• Clinical and Experimental Reproductive Medicine
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• v.27 no.3
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• pp.235-243
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• 2000

Objective: Our present studies were conducted to examine more effective isolating method of preantral follicles from mouse ovaries. Methods: ICR mice (3-6 weeks old) were sacrificed through cervical dislocation and their ovaries were removed and put into watch glasses containing Hams F-10 supplemented with 10% fetal bovine serum (FBS). Preantral follicles were isolated by three different methods; 1) enzymatical method and 2) mincing method, and 3) scraping method. Enzymatical method was carried out as following. Ovaries were bisected with a pair of fine 30G needles. Bisected ovaries were incubated at $37^{\circ}C$ and 5% $CO_2$ incubator in 2-well dish containing Hams F-10 supplemented with collagenase 600 lU/ml and DNAse 20 lU/ml. After 20 min., follicles were isolated by repeated pipetting. Isolated preantral follicles were collected, and the remnant of tissues was placed in incubator and previous procedure was repeated. Mincing method was carried out with a pair of fine 30G needles attached to 1 ml syringes and minced ovary. Scraping method was carried out with a pair of fine 30G needles and scratched to surface of ovary. The differences between isolating methods were analyzed using Student's t-test and Chi-square. Results were considered statistically significant when ${\rho}$ value was less than 0.05. Results: In handling time, mincing or scraping method ($28{\pm}3.42$ min or $16{\pm}1.58$ min) were significantly (p<0.00001) shorter than enzymatical method ($72{\pm}1.69$ min), and scraping method was significantly (p<0.01) shorter than mincing method. Total number of isolated follicles was significantly (p<0.0001) higher in enzymatical method ($49.8{\pm}3.91$) than in mincing or scraping method ($25.3{\pm}2.33$ or $20.5{\pm}1.75$). Isolated follicles in ${\leq}$90${\mu}m$ were significantly (p<0.005) higher in enzymatical method ($15{\pm}1.71$) than in mincing or scraping method ($7.8{\pm}0.98$ or $8.1{\pm}1.31$). In 91~130 ${\mu}m$, isolated follicles were significantly (p<0.0005) higher in enzymatical method ($33{\pm}3.27$) than in mincing or scraping method ($16.3{\pm}1.82$ or $10.7{\pm}1.38$). In ${\geq}$ 131 ${\mu}m$, isolated follicles were not significantly differences between all groups. In equal sizes, the rate of isolated follicles in ${\leq}$ 90 ${\mu}m$ was highest in scraping method (39.6% vs. enzymatical method: 30.1%, p<0.05; mincing method: 30.9%, p=0.11719, NS). Rate of follicles in $91{\sim}130$ ${\mu}m$ was significantly (p<0.05) lower in scraping method (52.7%) than in enzymatical or mincing method (66.3% or 64.5%). Rate of follicles in ${\geq}$131 ${\mu}m$ was highest in scraping method (8.3% vs. enzymatical or scraping method: 3.6%, p<0.05 or 4.6%, p=0.19053, NS). Conclusions: This study suggests that scraping method is simple and useful for isolation of preantral follicles, because this method reduced handling time and recovered enough follicles. The recovered rate of isolated follicles in diameter of 91 ~ 130 ${\mu}m$ was highest in all methods.

• Food Science and Preservation
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• v.22 no.1
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• pp.134-144
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• 2015

The anti-diabetic effects of Allium tuberosum Rottler extracts (ATE) and ATE fermented with lactic acid bacteria in db/db mice were evaluated. The electron donating activity of ATE fermented with Lactobacillus plantarum, and Lactobacillus casei, respectively, increased compared to that of ATE, but the superoxide radical scavenging activity of the ATE incubated with L. plantarum decreased. The superoxide radical scavenging activity of the ATE fermented with both L. plantarum and L. casei was similar to that of the ATE. Therefore, fermented ATE (FATE) was prepared for in vivo testing by incubating it with both L. plantarum and L. casei. The db/db mice were divided into six groups: normal (non-diabetic mice), diabetic control (DM), and four experimental groups administered 200 or 400 mg/kg/day ATE (ATE200 and ATE400) and 200 or 400 mg/kg/day FATE (FATE200 and FATE400). Weight gain was significantly inhibited in the FATE200 group compared with that in the other db/db mice groups (p<0.05). The areas under the curve of the ATE400 and FATE400 groups were significantly smaller than that of the DM group in the glucose tolerance evaluation. The serum glucagon-like peptide-1 levels in the ATE400 and FATE400 groups increased. These results indicate that administering ATE and FATE may be effective against anti-hyperglycemia by regulating insulin resistance. In particular, FATE may be beneficial for controlling obesity in type 2 diabetes.

