• Journal of Microbiology and Biotechnology
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• v.21 no.12
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• pp.1280-1286
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• 2011

A quantitative, real-time PCR method was developed to enumerate Lactobacillus plantarum IWBT B 188 during the malolactic fermentation (MLF) in Grauburgunder wine. The qRT-PCR was strain-specific, as it was based on primers targeting a plasmid DNA sequence, or it was L. plantarum-specific, as it targeted a chromosomally located plantaricin gene sequence. Two 50 l wine fermentations were prepared. One was inoculated with 15 g/hl Saccharomyces cerevisiae, followed by L. plantarum IWBT B 188 at $3.6{\times}10^6$ CFU/ml, whereas the other was not inoculated (control). Viable cell counts were performed for up to 25 days on MRS agar, and the same cells were enumerated by qRT-PCR with both the plasmid or chromosomally encoded gene primers. The L. plantarum strain survived under the harsh conditions in the wine fermentation at levels above $10^5$/ml for approx. 10 days, after which cell numbers decreased to levels of $10^3$ CFU/ml at day 25, and to below the detection limit after day 25. In the control, no lactic acid bacteria could be detected throughout the fermentation, with the exception of two sampling points where ca. $1{\times}10^2$ CFU/ml was detected. The minimum detection level for quantitative PCR in this study was $1{\times}10^2$ to $1{\times}10^3$ CFU/ml. The qRT-PCR results determined generally overestimated the plate count results by about 1 log unit, probably as a result of the presence of DNA from dead cells. Overall, qRT-PCR appeared to be well suited for specifically enumerating Lactobacillus plantarum starter cultures in the MLF in wine.

• Microbiology and Biotechnology Letters
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• v.14 no.5
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• pp.421-426
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• 1986

This study was attempted to find out effective storage methods of Lactobacillus casei YIT 9018, industrial strain for fermented mil k production, without severe bacterial death and activity deteriorations. The cryoprotective effect of the ammo acid mixture consisting of glycine and DL-g1utamic acid on the test strain were examined and also compared with those other protectants already reported. The apparent protective effect by the amino acid mixture was observed to controls. Both glycine and DL-glutamic acid prevented the freezing death of test strain and his effect of 1. casei YIT 9018 had reached stationary stage in MRS-broth 18h after inoculation. Cells harvested from stationary stage were most resistant to freezing damage. The viability of the test strain was affected by rehydration media and the recovery of viable cells was increased about threefold when amino acid mixture was used for rehydration. The presence of non-fat milk solid (NFMS), sucrose and lactose in amino acid mixture increased viability of the test strain up to 85％. In this case, optimal concentrations of NFMS, sucrose and lactose were 10％, 7.5-10％, 7.5-10％, respectively.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.34 no.4
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• pp.365-369
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• 2001

Streptococcus spp. of bacterial pathogen of fish were isolated from the cultured flounder (Paralichthys olivaceus) in fish farm of Jeju Island. Clinical signs of the infected flounder which are the most commons symptoms are as follows: erratic swimming, darkening of the body colour, unilateral or bilateral exophthalmia, corneal opacity, hemorrhages in the opercular and the bases of the fins, and the ulceration of the body surface. Biochemical characteristics of pathogenic fish Streptococcus spp, were gram positive, spherical form, catalase negative, oxidase negative and $\beta$-haemolytic, respectively, The viable cells counted from the tissue of the diseased flounder were the largest in the order of the ulcer, the kidney, the blood and the brain, The drugs used were ampicillin, ciprofloxacin, doxycycline, gentamycin, tetracycline, erythromycin, streptomycin and oxytetracycline, Streptococcus spp. were found to be sensitive to ampicillin, ciprofloxacin, doxycycline and gentamycin, but were resistant to tetracycline, erythromycin, streptomycin and oxytetracycline. The pathogenicity of Streptococcus spp. on the cultured flounder with an abdominal cavity injection was high. The haemolytic activity of the toxin against the sheep red blood cells reached the maximum after 30 min incubation at $37^{\circ}C$ or $50^{\circ}C$. The toxin showed highest activity at pH $5.5\sim6.5$.

