Objectives: This study was designed to investigate intestinal immune system activation and antimetastatic effect of Ojeok-san on cancer cells by oral administration. Methods: Cell viability of Ojeok-san was tested with colon 26-M3.1 carcinoma cells and Peyer's patch cells in vitro. Antimetastatic experiments were conducted in vivo mouse model by using colon 26-M3.1 carcinoma cell. To observe immunomodulating effects of Ojeok-san on Peyer's patch cells, we measured interleukin (IL)-4, GM-CSF. In addition to observing effects of Ojeok-san on hematopoiesis, we measured proliferation of bone marrow cells mediated by Peyer's patch cells in vitro. IgA induction activated in serum and intestinal content was measured to observe the effect of orally administered Ojeok-san on mucosal immune system. After administering Ovalbumin (OVA) with Ojeok-san, Proliferation of Peyer's patch cell was measured to investigate gut immunostimulatory effect. Results: in vitro cytotoxicity analysis, the inhibitory concentration $(IC)_{50}$ of the colon 26-M3.1 carcinoma cell was $890{\mu}g/ml$. $IC_{50}$ of the Peyer's patch cells with LPS was $990{\mu}g/ml$. We found that orally administered Ojeok-san significantly inhibited tumor metastasis in vivo. In addition, the amounts of IL-4 and GM-CSF in the culture supernatant of Peyer's patch cells were significantly increased compared to the control group. The proliferation of bone marrow cell was significantly up-regulated with Ojeok-san. These results indicate that oral administration of Ojeok-san enhances the secretion of hematopoietic growth factors such as GM-CSF and IL-4 from Peyer's patch cells, and these cytokines also act on modulator of bone marrow cell proliferation. After orally administering Ovalbumin (OVA) with Ojeok-san, IgA induction and Proliferation of peyer's patch cell was up-regulated with Ojeok-san. These results means orally administered Ojeok-san activates intestinal immune system and has an inhibitory effect on tumor metastasis. Conclusions: Orally administered Ojeok-san appears to have considerable activity on the anti-metastasis by activation of immune system.
The antigenecity of somatic substances of S. pullorum standard strain and variant strain extracted byheat treatment, acid treatment and their modification, ammonium sulfate saturation (60 per cent), trypsin digestion was tested by indirest hemagglutination test and precipitation test and following results were optained. 1. Teatment at $100^{\circ}C$ for an hour of the bacteria could extract the antigen of S. pullorum standard strain and variant strain which was demonstrable by hemagglutination reaction with the human a group and chicken red blood cell. 2. Trypsin digestion was more enhanced its antigenecity in acid extracted antigen of S. pullorum variant strain compare with the S. pullorum standard strain. 3. The extracted antigenic substances of S. pullorum standard strain existed chiefly in the elicited fraction of precipitate at the treatment of ammonium sulfate saturation and after trypsin digestion, its antigenecity was demonstrated by hemagglutination. 4. At the treatment of ammonium sulfate treatment, did not occur the precipicate in acid extracted antigens of S. pullorum variant strain, however, the heal extracted antigen, positive reactions were obtained in both of the precipitate and supernatant fraction of the S. pullorum variant strain by hemagglutination reaction. After trypsin digestion, these fraction also exhibited positive reactions. 5. Precipitation test also tested dub could not detect in any soft of the antigens.
This study is focused on establishing the optimal conditions to enhance the production of antilisterial substances by Lactobacillus paracasei BK57 isolated from Baikkimchi. In addition, the growth and in situ lactic acid and bacteriocin production of this strain were investigated during the manufacture of fresh cheese. And then the efficacy of using Lactobacillus starter as a protective culture to improve the safety of fresh cheese against Listeria monocytogenes KCTC 3569 was estimated. Maximum growth rate and activity of antibacterial substances were obtained in Lactobacilli MRS broth at $37^{\circ}C$ with controlled pH 6.0 after 30 h of incubation under aerobic condition. However, the growth rate and antimicrobial activity of bacteriocin produced in whole milk supplemented with yeast extract (2.0%) as a substrate were lower than those obtained in MRS broth. Live cells and cell-free culture supernatant of BK57 strain were effective in the suppression of L. monocytogenes in milk, whereas the inhibitory of the bacteriocin obtained from BK57 strain was higher in BHI broth than in milk. During storage at $4^{\circ}C$ and $15^{\circ}C$ for 6 days, no significant difference was found in the cell viability and antimicrobial activity of BK 57 strain in fresh cheese. In samples held at two temperatures, there was at least a 15% reduction in the numbers of the pathogen in fresh cheese artificially contaminated with approximately $10^5CFU/ml$ of L. monocytogenes within 6 days. Our results demonstrated the usefulness of L. paracasei BK57 having antilisterial activity as a biopreservative in the cheese making process.