• Food Science and Preservation
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• v.24 no.1
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• pp.13-20
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• 2017

This study was carried out to investigate the quality characteristics of fried fish paste prepared with different amounts of squeezed Aronia melanocarpa juice. Squeezed Aronia melanocarpa juice (AMJ) was incorporated into fish paste at different levels (containing 2, 7, and 12 g of Aronia melanocarpa juice in 2 AMJ, 7 AMJ, and 12 AMJ, respectively) based on the total weight of water. Sugar contents and total acidity increased with increasing AMJ content. With increasing amounts of AMJ in fried fish paste, L value inside and on the surface decreased, a value increased, and b value inside decreased, whereas b value on the surface increased. pH decreased with increasing levels of AMJ. As the result of textural properties, folding test in all samples showed that AA means good flexibility. The strength, hardness, and chewiness of fried fish paste with AMJ increased while cohesiveness was not significantly different. Total polyphenol contents increased with higher levels of AMJ. DPPH radical scavenging activity was significantly higher than those of the control. In the sensory evaluation, fried fish paste containing 7 AMJ received the highest score than both the control and other samples.

• Korean Journal of Food Science and Technology
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• v.37 no.5
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• pp.776-782
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• 2005

The effects of purification using ${\alpha}-amlyase$ (Termamyl 120L) on physicochemical properties of ${\beta}-glucan$ from oat bran were studied. Four fractions were selected as fraction A ($55^{\circ}C$, 15%, pH 6), fraction B ($45^{\circ}C$, 15%, pH 6), fraction C ($50^{\circ}C$, 0%, pH 7), and fraction D ($50^{\circ}C$, 10%, pH 5) from the result of physiological test, and three consecutive subfractions were obtained by repeated ${\alpha}-amlyase$ treatments on the each fractions. The contents of ${\beta}-glucan$, protein, and ash after purification were in 81.4-88.2%, 4.1-6.3% and 2.6-6.2%, respectively. The apparent viscosities of purified ${\beta}-glucan$ aqueous solutions were similar to those of hydroxy methyl cellulose. Glucose was a major monosaccharide of ${\beta}-glucan$ extracts, and xylose and arabinose were also detected as minor constituents on TLC. The average molecular weight ranged $2.0{\times}10^6-5.1{\times}10^6$ and was decreased after purification. From the result of the differential scanning calorimetry, the melting point ranged $130-140^{\circ}C$ with purification step and thermal transition enthalpy was increased. The ratio of ${\beta}-(1{\rightarrow}3)\;to\;{\beta}-(1{\rightarrow}4) $ linkages were 1:2.22-1:2.52, and increased up to 1:5.50 after purification.

• Korean Journal of Food Science and Technology
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• v.44 no.4
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• pp.428-433
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• 2012

Probiotics and their products, such as yogurt and cheese have been widely consumed in many countries with proven health benefits including anti-microbial activity and anti-diarrheal activity. LHFM (Lactobacillus helveticus - fermented milk) is a processed skim milk powder, fermented by a probiotics, L. helveticus IDCC3801. In the present study, we aimed to investigate the neuroprotective effects and the cognitive improvements of LHFM. LHFM itself did not show any cytotoxicity to the human neuroblastoma cell line, SH-SY5Y; however, it dose-dependently protected against glutamate-induced neuronal cell death. LHFM also attenuated scopolamine-induced memory deficit in Y-maze and Morris-water maze. In the analysis of hippocampus after a behavior test, LHFM significantly increased the acetylcholine level and also inhibited acetylcholine esterase activity. Therefore, the raised acetylcholine release partially contributes to the improvement of learning and memory by a treatment with LHFM. These results suggest that LHFM is an effective material for prevention or improvement of cognitive impairments caused by neuronal cell damage and central cholinergic dysfunction.

