• Journal of Microbiology and Biotechnology
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• v.29 no.7
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• pp.1165-1176
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• 2019

Botulinum neurotoxins (BoNTs), produced by Clostridium botulinum, are the most toxic substances known. However, the number of currently approved medical countermeasures for these toxins is very limited. Therefore, studies on therapeutic antitoxins are essential to prepare for toxin-related emergencies. Currently, more than 10,000 Halla horses, a crossbreed between the native Jeju and Thoroughbred horses, are being raised in Jeju Island of Korea. They can be used for equine antitoxin experiments and production of hyperimmune serum against BoNT/A1. Instead of the inactivated BoNT/A1 toxoid, Halla horse was immunized with the receptor-binding domain present in the C-terminus of heavy chain of BoNT/A1 (BoNT/A1-HCR) expressed in Escherichia coli. The anti-BoNT/A1-HCR antibody titer increased rapidly by week 4, and this level was maintained for several weeks after boosting immunization. Notably, $20{\mu}l$ of the week-24 BoNT/A1-HCR(-immunized) equine serum showed an in vitro neutralizing activity of over 8 international units (IU) of a reference equine antitoxin. Furthermore, $20{\mu}l$ of equine serum and $100{\mu}g$ of purified equine $F(ab^{\prime})_2$ showed 100% neutralization of 10,000 $LD_{50}$ in vivo. The results of this study shall contribute towards optimizing antitoxin production for BoNT/A1, which is essential for emergency preparedness and response.

• Journal of Physiology & Pathology in Korean Medicine
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• v.17 no.1
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• pp.1-24
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• 2003

This study was done to compile 104 experimental theses which are related to the antitumor and immuno-activating therapies between February 1990 through February 2002. Master's and doctoral theses were dassified by schools, degrees, materials, effects, experimental methods of antitumor and immunoactivity, and results. The following results were obtained from this study : 1. Classifying the theses by the school, 34.6% were presented by Daejeon University, 29.8% by Kyung-hee University and 11.5% by Won-kwang University. Of all theses, 51.0% were aimed for the doctoral degree and 43.3% were for the master's degree. All of three universities have their own cancer centers. 2. Classifying the theses by herb materials, complex prescription accounted for 60.3%, single herb accounted for 24.8% and herbal acupuncture accounted for 14.2%. Considering the key principles of the traditional medicine, complex prescription was much more thoroughly studied than single herb prescription. The results showed that the complex prescription had both antitumor activity and immuno-activating activity, which might reflects on multi-activation mechanisms by complex components. 3. Classifying the theses by the efficacy of herbs examined, in single herb, invigorating spleen and supplementing was 35.5%, expelling toxin and cooling was 29.0%, activating blood flow and removing blood stasis was 12.9%. In herbal acupuncture, invigorating spleen and supplementing was 52.9%, expelling toxin and cooling was 29.4%. In complex prescription, pathogen-free status was 41.9%, strengthening healthy qi to eliminate pathogen was 35.5%, strengthening healthy qi was 22.6%. It is presumed that the antitumor and immunoactivating therapy based on syndrome differentiation is the best way to develop oriental oncology. 4. Classifying the theses by antitumor experiments, cytotoxic effect was 48.1 %, survival time was 48.1 % and change of tumor size was 42.3%. Survival rate was not necessarily correlated with cytotoxicity. These data reflect the characteristic, wholistic nature of the oriental medicine which is based on BRM (biological response modifier). 5. Classifying the theses by immunoactivating experiments, hemolysin titer was 51.0%, hemagglutinin titer was 46.2% and NK cell's activity was 44.2%. In the future studies, an effort to elucidate specific molecular and cellular mechanisms of cytokine production in the body would be crucial. 6. Classifying the theses according to the data in terms of antitumor activity, 50% was evaluated good, 24.0% was excellent, and 15.5% have no effect. In an evaluation of immuno-activating activity, 35.9% was excellent and 18.0% showed a little effect. The index point, as described here, may helps to use experimental data for clinical trials. Changes in index points by varying dosage implicate the importance of oriental medical theory for prescription. 7. In 167 materials, IIP (immunoactivating index point, mean : 3.12±0.07) was significantly higher than AIP(antitumor index point, mean : 2.83±0.07). These data demonstrate that the effect of herb medicine on tumor activity depends more on immunoactivating activity than antitumor activity. This further implies that the development of herbal antitumor drugs must be preceded by the mechanistic understanding of immunoactivating effect. 8. After medline-searching tumor and herb-related articles from NCBI web site, we conclude that most of the studies are primarily focused on biomolecular mechanisms and/or pathways. Henceforth, we need to define the biomolecular mechanisms and/or pathways affected by herbs or complicated prescriptions. 9. Therefore, the most important point of oriental medical oncology is to conned between experimental results and clinical trials. For the public application of herbal therapy to cancer, it is critical to present the data to mass media. 10. To develop the relationship of experimental results and clinical trials, university's cancer clinic must have a long-range plan related to the university laboratories and, at the same time, a regular consortium for this relationship is imperative. 11. After all these efforts, a new type herbal medicine for cancer therapy which is to take care of the long-term administering and safety problem must be developed. Then, it would be expected that anti-tumor herbal acupuncture can improve clinical symptoms and quality of life (QOL) for cancer patients. 12. Finally, oriental medical cancer center must be constructed in NCC (National Cancer Center) or government agency for the development of oriental medical oncology which has international competitive power.

