New antitumor active polymers, poly(methacryloyl-2-oxy-1,2,3-propanetricarboxylic acid-co-exo-3,6-epoxy-l,2,3,6-tetrahydrophthalic acid) [poly(MTCA-co-ETAc)], poly(methacryloyl-2-oxy-l,2,3-propanetricarboxylic acid-co-hydrogen ethyl-exo-3,6-epoxy-l,2,3,6-tetrahydrophthalate) [poly(MTCA-co-HEET)], and poly(methacryloyl-2-oxy-l,2,3-propanetricarboxylic acid-co-a-ethoxy-exo-3,6-epoxy-1,2,3,6-tetrahydrophthaloyl-5-fluorouracil) [poly(MTCA-co-EETFU)] were synthesized and characterized. Their antitumor activity, inhibition of DNA replication and antiangiogenesis were examined. The structures of the polymers were identified by FT-IR, $^1H$ and $^{13}C$-NMR spectroscopy. The number average molecular weights of the fractionated polymers determined by GPC ranged from 9,400 to 14,900, and polydispersity indices were less than 1.7. The in vitro cytotoxicity of these polymers was determined and their antitumor activity was evaluated. The $IC_{50}$ values (the drug concentration at inhibition of 50% tumor growth) indicated that the synthesized polymers were much better inhibitors of cancer cells and showed lower cytotoxicity than the free 5-FU. The in vivo antitumor activity of the conjugates was examined using mice bearing the sarcoma 180 tumor cell line. The life spans (TIC) of the mice treated with the conjugates were higher than those treated with the free 5-FU. In addition, the synthesized conjugates showed excellent antiangiogenic activity based on an embryo chorioallantoic membrane assay.
Journal of the Korean Society of Food Science and Nutrition
/
v.31
no.3
/
pp.411-416
/
2002
Yogurt base was prepared from whole milk and skim milk added with 0.2∼1.0% (w/v) of Saururus chinensis(Lour.) Bail water extract (SCe) and fermented with lactic acid bacterias (the mixed strain of Streptococcas themophilus and Lactobacillus bulgaricus) at 37$\^{C}$ for 24 hr. Quality characteristics of the yogurt were evaluated in terms of acid production, number of viable cells, viscosity and sensory property during lactic acid fermentation. The composition of organic acids was also measured by HPLC. Addition of SCe stimulated the growth of lactic acid bacteria and remarkably enhanced the acid production. The viscosity and lactic acid content of yogurt were also increased by addition of SCe. The sensory score of yogurt added with 0.4% of SCe was significantly higher than other groups in taste and overall acceptability The storage abilities of yogurts added with SCe were relatively good at 5$\^{C}$ for 15 days.
Kim, Joung Hee;Lee, Syng-Ook;Do, Kook Bae;Ji, Won Dae;Kim, Sun Gun;Back, Young Doo;Kim, Keuk-Jun
Korean Journal of Clinical Laboratory Science
/
v.50
no.1
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pp.37-43
/
2018
Chamaecyparis obtusa (CO) has recently been attracting attention because of its beneficial effects on skin allergies, atopic dermatitis, and skin diseases, such as acne and eczema. In the present study, the extract from CO leaf grown in Jangseong gun, Jeollanam-do, Korea was evaluated for its anti-oxidant, anti-inflammatory, and anti-allergic effects in vitro. The total polyphenol content of the CO leaf extract was $25.89{\pm}0.31mg$ gallic acid equivalents (GAE)/g. Gas-chromatography mass-spectrometry (GC-MS) analysis revealed the presence of six compounds in the CO leaf extract: ${\alpha}-terpinene$ (3.03 mg/g), ${\alpha}-terpineol$ (9.48 mg/g), limonene (5.96 mg/g), borneol (59.78 mg/g), myrcene (4.85 mg/g), and sabinene (11.31 mg/g). The $RC_{50}$ values of the CO leaf extract for $H_2O_2$ and ABTS radical were $5.47{\pm}0.13mg/mL$ and $4.00{\pm}0.01mg/mL$, respectively. In addition, the CO leaf extract showed significant inhibitory effects on lipopolysaccharide-induced nitric oxide production in RAW 264.7 cells and IgE-induced release of ${\beta}-hexosaminidase$ (degranulation) in mast-cell like RBL-2H3 cells. The cell viability assay showed that the CO leaf extract ($100{\sim}800{\mu}g/mL$) did not affect the viability of human normal skin fibroblast CCD-986sk cells significantly. Overall, these results suggest that the CO leaf extract is a potential functional cosmetic ingredient that can exert anti-oxidant, anti-inflammatory, and anti-allergic effects.
