• Korean Journal of Food Science and Technology
• /
• v.48 no.4
• /
• pp.306-316
• /
• 2016

This study was conducted to evaluate the functional characteristics and safety properties of lactic acid bacteria isolated from salt-fermented anchovy, a putative probiotic candidate. The following isolates were identified by biochemical profiles, carbohydrate fermentation patterns, and 16S rRNA sequencing: Enterococcus faecium AJ06, Leuconostoc mesenteroides AJ13, Pediococcus halophilus AJ22, Lactobacillus sakei AJ29, and Pediococcus pentosaceus AJ35. The strains AJ06, AJ22, AJ29 exhibited high tolerance to simulated gastric and intestinal juices and were able to produce bile salt hydrolase on MRS agar plates supplemented with taurocholic acid and/or taurodeoxycholic acid. The strains AJ22 and AJ29, which demonstrated high adherence to Caco-2 cells and resistance to various antibiotics, effectively inhibited the growth of food-borne pathogens by the production of antimicrobial substances. These strains did not show ${\alpha}-$ or ${\beta}$-haemolysis on blood agar. Furthermore, biogenic amines in MRS broth containing the precursor amino acids were not mutagenic in Salmonella Typhimurium TA98 and TA100.

• The Journal of Korean Medicine
• /
• v.29 no.2
• /
• pp.182-192
• /
• 2008

Objective: The purpose of this study was to evaluate on the antimicrobial effect on the periodontal pathogens and anti-inflammatory effect of Eriobotryae folium. Eriobotryae folium are constituent herbs of Gagamgamroum, which has been used for a long time in oriental medicine as a herbal medicine for treating halitosis and toothache. Method: Eriobotryae folium was prepared by extracting medicinal herb with water. We investigated antimicrobial activity by the minimum inhibitory concentration (MIC) test. We also investigated inhibition of $IL-1{\beta}-induced$ collagenase (mmp-1), stromelysin-1 (mmp-3), interleukin-6 gene expression in human gingival fibroblasts using RTPCR analysis. Result: The antimicrobial effects of Eriobotryae folium was evaluated with MIC against periodontopathogens; Porphyromonas gingivalis 2561, W50, A7A1-28, 9-14K-1, Prevotella intermedia 28, and Actinobacillus actinomycetemcomitans Y4. MICs of Eriobotryae folium were 1.25 mg/ml, 2.5 mg/ml, 0.625 mg/ml, 1.25 mg/ml, 10 mg/ml and 10 mg/ml. The anti-inflammatory effect of Eriobotryae folium was evaluated with influence of herbs on the $IL-1{\beta}-induced$ expression of mmp-1, mmp-3, and interleukin-6. $IL-1{\beta}$ increased mmp-1, mmp-3, and interleukin-6 mRNA levels. Eriobotryae folium significantly inhibited $IL-1{\beta}-induced$ mmp-1, mmp-3, and interleukin-6 gene expressions in a dose-dependent manner. Conclusion: These results suggested that Eriobotryae folium might reduce the excessive proteolytic capacity of the gingival fibroblast during inflammation and could be developed as a new drug for periodontitis.

• Journal of Microbiology and Biotechnology
• /
• v.16 no.8
• /
• pp.1201-1209
• /
• 2006

The purpose of this study was to isolate probiotic lactic acid bacteria (LAB) that produce bile salt hydrolase (BSH), and to evaluate its effects on serum cholesterol level. One-hundred-twenty bacterial colonies were initially isolated from human feces, and five strains were selected after screening based on their resistance to acids, tolerance against bile salts, and inhibitory activity on Escherichia coli. The Lactobacillus plantarum strain with the highest level of BSH activity was identified using 16S rRNA sequences, and was named L. plantarum CK 102. L. plantarum CK 102 at a level of 1.36$\times$10$^8$cfu/ml survived in pH 2 buffer for 6 h and exhibited excellent tolerance for bile salt. Coculturing the strain with E. coli in MRS broth resulted in strong inhibition against growth of E. coli at 18 h. Furthermore, the potential effect of CK 102 on serum cholesterol level was evaluated in rats. Thirty-two rats [Sprague-Dawley (SD) male, 129$\pm$l g, 5 weeks old] were divided into four groups of eight each. For six weeks, Group 1 was fed a normal diet (negative control); Group 2 was fed a cholesterol-enriched diet (positive control); Group 3 was fed a cholesterol-enriched diet plus L. plantarum CK 102 at 1.0$\times$10$^7$cfu/ml; and Group 4 was fed a cholesterol-enriched diet plus L. plantarum CK 102 at 5.0$\times$10$^7$cfu/ml. Blood samples were collected, serum lipids were analyzed, and weights of the organs were measured. Total blood cholesterol level, triglyceride, LDL-cholesterol, and free-cholesterol values were lower in rats that were fed 1. plantarum CK 102 than in those not fed L. plantarum CK 102. This cholesterol lowering effect implies that L. plantarum CK 102 could be utilized as an additive for health-assistance foods. In conclusion, these results suggest that the 1. plantarum CK 102 isolated could be used commercially as a probiotic.

