• Title/Summary/Keyword: Invitro

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Effects of Steam Flaking of Corns imported from USA and India on the in vitro Fermentation Characteristic and the Mycotixin Contents of Logistic Processing Line (미국산과 인도산 옥수수의 steam flaking 처리가 공정라인별 mycotoxin 함량의 변화와 in vitro 발효 특성에 미치는 영향)

  • Lee, Shin-Ja;Lee, Ji-Hun;Shin, Nyeon-Hak;Han, Jung-Hun;Hyun, Jong-Hwan;Moon, Yea-Hwang;Lee, Sung-Sill
    • Journal of Life Science
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    • v.19 no.1
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    • pp.65-74
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    • 2009
  • The objective of this study was to examine the effects of steam flaking treatment of corn grains imported from USA and India on in vitro gas production, microbial growth and contents of aflatoxin $B_1$ and ochratoxin A. Each treatment was composed of total 4 treatments including (1) USCW (US com-whole type), (2) USCF (US corn-flaked type), (3) IDCW (India corn-whole type) and (4) IDCF (India orn-flaked type) with 4 replications $\times$ 6 incubation times (3, 6, 9, 12, 18 and 24 hr). Mycotoxin (aflatoxin $B_1$ & ochratoxin A) contents in test corns tended to increase gradually with increasing logistics periods from the harbor, hopper, silo to processing line. The contents of aflatoxin $B_1$ in India corn (IDCW) and US corn (USCW) were 11.71 and 1.78 ppb, respectively when measured at the hopper. After steam flaking, both contents of aflatoxin $B_1$ in USCW and IDCW were 0.00 ppb. It means that Aspergillus flavus could be decreased by steam flaking. However, this trend was not observed in ochratoxin content. The gas production rate of USA corns (USCW & USCF) was significantly (p<0.05) higher than India corns (IDCW & IDCF), and that of steam flaked corns (USCF & IDCF) was higher $1.5{\sim}2%$ than whole corns (USCW & IDCW) after 3 hr incubation in in vitro experiment. pH value was optimally maintained for microbial growth during whole incubation times with the value of 6.05 to 6.54, and was not significantly different between treatments, but USCF was somewhat lower than other treatments. pH value decreased following 12 hr of incubation but gas production increased rapidly during the same period. In addition, in vitro microbial growth rates also increased with up to 18 hr of incubation period, thereafter experienced a decrease with extended incubation time. In conclusion, US corn was superior to India corn by origin based on the results of in vitro and mycotoxin contents. And steam flaking process of imported corns tended to decrease mycotoxin contents such as aflatoxin $B_1$ and ochratoxin A as well as improve in vitro gas production and microbial growth rates.

In Vitro AChE Inhibition Tests of Insecticides Using Electric eel and Housefly AChE (전기뱀장어 및 집파리 AChE를 이용한 살충제의 In Vitro AChE 저해 시험)

  • 이시혁;이준호;조광연
    • Korean journal of applied entomology
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    • v.31 no.2
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    • pp.122-132
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    • 1992
  • Experiments were conducted to establish an in vitro AChE inhibition test system to evaluate the potency of AChE inhibition of new chemical compounds. For a fixed time inhibition test, optimal inhibition (incubation) time to evaluate their AChE inhibition potency was 10 min. for AChE inhibitors such as DFP, DDVP, and paraoxon. The concentration of new chemical compounds with an ester group for evaluation of their inhibition potency was 10 $\mu$M under 10 min. preincubation conditions. However, the stepwise inhibition test with higher concentrations seemed to be needed for other chemical compounds. For a progressive inhibition test to calculate inhibition constants such as $K_d$, $K_3$ and $K_i$, extremely low $K_d(1.3\times10-^85.6\times10^{-7})$ and $K_3$(0. 21-0.27 $min^{-1}$) were observed under lagged preincubation time (0.8-13.3 min) and low in¬hibitor concentrations $(1\times10-^92\times10-^6M)$. However, this method seemed to be useful for comparison of AChE inhibition potency among inhibitors. Differences in inhibition potency among DFP, paraoxon, and KH501 were due to the differences in $K_d$, in other words, differences in affinities between inhibitors and AChEs. Therefore, AntiChE screening should consist of two steps. The first step is to evaluate the potency of AChE inhibition based on $I_50$ valuse obtained from fixed time inhibition tests. The second step is to study inhibition patterns and characteristics of chemical compounds selected in the first step.