• IMMUNE NETWORK
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• v.12 no.2
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• pp.58-65
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• 2012

Background: A cell line with transfected Wilms' tumor protein 1 (WT1) is has been used for the preclinical evaluation of novel treatment strategies of WT1 immunotherapy for leukemia due to the lack of appropriate murine leukemia cell line with endogenous WT1. However, silencing of the transgene occurs. Regarding the effects of hypomethylating agents (HMAs) on reactivation of silenced genes, HMAs are considered to be immune enhancers. Methods: We treated murine WT1- transfected C1498 (mWT1-C1498) with increasing doses of decitabine (DAC) and azacitidine (AZA) to analyze their effects on transgene reactivation. Results: DAC and AZA decreased the number of viable cells in a dose- or time-dependent manner. Quantification of WT1 mRNA level was analyzed by real-time polymerase chain reaction after mWT1-C1498 treated with increasing dose of HMA. DAC treatment for 48 h induced 1.4-, 14.6-, and 15.5-fold increment of WT1 mRNA level, compared to untreated sample, at 0.1, 1, and $10{\mu}M$, respectively. Further increment of WT1 expression in the presence of 1 and $10{\mu}M$ DAC was evident at 72 h. AZA treatment also induced up-regulation of mRNA, but not to the same degree as with DAC treatment. The correlation between the incremental increases in WT1 mRNA by DAC was confirmed by Western blot and concomitant down-regulation of WT1 promoter methylation was revealed. Conclusion: The in vitro data show that HMA can induce reactivation of WT1 transgene and that DAC is more effective, at least in mWT1-C1498 cells, which suggests that the combination of DAC and mWT1-C1498 can be used for the development of the experimental model of HMA-combined WT1 immunotherapy targeting leukemia.

• Korean journal of food and cookery science
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• v.19 no.6
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• pp.701-707
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• 2003

Kimchi was prepared with the addition of 2.5％ and 5.0％ Monascu purpureu koji(MPK) paste (20％) and were fermented at 20$^{\circ}C$ for 18 days. The quality and sensory characteristics of the kimchi were evaluated by analyzing the pH, acidity, number of viable cells, the concentration of reducing sugar, and sensory properties during fermentation. The pH and titratable acidity of the kimchiprepared with MPK(MPK kimchi) were higher and lower, respectively, than those of the control kimchi. The MPK kimchi showed high 'L' and 'b' values during storage, but the 'a' values were low. The contents of the reducing sugar of the MPK kimchi tended to increase during fermentation, particularly after six days. The number of total microbial cells, lactic acid bacteria and yeast in the MPK kimchi were lower than those of the control kimchi until 3 days of fermentation. However, the number of these bacteria in the MPK kimchi and the control kimchi after six days of fermentation was similar. The sensory score of the kimchi with 2.5％ and 5.0％ added MPK paste were significantly higher than the control groups in terms of the sweetness and overall acceptability.

• Korean journal of food and cookery science
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• v.21 no.5
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• pp.717-724
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• 2005

Sensory characteristics and antimicrobial activity of Korean wheat noodle with pine pollen powder and antioxidant activity of pine pollen extracts were investigated to develop the health promoting product. Pine pollen powder was extracted with water and 70% ethanol and extracts were tested electron donating ability (EDA) and nitrite scavenging ability (NSA). Sensory characteristics were evaluated by its color, flavor, moisture, softness, texture and taste. Microbiological quality was tested by total viable cells. EDA was highest in 1,000 ppm for both water extract (67% of EDA) and ethanol extract (74% of EDA). NSA was highest in pH 1.2 for both water extract(46% of NSA) and ethanol extract (48% of NSA). Antimicrobial activity of Korean wheat noodle with 3% pine pollen powder were 0.5 log $cycle{\sim}1$ log cycle lower than that of control at 5days of storage in total vival cells and at 4days of storage in total fungus. From sensory evaluation, Korean wheat noodle with 3% pine pollen powder had significantly higher of all scores.

