Journal of the Korean Society of Food Science and Nutrition
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v.32
no.8
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pp.1245-1252
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2003
A starter formulation was developed to substitute a baker's yeast with natural starter when manufacturing bread products. To develop an active starlet, starter was formulated varying with types of wheat flours, level of water contents and various nutrients. Activities of starter were investigated in terms of viable counts of microbes and change of pH and total titratible acidity Domestic wheat flours contain 100 times more number of lactic acid bacteria than yeast regardless of types of wheat flours. The more protein contents in wheat flours, the more stable microbes in starter. This was considered to be the result of buffering effect of wheat proteins. The optimum level of protein content to ensure the activity of starter was more than 12.0%. Optimum level of water content in active starter was 110% based on strong flour. The more water or the less water had the tendency of decreasing viable counts of microbes. Addition of salt and sucrose had increased the activity of starters. However oligosaccharides did not affect the activity of starter. The optimum concentrations of salt and sucrose were 1.0% and 5.0% respectively. Bread with the starter was higher scored than breads with yeast in terms of all the quality and sensory characteristics except their volumes. In conclusion, a starter formulated with strong flour 100%, water 110%, salt 1% and sucrose 5% was considered to have high potential as a substitute of yeast in making natural bread.
Kim, Eunsung;Park, Jiyeong;Lee, Sanghoon;Kim, Yonggyun
Korean journal of applied entomology
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v.53
no.1
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pp.15-26
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2014
The black soldier fly, Hermetia illucens, larvae may depend on indigenous bacteria in the intestine to feed and digest diverse food sources. To prove this hypothesis, we isolated and identified the intestinal bacteria of the black soldier fly for their digestive and antimicrobial abilities. The last instar larvae had long digestive tracts, which were about seven times longer than its body length. An individual of H. illucens larvae possessed a total of $5.0{\pm}10^6$ bacteria in the whole intestine, of which more than 98% bacteria were located in the hindgut. Three different bacterial isolates cultured on nutrient agar (NA) medium were detected in the intestine and identified as Morganella morganii, Providencia rettgeri and Bacillus halodurans by Biolog microbial identification system. Analysis of 16S rDNA sequences of the intestinal bacteria detected the additional bacteria of Proteus mirabilis, Providencia alcalifaciens, and Providencia sp. These intestinal bacteria cultured on NA medium exhibited high resistance to 4 antibiotics and inhibited growth of other microbes which are mainly plant pathogens. Also, these bacteria exhibited catalytic activities to degrade cellulose, lipid, proteins, and carbohydrates. These results suggest that H. illucens larvae possess intestinal bacteria that may play crucial roles in their digestive physiology.
Salmonella infections cause the disease in pigs but also some zoonotic Salmonella serotypes can be transmitted to human through swine products, resulting in food poisoning. The objective of this study was to investigate the bacteriological prevalence and detection of invA gene using Salmonella specific polymerase chain reaction (PCR), the epidemiological characteristics related to Salmonella strains cultured from pig samples in Gangwon areas using serotyping, random amplified polymorphic DNA (RAPD) and pulsed field gel electrophoresis (PFGE) methods. During the period of November 2001 through April 2002, 1,174 ileocecal lymph node were collected from the slaughtered pigs raised in 38 farms located in Gangwon province. The samples were submerged in boiling water and macerated in saline and lymph node homogenates were inoculated into Tetrathionate broth with iodine (TTB, Difco, 0.5% iodine was added) for enrichment growth. Then additional tests were performed using several mediums, and suspects were identified by API 20E kit (BioMerieux) and PCR. Of total 1,174 samples from 38 farms, 44 (3.7%) were isolated as Salmonella spp from 13 farms (34.2%). Of 44 isolates, 31 were in Yangyang region, followed by 9 in Goseong, 2 in both Gangneung and Sokcho. However, there was no difference in regional isolation frequency. All isolates have a 521bp amplified product in Salmonella specific PCR with primer invA which encodes in proteins for invasion of epithelial cells. Of 44 recovered serotypes, 23 (52.3%) were S Eingedi, 10 (22.7%) S Schwarzengrund, 9 (20.5%) S Typhimurium, and 2 (4.5%) S Mbandaka. In RAPD analysis, there appeared to be unique bands distinguishing each serotype, although similarities exist between the different serotypes. Four serotypes of 44 Salmonella isolates appeared to fall into 14 different RAPD types. In PFGE analysis, 9 S Typhimurium were tested with XbaI enzyme and SpeI enzyme. The combination of results obtained with two enzymes subdivided the 9 S Typhimurium into 4 PFGE types.
