Heterogenous samples of locust bean gum (galactomannan) were prepared into homogeneous substances. Locust bean gum was fractioned using ammonium sulfate (14.11-23.08%, w/w). The intrinsic viscosity was obtained by extrapolating reduced viscosity versus concentration by using an Ubbelohde viscometer. The ranges of intrinsic viscosity for fractions that not included protein (F3-F6) and fractions that included protein (F1-F2) were 9.89-8.10 and 8.44-4.59, respectively. Values for Huggins' coefficient (k'), which depends on physical interactions, were 0.46-0.78. Increasing ammonium sulfate concentration was associated with a weak trend towards lower molecular weight and intrinsic viscosity by size-exclusion chromatography (SEC): $M_w$ ranged from 674 to 617 kg/mol and [${\eta}$] from 9.80 to 8.10 dL/g between F3 and F6. The evaluations of those fractions by using SEC and the Ubbelohde viscometer produced very similar values, as predicted. We verified the application of a gradient of ammonium sulfate to precipitate locust bean gum into fractions of different molecular size and show structural variations.
Purpose: This study was performed to investigate the effects of immune cell activation and the antitumor effect for the combination of treatment with X-irradiation and E/eutherococcus senticosus Maxim Root (ESMR) on mouse tumor cells. Materials and Methods: ESMR (250g) was extracted with 80% methanol, concentrated under decompression and lyophilized. To determine whether ESMR is able to activate the immune cells or not, the proliferation of splenocytes in vitro and the number of B cells and T cells in splenic lymphocytes in ESMR-pretreated mice were evaluated. X-irradiation was given to the mouse fibrosarcoma tumor cells (FSa II) by 250 kv X-irradiation machine. The cytotoxicity of ESMR was evaluated from its ability to reduce the clonogenecity of FSa II cells. In X-irradiation alone group, each 2, 4, 6 and 8 Gy was given to FSa II cells. In X-irradiation with ESMR group, 0.2 mg/ml of ESMR was exposed to FSa II cells for 1 hour before X-irradiation. Results: The proliferation of cultured mouse splenocytes and thymocytes were enhanced by the addition of ESMR in vitro. The number of B cells and T cells in mouse splenic lymphocytes was significantly increased in ESMR pretreated mice in vivo. In FSa II cells that received a combination of 0.2 mg/ml of ESMR with X-irradiation exposure, the survival fraction with a dose of 2, 4 and 6 Gy was $0.39{\pm}0.005$, $0.22{\pm}0.005$ and $0.06{\pm}0.007$, respectively. For FSa II cells treated with X-irradiation alone, the survival fraction with a dose of 2, 4 and 6 Gy was $0.76{\pm}0.02$, $0.47{\pm}0.008$ and $0.37{\pm}0.01$. The difference in the survival fraction of the mouse FSa II cells treated with and without ESMR was statistically significant (p<0.05). Conclusion: Treatment with ESMR increased cell viability of mouse splenocytes in vitro and especially the subpopulation of B cells and T cells in splenocytes in ESMR-pretreated mice. However, treatment with ESMR did not increase the level of Th and Tc subpopulations in the thymocytes. Treatment with the combination of ESMR and X-irradiation was more cytotoxic to mouse tumor cells than treatment with X-irradiation alone; this finding was statistically significant.
Beetle larvae have been used as a traditional medicine to treat various human liver diseases. To prove the liver protective function of Allomyrina dichotoma larvae (ADL), we induced liver damage by the intraperitoneal injection of a hepatotoxic reagent, diethylnitrosamine (DEN), to C3H/HeN male mice and orally administered freeze-dried ADL powder. ADL powder lessened DEN-induced hepatotoxicity considering the reduced signs of acute and chronic hepatotoxicities, such as the ALP level in the blood serum, TUNEL-positive hepatocytes, ductural reactions, steatotic hepatocytes, and collagen deposition of the Masson’s trichrome staining. In addition to hepatoprotection, the anti-cancer activity of ADL has been examined. The ADL powder was extracted with ethanol and then fractionated with hexane, ethyl acetate, and water by a solvent partition technique. The ethyl acetate fraction showed cytotoxicity to various cancer cells through induction of apoptosis and necrosis, as well as the perturbed metabolism of the cancer cell to trigger autophagy. Collectively, ADL contains bioactive substances that can protect hepatocytes from toxic chemicals and trigger cell death in cancer cells. Thus, further purification and analyses of ADL fractions could lead to the identification of novel bioactive compounds.