• Journal of Life Science
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• v.26 no.6
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• pp.633-639
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• 2016

Biomaterials including polymer, metal, ceramic, and composite have been widely applied for medical uses as medical fibers, artificial blood vessels, artificial joints, implants, soft tissue, and plastic surgery materials owing to their physicochemical properties. However, the biocompatibility and toxicity for film- and plate-form biomaterials is difficult to measure in mammalian cells because there is no appropriate incubation system. To solve these problems, we developed a novel mammalian cell culture system consisting of a silicone ring, top panel, and bottom panel and we applied two polymer films (PF) and one metal plate (MP). This system was based on the principal of sandwiching a test sample between the top panel and the bottom panel. Following the assembly of the culture system, SK-MEL-2 cells were seeded onto Styela Clava Tunic (SCT)-PF, NaHCO₃-added SCT (SCTN)-PF, and magnesium MP (MMP) and incubated at 37℃ for 24 hr and 48 hr. An MTT assay revealed that cell viability was maintained at a normal level in the SCT-PF culture group at 24 or 48 hr, although it rapidly decreased in the SCTN-PF culture group at 48 hr. Furthermore, the cell viability in the MMP culture group was very similar to that of the control group after incubation for 24 hr and 48 hr. Together, these results suggest the sandwich-type mammalian culture system developed here has the potential for the evaluation of the biocompatibility and toxicity of cells against PF- and MP-form biomaterials.

• Journal of Life Science
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• v.17 no.10
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• pp.1447-1451
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• 2007

Through the screening of marine natural compounds that inhibit cancer cell proliferation, we previously reported that pectenotoxin-2 (PTX-2) isolated from marine sponges exhibits selective cytotoxicity against several cell lines in p53-deficient tumor cells compared to those with functional p53. However, the molecular mechanisms of its anti-proliferative action on malignant cell growth are not completely known. To further explore the mechanisms of its anti-cancer activity and to test whether the status of p53 in liver cancer cells correlates with their chemo-sensitivities to PTX-2, we used two well-known hepatocarcinoma cell lines, p53-deficient Hep3B and p53-wild type HepG2. We have demonstrated that PTX-2 markedly inhibits Hep3B cell growth and induces apoptosis whereas HepG2 cells are much more resistant to PTX-2 suggesting that PTX-2 seems to act by p53-independent cytotoxic mechanism. The apoptosis induced by PTX-2 in Hep3B cells was associated with the modulation of DNA fragmentation factor (DFF) family proteins, up-regulation of pro-apoptotic Bcl-2 family members such as Bax and Bcl-xS and activation of caspases (caspase-3, -8 and -9). Blockade of the caspase-3 activity by caspase-3 inhibitor, z-DEVD-fmk, prevented the PTX-2-induced growth inhibition in Hep3B cells. Moreover, treatment with PTX-2 also induced phosphorylation of AKT and extracellular-signal regulating kinase (ERK), but not c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase (MARK). Specific inhibitors of PI3K inhibitor (LY294002) and ERK1/2 inhibitor (PD98059) significantly blocks PTX-2-induced-anti-proliferative effects, whereas a JNK inhibitor (SP600125) and a p38 MAPK inhibitor (SB203580) have no significant effects demonstrating that the pro-apoptotic effect of PTX-2 mediated through activation of AKT and ERK signal pathway in Hep3B cells.

