• Microbiology and Biotechnology Letters
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• v.46 no.3
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• pp.288-299
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• 2018

Food bio preservation is of major interest in the food industry. Many types of antimicrobial compounds can be produced by lactic acid bacteria (LAB), including bacteriocins. Bacteriocins increase the shelf-life of food by decreasing some food-borne diseases. In this study, a multi-coding sequence of bacteriocin genes was used for primer design to produce bacteriocin genes in Enterococcus faecium AH2 strain and Pediococcus pentosaceus AH1. Multi-coding sequences were aligned to detect conserved sequences in the bacteriocin gene. Eight genes encoding proteins involved in bacteriocin production were isolated and sequenced, including six from E. faecium AH2 (entA, entI, entF, entR, orfA2, orfA3) and two from P. pentoceseus AH1 (papA, pedB), and all gene sequences were deposited in the Gen Bank database under accession numbers LC064146-LC064151, LC101300, and LC101789, respectively. P. pentosaceus AH1 and E. faecium AH2 strains displayed bacteriocin activities of $2610AU\;mL^{-1}$ and $690AU\;mL^{-1}$ and inhibition zones of 26 mm and 19 mm, respectively. Overexpression of entA in E. faecium AH2 increased the bacteriocin and antimicrobial activities.

• Journal of Food Hygiene and Safety
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• v.31 no.3
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• pp.194-200
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• 2016

Aflatoxins and ochratoxin A (AFTs and OTA) are secondary fungal metabolites produced by several moulds, mainly by Aspergillus flavus by Aspergillus ochraceus and Penicillium verrucosum, and these toxins can be transferred to animals and humans through the ingestion of contaminated feed and food. This study was to develop the analytical method for determination the levels of AFTs ($B_1$, $B_2$, $G_1$ and $G_2$) and OTA in pork. The AFTs and OTA were analyzed simultaneously by electrospray ionization in positive ion mode and mass reaction monitoring (MRM) after solid phase extract (SPE) columns clean-up. Performance characteristics, such as accuracy, precision, linear range, limit of detection (LOD) and quantification (LOQ), were also determined. Matrix-matched standard calibration was used for quantification, obtaining the recoveries in the range of 67.3~108.2% with the relative standard deviations of < 20%. Limits of detection and quantification were also estimated, obtaining the limits of quantification ranged in $0.7{\sim}1.3{\mu}g/kg$. The results of the inter-day study, which was performed with pork samples for 3 days, showed an accuracy of 92.0~109.9%. The precisions (expressed as relative standard deviation values) for the inter day variation were 2.6~17.8%. The method developed in this study was able to carry out the analysis with the satisfactory intensity and accuracy.

• Journal of Ginseng Research
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• v.43 no.1
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• pp.86-94
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• 2019

Background: Ginseng is believed to have antitumor activity. Autophagy is largely a prosurvival cellular process that is activated in response to cellular stressors, including cytotoxic chemotherapy; therefore, agents that inhibit autophagy can be used as chemosensitizers in cancer treatment. We examined the ability of Korean Red Ginseng extract (RGE) to prevent autophagic flux and to make hepatocellular carcinoma (HCC) cells become more sensitive to doxorubicin. Methods: The cytotoxic effects of total RGE or its saponin fraction (RGS) on HCC cells were examined by the lactate dehydrogenase assay in a dose- or time-dependent manner. The effect of RGE or RGS on autophagy was measured by analyzing microtubule-associated protein 1A/1B-light chain (LC)3-II expression and LC3 puncta formation in HCC cells. Late-stage autophagy suppression was tested using tandem-labeled green fluorescent protein (GFP)-monomeric red fluorescent protein (mRFP)-LC3. Results: RGE markedly increased the amount of LC3-II, but green and red puncta in tandem-labeled GFP-mRFP-LC3 remained colocalized over time, indicating that RGE inhibited autophagy at a late stage. Suppression of autophagy through knockdown of key ATG genes increased doxorubicin-induced cell death, suggesting that autophagy induced by doxorubicin has a protective function in HCC. Finally, RGE and RGS markedly sensitized HCC cells, (but not normal liver cells), to doxorubicin-induced cell death. Conclusion: Our data suggest that inhibition of late-stage autophagic flux by RGE is important for its potentiation of doxorubicin-induced cancer cell death. Therapy combining RGE with doxorubicin could serve as an effective strategy in the treatment of HCC.