Ameloblasotma is slowly growing, locally invasive neoplasm with a potentially destructive behavior. The epithelium of ameloblastoma is thought to have an intrinsic growth potential and has been shown to present a higher rate of proliferation as compared to odontogenic cysts with low local recurrence rate. The molecular mechanisms that regulate the cell growth and invasion of ameloblastoma cells are unknown. Bcl-2 protein, which prevent apoptosis, is expressed in immortalized ameloblastoma cell line(AM-1)(Harada et al 1998). Expression of bcl-2 protein occurs in tooth germs, whose epithelial component may act as the histogenic precursor of ameloblastoma. Bax is considered as a main effector of apoptosis. Bax forms homodimers and also heterodimers with bcl-2. p53 tumor supressor gene participates not only in cell proliferation control but also in induction of apoptosis. The objective of the present study was to evaluate the apoptosis related protein expression in odontogenic cyst and ameloblastoma. A total of 10 dentigerous cysts and 16 ameloblastomas were used in the present study. Dentigerous cyst showed negative or slight positive for p53 and bcl-2 but strongly positive for bax, ameloblastoma, on the other hand, strongly positive for p53 and bcl-2 but weekly positive for bax. Bcl-2 was expressed for ameloblastoma mainly in outer layer or whole layer of epithelium and for dentigerous cyst mainly in basal layer. The difference in expression of apoptosis related protein in dentigerous cyst and ameloblastoma might explain the peculiar aggressive growth pattern of ameloblastoma.
Kim, Yu-Na;Lee, Jae-Ran;Kim, Mu-Chan;Kim, Sung-Bae;Chang, Yong-Keun;Hong, Soon-Kwang;Kim, Chang-Joon
Korean Chemical Engineering Research
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v.49
no.6
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pp.840-845
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2011
Degradation products of agarose are biologically active and thus used as an ingredient in pharmaceuticals or functional cosmetics. Furthermore, it has been strongly considered as a substrate for bio-ethanol fermentation. Recently, we isolated new agarase-producing microorganism, Pseudoalteromonas sp. from south sea of Korea. In this study, we aimed to separate and purify the agarase from culture broth of this strain. Separation of agarase was performed by ion- exchange chromatography on DEAE-Sepharose resin. Equilibrium pH and volume ratio of resin to the amount of protein were optimized for the efficient adsorption of protein. 410 ${\mu}g$ of protein was completely adsorbed to 3 mL of resin at pH 7.5. The total amount of eluted protein increased as NaCl concentration increased to 400 mM at isocratic elution. Agarase was separated by linear gradient elution of NaCl (0~1,000 mM). Three major protein peaks were observed and the presence or absence of agarase in these eluted proteins was measured by Lugol's staining technique. Only six eluted protein fractions showed strong agarase activity.
