• Journal of the Korean Society of Food Science and Nutrition
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• v.44 no.4
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• pp.592-601
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• 2015

Rubus coreanus is known as Korean black raspberry, native to Korea, Japan, and China. Preliminary studies evaluating their potential for cancer treatment in mammalian test systems are ongoing. In recent years, interest has been renewed due to their high levels of anthocyanins. Anthocyanins in black raspberry are important due to their potential health benefits as dietary antioxidant, anti-inflammatory compound, and as a chemopreventive agent. In the present study, Saccharomyces cerevisiae BA29 was isolated from black raspberry fruit and fruit juice as a biogenic amine non-producing strain for manufacturing of black raspberry wine, after which we investigated its characteristics: biogenic amine-producing ability, cell growth ability, alcohol-fermentation ability, and resistance to alcohol, glucose, and sulfur dioxide. Based on preliminary experiments, we optimized culture medium compositions for improving dried cell weight of S. cerevisiae BA29 by response surface methodology (RSM) as a statistical method. Design for RSM used a central composite design, and molasses with the industrial applicability was used as a carbon source. Through statistical analysis, we obtained optimum values as follows: molasses 200 g/L, peptone 30 g/L, and yeast extract 40 g/L. For the model verification, we confirmed about 3-fold improvement of dried cell weight from 6.39 to 20.9167 g/L compared to basal yeast peptone dextrose medium. Finally, we manufactured black raspberry wine using S. cerevisiae BA29 and produced alcohol of 20.33%. In conclusion, S. cerevisiae isolated from black raspberry fruit and juices has a great potential in the fermentation of black raspberry wine.

• Journal of Life Science
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• v.20 no.4
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• pp.467-473
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• 2010

Gangliosides are a major component of the plasma membrane of mammalian cells, which are directly involved in a variety of immunological events, including cell-to cell or cell-to-protein interactions. In this study, we investigated whether gangliosides, sialic acid-containing glycosphingolipids, are related to rejection during the xenotransplantation of NIH-miniature pig livers and hearts to humans. Both high performance thin-layer chromatography and immunohistochemistry analyses revealed that the expression of gangliosides in the liver tissue of NIH-miniature pigs was higher than that in the heart. Gangliosides GD3, GD1a, GD1b, GT1b and GQ1b were observed in both the liver and heart, whereas GQ1b was detected only in the liver, indicating that the ganglioside expression profiles are tissue specific. Moreover, other ganglio-series gangliosides, including GM3, were not detected in the livers and hearts of NIH-miniature pigs. Taken together, these results suggest that gangliosides may play important roles in immune responses in clinical xenotransplants of pig livers and hearts.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1994.04a
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• pp.186-186
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• 1994

Many agonists have been known to activate the hydrolysis of membrane phospholipids through the bindings with corresponding receptors on the various cells. Diacylglycerol and inositol 1,4,5-trisphosphate(IP3) generated by the action of phosphoinositide-specific phospholipase C (PI-PLC) are well known second messengers for the activation of protein kinase C and the mobilization of Ca2+ in many cells. Three types of PI-PLC isozyme (${\alpha}$,${\gamma}$, and $\delta$) and several subtrpes for each type have been identified from mammalian sources by purification of enzymes and cloning of their cDNAs. Each type PI-PLC isozyme is coupled to different receptors and mediators, for example, ${\beta}$-types are coupled to the seven-transmembrane-receptors via Gq family of G-proteins and ${\beta}$-types directly to the receptor tyrosine kinases. Specific modulators for the signaling pathway through each type of PI-PLC should be very useful as potential potential candidates for lend substances in developing novel drugs. To establish the sensitive and convenient screening systems for searching modulators on PI-PLC mediated signaling, two kinds of approaches have been tried. (1) Establishment of in vitro assay condition for each type of PI-PLC isozyme: Overexpression by using vaccinia virus and purification of each isozyme was carried out for the preparation of large amounts of enaymes. Optimum and sensitive assay condition for the measurements of PI-ELC activities were established. (2) Development of the cell lines in which each type of PI-PLC is permanently overexpressed: A fibroblast cell line (3T3${\gamma}$1-7) in which PI-PLC-${\gamma}$1 was overexpressed by using pZip-neo expression vector was developed and used for the measurement of PDGF-induced IP3 formation. The responses for IP3 formed in 3T3${\gamma}$1-7 cells by the treatment of PDGF is 8 times more sensitive than those in control cells. 3T3${\gamma}$l-7 cell is useful for the screening of the inhibitors on the PDGF-induced cellular responses from large number of samples in a small volume(50 ${\mu}$l) and short time(5-15 min). Using these systems, we screened hundreds of herb-extracts for the inhibition of PDGF-induced IP3 formation and selected several extracts that showed the inhibition as the candidates for isolation and characterization of active substances. The determination of the acting point of selected extracts or fractions in the PDGF signaling pathway has been analyzing.