• Journal of Microbiology and Biotechnology
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• v.21 no.7
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• pp.719-727
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• 2011

Botulism is a neuroparalytic disease caused by Clostridium botulinum, which produces seven (A-G) antigenically diverse neurotoxins (BoNTs). BoNTs are the most poisonous substances known to humans, with a median lethal dose ($LD_{50}$) of approximately 1 ng/kg of body weight. Owing to their extreme potency and lethality, they have the potential to be used as a bioterrorism agent. The mouse bioassay is the gold standard for the detection of botulinum neurotoxins; however, it requires at least 3-4 days for completion. Attempts have been made to develop an ELISA-based detection system, which is potentially an easier and more rapid method of botulinum neurotoxin detection. The present study was designed using a synthetic gene approach. The synthetic gene encoding the catalytic domain of BoNT serotype B from amino acids 1-450 was constructed with PCR overlapping primers (BoNT/B LC), cloned in a pQE30 UA vector, and expressed in an E. coli M15 host system. Recombinant protein production was optimized at 0.5 mM IPTG final concentration, 4 h post induction, resulting in a maximum yield of recombinant proteins. The immunogenic nature of the recombinant BoNT/B LC protein was evaluated by ELISA. Antibodies were raised in BALB/c mice using various adjuvants. A significant rise in antibody titer (p<0.05) was observed in the Alum group, followed by the Titermax Classic group, Freund's adjuvant, and the Titermax Gold group. These developed high-titer antibodies may prove useful for the detection of botulinum neurotoxins in food and clinical samples.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.1
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• pp.102-106
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• 2005

This study was carried out to find the attributes for liquid-type yogurt with Lactobacillus casei 911LC during 72 h fermentation at $37^{\circ}C$. The pH decreased up to 32 h and plauteaued thereafter, and the titratable acidity increased up to 40 h. The growth of lactic acid bacteria sharply increased with $3.5{\times}10^7$ cfu/ml up to 40 h of fermentation and slowly decreased thereafter. The free amino acids produced during fermentation reached the maximum value at 44 h and gradually decreased thereafter. Bitterness sensory scores were the highest at 44 h of fermentation. In the result of electrophoresis, the band mostly disappeared at 72 h fermentation. The present data showed that the range of optimum fermentation time for liquid-type yogurt using Lactobacillus casei 911LC was from 40 to 44 h.

• 한국생물공학회:학술대회논문집
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• 2002.04a
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• pp.103-106
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• 2002

A gradient LC/MS system was constructed and applied for separation of biological samples. For example, a rapid and simple analytical method without pretreatment based on gradient ${\mu}LC/MS$ with a disposable microcolumn has been developed to determine B group vitamins in urine. Urine samples were directly injected to the disposable home-made microcolumn. The microcolumn can be emptied after being used for a series of urine samples, and repacked with fresh stationary phase. An overdose of vitamin pills were swallowed by healthy volunteers and the urine samples were taken 1,2,3,5, and 8 hours after swallowing. Vitamins immediately showed up in urine, hit the maximum, and disappeared swiftly. This technique is expected to have some application for clinical purposes.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.9
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• pp.1253-1259
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• 2001

Three cassava varieties, namely MM 92 (MM), Black Twig (BT) and Local (LC), were arranged in a randomized complete block design to evaluate their dry fodder and crude protein (CP) productivity as well as growth persistency. Cassava plants grown in small plots of $5m{\times}10m$ at a planting distance of $25cm{\times}25cm$ were harvested every 6 weeks starting from 3 months after planting. Dry fodder yields of MM, BT and LC over the 8 harvests were 8.55, 8.01 and 6.15 t/ha, respectively. All varieties produced more leaves than stems with average leaf:stem ratios of 5, 5.9 and 4.8 for MM, BT and LC, respectively. In terms of CP production, MM was the highest yielder (272 kg/ha/harvest), followed by BT and LC (238 and 184 kg/ha/harvest, respectively). The total accumulative CP amounts over the 8 harvests were 2179, 1903 and 1474 kg/ha for MM, BT and LC, respectively. The mortality rates were 9.91, 14.01 and 13.98% for MM, BT and LC, respectively. Phosphorus content was more stable than potassium content during defoliation. MM, BT and LC had whole plant phosphorus contents of 0.41, 0.41 and 0.39%, respectively; whole plant potassium contents were 1.25, 1.38 and 1.20%.