Chung, Mi Yeon;Kwon, Eun-Young;Hwang, Jae-Sam;Goo, Tae-Won;Yun, Eun-Young
Journal of Life Science
/
v.23
no.3
/
pp.426-431
/
2013
With the objective of developing a functional food source, we established optimal processing conditions for the larvae of Allomyrina dichotoma, which have been used in traditional medicine to treat hepatic disorders in Korea. Without suitable processing, the larvae are difficult to consume as a food because of their disgusting taste and smell; moreover, in this form they might be a potential microbial hazard. In this study, we investigated the effect of feeding material, sterilization, and powdering after freeze-drying on the food quality of the larvae of A. dichotoma and on cytotoxicity against Raw 264.7 cells. Three to five days feeding with the sawdust from discarded oak-trees is sufficient for the breeding process. The sawdust was sterilized by vapor for five minutes. Sterilization of the larvae at a high temperature ($115^{\circ}C$ for 5 min, 0.9 $kgf/cm^2$) is necessary to eliminate pathogenic bacteria and fungi. The results of the cytotoxicity assay showed no toxicity in the prepared extract from larvae of A. dichotoma. In addition, to prepare the larvae for human consumption, various feeds were used and the smell, color, and taste were evaluated. Our results suggested that larvae of A. dichotoma could be developed as food source when a suitable processing method is established.
Storage characteristics of fresh and thawed sliced raw fish (flounder and rockfish) washed in different solutions (tap water, A; jade water, B; 0.2% chitosan-ascorbate (CA), C; 0.03 ppm ozone water, D; 1.5% vinegar containing jade water, E; 1.5% vinegar and 0.2% CA containing jade water, F) at $10^{\circ}C$ were investigated. Changes in pH and acidity of thawed sliced raw fish (TS) during storage were lower than for fresh sliced raw fish (DS). The total microbial content (log cfu/g) of A stored for 3 days in DS was 6.7 (which represented an increase of 1 log cycle compared with day zero), but was 5.50 in B, 3.23 in C, 4.90 in D, 2.40 in E and 1.77 in F, the latter similar to counts at day zero. The degree of microbial contamination of DS and TS followed the order F > E > D > C > B > A in flounder, and F > E > C > D > B > A in rockfish. In general the hardness and chewiness of TS was less than for DS. While the effect of CA on TS texture was not significant in flounder, the effect showed in rockfish. For DS, the appearance in B, C and D was relatively good, as was freshness. Fishiness of flavor was in the order A > B > F > E > D > C. Overall acceptability of flounder and rockfish treated with C was better than treatment with the other washing agents. For TS the appearance of flounder and rockfish were good in B and C. The freshness of flounder and rockfish were in the order of D > C > B > A > E > F and D > C > B > A > E > F, respectively. Fishiness of the flavor of sliced raw fish was lowest in D, which also provided the best overall acceptability.
Purpose: Microsatellite instability(MSI) is frequently used as an indicator of microsatellite mutator phenotype(MMP) tumors. MSI has been observed in a percentage of non-small cell lung cancer(NSCLC). However, its role in tumorigenesis of NSCLC remains unknown. The frequency and pattern of MSI in NSCLC were evaluated and clinical parameters of MSI-positive tumors with those of MSS(microsatellite stable) tumors were compared. Materials and Methods: Twenty surgically resected NSCLCs were analyzed for 15 microsatellite markers located at chromosomes 3p and 9p. The peripheral blood lymphocytes of patients were used as the source of the normal DNA. Results: 1) Of 20 cases, 8(40%) demonstrated MSI. 2) Instability was observed more frequently in tri- and tetra-nucleotide repeats than in dinucleotide repeats. In all cases, instability appeared as a shift of individual allelic bands. 3) LDH was observed in 10(50%) of 20 tumors analyzed. 4) Of 20 cases, MSI-H tumor(showing MSI in the majority of markers) was absent. There were 5 MSI-L tumors(showing MSI in a greater than 10% of markers). 5) No significant difference was observed between MSI-L tumors and MSI-negative tumors in terms of clinicopathologic features such as pack-year history of smoking, histologic subtype, and(delete) stage of disease. There was also no significant difference in the incidence of LDH in relation to the status of MSI. Conclusion: These data strongly suggest that MSI plays different roles in lung and colon cancer. MMP pathway appears to be far less important in the tumorigenesis of NSCLC, caused mainly by cigarette smoke, with little familial tendency.
Kim, Seok-Young;Kim, Hee-Sun;Kim, Jin-Se;Han, Gwi-Jung
Food Science and Preservation
/
v.24
no.6
/
pp.746-757
/
2017
This study was performed to evaluate the effect of different freezing conditions and storage periods on physicochemical and microbial characteristics of garlic. Garlics were washed, dried, sliced to 0.3 cm then packed in LDPE+LLDPE film bags. They were treated with still-air freezing at $-20^{\circ}C$ (SAF20), $-40^{\circ}C$ (SAF40) and immersed-liquid freezing at $-40^{\circ}C$ (ILF40). Frozen garlics were stored under frozen storage conditions for 7 months at $-20^{\circ}C$ and quality characteristics were measured monthly during the frozen storage. Freezing rate of garlic was the fastest in ILF 40 (10 min), SAF40 (40 min) and SAF20 (1,600 min) sequentially. In ILF40, drip loss, cutting force, total aerobic bacteria count and pH were the lowest, whereas pyruvic acid and allicin content were the highest (p<0.05) during frozen storage, these results were the most similar characteristics with the fresh garlic. During frozen storage, drip loss, color difference and total organic acid content were significantly fluctuated in SAF20 (p<0.05), while they were not changed in ILF40. Overall, total aerobic bacteria count and pH decreased, cutting force, pyruvic acid and allicin content remained unchanged in all groups. In conclusion, the optimal freezing conditions for garlic with the least quality changes was considered to be ILF40 (immersed liquid freezing), keeping quality characteristics up to 7 months by freezing storage.
