Kim, Min-Ju;Kwak, Jung-Hyun;Baek, Seung-Han;Yeo, Hyun-Yang;Song, Ju-Hyun;Cho, Bong-Jun;Kim, Oh-Yoen
Preventive Nutrition and Food Science
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v.15
no.2
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pp.137-142
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2010
Previous studies report that organo-sulfur compounds derived from garlic inhibited smooth muscle cell (SMC) proliferation and induced apoptosis of cancer cells. Recently, lipid-soluble compounds such as diallyl sulfides (DAS) and diallyl disulfides (DADS) have been reported to more effectively suppress tumor cell proliferation. However, there were few studies on the suppressive effects of lipid-soluble garlic sulfur compounds on the proliferation and migration of vascular smooth muscle cells (VSMC). Therefore, this study investigated the effect of DAS and DADS on VSMC proliferation/migration induced by oleic acid (OA), a principal fatty acid in circulating triglyceride of blood stream. Assays performed include a tetrazole (MTT) assay, a wound healing assay and a Western blots. VSMC proliferations were enhanced by OA in a dose-dependent manner at concentrations of $10{\sim}50\;{\mu}M$ and inhibited by DAS and DADS compared to non-treated control. OA-induced proliferations were also attenuated by DAS and DADS. OA-induced cell migrations were 2.5 times higher than non-treated control, and they were significantly attenuated by DAS (32% at $150\;{\mu}M$ and 50% at $200\;{\mu}M$) and DADS (40% at $150\;{\mu}M$ and 46% at $200\;{\mu}M$). OA-induced cell migration was also attenuated by PD98059 (ERK inhibitor), SB203580 (P38 inhibitor) and particularly by LY204002 (PI3K inhibitor) and SP600125 (JNK2 inhibitor). Additionally, Western blot assays showed that OA-induced JNK1/2-phosphorylation was down-regulated after treatment with DAS and DADS. In conclusion, the findings of our study support the idea that DAS and DADS may have a suppressive effect on the proliferation and migration of OA-induced VSMC and that this effect may be partly associated with PI3K and JNK2 pathways.
Purpose: To study the expression of angiogenin-2 (Ang-2) and its receptor Tie-2 in colorectal cancer and discuss the possible mechanisms behind this process. Materials and Methods: Using the streptavidin-peroxidase (SP) immunohistochemical method, paraffin sections from 100 colorectal cancer samples and 10 samples from tumor-adjacent normal tissue (> 2 cm from the edge of the gross tumor) were tested for protein expression of Ang-2, Tie-2, PI3K, and AKT. Reverse transcription-polymerase chain reaction and Western blots were further used to measure expression of the 4 genes and proteins in 20 freshly-resected colorectal cancer samples and tumor-adjacent normal tissues. Results: In colorectal cancer tissues, the expression of the Ang-2, Tie-2, PI3K, and AKT genes and their proteins was significantly higher than in tumor-adjacent normal tissues. Protein expression in poorly-differentiated adenocarcinoma was higher than that in well and moderately differentiated adenocarcinoma. According to Duke's classification, the protein expression in Stages C and D was significantly higher than that in Stages A and B. In the group with lymphatic metastasis, the protein expression was higher than that without lymphatic metastasis. Conclusions: In colorectal cancer, the expression of the Ang-2, Tie-2, PI3K, and AKT genes and their proteins is markedly higher than those in tumor-adjacent normal tissues. No correlation was observed between protein expression and gender, location, or histologic type. Correlations did exist between protein expression and differentiation level, stage of Duke's classification, and lymphatic metastasis; in colorectal cancer tissues with lower differentiation levels, higher stages of Duke's classification, and lymphatic metastasis, the expression of all 4 proteins was higher. The study of their expression patterns and relationships with aggression and metastasis will provide a valuable experimental foundation for assessing prognosis and targeted therapy of colorectal cancer.