Lactic acid fermentation of black raspberry (Rubus occidentalis) juice was carried out by using the Lactobacillus plantarum GBL17 strain. The sterilized black raspberry juice was fermented using the L. plantarum GBL17 strain at $30^{\circ}C$ for 72 hours after which the total acidity increased and the pH value decreased. In addition, the highest total acidity content (2.38%) was reached, the lowest pH value (3.22) was observed, and the sugar content decreased by $9.8^{\circ}Brix$ after the 72 hour fermentation. The number of viable cells rapidly increased up until 24 hours, after which it gradually decreased. HPLC analysis of the organic acids showed 14.51 mg/g of lactic acid content in the fermented black raspberry juice, which was not detected in the non-fermented black raspberry juice (control). The content of fructose and glucose slightly decreased after fermentation. The total polyphenol and flavonoid contents of the fermented black raspberry juice increased significantly after fermentation. The DPPH radical scavenging activity of fermented black raspberry juice (70.92%) was higher than that of the control (62.96%). After lactic acid fermentation, there was no significant increase in ABTS radical scavenging activity. These results confirm that lactic acid bacteria, such as L. plantarum GBL17, showed generally higher activities with a potential as a functional beverage.
Purpose: Radiolabeled CEA79.4 antibody has a possibility to be used in radioimmunoscintigraphy or radioimmunotherapy of cancer. We investigated the in vitro properties and biodistribution of CEA79.4 antibody labeled with Re-188 or Tc-99m. Materials and Methods: CEA79.4 was reduced by 2-mercaptoethanol to produce-SH residue, and was labeled with Re-188 or Tc-99m. For direct labeling of Tc-99m, methylene-diphosphonate was used as transchelating agent. CEA79.4 in 50 mM Acetate Buffered Saline (ABS, pH 5.3) was labeled with Re-188, using stannous tartrate as reducing agent. In order to measure immunoreactivity and the affinity constant of radiolabeled antibody, cell binding assay and Scatchard analysis using human colon cancer cells SNU-C4, were performed. Biodistribution study of labeled CEA79.4 was carried out at 1, 14 and 24 hr in ICR mice. Results: Labeling efficiencies of Tc-99m and Re-188 labeled antibodies were $92.4{\pm}5.9%$ and $84.7{\pm}4.6%$, respectively, In vitro stability of Tc-99m-CEA79.4 in human serum was higher than Re-188-CEA79.4. Immunoreactivity and affinity constant of Tc-99m-CEA79.4 were 59.2% and $6.59{\times}10^9\;M^{-1}$, respectively, while those of Re-188-CEA79.4 were 41.6% and $4.2{\times}10^9\;M^{-1}$, respectively. After 24 hr of administrations of Re-188 and Tc-99m labeled antibody, the remaining antibodies in blood were 6.32 and 9.35% ID/g respectively. The biodistribution of each labeled antibody in other organs was similar because they did not accumulate in non-targeted organs. Conclusion: In vitro properties and biodistribution of Re-188-CEA79.4 were similar to those of Tc-99m-CEA79.4. It appears that Re-188-CEA79.4 can be used as a suitable agent for radioimmunotheraphy.