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Transformation is Mechanism of Gene Transfer in Soil (토양에서 Transformation에 의한 유전자 전이)

  • ;Stotzky, G.
    • Korean Journal of Microbiology
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    • v.28 no.3
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    • pp.210-218
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    • 1990
  • The survival and transfer of chromosomal genes coding for the synthesis of amino acids (threonine, tryptophan, histidine, leucine, methionine) and of plasmid-borne genes coding for resistance to antibiotics (chloramphenicol, kanamycin, erythromycin) by transformation in sterile and nonsterile soil (the soil was amended to 12% vol/vol with the clay mineral, montmorillonite) was studied. In pure culture, the numbers of vegetative cells of the Bacillus subtilis strains decreased by 1 to 1.5 orders of magnitude within one week, but spores of each strain showed lesser decreases. In sterile soil, the populations of vegetative cells and spores decreased by 1.5 to 3 orders of magnitude within 2 to 4 days and then showed little additional decreased. The transformation frequencies (number of transformants/numbers of donors and recipients) of individual amino acid-genes invitro ranged from $1.3{\pm}0.6{\times}10^{-6}$ to $6.0{\pm}2.36{\times}10^{-6}$, of two amino acid-genes from $8.5{\pm}0.7{\times}10^{-8}$ to $3.1{\pm}0.6{\times}10^{-7}$, and of the antibiotic-resistance genes from $1.5{\pm} 0.2{\itmes} 10^{-7}$ to $1.4{\pm} 0.4{\times} 10^{-5}$ . In sterile soil, the frequencies of transfer of individual amino acid-genes ranged from $2.0{\times} 10^{-7}$ to $2.0{\times} 10^{-5}$ and of the antibiotic-resistance genes from $2.0{\times} 10^{-7}$ to $9.4{\pm} 4.7{\times} 10^{-6}$. The transfer of two amino acid-genes in sterile soil was detected at a frequency of $2.0{\times} 10^{-6}$ to $4.5{\times} 10^{-6}$, but only in three instances. The transformation frequencies of antibiotic-resistance genes in nonsterile soil were essentially similar to those in sterile soil. However, to detect transformants in nonsterile soil, higher concentrations of antibiotics were needed, as the result of the large numbers of indigenous soil bacteria resistant to the concentration of antibiotics used in the sterile soil and in vitro studies. The results of these studies show that genes can be transferred by transformation in soil and that this mechanism of transfer must be considered in risk assessment of the release of genetically engineered microorganisms to the environment.

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In vitro Anti-tumor Effect of an Engineered Vaccinia Virus in Multiple Cancer Cells and ABCG2 Expressing Drug Resistant Cancer Cells (재조합 백시니아 바이러스의 다양한 암세포 및 ABCG2 과발현 내성 암세포에 대한 항 종양 효과 연구)

  • Park, Ji Hye;Yun, Jisoo;Heo, Jeong;Hwang, Tae Ho;Kwon, Sang Mo
    • Journal of Life Science
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    • v.26 no.7
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    • pp.835-846
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    • 2016
  • Chemo-resistance is the biggest issue of effective cancer therapy. ABCG2 is highly correlated with multi-drug resistance, and represent a typical phenotype of multiple cancer stem-like cells. Accumulating evidence recently reported that oncolytic viruses represent a new strategy for multiple aggressive cancers and drug resistant cancers including cancer stem cell-like cells and ABCG2 expressing cells. In this study, we generated an evolutionally engineered vaccinia virus, SLJ-496, for drug-resistant cancer therapy. We first showed that SLJ-496 treatment enhanced tumor affinity using cytopathic effect assay, plaque assay, as well as cell viability assay. Next, we clearly demonstrated that in vitro SLJ-496 treatment represents significant cytotoxic effect in multiple cancers including colorectal cancer cells (HT-29, HCT-116, HCT-8), gastric cancer cells (AGS, NCI-N87, MKN-28), Hepatocellular carcinoma cells (SNU-449, SNU-423, SNU-475, HepG2), as well as mesothelioma cell (NCI-H226, NCI-H28, MSTO-221h). Highly ABCG2 expressing HT-29 cells represent cancer stem like phenotype including stem cell marker expression, and self-renewal bioactivities. Interestingly, we demonstrated that in vitro treatment of SLJ-496 showed significant cytotoxicity effect, as well as viral replication capacity in ABCG2 overexpressing cell. In addition, we also demonstrated the cytotoxic effect of SLJ-496 in Adriamycin-resistant cell lines, SNU-620 and ADR-300. Taken together, these findings provide us a pivotal clue that cancer therapy using SLJ-496 vaccinia virus might be new therapeutic strategy to overcome ABCG2 expressing cancer stem-like cell and multiple chemo-resistance cancer cells.