• Korean Journal of Animal Reproduction
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• v.18 no.2
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• pp.87-94
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• 1994

The objective of this study is to evaluate the developmental ability of mouse embryo in the presence of human amniotic fluid (hAF), The highest development rate was found in the culture media supplemented with 20% mid-term hAF but this rate was concomitantly reduced with more than 20% hAF. Furthermore, mouse two-cell embryos cultured in 20% mid-term hAF were developed more consistently to the expanded and hatched blastocyst stages compared to those cultured in simple medium. However, no significant differences in the embryo development rates were observed among the supplemented effects of 20% mid-term hAF, 0.3% bovine serum albumin (BSA), and 10% fetal calf serum (FCS), Development rates of two-ceiI mouse embryos cultured in 20% full-term hAF were declined compared to 20% mid-term hAF. Outgrowth of hatched blastocysts were observed when the embryos were cultured in medium containing 20% mid-term hAF or 10% FCS. But two-cell mouse embryos cultured in the presence of 20% full-term hAF or O.3% BSA was not observed their outgrowth. The kinetics of outgrowth processes in the presence of hAF were similar to those with 10% FCS. However, embryos with FCS showed a considerably greater extents of trophetodermal cell proliferation and outgrowth. Taken together, these data suggest that mid-term hAF may have a suitability for the mammalian embryos and induce embryonic outgrowth.

• Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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• 2018.05a
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• pp.588-591
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• 2018

Baculovirus was originally isolated from the alfalfa looper and contains a 134-kbp genome with 154 open reading frames (ORF). The major capsid protein VP39 together with some minor proteins forms the nucleocapsid ($21nm{\times}260nm$) that encloses the DNA with p6.9 protein. They are double-stranded, circular, supercoiled DNA molecules in a rod-shaped capsid. Wild-type baculoviruses exhibit both lytic and occluded life cycles that develop independently throughout the three phases of virus replication. Recombinant baculoviruses can transfer their vectors and express their recombinant proteins in a wide range of mammalian cell types. Especially, inclusion of a dominant selectable marker in these baculoviral vectors can express diverse recombinant genes in many cells. Baculoviral vectors were reconstructed with cytomegalovirus (CMV) promoter,uroplakin II promoter, polyhedron promoter, vesicular stomatitis virus G (VSVG), enhanced green fluorescent protein (EGFP), protein transduction domain (PTD) gene and so on. These reconstructed vectors were infected into various cell and cell lines. We performed transfection and expression of these recombinant vectors comparison with other control vectors. From this study, we knew that transfection and expression of these recombinant vectors have higher efficacy than any control vector. This work was supported by a grant from Mid-Career Researcher Program(NRF-2016R1A2B4016552) through the National Research Foundation of Korea(NRF) funded by the Ministry of Science, ICT & Future Planning(MSIP).