The present study focused on establishing the effects of interferon-gamma (IFN-${\gamma}$) on interleukin-18 (IL-18) expression patterns and pregnancy outcomes in pregnant rats. Pregnant rats at the post-implantation stage were randomized into control, low IFN-${\gamma}$ (L-IFN-${\gamma}$) and high IFN-${\gamma}$ groups (H-IFN-${\gamma}$) that received normal saline, 100 IU/g of IFN-${\gamma}$ and 500 IU/g of IFN-${\gamma}$ vaginal muscular injection, respectively. The effects of IFN-${\gamma}$ on IL-18 expression and pregnancy outcomes were assessed systematically using several methods, including immunohistochemistry streptavidin-perosidase (SP), image pattern analysis, enzyme-linked immune-sorbent assay (ELISA), whole blood count (WBC) count, microscopy and visual observation. IL-18 was detected in the uteri of all pregnant rats, and mainly distributed in the endometrium, decidual cells, vascular endothelium and myometrium. Immunohistochemistry and image pattern analyses revealed significantly lower IL-18 expression in the H-IFN-${\gamma}$ group compared to the L-IFN-${\gamma}$ and control groups (p<0.01), indicating that high doses of IFN-${\gamma}$ induce downregulation of IL-18 in the uterus of pregnant rats. ELISA results disclosed that IL-18 expression in peripheral blood of the H-IFN-${\gamma}$ group was lower than that of the L-IFN-${\gamma}$ group (p<0.05), and significantly reduced compared to the control group (p<0.01). Moreover, the number of peripheral leukocytes in the H-IFN-${\gamma}$ group was significantly higher than those in the control and L-IFN-${\gamma}$ groups (p<0.01). Morphology analysis showed no evident differences between the L-IFN-${\gamma}$ and control groups. However, for the H-IFN-${\gamma}$ group, uterine mucosa bleeding, necrosis and excoriation were observed using microscopy. Visual observation revealed marroon, swelling, crassitude and no embryo in the uterus, which are obvious indicators of abortion. These results indicate that IFN-${\gamma}$ plays a regulatory role in IL-18 expression in the uterus and peripheral blood of pregnant rats at the post-implantation stage. Moreover, high levels (500 IU/g) of IFN-${\gamma}$ influence normal pregnancy at the early stages in rats by downregulating IL-18 expression in the uterus and peripheral blood and increasing the number of peripheral leukocytes, consequently triggering termination of pregnancy.
In this work, the fragrant oil release behavior of poly($\varepsilon$-caprolactone) (PCL) microcapsules containing SiO$_2$ was investigated. The SiO$_2$ was chemically treated in 10, 20, and 30 wt% hydrochloric acid and sodium hydroxide. The acid and base values were determined by Boehm's titration technique and $N_2$/77 K adsorption isotherm characteristics, the specific surface area and total pore volume were studied by BET. The PCL microcapsules containing SiO$_2$ and fragrant oil were prepared by oil-in-water (o/w) emulsion solvent evaporation method. The shape and surface of PCL microcapsules were observed using image analyzer and scanning electron microscope (SEM). The fragrant oil release behavior of PCL microcapsules was characterized using UV/vis. spectra. The average diameters of PCL microcapsules were decreased from 35 to 21 $\mu$m with increasing stirring rate. It was found that in the case of acidic treatment the fragrant oil adsorption capacity and release rate were increased due to the increase of specific surface area and acid value. In the case of basic treatment, the fragrant oil adsorption capacity and release rate were decreased due to the decrease of sp ecific surface area and the increase of acid-base interactions between SiO$_2$-NaOH and fragrant oil with increasing base value of SiO$_2$.