Physiological plasticity of insects can be closely related with their epigenetic change. This hypothesis was tested using a polyphagous lepidopteran insect, Spodoptera exigua, by assessing the effects of different diets on development and DNA methylation. Three different diets (Welsh onion (WO), Chinese cabbage (CC), artificial diet (AD)) were assessed by feeding a cohort of larvae from neonate to last instar. There were significant differences in larval developmental rate, pupal weight and adult emergence according to diet treatments. AD-fed larvae exhibited the fastest developmental rate along with the highest pupal weight and adult emergence. Among natural hosts, WO was more favorable for development of S. exigua than CC. Total hemolymph proteins and sugars in the last instar larvae were varied among different diets. Gene expression of an insulin-like peptide (SeILP1) presumably associated with development was also varied among diets. Cytosine methylation of genomic DNA was assessed using a monoclonal antibody. Genomic DNA of S. exigua larvae was methylated. DNA methylation was apparently varied among different diet-fed larvae. The facts that a cohort of S. exigua was differentiated in developmental rate and cytosine methylation by different diets suggest that epigenetic factor(s) may play a crucial role in the physiological plasticity.
Pure Bombus ignitus venom samples were submitted to two-dimensional gel electrophoresis. A total of 64 excised spots were analyzed by mass spectrometry. Three main proteins resulted in the identification have not been described in other bee venoms before. Dose-dependence against human carcinoma (Hep3B, BT-20, A549 and AGS) were observed from 1ng/ml to 100ng/ml. Expecially, the treatment of 100ng/ml B. ignitus venoms showed the highest cytotoxicity with 55% against hepatocellular carcinoma (Hep3B). The B. ignitus venoms showed strong antimicrobial activities against Enterococcus faecium and Shigella sonnei, and practically antimicrobial activity against the other microorganisms tested. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of E. faecium and S. sonnei, were 0.256ug/ml, respectively.
The uptake of glucose for metabolism and growth is essential to most animal cells and is mediated by glucose-transporter (GLUT) proteins. The aim of this study was to determine which class of glucose transporter molecules was responsible for uptake of glucose in the mouse early embryo and at which stage the corresponding genes were expressed. In addition, co-culture system with vero cell was used to investigate the effect of the system on GLUT expression. Two-cell stage embryos were collected from the superovulated ICR female and divided into 3 groups. As a control, embryos were cultured in 0.4% BSA-T6 medium which includes glucose. For the experimental groups, embryos were cultured in either co-culture system with vero cells or glucose-free T6 medium supplemented with 0.4% BSA and pyruvate as an energy substrate. 2-cell to blastocyst stage embryos in those groups were respectively collected into microtubes (50 embryos/tube). Total RNA was extracted and RT-PCR was performed. The products were analysed after staining ethidium bromide by 2% agarose gel electrophoresis. Blastocysts were collected from each group at l20hr after hCG injection. They were fixed in 2.5% glutaraldehyde, stained with hoechst, and mounted for observation. In control, GLUT1 was expressed from 4-cell to blastocyst. GLUT2 and GLUT3 were expressed in morula and blastocyst. GLUT4 was expressed in all stages. When embryos were cultured in glucose-free medium, no significant difference was shown in the expression of GLUT1, 2 and 3, compared to control. However GLUT4 was not expressed until morular stage. When embryos were co-cultured with vero cell, there was no significant difference in the expression of GLUT1, 2, 3 and 4 compared to control. To determine cell growth of embryos, the average cell number of blastocyst was counted. The cell number of co-culture ($93.8{\pm}3.1$, n=35) is significantly higher than that of control and glucose-free group ($76.6{\pm}3.8$, n=35 and $68.2{\pm}4.3$, n=30). This study shows that the GLUT genes are expressed differently according to embryo stage. GLUTs were detectable throughout mouse preimplantation development in control and co-culture groups. However, GLUT4 was not detected from 2- to 8-cell stage but detected from morula stage in glucose-free medium, suggested that GLUT genes are expressed autocrinally in the embryo regardless of the presence of glucose as an energy substrate. In addition, co-culture system can increase the cell count of blastocyst but not improve the expression of GLUT. In conclusion, expression of GLUT is dependent on embryo stage in preimplantation embryo development.