• Journal of Korean Ophthalmic Optics Society
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• v.20 no.3
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• pp.391-396
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• 2015

Purpose: The objective of this study was to investigate the visual system of the greater horseshoe bat (Rhinolophus ferrumequinum) by location analysis of some major neurotransmitters glutamate, ${\gamma}$-aminobutyric acid (GABA), acetylcholine, and their receptors, and $m{\ddot{u}}ller$ glial cells in retina. Methods: Standard immunocytochemical techniques were used after vibratome section of retinal tissues of adult greater horseshoe bat for this study. Immnoreactions in immunofluorescence images were analyzed using confocal microscope. Results: Anti-glutamate-immunoreactive neurons were mainly localized in the ganglion cell layer (GCL). The majority of anti-GABA-immunoreactive cells distributed in the inner nuclear layer (INL), and GABAA receptors were localized in the inner plexiform layer (IPL). Anti-choline acetyltransferase-immuoreactive cholinergic neurons were mainly located in the INL and GCL, and most of nicotinic acetylcholine receptors were localized in the IPL. The $m{\ddot{u}}ller$ cells in the retina of the greater horseshoe bat stretched theirs range from the GCL to outer nuclear layer (ONL). Conclusions: This study revealed that the retinas of the greater horseshoe bats contain the same major neurotransmitters and receptors, and glial cell in visually functional mammalian retinas. The present results may suggest that the greater horseshoe bats have the functional retinas for visual analysis through the organized retinal neural circuits.

• YAKHAK HOEJI
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• v.12 no.1_2
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• pp.1-15
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• 1968

The levels of activities of amylase, lipase and trypsin in both the pancreatic tissue and serum of 18 species of vertebrate animals were measured and enzymologically compared with each other. 1) The value of amylase in the pancreas of experimental mammalia has been found decreasing in the order pig, rat, dog, cat, rabbit and cow; that of pancreatic lipase has been found decreasing in the order of pig, dog, cat, rat, rabbit and cow; and that of trypsin has been found decreasing in the order of pig, cow, dog, rat, rabbit. Thus the value of all the above three kinds of enzymes were observed highest in pig, but in cow amylase and lipase were observed lowest while trypsin were observed considerably high. 2) In view of diets, the comparatively high values of pancreatic enzyme were observed in the ommivorous animals such as pig, rat, dog, while the values observed low in the herbivorous animals, such as cow and rabbit. 3) In the bovine, the values were observed moderately high except lipase which were found comparatively low. 4) In the Reptilia and Amphibia such a mud turttle and frog, the values were shown in similar measure with each other, that is, the pancreatic amylase and trypsin were observed considerably high while the lipase was found low. 5) In the species of Reptilia such as a viper and snake, the activities of pancreatic enzymes were not detected. But in the tissue of liver, stomach, activities of the enzymes were found considerably high. Lacertilia animals such as lizard the values of pancreatic enzymes were little observed. 6) In the fish in which the pancreatic tissue is scattered in the liver, the pancreatic enzymes were found in the liver tissue considerably higher than in the other tissues but lower than in the warm-blooded animals, especially the lipase was lower. 7) In generally the values of serum amylase and lipase were observed higher than those of man; and even in the cold-blooded animals in which the values of pancreatic enzymes were shown low or none, the values were also observed high. 8) The above three kinds of pancreatic enzyme values of those experimental animals have shown a tendency of higher degree in higher taxa than in lower taxa according to taxonomical order. 9) In view of tissue, the pancreatic cell was observed large in the mammalian animals such as rat and pig and cytoplasm was also abundantly contained in the acinous cell; and the bovine and the snake haave the pancreatic cells of the similar rosette form the comparatively large acinous cells of long rhombic form in the comparatively large acinous cells of long rhombic form in which the spindle shaped neucleus and the abundant cytoplasm were contained. In the fish the pancreatic cell were found scattered in the liver in which the very large pancreatic islet were found.