Kim, Yong-Sang;Jeong, Jin-Oh;Cho, Sung-Ho;Jeong, Do-Yeon;Uhm, Tai-Boong
Korean Journal of Microbiology
/
v.48
no.2
/
pp.163-170
/
2012
In order to inhibit the growth of pathogens and degrade biogenic amines during the fermentation of soybean products, an isolate with antimicrobial activity against pathogens and biogenic amine-degrading property was obtained from 83 traditionally fermented soybean products. The morphological and biochemical tests and the phylogenetic relationship among 16S rRNA gene sequences indicated that the isolate named as the strain SCK B11 was most closely related to Bacillus licheniformis. The cell-free supernatant of two day cultures was active against several pathogens including Enterococcus faecalis, Listeria monocytosis, Micrococcus luteus, Pseudomonas aeruginosa, Bacillus cereus, and Staphylococcus aureus. PCR analysis was conducted to determine relatedness to antimicrobial lantibiotics and biosurfactants produced by Bacillus spp., but showed negative for the genes encoding surfactin, lichenysin, and lichenicidine. Electron microscopic observation indicated that the antimicrobial agent seemed to attack the membrane of the pathogens, leaving the ghost or shrunken cells. The strain was found to degrade histamine by 72% and tyramine by 66% in the cooked soybean containing 5.3% of biogenic amine over 10 days of fermentation time. The use of selected strain would be a potential control measure in manufacturing traditionally fermented soybean products that are difficult to control pathogens and biogenic amine levels.
Valizadeh, Armita;Ahmadzadeh, Ahmad;Saki, Ghasem;Khodadadi, Ali;Teimoori, Ali
Asian Pacific Journal of Cancer Prevention
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v.16
no.18
/
pp.8533-8539
/
2016
Background: B-cell chronic lymphocytic leukemia B (B-CLL), the most common type of leukemia, may be caused by apoptosis deficiency in the body. Adipose tissue-derived mesenchymal stem cells (AD-MSCs) as providers of pro-apoptotic molecules such as tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), can be considered as an effective anti-cancer therapy candidate. Therefore, in this study we assessed the role of tumor necrosis factor-producing mesenchymal stem cells oin apoptosis of B-CLL cells resistant to fludarabine-based chemotherapy. Materials and Methods: In this study, after isolation and culture of AD-MSCs, a lentiviral LeGO-iG2-TRAIL-GFP vector containing a gene producing the ligand pro-apoptotic with plasmid PsPAX2 and PMDG2 virus were transfected into cell-lines to generate T293HEK. Then, T293HEK cell supernatant containing the virus produced after 48 and 72 hours was collected, and these viruses were transduced to reprogram AD-MSCs. Apoptosis rates were separately studied in four groups: group 1, AD-MSCs-TRAIL; group 2, AD-MSCs-GFP; group 3, AD-MSCs; and group 4, CLL. Results: Observed apoptosis rates were: group 1, $42{\pm}1.04%$; group 2, $21{\pm}0.57%$; group 3, $19{\pm}2.6%$; and group 4, % $0.01{\pm}0.01$. The highest rate of apoptosis thus occurred ingroup 1 (transduced TRAIL encoding vector). In this group, the average medium-soluble TRAIL was 72.7pg/m and flow cytometry analysis showed a pro-apoptosis rate of $63{\pm}1.6%$, which was again higher than in other groups. Conclusions: In this study we have shown that tumor necrosis factor (TNF) secreted by AD-MSCs may play an effective role in inducing B-CLL cell apoptosis.
Kim, Seogku;Kim, Jahyun;Lim, Junhyuk;Lee, Jeakun;Lee, Taeyoon
Journal of the Korean GEO-environmental Society
/
v.15
no.10
/
pp.15-21
/
2014
In this study, biochemical methane potential test was conducted to estimate ultimate methane and carbon dioxide yield for anaerobic digestion and pretreatment with sewage sludge cake. Two of 0.2 % TS of sewage sludge cakes were treated with 5M NaOH or sonication of 0.51 W/mL during 30 min respectively. Another sample was treated simultaneously with NaOH and sonication in same condition. Then, initial soluble COD increased from 33.1 mg/L to 494 mg/L. After BMP test, methane production ranged from 3.12 and 84.2 mL $CH_4$ per g of Volatile Solid (VS) and 9.2 and 13.5 mL $CO_2$ per g of Volatile Solid (VS) for carbon dioxide. In other tests, injection of nutrient media or sludge supernatant produced 73.1 and 73.8 mL $CH_4$ per g of Volatile Solid (VS) and 11.2 and 13.6 mL $CO_2$ per g of Volatile Solid (VS) respectively. When BMP test finished, 62 % of initial volatile solids decreased to 33.8~45.4 %. Simultaneous pretreatment increased soluble COD, reduction rate of volatile solids and digestion efficiency than those for alkaline and ultrasonic pretreatment.