• Food Science of Animal Resources
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• v.24 no.1
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• pp.97-102
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• 2004

The objective of this study was to control Kimchi fermentation using pH sensitive bacteriocin entrapping liposome(bacteriocin-liposome). The liposomes were prepared by the reverse-phase evaporation method from a mixture of DPPC(dipalmitoyl phosphatidylcholine, DPPE(dipalmitoyl phosphatidylethanolamine), DOPC(dioleoyl phosphatidylcholine) and cholesterol in a molar ration of 4:2:1:4. The bacteriocin-liposome was disruptured at pH 4 of buffer and was stable at alkaline pHs(6 and 7). Irrespective of the addition of the bacteriocin-liposomes, the pH of every Kimchi sample decreased to 5 during 5 days storage at 5$^{\circ}C$. Kimchi samples treated with bacteriocin-liposomes maintained pH 4 or higher, while Kimchi samples not treated with bacteriocin-liposomes exhibited pH 3.58 or lower. In general, the pH of Kimchi samples stored at 20$^{\circ}C$ decreased faster, compared to that of Kimchi samples stored at 5$^{\circ}C$. The pH of Kimchi samples treated with the bacteriocin-liposomes was 3.9 during 90 days storage, while that of the samples not treated with the bacteriocin-liposomes was 3.68 and 3.32 during 30 days and 90 days storages, respectively. Lactic acid bacteria in Kimchi treated with the bacteriocin-liposome grew relatively slow at 5$^{\circ}C$. The viable cell number of lactic acid bacteria increased up to 4${\times}$10$\^$7/ cells/ml and then decreased to 8${\times}$10$\^$6/ cells/ml during 90 days storage at 5$^{\circ}C$.

• Journal of the East Asian Society of Dietary Life
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• v.17 no.5
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• pp.678-688
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• 2007

In this study, we investigated the effects of ripened tomato on the Baechukimchi fermentation, by measuring sensory, physicochemical, and microbiological properties during fermentation up to 28 days. The Baechukimchi, with various levels [0, 10, 20, 30%(w/v)] of ripened tomato, was fermented at $10^{\circ}C$. The product containing 20% ripened tomato showed the highest scores for overall acceptability, sour taste, texture, sour smell, and color. The intensity characteristic scores related to sour smell and sour taste were the highest in the control, and the texture score was highest in the 20% ripened tomato sample. During fermentation, total acidity increased while pH gradually decreased. The reducing sugar and total vitamin C contents of the experimental groups with added ripened tomato were higher than those of the control. Also, the time periods to reach maximum levels of lactic acid bacteria, by log numbers of cells and total viable cells, were more delayed in the experimental groups with added ripened tomato than in the control. Overall, the results indicate that a 20% addition of ripened tomato to Baechukimchi during fermentation can enhance sensory qualities and extended the palatable period of the product.

• Clinical and Experimental Reproductive Medicine
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• v.25 no.2
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• pp.115-122
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• 1998