• Journal of Microbiology and Biotechnology
• /
• v.27 no.3
• /
• pp.460-470
• /
• 2017

Mussels are major fouling organisms causing serious technical and economic problems. In this study, antifouling activity towards mussel was found in three compounds isolated from a marine bacterium associated with the sea anemone Haliplanella sp. This bacterial strain, called PE2, was identified as Vibrio alginolyticus using morphology, biochemical tests, and phylogenetic analysis based on sequences of 16S rRNA and four housekeeping genes (rpoD, gyrB, rctB, and toxR). Three small-molecule compounds (indole, 3-formylindole, and cyclo (Pro-Leu)) were purified from the ethyl acetate extract of V. alginolyticus PE2 using column chromatography techniques. They all significantly inhibited byssal thread production of the green mussel Perna viridis, with $EC_{50}$ values of $24.45{\mu}g/ml$ for indole, $50.07{\mu}g/ml$ for 3-formylindole, and $49.24{\mu}g/ml$ for cyclo (Pro-Leu). Previous research on the antifouling activity of metabolites from marine bacteria towards mussels is scarce. Indole, 3-formylindole and cyclo (Pro-Leu) also exhibited antifouling activity against settlement of the barnacle Balanus albicostatus ($EC_{50}$ values of 8.84, 0.43, and $11.35{\mu}g/ml$, respectively) and the marine bacterium Pseudomonas sp. ($EC_{50}$ values of 42.68, 69.68, and $39.05{\mu}g/ml$, respectively). These results suggested that the three compounds are potentially useful for environmentally friendly mussel control and/or the development of new antifouling additives that are effective against several biofoulers.

• Journal of Applied Biological Chemistry
• /
• v.55 no.4
• /
• pp.227-234
• /
• 2012

We observed that oil began to accumulate at 25 seed days after flowering (DAF) and reached the maximum potential at 35 seed DAF of oilseed rape, and the greatest weight of 100 seeds was obtained at 35 seed DAF. To survey a broad analysis of gene expression in developing embryos of Brassica napus, the Bn 300k microarray have been constructed. The Bn 300k Microarrary was designed from 80,696 unigenes clustered from 543,448 ESTs and 780 cDNA at NCBI. These arrays have been hybridized in a series of experiments with probes derived from seeds and leaf of B. napus. Approximately 8.5% of the 7,000 genes were expressed as ratios 2-fold higher in seed (25 DAF) than leaves and 0.4% at ratios 10. Also we observed that storage and cell differentiation-related genes were highly expressed at 10 DAF, whereas energy-related genes including fatty acid metabolism were increased up depending on seed maturation using Microarray, which was confirmed by reverse transcriptase polymerase chain reaction. These results suggest that B. napus arrays provide a very useful data set of seed-specific expression that can be further analyzed by examination of the promoter regions of these genes and help our understanding of the complex regulatory network in developing seeds.

• Reproductive and Developmental Biology
• /
• v.34 no.1
• /
• pp.47-53
• /
• 2010