Optimal medium composition, culture conditions, characteristics of phase variation and activity of insecticidal toxin by Xenorhabdus nematophilus isolated and identified from Korean entomopathogenic nematode Steinernema carpocapsae were examined. Optimal medium composition of this strain was 50-70 g/L yeast extract, 3 g/L $K_{2}HPO_{4}$, 1g/L $NH_{4}H_{2}PO_{4}$, 2g/L ${MgSO}_4$$\cdot$${7H}_{2}O$, 10g/L NaCl and, these, yeast extract was found as a limiting nutrient for cell growth. When Monod equation was applied, maxmum specific growth rate and Monod constant were estimated as 0.13 $hr^{-1}$ and 20g/L, respectively. The pH of culture medium increased up to 8.5-9.5 regardless of initial pH 6-7 as the cells continued to grow. The specific growth rate in a 7 L fermentor was 0.18 $hr^{-1}$, which was enhancement 1.4 fold compared to a flask culture. In case of phase variation, phase I fraction was maintained above 90% at the stationary phase for both flask and fermentor cultures. According to oral toxicity test of Gallena mellonella by Xenorhabdus nematophilus, the addition of cell pellets into feed inhibited normal growth of insect larvae and killed completely then after 20 days cultivation. When culture supernatant of this strain was injected into hemolymph of insect larva, the toxicity was strongest at 24hr cultivation in the early exponential phase and gradually decreased as the culture time proceeded.
Kim Dong Kkun;Kim Ji Seong;Kim Ho Il;Lee Yu Na;Pak Dae Won
한국생물공학회:학술대회논문집
/
2004.07a
/
pp.21-33
/
2004
In this study, Anaerobic sewage sludge in a batch reactor operating at $35^{\circ}C$ was used as the seed to investigate the effect of pretreatments of waste activated sludge and to evaluate its hydrogen production potential by anaerobic fermentation. Various pretreatments including physical, chemical and biological means were conducted to utilize for substrate. As a result, SCODcr of alkali and mechanical treatment was 15 and 12 times enhanced, compared with a supernatant of activated sludge. And SCODcr was 2 time increase after re-treatment with biological hydrolysis. Those were shown that sequential hybridized treatment of sludge by chemical & biological methods is most efficient process for sludge treatment. The pre-treatment activated sludge was tested to conform hydrogen production potential in batch experiments. When buffer solution was added to the activated sludge, hydrogen production potential increased as compare with no addition.
Journal of the Korean Society of Food Science and Nutrition
/
v.18
no.2
/
pp.167-174
/
1989
AS a part of investigation to use anchovy more effectively as food source, this work was undertaken the processing conditions of extracts from rapid fermented anchovy sauce. To prepare rapid fermented products, the chopped anchovy was mixed with 50% water (w/w), and then hydrolyzed by autolysis or addition of koji under different conditions of hydrolysis. The optimal conditions for hydrolysis of chopped anchovy were revealed $50^{\circ}C$, 6 hours, pH 8.0 by autolysis, and ,$50^{\circ}C$, 6 hours, pH 7.0 by addition of 10% koji, respectively. But, pH control was not much beneficial in increasing yield. The effect of soybean protein isolote for improvement of bitter taste was also tested. The reasonable amount of added soybean protein isolate was revealed 5% on the weight basis of the chopped anchovy. The reaction mixture hydrolyzed under optimal conditions were added with 1% onion powder (w/w), 1% garlic powder(w/w) and 1% red pepper powder(w/w) for masking fishy odor, inactivated for 20 min at $100^{\circ}C$, and then centrifuged for 20 min at 4,000 rpm. The supernatant liquor was filtrated and evaporated to 50%(v/v). finally, table salt was added for bateriostatic effect and characteristic taste of rapid fermented products. the reasonable amount of added table salt was reversed 15% on the volume basis of the evaporated liquor. The hydrolysis ratio of product made by addition of water, product made by addition of koji and water, and product made by addition of soybean protein isolate, koji and water hydrolyzed under optimal conditions were 58.4%, 82.1% and 86.2%, respectively.
Studies of immune responses in insects have focused on mechanisms that interact directly with invading microorganisms. However, few studies have examined the immune response to various metabolites produced by microorganisms after they enter the host. Here, we examined immune responses in Protaetia brevitarsis seulensis larvae induced by metabolites produced by symbiotic and pathogenic bacteria. The two types of bacteria were cultured under the same conditions. The bacteria were then removed and the remaining culture supernatant was injected into the larvae. The larvae injected with culture medium (Ch-medium) from symbiotic bacteria remained relatively healthy and did not develop an immune response, whereas more than 60% of the larvae injected with pathogen culture medium (Ec-medium) died after 150 hours and dark brown patches of melanin were observed at the injection site. This immune response was confirmed by the finding of activated lysosomes in insect granulocytes. More than 50% of lysosomes in larvae injected with pathogen culture medium were strongly stained after 12 h, but less than 5% of those injected with symbiotic culture media were stained. Therefore, it is assumed that symbiotic bacteria produce few (if any) substances that induce host immune responses.
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