The maintenance of a viable pregnancy has long been viewed as an immunological paradox. The deveolping embryo and trophoblast are immunologically foreign to the maternal immune system due to their maternally inherited genes products and tissue-specific differentiation antigens (Hill & Anderson, 1988). Therefore, speculation has arisen that spontaneous abortion may be caused by impaired maternal immune tolerance to the semiallogenic conceptus (Hill, 1990). Loss of recall antigen has been reported in immunosuppressed transplant recipients and is associated with graft survival (Muluk et al., 1991; Schulik et al., 1994). Progesterone $(10^{-5}M)$ has immunosuppressive capabilities (Szekeres-Bartho et al., 1985). Previous study showed that fertile women, but not women with unexplained recurrent abortion (URA), lose their immune response to recall antigens when pregnant (Bermas & Hill, 1997). Therefore, we hypothesized that immunosuppressive doses of progesterone may affect proliferative response of lymphocytes to trophoblast antigen and alloantigen. Proliferative responses using $^3H$-thymidine ($^3H$-TdR) incorporation of peripheral blood mononuclear cells (PBMCs) to the irradiated allogeneic periperal blood mononuclear cells as alloantigen, trophoblast extract and Flu as recall antigen, and PHA as mitogen were serially checked in 9 women who had experienced unexplained recurrent miscarriage. Progesterone vaginal suppositories (100mg b.i.d; Utrogestan, Organon) beginning 3 days after ovulation were given to 9 women with unexplained RSA who had prior evidence of Th1 immunity to trophoblast. We checked proliferation responses to conception cycle before and after progesterone supplementation once a week through the first 7 weeks of pregnancy. All patients of alloantigen and PHA had a positive proliferation response that occmed in the baseline phase. But 4 out of 9 patients (44.4%) of trophoblast antigen and Flu antigen had a positive proliferative response. The suppression of proliferation response to each antigen were started after proliferative phase and during pregnancy cycles. Our data demonstrated that since in vivo progesterone treated PBMCs suppressed more T-lymphocyte activation and $^3H$-TdR incorporation compare to PBMCs, which are not influenced by progesterone. This data suggested that it might be influenced by immunosuppressive effect of progesterone. In conclusion, progesterone may play an important immunological role in regulating local immune response in the fetal-placental unit. Furthermore, in the 9 women given progesterone during a conception cycle, Only two (22%) repeat pregnancy losses occured in these 9 women despite loss of antigen responsiveness (one chemical pregnancy loss and one loss at 8 weeks of growth which was karyotyped as a Trisomy 4). These finding suggested that pregnancy loss due to fetal aneuploidy is not associated with immunological phenomena.

• Bulletin of the Korean Chemical Society
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• v.35 no.2
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• pp.505-512
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• 2014

A new donor-accepter-donor-accepter-donor (D-A-D-A-D) type 2,1,3-benzothiadiazole-thiophene-based acceptor unit 2,5-di(4-(5-bromo-4-octylthiophen-2-yl)-2,1,3-benzothiadiazol-7-yl)thiophene ($DTBTTBr_2$) was synthesized. Copolymerized with fluorene and indeno[1,2-b]fluorene electron-rich moieties, two alternating narrow band gap (NBG) copolymers PF-DTBTT and PIF-DTBTT were prepared. And two copolymers exhibit broad and strong absorption in the range of 300-700 nm with optical band gap of about 1.75 eV. The highest occupied molecular orbital (HOMO) energy levels vary between -5.43 and -5.52 eV and the lowest unoccupied molecular orbital (LUMO) energy levels range from -3.64 to -3.77 eV. Potential applications of the copolymers as electron donor material and $PC_{71}BM$ ([6,6]-phenyl-$C_{71}$ butyric acid methyl ester) as electron acceptors were investigated for photovoltaic solar cells (PSCs). Photovoltaic performances based on the blend of PF-DTBTT/$PC_{71}BM$ (w:w; 1:2) and PIF-DTBTT/$PC_{71}BM$ (w:w; 1:2) with devices configuration as ITO/PEDOT: PSS/blend/Ca/Al, show an incident photon-to-current conversion efficiency (IPCE) of 2.34% and 2.56% with the open circuit voltage ($V_{oc}$) of 0.87 V and 0.90 V, short circuit current density ($J_{sc}$) of $6.02mA/cm^2$ and $6.12mA/cm^2$ under an AM1.5 simulator ($100mA/cm^2$). The photocurrent responses exhibit the onset wavelength extending up to 720 nm. These results indicate that the resulted narrow band gap copolymers are viable electron donor materials for polymer solar cells.