• Journal of Food Hygiene and Safety
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• v.33 no.5
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• pp.404-411
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• 2018

To evaluate bacteriological and toxicological safety hygienic indicator bacterium and paralytic and diarrhetic shellfish toxins in the shellfish produced in Hansan Geojeman 2013-2017 were investigated. Fecal coliforms were < 18~330 MPN/100 g in 404 oyster samples. But all samples tested, did not exceed 230 E. coli MPN/100 g. Geometric mean of E. coli for oyster samples collected during major shellfish production period was 24.3 MPN/100 g, considerde stable results. Bacteriological quality of oysters collected from Hansan Geojeman meets the standard value based on shellfish hygiene of the Food Sanitation Act of Korea and also meets Grade A, according to classification of shellfish harvesting areas of the European Union. For toxicological evaluation of Hansan Geojeman, 532 oyster samples and 268 mussel samples as an indicator, were analyzed. Paralytic shellfish toxins were detected in the range of 0.42~2.29 mg/kg in eight mussel samples, and exceeded criteria in three samples from early to late April 2013. Diarrhetic shellfish toxin was detected in three of 120 samples, but it was revealed to be under regulation value (0.16 mg Okadaic Acid equ./kg). As a result of toxicological evaluation, paralytic and diarrhetic shellfish toxins were not detected in oyster samples, but it was found that mussel as an indicator species, exceeded the threshold value of paralytic shellfish toxin. Accordingly, sanitary surveys were continuously requested for food safety management of shellfish.

• Korean Journal of Microbiology
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• v.51 no.3
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• pp.300-307
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• 2015

For selecting Bacillus strains producing high-quality Cheonggukjang, 8 strains were isolated from the different Cheonggukjang samples. Seven of them exhibited the highest 16S rRNA gene sequence similarity value of over 99.9% to Bacillus subtilis subsp. subtilis and one of them showed the similarity to B. licheniformis. All the strains showed positive activities for amylase, cellulase, protease and lipase, and 6 strains are positive for fibrinolytic activity. To confirm the safety of the strains isolated from the samples of Cheonggukjang which are manufactured by traditional method, strains were analyzed for the presence of seven toxin genes of Bacillus cereus and results were found negative. And 7 strains did not produce at all or merely produce both histamine and tyramine, the representative biogenic amines. Biogenic amine degradation analysis by HPLC revealed that, most of them exhibited tyramine degradation activity. For Cheonggukjang fermented by artificial inoculation of selected strains, fermentation property, sensory test, volatile basic nitrogen production and metabolic profiles by $^1H-NMR$ were tested. Seven strains were confirmed to make high-quality Cheonggukjang.