• Journal of Life Science
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• v.19 no.7
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• pp.956-962
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• 2009

The aim of this study was to assess the effects of dentifrice-contatning grapefruit seed extract (GSE) and processed sulfur solution (PSS) on antimicrobial effects against oral pathogens. We first evaluated the antimicrobial effects of GSE and PSS against oral microbes: Streptococcus mutans (Sm), Prevotella intermedia (Pi), Porphyromonas gingivalis (Pg) and Candida albicans (Ca). When antimicrobial activity against Sm, Pi, Pg and Ca was tested, at 40 $\mu$l/disk, the inhibition zones of GSE were 11.0, 9.5, 8.0 and 9.0 mm, respectively. With the same method, the inhibition zones of PSS were 2.0, 3.5, 0.0 and 1.5 mm, respectively. In the micro broth dilution method, the MIC values of GSE against Sm, Pi, Pg and Ca were 0.24, 0.06, 0.10 and 15.63 $\mu$l/rnl, respectively. The MIC values of PSS were 0.12, 3.91,>125 and 7.81 $\mu$l/ml, respectively. When pH, refractive index, viscosity and color value of dentifrice-containing GSE and PSS were measured, there were no significant changes in these physical properties compared to the control samples. Antimicrobial activities of dentifrice products containing 0.5% GSE and 0.5% PSS against oral pathogens were 7.3, 4.3, 2.2 and 1.5 mm, respectively. According to these results, we conclude that there may be a role for GSE and PSS in the development of new oral supplies.

• Journal of Life Science
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• v.19 no.7
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• pp.949-955
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• 2009

Mucosal epithelia sense external stress signals and transmit them to the intracellular cascade responses. Ribotoxic stress-producing chemicals such as deoxynivalenol (DON) or other trichothecene mycotoxins have been linked with gastrointestinal inflammatory diseases by Fusarium-contamination. The purpose of this study was to test the hypothesis that DON evokes the epithelial sentinel signals of RNA-dependent protein kinase (PKR) and early growth response gene 1 (EGR-1), which together contribute to the pro-inflammatory cytokine interleukin 8 (IL-8) in human intestinal epithelial cells. PKR suppression by the dominant negative PKR expression attenuated DON-stimulated interleukin-8 production. Moreover, 1L-8 transcriptional activation by DON was also reduced by PKR inhibition in the human intestinal epithelial cells. Treatment with the PKR inhibitor also suppressed EGR-1 promoter activity, mRNA and protein induction, although mitogen-activated protein (MAP) kinases such as extracellular signal-regulated protein kinases (ERK) 1/2, p38, c-Jun N-terminal Kinase (INK) were little affected or even enhanced in presence of a PKR inhibitor. These patterns were also compared in the EGR-1-suppressed cells, which showed much more suppressed production of 1L-8. All things taken into consideration, DON-activated sentinel signals of EGR-1 via PKR mediated interleukin-8 production in human intestinal epithelial cells, which provide insight into the possible general mechanism associated with mucosal inflammation as an intestinal toxic insult by ribotoxic trichothecene mycotoxins.

• Journal of Life Science
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• v.20 no.7
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• pp.1078-1085
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• 2010

Polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs) and dioxin-like polychlorinated biphenyls (dl-PCBs) are bioaccumulative chemicals that are considered to be toxic contaminants based on several epidemiological studies. These chemicals in colostrum were investigated and estimated for their residual consistencies by maternal characteristics like parity and maternal ages. Test subjects were healthy primipara and multipara mothers with a mean age of 31.5 (S.D=3.6) in 2007. Seven isomers of PCDDs, 10 of PCDFs, 4 of non-orthopolychlorinated biphenyls(non-ortho PCBs) and 8 of mono-orthochlorinated polychlorinated biphenyls (mono-ortho PCBs) were analyzed by HRGC/HRMS. From the analyzed data, the mean level of total WHO-TEQs was 9.41 pg TEQ/g lipid, which is significantly lower than the level found in individuals from other countries. The main contributors to the total WHO-TEQs with increasing percentages were 2,3,4,7,8-PeCDF, 1,2,3,7,8-PeCDD and 3,3',4,4',5-PCB (#126), and they accounted for more than 60% of the total WHO-TEQs. PCDFs concentrations and total WHO-TEQs were negatively associated with parity (p<0.05), and maternal age was positively associated with total WHO-TEQs (p<0.01). However, the associations with body mass index (BMI) and fish intake during pregnancy were not significant. These results were suggested that parity and maternal age are an important factor affecting the concentrations of PCDD/DFs and dl-PCBs in these specimens.