Effects of Non-ionic Surfactant Tween 80 on the in vitro Gas Production, Dry Matter Digestibility, Enzyme Activity and Microbial Growth Rate by Rumen Mixed Microorganisms (비이온성 계면활성제 Tween 80의 첨가가 반추위 혼합 미생물에 의한 in vitro 가스발생량, 건물소화율, 효소활력 및 미생물 성장율에 미치는 영향)

  • Lee, Shin-Ja;Kim, Wan-Young;Moon, Yea-Hwang;Kim, Hyeon-Shup;Kim, Kyoung-Hoon;Ha, Jong-Kyu;Lee, Sung-Sil
    • Journal of Life Science
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    • v.17 no.12
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    • pp.1660-1668
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    • 2007
  • The non-ionic surfactant (NIS) Tween 80 was evaluated for its ability to influence invitro cumulative gas production, dry matter digestibility, cellulolytic enzyme activities, anaerobic microbial growth rates, and adhesion to substrates by mixed rumen microorganisms on rice straw, alfalfa hay, cellulose filter paper and tall fescue hay. The addition of NIS Tween 80 at a level of 0.05% increased significantly (P<0.05) in vitro DM digestibility, cumulative gas production, microbial growth rate and cellulolytic enzyme activity from all of substrates used in this study. In vitro cumulative gas production from the NIS-treated substrates; rice straw, alfalfa hay, filter paper and tall fescue hay was significantly (P<0.05) improved by 274.8, 235.2, 231.1 and 719.5% compared with the control, when substrates were incubated for 48 hr in vitro. The addition of 0.05% NIS Tween 80 to cultures growing on alfalfa hay resulted in a significant increase in CMCase (38.1%), xylanase (121.4%), Avicelase (not changed) and amylase (38.2%) activities after 36 h incubation. These results indicated that the addition of 0.05% Tween 80 could greatly stimulate the release of some kinds of cellulolytic enzymes without decreasing cell growth rate in contrast to trends reported with aerobic microorganism. Our SEM observation showed that NIS Tween. 80 did not influence the microbial adhesion to substrates used in the study. Present data clearly show that improved gas production, DM digestibility and cellulolytic enzyme activity by Tween 80 is not due to increased bacterial adhesion on the substrates.

Optimization of GFR value according to Kidney Depth Measurement Methods (신장 Depth 측정 방법에 따른 GFR 값의 최적화)

  • Kwon, Hyeong-Jin;Moon, Il-Sang;Noh, Gyeong Woon;Kang, Keon Wook
    • The Korean Journal of Nuclear Medicine Technology
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    • v.23 no.2
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    • pp.25-28
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    • 2019
  • Purpose In patients with unusual kidney position after $^{99m}Tc-DTPA$ renal dynamic imaging study, the GFR(Glomerular Filtration Rate) values are significantly different according to the depth of the kidney. Thus, we tried to compare the difference of the GFR values between the depth measurement methods and in-vitro test. 30 adult patients who were subjected to renal study. 27 patients were in usual position and 3 patients were in unusual. $555{\pm}37MBq$ of $^{99m}Tc-DTPA$ was administrated to all patients. GE infinia gamma camera was used. GFR values were obtained in-vivo(gates method) and in-vitro(blood). The kidney depth in-vivo was calculated by three methods(tonnensen, manual, taylor). In-vitro, GFR was performed by blood test. Differences in the mean values of GFR and correlation between depth and GFR values were evaluated using the SPSS 12.0 statistical program. The GFR values for 27 patients with kidney in the usual position are as follows(1.tonnensen 2.manual 3.taylor 4.invitro); $69.3{\pm}4.2$, $88.2{\pm}5.6$, $77.8{\pm}4.3$, $82.2{\pm}5.8ml/min$. The three unusual cases are as follows, first(congenital renal anomaly): 66.4, 101.24, 69.07, 94.8 ml/min. second(transplantation kidney): 12.22, 29.99, 19.36, 23.5 ml/min. third(horseshoe kidney): 37.37, 93.54, 35.9, 92.5 ml/min. There was a difference between tonnensen and manual in the usual position of the kidney(p<0.05). There was no significant difference between the other methods. However, there was a significant difference in case of the unusual position of the kidneys. Correlation analysis between both kidney depth and GFR value shows person correlation as follows; Rt kidney: 0.298, Lt kidney: 0.322. When compared with the GFR values in-vitro test, it was useful to calculate the GFR value by measuring the kidney depth using a manual formula in the unusual position of the kidneys. GFR values and kidney depth were significantly related.