Effects of dietary calcium lactate (CaL-A) and Chungkukjang (Korean native fermented soybean) on bone mass, calcium status, body weight, serum glucose and cholesterol levels in young male rats were investigated. Chungkukjang was fermented by mixing 4 types of Bacillus sp., and then dried at 45$^{\circ}C$. Calcium lactate was prepared from the ash of black snail. The rats were fed a commercial rat diet for 1 week and then the experimental diets for 4 weeks. Animals were divided into four dietary groups: one calcium-deficient diet (Ca-De) and one of three with calcium supplemented diets (5 g calcium/kg diet) with either calcium phosphate (Ca-P), CaL-A, or CaL-A + Chuntkukjang (CaL-AC). Calcium supplemented diets contained 39 g Ca-P/kg diet and 28 g/kg of calcium lactate in the CaL-A and CaL-AC diets. Body weight gains during the 4 weeks in the Ca-P, CaL-A, CaL-AC and Ca-De groups were 130.45 g,112.50 g, 143.40 g and 10.20 g, respectively. Feed consumption of the groups from high to low was CaL-AC > Ca-P > CaL-A > Ca-De. The Ca-De group had low femur weights and low serum calcium concentrations, while they were comparatively high in CaL-AC, Ca-P and CaL-A groups. The Ca-De groups excreted less calcium in urine than did the other rats, probably due to increased absorption of the mineral in Ca-P, CaL-A and CaL-AC groups. Microscopic observations revealed that there were many regularly spaced holes in the femur of Ca-De group, while there were much smaller regularly spaced holes in Ca-P group. However, no holes in femur were observed in the CaL-A and CaL-AC groups. Bone surfaces were especially smooth and clean in the CaL-AC group. Serum concentrations of glucose and total cholesterol were remarkably lower in the CaL-AC group than in the other supplemented groups. These results suggest that calcium from CaL-A has higher bioavailability than from Ca-P, and dietary Chungkukjang may have a beneficial effect on calcium metabolism.
Postharvest decay of onion bulbs was examined by inspecting the commercial packages in the market or in storage. Bulb rot incidence was unexpectedly high, and onion bulbs with 1st quality grade were rotten most severely by 51%, followed by 32% for 2nd and 21% for 3rd grades. This indicates that larger bulbs had higher incidences of bulb rots. Major pathogens associated with basal and neck rots were Fusarium oxysporum and Aspergillus sp. or Botrytis allii, respectively, of which basal rot was most prevalent and damaging during storage. Among the epiphytic microorgani는 from onion plants, several Bacillus and Paenibacillus spp. and previously selected Pseudomonas putida and Trichoderma harzianum had inhibitory efficacy against bulb rot pathogens. Among these B. amyloliquefaciens BL-3, Paenibacillus polymyxa BL-4, and P. putida Cha 94 were highly inhibitory to conidial germination of F. oxysporum and B. allii. P. putida Cha 94, B. amyloliquefaciens BL-3, P. polymyxa BL-4, and T. harzianum TM were applied in the rhizoplane of onion at transplanting. Initially antagonist populations decreased rapidly during the first one month. However, among these antagonists, rhizoplane population densities of BL-3, Cha 94, and TM were consistently high thereafter, maintaining about 10$^4$-10$^{5}$ cells or spores per gram of onion root up to harvest time. The other bacterial antagonist BL-4 survived only for two months. TM was the most effective biocontrol agent against basal rot, with the number of rotten bulbs recorded at 4%, while that of the control was 16%. Cha 94 was effective for the first 20 days, but basal rot increased thereafter and had about the same control efficacy as that of BL-3 and BL-4. When the antagonists were applied to the topping areas of onion bulbs at harvest, TM was the most effective in protecting the stored onion bulbs from neck rotting. The second effective antagonist was BL-3. TM and BL-3 completely suppressed the neck rot in another test, suggesting that biocontrol of postharvest decay of onion using these microorganisms either at the time of transplanting or at harvesting may be promising.
The purification and characteristics of the biosynthesis enzyme of vitamin C from microorganisms related with kimchi fermentation were investigated to define vitamin C biosynthetic pathways in yeast. A yeast strain (Pichia onychis 16-4) which synthesizes vitamin C with galacturonic acid as substrate at high rate was isolated from kimchi. The enzyme $L-galactono-{\gamma}-lactone$ oxidase isolated from the yeast was purified and characterized. The specific activity of the crude enzyme was 7.26 unit/mg protein, which increased to 4,698 unit/mg protein with a chromatography of Sephacryl S-200HR; indicating a 647.1-fold level of purification. The molecular weights of the dissociated enzymes were estimated to be 31,000, 39,000, and 50,000 KD. Among the substrates tested, $L-galactono-{\gamma}-lactone$ was the most effective. The enzyme showed optimum activity ah pH 7.8 and 35c. The purified enzyme uses $O_2$ as the electron acceptor for oxidation of $L-galactono-{\gamma}-lactone$.