• Archives of Pharmacal Research
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• v.27 no.12
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• pp.1202-1206
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• 2004

A rapid, sensitive and selective liquid chromatography-tandem mass spectrometric (LC-MS/ MS) method for the determination of lithospermic acid B (LSB) in rat serum was developed. LSB and internal standard, 7-hydroxy-3-phenyl-chromen-4-one (HPC) were extracted from rat serum with methyl-tert-butyl ether at acidic pH and analyzed on a Luna $C_8$ column with the mobile phase of acetonitrile-ammonium formate (10 mM, pH 6.5) (50:50, v/v). The analytes were detected using a negative electrospray ionization tandem mass spectrometry in the multiple- reaction-monitoring mode. The standard curve was linear $(r^2 = 0.997)$ over the concentration range of 10.0-500 ng/mL. The coefficient of variation and relative error for intra- and interassay at three QC levels were 1.1~6.2% and -10.3~-2.7%, respectively. The recovery of LSB from serum sample ranged from 73.2 to 79.5%, with that of HPC (internal standard) being 75.1 %. The lower limit of quantification for LSB was 10 ng/mL using 50 ${\mu}L$ of serum sample.

• Mass Spectrometry Letters
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• v.7 no.4
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• pp.106-110
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• 2016

Ginseng, a traditional herbal drug, has been used in Eastern Asia for more than 2000 years. Various ginsenosides, which are the major bioactive components of ginseng products, have been shown to exert numerous beneficial effects on the human body when co-administered with drugs. However, this may give rise to ginsenoside-drug interactions, which is an important research consideration. In this study, acassette assay was performed the inhibitory effects of 12 ginsenosides on seven cytochrome P450 (CYP) isoforms in human liver microsomes (HLMs) using LC-MS/MS to predict the herb-drug interaction. After incubation of the 12 ginsenosides with seven cocktail CYP probes, the generated specific metabolites were quantified by LC-MS/MS to determine their activities. Ginsenoside Rb1 and F2 showed strong selective inhibitory effect on CYP2C9-catalyzed diclofenac 4'-hydroxylation and CYP2B6-catalyzed bupropion hydroxylation, respectively. Ginsenosides Rd showed weak inhibitory effect on the activities of CYP2B6, 2C9, 2C19, 2D6, 3A4, and compound K, while ginsenoside Rg3 showed weak inhibitory effects on CYP2B6. Other ginsenosides, Rc, Rf, Rg1, Rh1, Rf, and Re did not show significant inhibitory effects on the activities of the seven CYPs in HLM. Owing to the poor absorption of ginsenosides after oral administration in vivo, ginsenosides may not have significant side effects caused by interaction with other drugs.

• Journal of the Korea Institute of Information and Communication Engineering
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• v.8 no.7
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• pp.1429-1435
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• 2004

In this work, high performance broadband amplifier MMIC including all the matching and biasing components, and electrostatic discharge (ESD) protection circuit was developed for K/Ka band applications. Therefore, external biasing or matching components were not required for the operation of the MMIC. STO (SrTiO3) capacitors were employed to integrate the DC biasing components on the MMIC, and miniaturized LC parallel ESD protection circuit was integrated on MMIC, which increased ESD breakdown voltage from 10 to 300 V. A pre-matching technique and RC parallel circuit were used for the broadband design of the amplifier MMIC. The amplifier MMIC exhibited good RF performances and good stability in a wide frequency range. The chip size of the MMICs was $1.7{\pm}0.8$ mm2.