Effects of refrigeration temperature and its fluctuation range on the growth of psychrotrophic microorganisms and the quality of refrigerated foods such as apple, fish and oyster were evaluated to find optimum storage conditions for a domestic refrigerator. Refrigeration temperature was $2^{\circ}C$ or $4^{\circ}C$, and fluctuation ranges were varied: ${\pm}0.3,\;{\pm}1.0,\;{\pm}1.2,\;or\;{\pm}4.0^{\circ}C$. Changes in hardness of apples stored at $2{\pm}0.3^{\circ}C$ were much slower than those of apples stored at $4{\pm}1.2^{\circ}C$. Freshness of fish and oyster also lasted much longer at low temperature such as $2{\pm}0.3^{\circ}C$. The growth of Listeria monocytogenes inoculated on sliced ham was inhibited for 1 month at $2{\pm}0.3^{\circ}C$, but the cells at $4{\pm}1.2^{\circ}C$ began to grows as time elapsed. Therefore, it was expected that shelf-life of certain food stored in a domestic refrigerator could be extended by lowering temperature to $2^{\circ}C$ and by reducing fluctuation range of refrigerator.
Recently Carter(1952) reported the capsule antigens of Pasteurella multocida could be divided into four serological types A,B,C and D by means of precipitation tests. Subsequently he showed that the most sensitive for identification of these types involved the use of capsule substance adsorbed by erythrocytes in hemagglutination test. It may be somewhat difficult to conduct the hemagglutination test in small laboratory, because relatively large amounts of antisera and erythrocytes of the human O type are required for the test. A simple method for serological typing of P. multocida was the slide agglutination test employed by Little et al. (1943) and Namioka et al. (1962), but this method is still in controversy. The author tried adapting Carter's hemagglutination method to the slide method so called "micromethod technique", and studied on the stabilization of erythrocytes for use of slide hemagglutination to P. multocida although many invesigators reported the stabilization of erythrocytes. The results obtained are summarized as follows: 1. A simplified method (slide method) for capsule typing of the organism was developed by adapting Carter's hemagglutination reaction(tube method). Antibody-containing serum can be diluted serially on Boerner's microtest slide with capillary or serological pipetts with a considerable accuracy. The slide reaction can be carried out with case on the slide by adding $0.05m{\ell}$ of antigen-sensitized erythrocytes suspension diluted to one percent on $0.05m{\ell}$ of serially diluted antibody-containing sera, and the final result can be read after 60 minutes at the room temperature ($15^{\circ}C$). 2. It is difficult to determine superiority of inferiority between the slide method and the tube method on the pattern of the reaction of hemagglutination. 3. The pH range of 6.6 to 8.3 is optimal for the slide hemagglutination reaction. 4. The antigen-sensitization against erythrocytes at $37^{\circ}C$ is optimal for the slide hemagglutination. 5. Both the doses and concentration of antigen do not influence the antigen-adsorbing capacity of erythrocytes. 6. The reduction of antigen-sensitizing hours does not influence the antigen-adsorbing capacity of erythrocytes even 30 minutes. 7. The tannic acid treatment against formalinized and non-formalinized erythrocytes showed no effect on the reaction of hemagglutination. 8. The erythrocytes preserved at $4^{\circ}C$ in the ACD solution do not decrease the reactivity on the reaction of hemagglutination for 60 days, while they begin slight hemolysis 30 days after preserving. 9. The stable preparation of erythrocytes can be obtained by treating the cells at $37^{\circ}C$ for 20 hours with from 4 to 8 percent of formalin in saline or buffer. These cells can be preserved at $4^{\circ}C$ for more than 8 months experimented without hemolysis. With low concentration of formalin, the cells were not sufficiently stabilized resulting in the hemolysis after short period of preservation at $4^{\circ}C$. 10. The erythrocytes treated with 16 percent of formalin remain constantly or increase the reactivity for the reaction of hemagglutination. On the contrary, the cells treated with I to 8 percent of formalin decrease the reactivity. 11. There is no difference between nontreated fresh erythrocytes and the erythrocytes preserved in the ACD solution on the reactivity against the hemagglutination, and the erythrocytes treated with 16 percent of formalin showed the reactivity of higher level than that of the above two kinds of erythrocytes. 12. There is no difference between the saline and the isotonic buffer solution on the reaction of hemagglutination.