Glycoprotein hormones have a common $\alpha$-subunit that is involved in the signaling pathway together with G protein, adenylcyclase and cAMP induction; however, it is an unclear how this common structure is related to hormonal action. To determine the biological functions of the COOH-terminal amino acids in the $\alpha$-subunit of these glycoprotein hormones, a tethered-molecule was constructed by fusing the $NH_2$-terminus of the $\alpha$-subunit to the COOH-terminus of the $\beta$-subunit of equine chorionic gonadotropin (eCG). The following deletion mutants were created by PCR; Ile was inserted at position 96 to form ${\Delta}96$, Lys was substituted at position 95 to form ${\Delta}95$, His was inserted at position 93 to form ${\Delta}93$ and Tyr was substituted at position 87 to form ${\Delta}87$. Each mutant was transfected into CHO-K1 cells. Tethered-wt eCG, and ${\Delta}96$, ${\Delta}95$, and ${\Delta}93$ mutants were efficiently secreted into the medium but the ${\Delta}87$ mutant was not secreted. Interestingly, the RT-PCR, real-time PCR, and northern blot analyses confirmed that the RNA was transcribed in the ${\Delta}87$ mutant. However, the ${\Delta}87$ mutant protein was not detected in the medium or the intracellular fraction of the cell lysates. The LH- and FSH-like activities of the recombinant proteins were assayed in terms of cAMP production using rat LH/CG and rat FSH receptors. The metabolic clearance rate (MCR) was determined by injecting rec-eCG (2 IU) into the tail vein. The ${\Delta}95$ and ${\Delta}93$ mutants were completely inactive in both the LH- and FSH-like activity assays. The ${\Delta}96$ mutant showed slight activity in the LH-like activity assay. In comparison to the wild type, the activity of the ${\Delta}96$ mutant in the FSH-like activity assay was the highest among all the mutants. The MCR assay in which rec-eCG was injected showed a peak at 10 min in all the treatment groups, which disappeared 4 h after injection. These results imply a direct interaction between the receptor and the COOH-terminal region of the a-subunit. The data also reveal a significant difference in the mechanism by which the eCG hormone interacts with the rLH and rFSH receptors. The COOH-terminal region of the $\alpha$-subunit is very important for the secretion and functioning of this hormone.

• Journal of the Korea Academia-Industrial cooperation Society
• /
• v.12 no.1
• /
• pp.312-317
• /
• 2011

Vanilloid receptor TRPV1 (known as capsaicin channel, transient receptor potential vanilloid 1) is known to be a key protein in the pain signal transduction. However, the proteins controlling the activity of the channel are not much known yet. Recently mouse Rab11-FIP3 (Rab11-family interaction protein 3) was found and reported to interact with rat TRPV1. Rab11 has been shown to play a key role in a variety of cellular processes including plasma membrane recycling, phagocytosis, and transport of secretory proteins from the trans-Golgi network. Therefore, Rab11-FIP3 was proposed to be involved in the membrane trafficking of TRPV1. In this study, the unreported rat Rab11-FIP3 was yet cloned in order to show the specific interaction of the TRPV1 and Rab11-FIP3 in the same species of rat and to examine the membrane trafficking of TRPV1. The result showed that rat Rab11-FIP3 is expected to have 489 amino acids and showed 80% identity with that of human and over 90% identity with that of mouse. Rab11-FIP3 was found to be expressed in heart, brain, kidney, testis using northern and western blot analyses. We also found that rat Rab11-FIP3 was colocalized with rat TRPV1 but not with TRPV2 of same family in the rat brain by using immunohistochemistry showing that two proteins interact specifically, suggesting the role of Rab11-FIP3 in the membrane trafficking.

• Asian-Australasian Journal of Animal Sciences
• /
• v.21 no.2
• /
• pp.277-290
• /
• 2008

The process of wound repair involves an ordered sequence of events such as overlapping biochemical and cellular events that, in the best of circumstances, result in the restoration of both the structural and functional integrity of the damaged tissue. An important event during wound healing is the contraction of newly formed connective tissues by fibroblasts. The polypeptide growth factors, like transforming growth factor-${\beta}$(TGF-${\beta}$, insulin-like growth factor I (IGF- I) and epidermal growth factor (EGF), play very important mediator roles in the process of wound contraction. Deer antlers, as models of mammalian regeneration, are cranial appendages that develop after birth as extensions of a permanent protuberance (pedicle) on the frontal bone. Antlers contain various growth factors which stimulate dermal fibroblast growth. They are involved in digestion and respiration and are necessary for normal wound healing and skin health. In order to investigate and evaluate the effects of red deer antlers on skin wound site, the speed of full-thickness skin wound healing and the expression of IGF-I, TGF-${\beta}$ and EGF in skin wounds, three groups of skin full-thickness rat models with a high concentration of antler ointment, a low concentration of antler ointment and without antler ointment were compared. At post-injury days 0, 2, 4, 8, 16, 20, 32, 40 and 60, the skin wound area was measured, the expressions of IGF-I, TGF- ${\beta}$ and EGF mRNA were detected by reverse transcriptase polymerase chain reaction (RT-PCR) and collagen formation by sirius red dye and the localization of IGF-I, TGF-${\beta}$ and EGF peptides were inspected by histological immunohistochemical techniques. Wound healing was significantly more rapid in antler treated skins. In addition, the wound treated with a high concentration antler ointment, a low concentration antler ointment, and the control closed completely at post-injury day 40, day 44 and day 60, respectively. Via RT-PCR, the expressions of IGF-I (day 8 and day 16), TGF-${\beta}$(day 8, day 16 and day 20) and EGF (day 4, day 8, day 16, and day 32) were obviously up-regulated in high concentration antler-treated skins compared to control skins. Similar results could be seen in the histological detection of collagen dye and immunohistochemical methods using the corresponding polyclone antibodies of IGF-I, TGF-${\beta}$ and EGF. These results illustrate that antlers stimulate and accelerate the repair of cutaneous wounds.