Two kinds of Korean rice-wine (Yakju) with different process and ingredients, and Japanese rice-wine (Sake) were chosen for this study, and throughly dried and solubilized in water or cell culture medium. In vitro cytotoxicity assays of the solubilized wine solids exhibited that maximum dilution factors for inhibition of B 16BL6 mouse melanoma cell growth were 16X for herbal medicine-added rice-wine (Korean rice-wine I) and typical Korean rice-wine (Korean rice-wine II), and 8X for Japanese rice-wine. Their cytotoxic effects on HRT18 human colon adenocarcinoma cells were even lower than those on B16BL6 cells. The morphology of the tumor cells were changed by addition of the solubilized wine solids. Inhibitory effect of the rice-wine on in vivo tumor growth and metastasis were monitored after implantation of B16BL6 cells into C57BL/6 mice with daily feeding the solubilized wine solids. Compared to non-fed control groups, B16BL6 tumor growth and metastasis to lung were clearly inhibited by feeding the wine solids, in order of Korean rice-wine I > Korean rice-wine II > Japanese rice-wine. The data of in vitro cytotoxicity and the cell shape changes indicate that the inhibitory effect of tumor progression may be attributed to tumor cell differentiation or immune stimulation induced by certain components in the rice-wine, rather than direct cytotoxicity of the components.
This study aimed increase the quality during ripening of Cheddar cheese made with proteinase-negative mutant of Streptococcus lactis KCTC 1913 selected by curing. The degradation of protein during cheese ripening were investigated by electrophoresis and chromatography. The results were summarized as follow ; 1. The number of lactic acid bacteria decreased with the ripening stage, and that of the control cheese decreased faster than that of the cheese made with mutant. 2. Polyacrylamide gel electrophoretic analysis of cheese caseins revealed no difference between the cheese made with mutant and the control cheese, but differences along with the ripening stage were evident. 3. On Sephadex G-25 column chromatography, the extracts of bitter components from the green cheese and 3 month ripended cheese were fractionated into 3 fractions. With the progress of ripening, bitter peptides were degraded to rather small peptides or free amino acids. 4. Sensory evaluation of the 3 month ripended Cheddar cheese found no significant differences in color but the cheese made with mutant evidenced higher palatability in flavor and better texture than the control cheese. 5. The yields of the cheddar cheese made with mutant was 0.14% higher than that of the control cheese.
Objectives: The objective of this study was to evaluate the effects of cytotoxicity, skin regeneration, anti-wrinkle, whitening and skin moisturizing of Oncheongeum (OCE).Methods: The cytotoxicity of OCE lyophilized aqueous extracts (yield=13.82%) was observed against human normal fibroblast cells and B16/F10 murine melanoma cells by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium Bromide (MTT) assay, and skin regeneration and anti-wrinkle effects were also evaluated through the assay of collagen type I synthesis compared to the transformation of the growth factor (TGF)-β1, hyaluronidase, collagenase and matrix metalloproteinase (MMP)-1 inhibitory assays compared to oleanolic acid (OA), and elastase inhibitory effects compared to phosphoramidon disodium salt (PP). In addition, OCE’s whitening effects were measured by a tyrosinase inhibitory assay and melanin formation test in B16/F10 murine melanoma cells compared to arbutin, and skin moisturizing effects were observed through a mouse skin water content test, respectively. Results: No OCE treatment-related cytotoxic effects appeared on human normal fibroblasts and B16/F10 murine melanoma cells. OCE concentration-dependently increased the collagen Type I synthesis on human normal fibroblast cells, and also effectively inhibited hyaluronidase, elastase, collagenase and MMP-1 activities. In addition, OCE inhibited melanin production of B16/F10 murine melanoma cells and activity of tyrosinase. And significant and dose-dependent increases of skin water content were detected in OCE-treated mouse skin compared to vehicle control skins. Conclusions: OCE showed favorable and sufficient effects in skin regeneration, anti-wrinkle, whitening and skin moisturizing in this experiment. But more detail mechanisms and studies on the skin protective efficiency of in vivo are needed with the screening of active biological compounds in individual OCE herbs.
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