• The Korean Journal of Physiology and Pharmacology
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• v.5 no.4
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• pp.287-297
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• 2001

Contraction of smooth muscle is initiated by an increase in cytosolic $Ca^{2+}$ leading to activation of $Ca^{2+}$/ calmodulin-dependnet myosin light chain (MLC) kinase and phosphorylation of MLC. The types of contraction and signaling mechanisms mediating contraction differ depending on the region. The involvement of these different mechanisms varies depending on the source of $Ca^{2+}$ and the kinetic of $Ca^{2+}$ mobilization. $Ca^{2+}$ mobilizing agonists stimulate different phospholipases $(PLC-{\beta},\;PLD\;and\;PLA_2)$ to generate one or more $Ca^{2+}$ mobilizing messengers $(IP_3\;and\;AA),$ and diacylglycerol (DAG), an activator of protein kinase C (PKC). The relative contributions of $PLC-{\beta},\;PLA_2$ and PLD to generate second messengers vary greatly between cells and types of contraction. In smooth muscle cell derived form the circular muscle layer of the intestine, preferential hydrolysis of $PIP_2$ and generation of $IP_3$ and $IP_3-dependent\;Ca^{2+}$ release initiate the contraction. In smooth muscle cells derived from longitudinal muscle layer of the intestine, preferential hydrolysis of PC by PLA2, generation of AA and AA-mediated $Ca^{2+}$ influx, cADP ribose formation and $Ca^{2+}-induced\;Ca^{2+}$ release initiate the contraction. Sustained contraction, however, in both cell types is mediated by $Ca^{2+}-independent$ mechanism involving activation of $PKC-{\varepsilon}$ by DAG derived form PLD. A functional linkage between $G_{13},$ RhoA, ROCK, $PKC-{\varepsilon},$ CPI-17 and MLC phosphorylation in sustained contraction has been implicated. Contraction of normal esophageal circular muscle (ESO) in response to acetylcholine (ACh) is linked to $M_2$ muscarinic receptors activating at least three intracellular phospholipases, i.e. phosphatidylcholine-specific phospholipase C (PC-PLC), phospholipase D (PLD) and the high molecular weight (85 kDa) cytosolic phospholipase $A_2\;(cPLA_2)$ to induce phosphatidylcholine (PC) metabolism, production of diacylglycerol (DAG) and arachidonic acid (AA), resulting in activation of a protein kinase C (PKC)-dependent pathway. In contrast, lower esophageal sphincter (LES) contraction induced by maximally effective doses of ACh is mediated by muscarinic $M_3$ receptors, linked to pertussis toxin-insensitive GTP-binding proteins of the $G_{q/11}$ type. They activate phospholipase C, which hydrolyzes phosphatidylinositol bisphosphate $(PIP_2),$ producing inositol 1, 4, 5-trisphosphate $(IP_3)$ and DAG. $IP_3$ causes release of intracellular $Ca^{2+}$ and formation of a $Ca^{2+}$-calmodulin complex, resulting in activation of myosin light chain kinase and contraction through a calmodulin-dependent pathway.

• Asian-Australasian Journal of Animal Sciences
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• v.26 no.11
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• pp.1592-1597
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• 2013

Type of dietary direct-fed microbials (DFMs) or poultry litter could directly influence the composition of gut microbiota. Gut microbiota plays an important role in shaping the developing immune system and maintaining the homeostasis of the mature immune system in mammal and chickens. The present study was carried out to investigate the interaction among litter, DFMs and immunity in broiler chickens exposed to a field-simulated environment. Immune status of broiler chickens was assessed by serum antibodies against Eimeria spp. and Clostridium spp. and intestinal cytokine mRNA expression. The current experimental design had a $3{\times}2$ factorial arrangement of treatments with three types of litter, i.e., fresh litter or used litter that was obtained from a farm with no disease outbreak (used litter) or a farm with history of a gangrenous dermatitis outbreak (GD litter), and two dietary treatments with or without DFMs. It was found that either DFM addition or type of litter significantly affected anticoccidial antibody levels of broiler chickens at d 42. In general, dietary DFMs increased the anticoccidial antibodies in the fresh-litter raised chickens, but lowered the levels in the GD-litter raised chickens. Serum antibodies against Clostridium perfringens ${\alpha}$-toxin were significantly (p<0.05) higher in chickens raised on GD litter compared with those raised on fresh litter. Cytokine mRNA expression was significantly (p<0.05) altered by either the type of litter or DFMs. Of interest, dietary DFMs lowered interferon-${\gamma}$, interleukin 1beta, and CXCLi2 cytokine mRNA expression in chickens raised on fresh litter but increased them in GD-litter raised chickens. In conclusion, dietary DFMs modulate various immune parameters of broiler chickens, but the DFM-mediated effects were dependent upon the type of litter on which chickens were raised.