• Development and Reproduction
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• v.2 no.1
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• pp.29-37
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• 1998

To investigate the role of oviductal environment in early mammalian development, we examined the effects of bovine oviductal fluid (bOF) on the development of mouse 2-cell embryos in vitro. All of the embryos cultured in medium containing 5% or more of bOF underwent degeneration after 48 hr, whereas only 5% of embryos cultured in the absence of bOF degenerated. When bOF was heated at 65 \circ C for 30 min and then added to the culture medium, the embryotoxic effect of bOF was not removed at all such that none of the embryos remained alive after 48 hr. However, when bOF heated at 90 \circ C for 30 min was added to the culture, nearly most (95%) of embryos was alive. Similarly, pretreatment of bOF with 0.1% chymotrypsin for 1 hr or overnight following heating at 65 \circ C resulted in the development of 95.5% of mouse 2-cell embryos to early blastula after 48 hr culture in the presence of treated bOF. Interestingly addition of an anti-oxidant removed the evbryotoxic effect of bOF so that 91.0% of 2-cell embryos developed to morulae or blastulae in the presence of both 5% bOF and 10 mM of glutathione (GSH) after 48 hr culture. Neither oxidized form of GSH (GSSG) nor other antioxidants, however, could support the embryonic development in the presence of bOF. From these results, it is suggested that bOF contains a protein-like factor(s) which becomes embryotoxic by exposing in vitro, probably via oxidation reaction.

• Korean Journal of Food Science and Technology
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• v.45 no.2
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• pp.137-141
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• 2013

Flavonoids have various biological activities; however, their cellular uptake mechanism is beginning to be understood only recently. This review focuses on cellular flavonoids transport mechanisms in both plants and animals. In plants, flavonoids exist in various cellular compartments, providing a specialized transport system. Newly synthesized flavonoids can be transported from the endoplasmic reticulum to the vacuoles or extracellular space via cellular trafficking pathway. Among membrane transporters, ATP binding cassette, multidrug and toxic extrusion, bilitranslocase homologue transporters play roles in both the influx and efflux of cellular flavonoids across the cell membrane. In recent years, extensive researches have provided a better understanding on the cellular flavonoid transport in mammalian cells. Bilitranslocase transports flavonoids in various tissues, including the liver, intestine and kidneys. However, other transport mechanisms are largely unknown and thus, further investigation should provide detailed mechanisms, which can potentially lead to an improved bioavailability and cellular function of flavonoids in humans.

• Journal of Life Science
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• v.16 no.5
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• pp.716-722
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• 2006

Superoxide dismutase (SOD) is physiologically important in regulating cellular homeostasis and apoptotic cell death, and its mutations are the cause of familial amyotrophic lateral sclerosis (FALS). Mitochondrial serine protease HtrA2 has a pro-apoptotic function and has known to be associated with neurodegenerative disorders. To investigate the relationship between genes associated with apoptotic cell death, such as HtrA2 and SOD1, we utilized the pGEX expression system to develop a simple and rapid method for purifying wild-type and ALS-associated mutant SOD1 proteins in a suitable form for biochemical studies. We purified SOD1 and SOD1 (G93A) proteins to approximately 90% purity with relatively high yields (3 mg per liter of culture). Consistent with the result in mammalian cells, SOD1 (G93A) was more insoluble than wild-type SOD1 in E. coli, indicating that research on the aggregate formation of SOD1 may be possible using this pGEX expression system in E. coli. We investigated the HtrA2 serine protease activity on SOD1 to assess the relationship between two proteins. Not only wild-type SOD1 but also ALS-associated mutant SOD1 (G93A) were cleaved by HtrA2, resulting in the production of the 19 kDa and 21 kDa fragments that were specific for anti-SOD1 antibody. Using protein gel electrophoresis and immunoblot assay, we compared the relative molecular masses of thrombin-cleaved GST-SOD1 and HtrA2-cleaved SOD1 fragments and can predict that the HtrA2-cleavage sites within SOD1 are the peptide bonds between leucine 9-lysine 10 (L9-K10) and glutamine 23-lysine 24 (Q23-K24). Our study indicates that SOD1 is one of the substrate for HtrA2, suggesting that both HtrA2 and SOD1 may be important for modulating the HtrA2-SOD1-mediated apopotic cell death that is associated with the pathogenesis of neurodegenerative disorder.