabilities of differentiation into chondrocytes in vitro. The mesenchymal progenitor cells (MPCs) collected from UCB were cultured in a low glucose DMEM medium with 10% FBS, L-glutamine and antibiotics. The human MSC colonies were positively stained by PAS reaction. When the immunophenotypes of surface antigens on the MSCs were analyzed by fluorescence-activated cell sorter (FACS) analysis, these cells expressed positively MSC-related antigens of CD 29, CD44, CD 90 and CD105, whereas they did not express antigens of CD14, CD31, CD34, CD45, CD133 and HLA-DR. Following induction these MSCs into chondrocytes in the chondrogenic differentiation medium for 3 weeks or more, the cells were stained positively with safranin O. We clearly confirmed that human MSCs were successfully differentiated into chondrocytes by RT-PCR and immunofluorescent stain of type-II collagen protein. These data also indicate that the isolation, proliferation and differentiation of the hUCB-derived MSCs in vitro can be used for elucidating the mechanisms involved in chondrogenesis. Moreover this differentiation technique can be applied to developing cell-based tissue regeneration or repair damaged tissues.