• Journal of Life Science
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• v.27 no.8
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• pp.873-878
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• 2017

We previously reported that NAD(P)H:quinone oxidoreductase 1 (NQO1)-knockout (KO) mice exhibited spontaneous inflammation in the gut. We also found that NQO1-KO mice showed highly increased inflammatory responses compared with NQO1-WT control mice when subjected to DSS-induced experimental colitis. In a Clostridium difficile toxin-induced mouse enteritis model, NQO1-KO mice were also sensitive compared with NQO1-WT mice. Moreover, numerous studies have shown that NQO1 is functionally associated with immune regulation. Here, we assessed whether NQO1 defects can alter macrophage activation. We found that peritoneal macrophages isolated from NQO1-KO mice produced more IL-6 and $TNF-{\alpha}$ than those isolated from NQO1-WT mice. Moreover, the dicumarol-induced inhibition of NQO1 significantly increased IL-6 and $TNF-{\alpha}$ production in peritoneal macrophages isolated from NQO1-WT mice, as well as in the cultured mouse macrophage cell line, RAW264.7. These results indicate that NQO1 may negatively regulate the activation of macrophages. Knockout or chemical inhibition of NQO1 markedly reduced the expression of $I{\kappa}B$ (inhibitor of $NF{\kappa}B$) in both mouse peritoneal macrophages and RAW264.7 cells. Finally, RAW264.7 cells treated with dicumarol exhibited morphological changes reflecting macrophage activation. Our results suggest that NQO1 may suppress the $NF{\kappa}B$ pathways in macrophages, thereby suppressing the activation of these cells. Thus, immunosuppressive activity may be among the many possible functions of NQO1.

• Korean Journal of Soil Science and Fertilizer
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• v.47 no.6
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• pp.510-517
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• 2014

The purpose of this study was to analyze microbial hazards for cultivation environments and personal hygiene of strawberry and tomato farms at the growth and harvesting stage. Samples were collected from thirty strawberry farms and forty tomato farms located in Korea and tested for Staphylococcus aureus and Bacillus cereus. To investigate the change in the distribution of the S. aureus and B. cereus, a total of 4,284 samples including air born, soil or medium, mulching film, harvest basket, groves and irrigation water etc. were collected from eight strawberry farms and nine tomato farms for one year. As a result, total S. aureus and B. cereus in all samples were detected. Among the total bacteria of strawberry farms, S. aureus (glove: $0{\sim}2.1Log\;CFU/100cm^2$, harvest basket: $0{\sim}3.0Log\;CFU/100cm^2$, soil or culture media: 0~4.1 Log CFU/g, mulching film: $0{\sim}3.8Log\;CFU/100cm^2$), B. cereus (glove: $0{\sim}2.8Log\;CFU/100cm^2$, harvest basket: $0{\sim}4.8Log\;CFU/100cm^2$, soil or culture media: 0~5.3 Log CFU/g, mulching film: $0{\sim}4.5Log\;CFU/100cm^2$) were detected in all samples. The total bacteria of tomato farms, S. aureus (glove: $0{\sim}4.0Log\;CFU/100cm^2$, harvest basket: $0{\sim}5.0Log\;CFU/100cm^2$, soil or culture media: 0~6.1 Log CFU/g, mulching film: $0{\sim}4.0Log\;CFU/100cm^2$), B. cereus (glove: $0{\sim}4.0Log\;CFU/100cm^2$, harvest basket: $0{\sim}4.3Log\;CFU/100cm^2$, soil or culture media: 0~5.9 Log CFU/g, mulching film: $0{\sim}4.7Log\;CFU/100cm^2$) were detected in all samples. The contamination of S. aureus and B. cereus were detected in soil, mulching film and harvest basket from planting until harvest to processing, with the highest count recorded from the soil. But S. aureus and B. cereus were not detected in irrigation water samples. The incidence of S. aureus and B. cereus in hydroponics culture farm were less than those in soil culture. The amount of S. aureus and B. cereus detected in strawberry and tomato farms were less than the minimum amount required to produce a toxin that induces food poisoning. In this way, the degree of contamination of food poisoning bacteria was lower in the production environment of the Korea strawberry and tomato, but problems can be caused by post-harvest management method. These results will be used as fundamental data to create a manual for sanitary agricultural environment management, and post-harvest management should be performed to reduce the contamination of hazardous microorganisms.