Journal of the Korean Association of Oral and Maxillofacial Surgeons
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v.32
no.4
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pp.295-307
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2006
Styela clava, called non-native tunicate or sea squirt, is habitat which include bays and harbors in Korea and several sites in the sea faced world. We fabricate cellulose membrane nerve conduit (CMNC) from this native sea squirt skin, and evaluate the capacity of promoting peripheral nerve regeneration in the rat sciatic nerve defect model. After processing the pure cellulose membrane from the sea squirt skin as we already published before, CMNC was designed as a non-tubular sheet with 14 mm length and 4 mm width. Total eleven male Spraque-Dawley rats (12 weeks, weighing 250 to 300g) were divided into sham group (n=2), silicone tube grafted control group (n=3) and experimental group (n=6). Each CMNC grafted nerve was evaluated after 4, 8 and 12 weeks in the experimental group, and after 12 weeks, sciatic function was evaluated with sciatic function index (SFI) and gait analysis, and histomorphology of nerve conduit and the innervated tissues of sciatic nerve were all examined using image analyzer and electromicroscopic methods in the all groups. The regenerated axon and nerve sheath were found only in the inner surface of the CMNC after 4 weeks and became more thicker after 8 and 12 weeks. In the TEM study, CMNC grafted group showed more abundant organized myelinated nerve fibers with thickened extracellular matrix than silicone conduit grafted group after 12 weeks. The sciatic function index (SFI) and ankle stance angle (ASA) in the functional evaluation were $-47.2{\pm}3.9$, $35.5^{\circ}{\pm}4.9^{\circ}$ in CMNC grafted group (n=2) and $-80.4{\pm}7.4$, $29.2^{\circ}{\pm}5.3^{\circ}$ in silicone conduit grafted group (n=3), respectively. And the myelinated axon was 41.59% in CMNC group and 9.51% in silicone conduit group to the sham group. The development of a bioactive CMNC to replace autogenous nerve grafts offers a potential and available approach to improved peripheral nerve regeneration. As we already published before, small peptide fragment derived from the basement membrane matrix proteins of squirt skin, which is a kind of anchoring protein composed of glycocalyx, induced the effective axonal regeneration with rapid growth of Schwann cells beneath the inner surface of CMNC. So the possibilities of clinical application as a peripheral nerve regeneration will be able to be suggested.
Background: Ginsenoside Rh2 (G-Rh2) is a ginseng saponin that is widely investigated because of its remarkable antitumor activity. However, the molecular mechanism by which (20S) G-Rh2 triggers its functions and how target animals avoid its cytotoxic action remains largely unknown. Methods: Phage display was used to screen the human targets of (20S) G-Rh2. Fluorescence spectroscopy and UV-visible absorption spectroscopy were used to confirm the interaction of candidate target proteins and (20S) G-Rh2. Molecular docking was utilized to calculate the estimated free energy of binding and to structurally visualize their interactions. MTT assay and immunoblotting were used to assess whether human serum albumin (HSA), bovine serum albumin (BSA), and bovine serum can reduce the cytotoxic activity of (20S) G-Rh2 in HepG2 cells. Results: In phage display, (20S) G-Rh2-beads and (20R) G-Rh2-beads were combined with numerous kinds of phages, and a total of 111 different human complementary DNAs (cDNA) were identified, including HSA which had the highest rate. The binding constant and number of binding site in the interaction between (20S)-Rh2 and HSA were $3.5{\times}10^5M^{-1}$ and 1, and those in the interaction between (20S) G-Rh2 and BSA were $1.4{\times}10^5M^{-1}$ and 1. The quenching mechanism is static quenching. HSA, BSA and bovine serum significantly reduced the proapoptotic effect of (20S) G-Rh2. Conclusion: HSA and BSA interact with (20S) G-Rh2. Serum inhibited the activity of (20S) G-Rh2 mainly due to the interaction between (20S) G-Rh2 and serum albumin (SA). This study proposes that HSA may enhance (20S) G-Rh2 water solubility, and thus might be used as nanoparticles in the (20S) G-Rh2 delivery process.
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