• Reproductive and Developmental Biology
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• v.38 no.4
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• pp.147-158
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• 2014

Differentiated nuclei can experimentally be returned to an undifferentiated embryonic status after nuclear transfer (NT) to unfertilized metaphase II (MII) oocytes. Nuclear reprogramming is triggered immediately after somatic cell nucleus transfer (SCNT) into recipient cytoplasm and this period is regarded as a key stage for optimizing reprogramming. In a recent study (Dai et al., 2010), use of m-carboxycinnamic acid bishydroxamide (CBHA) as a histone deacetylase inhibitor during the in vitro early culture of murine cloned embryos modifies the acetylation status of somatic nuclei and increases the developmental competence of SCNT embryos. Thus, we examined the effects of CBHA treatment on the in vitro preimplantation development of porcine SCNT embryos and on the acetylated status of histone H3K9 on cloned embryos at the zygote stage. We performed the three groups SCNT: SCNT (NT), CBHA treatment at the porcine fetus fibroblast cells (PFFs) used as donor cells prior to SCNT (CBHA-C) and CBHA treatment at the porcine SCNT embryos during the in vitro early culture after oocyte activation (CBHA-Z). The PFFs were treated with a $15{\mu}M$ of CBHA (8 h) for the early culture and the porcine cloned embryos were treated with a $100{\mu}M$ concentration of CBHA during the in vitro early culture (10 h). Cleavage rates and development to the blastocyst stage were assessed. No significant difference was observed the cleavage rate among the groups (82.6%, 76.4% and 82.2%, respectively). However, the development competence to the blastocyst stage was significantly increased in CBHA-Z embryos (22.7%) as compared to SCNT and CBHA-C embryos (8.6% and 4.1%)(p<0.05). Total cell numbers and viable cell numbers at the blastocyst stage of porcine SCNT embryos were increased in CBHA-Z embryos as compared to those in CBHA-C embryos (p<0.05). Signal level of histone acetylation (H3K9ac) at the zygote stage of SCNT was increased in CBHA-Z embryos as compared to SCNT and CBHA-C embryos. The results of the present study suggested that treatment with CBHA during the in vitro early culture (10 h) had significantly increased the developmental competence and histone acetylation level at the zygote stage.

• Korean journal of food and cookery science
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• v.13 no.2
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• pp.106-112
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• 1997

The effect of low concentrations of allspice (Pimenta dioica L.) in culture broth as an antibacterial agent against Escherichia coli 0157:H7 and Staphylococcus aureus 196E was tested at 35,5 and -20$^{\circ}C$. Tryptic soy broth (TSB) containing 0∼2% (w/v) of allspice was inoculated with 10$\^$5/∼10$\^$6/ cells/$m\ell$ of E. coli and S. aureus and incubated at each temperature. The growth of E. coli was not inhibited at 0.1∼1.0% allspice and growth occured at 2％ allspice but only after a prolonged lag period. Growth of S. aureus was inhibited with increasing concentration of allspice at 35$^{\circ}C$. Growth of S. aureus occured at the presence of 0.1∼0.3% allspice but the viability of S. aureus at 0.5∼2.0% allspice was decreased during storage at 35$^{\circ}C$. During refrigerated storage at 5$^{\circ}C$, inhibition of E. coli and S. aureus was increased with the progress of time and increasing spice concentration. During frozen storage at -20$^{\circ}C$, antibacterial activity of allspice against E. coli was increased with increasing storage time and spice concentration while that activity against S. aureus was effective during early period of storage. There was no major changes in population of S. aureus in TSB with different concentration of spice frozen at -20$^{\circ}C$. Viable counts of E. coli and S. aureus at 0.l％ of allspice was less than that of control during frozen storage.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.28 no.2
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• pp.209-218
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• 1995

Bacteriophage T7 gene 2.5 protein, a single-stranded DNA binding protein, has been implicated in T7 DNA replication, recombination, and repair. Purified gene 2.5 protein has been shown to interact with the phage encoded gene 5 protein (DNA polymerase) and gene 4 proteins (helicase and primase) and stimulates their activities. Genetic analysis of T7 phage defective in gene 2.5 shows that the gene 2.5 protein is essential for T7 DNA replication and growth. T7 phage that contain null mutants of gene 2.5 were constructed by homologous recombination. These mutant phage $(T7\Delta2.5)$ cannot grow in Escherichia coli. After infection of E. coli with $T7\Delta2.5$, host DNA synthesis is shut off, and $T7\Delta2.5$ DNA synthesis is reduced to less than $1\%$ of wild-type phage DNA synthesis (Kim and Richardson, 1993, Proc. Natl. Aca. Sci. USA, 90, 10173-10177). A truncated gene 2.5 protein $(GP2.5-\Delta21C)$ deleted the 21 carboxyl terminal amino acids was constructed by in vitro mutagenesis. $GP2.5-\Delta21C$ cannot substitute for wild-type gene 2.5 protein in vivo; the phage are not viable and exhibit less than $1\%$ of the DNA synthesis observed in wild-type phage-infected cells. $GP2.5-\Delta21C$ has been purified to apparent homogeneity from cells overexpressing its cloned gene. Purified $GP2.5-\Delta21C$ does not physically into「act with T1 gene 4 protein as measured by affinity chromatography and immunoblot analysis. The mutant protein cannot stimulate T7 gene 4 protein activity on RNA-primed DNA synthesis and primer synthesis. These results suggest that C-terminal domain of gene 2.5 protein is essential for protein-protein interactions.

• Journal of Food Hygiene and Safety
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• v.19 no.3
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• pp.151-160
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• 2004

Enterococcus faecium MJ5-14 isolated from Meju produced a bacteriocin, which was antagonistic towards Listeria monocytogenes. Bacteriocin activity reached a maximum (640 BU/mL) after incubation for 12 hr, the early stationary phase, then dropped after the late stationary phase. Bacterocin of E. faecium MJ5-14 was extremely active against a wide range of Listeria species, including L. monocytogenes with sensitives up to about 640 BU/mL. In case of mixed culture with 105 CFU/mL L. monocytogenes and 105 CFU/mL E. faecium MJ5-14, the inhibitory effect against L. monocytogenes at $37^{\circ}C$ was higher than at $25^{\circ}C$. The mode of action was identified as bactericidal, because the addition of 100 BU/mL this bacteriocin to cell suspensions of L. monocytogenes KCTC 3569, led to a marked decrease in the number of viable cells. Further, when held in contact with bacteriocin of E. faecium MJ15-14 for 12 hr, L. monocytogenes KCTC 3569 displayed the disruption of the cells and an important efflux of the intracellular material.

• 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.