• The Korean Journal of Pesticide Science
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• v.18 no.4
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• pp.358-363
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• 2014

We was investigated parasporal inclusion proteins change to use industrial medium of new strain Bacillus thuringiensis CAB 565, CAB 566. To confirm medium's oxygen efficient consist of glucose and yeast extract, we was conducted oxygen transfer coefficients (KLa) of medium's concentration and impeller in 20 l-Jar fermentor. When to increase air flow rate and medium concentration, KLa rate is rise. Also it is effective on agitation rate 200 rpm, but KLa rate is decrease when to rise agitation rate. To hold dissolved oxygen rate (upper 50%), Air flow rate is steadily increase on culture to use microsparger. When 16 hour of culture stage, B.t. CAB 565 and B.t. CAB 566 harvested respectively $2.3{\times}10^{10}$, $1.8{\times}10^{10}$ viable cell/ml. When 54 hour, B.t. CAB565, 566 harvested respectively $1.9{\times}10^{10}$, $1.4{\times}10^{10}spore/ml$. To resulting carbon's concentration, It is the most effective that glucose concentration is contained 5% in medium.

• The Korean Journal of Physiology and Pharmacology
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• v.1 no.4
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• pp.413-423
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• 1997

The importance of the kidney in the development of hypertension was first demonstrated by Goldblatt and his colleagues more than fifty years ago. Many hormones and other regulatory factors have been proposed to play a major role in the development of hypertension. Among these factors angiotensia II (ANG II) is closely involved in renal hypertension development since it directly regulates $Na^+$ reabsorption in the renal proximal tubule. Thus the aim of the present study was to examine signaling pathways of low dose of ANC II on the $Na^+$ uptake of primary cultured rabbit renal proximal tubule cells (PTCs) in hormonally defined seum-free medium. The results were as follows: 1) $10^{-11}$ M ANG II has a significant stimulatory effect on growth as compared with control. Alkaline phosphatase exhibited significantly increased activity. However, leucine aminopeptidase and ${\gamma}-glutamyl$ transpeptidase activity were not significant as compared with control. In contrast to $10^{-11}$ M ANG II stimulated $Na^+$ uptake $(108.03{\pm}2.16% of that of control)$, $10^{-9}$ M ANG II inhibited ($92.42{\mu}2.23%$ of that of control). The stimulatory effect of ANG II on $Na^+$ uptake was amiloride-sensitive and inhibited by losartan (ANG II receptor subtype 1 antagonist) and not by PD123319 (ANG II receptor subtype 2 antagonist). 2) Pertussis toxin (PTX) alone inhibited $Na^+$ uptake by $85.52{\pm}3.52%$ of that of control. In addition, PTX pretreatment prevented the AMG II-induced stimulation of $Na^+$ uptake. 8-Bromoadenosine 3',5'-cyclic monophosphate (8-Br-cAMP), forskolin, and isobutylmethylxanthine (IBMX) alone inhibited $Na^+$ uptake by $88.79{\pm}2.56,\;80.63{\pm}4.38,\;and\;84.47{\pm}4.74%$ of that of control, respectively, and prevented the ANG II-induced stimulation of $Na^+$ uptake. However, $10^{-11}$ M ANG II did not stimulate cAMP production. 3) The addition of 12-O-te-tradecanoylphorbol-13-acetate (TPA, 0.01 ng/ml) to the PTCs produced significant increase in $Na^+$ uptake ($114.43{\pm}4.05%$ of that of control). When ANG II and TPA were added together to the PTCs, there was no additive effect on $Na^+$ uptake. Staurosporine alone had no effect on $Na^+$ uptake, but led to a complete inhibition of ANG II- or TPA-induced stimulation of Na'uptake. ANG II treatment resulted in a $111.83{\mu}4.51%$ increase in total protein kinase C (PKC) activity. In conclusion, the PTX-sensitive PKC pathway is the main signaling cascade involved in the stimulatory effects of ANG II on $Na^+$ uptake in the PTCs.