• Asian Pacific Journal of Cancer Prevention
• /
• v.15 no.12
• /
• pp.4969-4976
• /
• 2014

Background: miR-152 is involved in the genesis and development of several malignancies. However, its role in HCC has not been fully clarified. The aim of this study was to investigate the clinicopathological significance of miR-152 and its effect on the malignant phenotype of HCC cells. Methods: miR-152 expression was detected using real-time quantitative RT-PCR in 89 pairs of HCC formalin-fixed paraffin-embedded and their adjacent tissues. Functionally, in vitro effects and mechanisms of action of miR-152 on proliferation, viability, caspase activity, apoptosis and motility were explored in HepG2, HepB3 and SNU449 cells, as assessed by spectrophotometry, fluorimetry, fluorescence microscopy, wound-healing and Western blotting, respectively. Results: miR-152 expression in HCC was downregulated remarkably compared to that in adjacent hepatic tissues. miR-152 levels in groups of advanced clinical stage, larger tumor size and positive HBV infection, were significantly lower than in other groups. A miR-152 mimic could suppress cell growth, inhibit cell motility and increase caspase activity and apoptosis in HCC cell lines. Furthermore, Western blotting showed that the miR-152 mimic downregulated Wnt-1, DNMT1, ERK1/2, AKT and TNFRS6B signaling. Intriguingly, inverse correlation of TNFRF6B and miR-152 expression was found in HCC and bioinformatics confirmed that TNFRF6B might be a target of miR-152. Conclusions: Underexpression of miR-152 plays a vital role in hepatocarcinogenesis and lack of miR-152 is related to the progression of HCC through deregulation of cell proliferation, motility and apoptosis. miR-152 may act as a tumor suppressor miRNA by also targeting TNFRSF6B and is therefore a potential candidate biomarker for HCC diagnosis, prognosis and molecular therapy.

• The Korean Journal of Physiology and Pharmacology
• /
• v.9 no.6
• /
• pp.333-339
• /
• 2005

B/K protein is a novel protein containing double C2-like domains. We examined the specific signaling pathway that regulates the transcription of B/K in PC12 cells. When the cells were treated with forskolin ($50{\mu}M$), B/K mRNA and protein levels were time-dependently decreased, reaching the lowest level at 3 or 4 hr, and thereafter returning to the control level. Chemicals such as dibutyryl-cAMP, cellpermeable cyclic AMP (cAMP) analogue and CGS21680, adenosine receptor $A_{2A}$ agonist, also repressed the B/K transcription. However, 1,9-dideoxyforskolin did not show inhibitory effect on B/K transcription, suggesting direct involvement of cAMP in the forskolin-induced inhibition of B/K transcription. Effect of forskolin, dibutyryl cAMP and CGS21680 was significantly reduced in PKA-deficient PC12 cell line (PC12-123.7). One cAMP-response element (CRE)-like sequence (B/K CLS) was found in the promoter region of B/K DNA, and electrophoretic mobility shift assay indicated its binding to CREM and CREB. Forskolin significantly suppressed the promoter activity in CHO-K1 cells transfected with the constructs containing B/K CLS, but not with the construct in which B/K CLS was mutated (AC：TG). Taken together, we suggest that the transcription of B/K gene in PC12 cells may be regulated by PKA-dependent mechanism.