During a study to document more fully the parasitic ciliates of Korean freshwater fishes, we found numerous small ciliates in the gills and skin of crucian carp, Carassius carassius. Cursory examination indicated that the ciliate were very similar with Chilodonella sp. In this study, we described the morphometrics of this ciliate in detail and compared with other species of Chilodonella in the world. Numerous small ciliates were observed in the body surface, fins and gills of infected fish and excessive mucus production were seen in those fish. Sometimes ulcer was observed in the body of moribund fish. From the scrutinized observation of the parasite, it was identified as Chilodonella hexasticha. The parasite was dorsoventrally flattened body, without a notch in the posterior margin, and it measured 30-$45{\mu}m$ long and 15-$30{\mu}m$ wide. In number of kineties, the right band ciliature was 3-5 and the left band ciliature was 3-5. The right ventral ciliary band shaped arch and was longer than the left, straight band. It had a single oval macronucleus, 8-$15{\mu}m$ in diameter, a single micronucleus, 2-$4{\mu}m$ in diameter. The cytopharynx was reinforced by 10-16 conspicuous nematodesmata, which shaped like a tube and curved at its inner end. Two contractile vacuoles, one anteriorly at right and the second posteriorly at left were observed in wet mounts. This parasite reproduced by a simple division at $22{\pm}1^{\circ}C$.
Park, In-Cheol;Seok, Soon-Ja;Kim, Jeong-Seon;Yoo, Jae-Hong;Ahn, Jae-Hyung
The Korean Journal of Mycology
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v.43
no.4
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pp.239-246
/
2015
Two fungal strains were isolated from rods of Quercus sp. (NAAS02335) and Pinus densiflora (NAAS05299) in Korea. These strains were identified as Gyrodontium sacchari by their morphological and mycological characteristics. The optimal growth temperature of NAAS02335 and NAAS05299 are $25^{\circ}C$ and $30^{\circ}C$, respectively. Production of cellulase, xylanase, and ligninase was tested on agar media supplemented dyes or substrates. Production of cellulase and xylanase of NAAS05299 was higher than those of NAAS02335, however ligninase activity of NAAS02335 was higher than that of NAAS05299. The activities of cellulase, xylanase, and amylase of strain NAAS05299 were estimated at 6.7~10.2 times higher than that of NAAS02335. Laccase activity was only estimated by strain NAAS02335. The lignocellulytic enzymes are induced by substrates such as rice straw, wooden chips of pine, oak, and poplar. The NAAS05299 was able to degrade filter paper completely after 4 weeks of culturing in liquid media containing a piece of filter paper at $28^{\circ}C$ with continuous shaking. NAAS05299 was able to degrade rice straw, pine chips, and oak chips after 4 months in solid culture, however NAAS02335 decomposed only rice straw among tested 4 kinds of biomass.
Background: Ginseng extracts are known to have angiogenic effects. However, to date, only limited information is available on the molecular mechanism underlying the angiogenic effects and the main components of ginseng that exert these effects. Human umbilical-vein endothelial cells (HUVECs) are used as an in vitro model for screening therapeutic agents that promote angiogenesis and wound healing. We recently isolated gintonin, a novel ginseng-derived lysophosphatidic acid (LPA) receptor ligand, from ginseng. LPA plays a key role in angiogenesis and wound healing. Methods: In the present study, we investigated the in vitro effects of gintonin on proliferation, migration, and tube formation of HUVECs, which express endogenous LPA1/3 receptors. Results: Gintonin stimulated proliferation and migration of HUVECs. The LPA1/3 receptor antagonist, Ki16425, short interfering RNA against LPA1 or LPA3 receptor, and the Rho kinase inhibitor, Y-27632, significantly decreased the gintonin-induced proliferation, migration, and tube formation of HUVECs, which indicates the involvement of LPA receptors and Rho kinase activation. Further, gintonin increased the release of vascular endothelial growth factors from HUVECs. The cyclooxygenase-2 inhibitor NS-398, nuclear factor kappa B inhibitor BAY11-7085, and c-Jun N-terminal kinase inhibitor SP600125 blocked the gintonin-induced migration, which shows the involvement of cyclooxygenase-2, nuclear factor kappa B, and c-Jun N-terminal kinase signaling. Conclusion: The gintonin-mediated proliferation, migration, and vascular-endothelial-growth-factor release in HUVECs via LPA-receptor activation may be one of in vitro mechanisms underlying ginsenginduced angiogenic and wound-healing effects.
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