Lactic acid fermentation of peach juice was carried out by using Lactobacillus plantarum KLAB21, a strain with a high level of antimutagenic activity, When the fermentation was carried out at 25, 30, 37 and $40^{\circ}C$, the highest level in the viable counts and acid production was obtained at $37^{\circ}C$. The sterilized peach juice showed a higher level of viable counts and acid production than the non-sterilized juice. And more viable counts and acid production were observed in the juice fermented by L. plantarum KLAB21 only than that obtained by a mixed culture of L. plantarum KLAB21 and Leuconostoc mesenteroides cells. When the lactic acid fermentation was performed for 5 days, the first 3 days of fermentation resulted in an increase of the viable counts from 8.2 to of 9.2 of log cfu/mL which is the highest level, as well as a decrease of the residual reducing sugar content from 5.6 to 0.1 % Decrease in the viable counts and m significant changes in the residual reducing sugar content were observed for further fermentation up to 5 days. However, the titratable acid content increased and the pH value decreased during the fermentation for 5 days to reach the highest titratable acid content (1,98%) and the lowest pH value (3.14) after 5 days of fermentation. HPLC analysis of the organic acids showed 1,236 mg% of lactic acid and 841 mg% of galacturonic acid contents in the fermented juice which were not detected in the fresh juice before fermentation. Antimutagenic effects of $100\;{\mu}L$ of the fermented peach juice supernatant were shown to be 97.7% against MNNG(N-methyl-N'-nitro-N-nitrosoguanidine), and 58.3% against NPD(4-nitro-O-phenylenediamine) in Salmonella enterica serovar Typhimurium TA100.
We tested Trichoderma harzianum as a biocontrol agent for Rhizoctonia solani AG2-1, using six natural antifungal materials to improve its efficacy. Among the six materials tested, peony (Paeonia suffruticosa) root bark (PRB) showed the strongest antifungal activity against R. solani AG2-1, and was not antagonistic to T. harzianum. Scanning electron microscopy showed that treatment with PRB extract resulted in shortened and deformed R. so/ani AG2-1 hyphal cells. The control of radish damping-off caused by R. so/ani AG2-1 was greatly increased by combined treatments of T. harzianum and PRB, as compared with either of the two treatments alone, with the control effect increased from 42.3-51.5% to 71.4-87.6%. The antifungal compound in PRB, which was isolated in chloroform and identified as paeonol by mass spectrometry, $^1H$ NMR, and $^{13}C$ NMR analyses, inhibited the growth of R. so/ani AG2-1 but not that of T. harzianum. Thus, PRB powder or extract may be used as a safe additive to T. harzianum to improve the control of the soil borne diseases caused by R. so/ani AG2-1.
Kong, Min-Ho;Do, Duc-H.;Miyazaki, Masashi;Wei, Feng;Yoon, Sung-Hwan;Wang, Jeffrey C.
Journal of Korean Neurosurgical Society
/
v.44
no.5
/
pp.327-333
/
2008
Objective: The purpose of this study is to verify the usefulness of the rabbit model for disc degeneration study. Materials: The L1-L2, L2-L3, L3-L4. or L4-L5 lumbar intervertebral disc (IVD) of 9 mature male New Zealand White rabbits were injured by inserting a 16-gauge needle to a depth of 5 mm in the left anterolateral annulus fibrosus while leaving L5-L6 IVD uninjured. Three other rabbits also received intradiscal injections of rabbit disc cells transfected with adenovirus and bone morphogenetic protein-2 (ad-BMP-2) at L4-L5 in addition to injury by 16-gauge needle at the L1-L2 level. Using digitized radiographs, measurements of IVD height were made and analyzed by using the disc height index (DHI). Magnetic resonance imaging (MRI) scans of the injured discs, injected discs, and uninjured L5-L6 discs were performed at 15 weeks post surgery and compared with preoperative MRI scans. Results: All twelve rabbits showed consistent results of disc degeneration within 15 weeks following annular puncture. DHIs of injured discs were significantly lower than that of the uninjured L5-L6 discs (p<0.05). The mean value of disc degeneration grade of injured discs was significantly higher than that of uninjured discs (p<0.05). The injection of disc cell transfected with ad-BMP-2 did not induce disc regeneration at 15 weeks after injection. Conclusion: This study showed that the injured disc had a significant change in DHI on simple lateral radiograph and disc degeneration grade on MRI scans within 15 weeks in all rabbits. Rabbit annular puncture model can be useful as a disc degeneration model in vivo.
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