This study was carried out to evaluate major agronomical characterization and phenol compound contents, Xanthine oxidase inhibitory activity (XO), Catalase activity, Superoxide dismutase activity (SOD) and 1,1-Diphenyl-2-picryl-hydrazyl (DPPH) radical sacvenging activity were analyzed in colored waxy corns. The mean of stem height and ear length were 248.8 cm and 18.6 cm, respectively. The pericarp thickness in CNU108 $(30.3{\mu}m)$ was thinner than other hybrids. The period of tasseling days in CNU69 and CNU202 were very shorter than other hybrids (59 days). 100-kernel weight of CNU50 was 35.6 g and heavier than the others. The antioxidant activities such as xanthin oxidase (XO), catalase and superoxide dismutase (SOD) were higher in CNU70 and CNUI 38. Especially highest EDA (electron donating ability) in DPPH radical scavenging effect was 94.8% and 94.6% in CNU160 and CNU193, respectively. In the results, the antioxidant enzyme activity and antioxidant acticity were higher in CNU109 and CNU 34 hybrids. The hybrids, CNU34, CNU70, CNU108, CNU 138 and CNU193 may be considered higher functional color waxy corn.
This study has been designed in order to examine a physiological role of $Ca^{2+}$ which has been known as an essential factor for capacitation, to confirm whether the enzyme activity of $Ca^{2+}$-ATPase on capacitation is important or not, and to clarify relationship between various levels of the $Ca^{2+}$ concentration and $Ca^{2+}$-ATPase which has been known to be an important factor of the plasma membranes. In the present study applying quercetin, a $Ca^{2+}$-ATPase inhibitor, the enzymatic effect of $Ca^{2+}$-ATPase on capacitation was found to be remarkable: a significant increase of the transition from the original type (type A) to the type B and the type AR of the spermatozoa. This finding suggests that $Ca^{2+}$-ATPase plays an important role in the efflux and the influx of the $Ca^{2+}$ which has been known to be an essential factor the capacitation and acrosome reaction, and that the inhibitory action of the $Ca^{2+}$-ATPase might be a prerequsite step toward the acrosome reaction. The conclusion reached can be deduced as follows: increment of the intracelluar $Ca^{2+}$ concentration occurred by controlling the slope of $Ca^{2+}$ concentration through $Ca^{2+}$-ATPase activities in both the intra- and extracelluar fluid may be an important procedure for capacitation and acrosome reaction, and ultimately for fertilization of the spermatozoa and the ova.
Objectives: The aim of this experimental study was to evaluate the anti-neoplastic and anti-inflammatory effects of single and mixed extracts of Ulmus davidiana (UD) and Oldenlandia diffusa (OD) on azoxymethane/dextran sodium sulfate (AOM/DSS)-induced colonic neoplasms in mice. Methods: AOM/DSS induces colitis-associated colonic neoplasms in mice. Mice were divided into seven groups: normal-no inducement and no treatment; control-colonic neoplasms with no treatment; UD-colonic neoplasms and treatment with UD; OD-colonic neoplasms and treatment with OD; UD1+OD1-colonic neoplasms and treatment with UD1 and OD1. UD1+OD2-colonic neoplasms and treatment with UD1 and OD2; UD2+OD1-colonic neoplasms and treatment with UD2 and OD1. Single and mixed preparations of UD and OD were applied to mice for six weeks. The colon length and weight and histopathologic changes of colon tissue were observed. Serum pro-inflammatory cytokines, including tumor necrosis factor-alpha ($TNF-{\alpha}$) and interleukin-6 (IL-6), were determined by enzyme-linked immunosorbent assay. The mRNA expression levels of Bax, Bcl-2, and interferon-gamma ($INF-{\gamma}$) were measured by RT-PCR. Results: The colon length was significantly increased in OD, UD1+OD2, and UD2+OD1 mice, and the colon weight was significantly decreased in OD and UD1+OD2 mice. The morphological change of colon epithelial cells was more suppressed in complex-treatment groups than in single-treatment groups. The inhibitory effect on inflammatory cell invasion was especially shown in UD1+OD2 mice. The serum level of the pro-inflammatory $TNF-{\alpha}$ was decreased in all complex-treatment groups, and the IL-6 level was decreased in UD1+OD1 mice. Single-treatment groups had an increase in the mRNA expression of the pro-apoptosis regulator Bax, and UD2+OD1 decreased the mRNA expression of the anti-apoptosis regulator Bcl-2. The mRNA expression of $INF-{\gamma}$ associated with inflammation was decreased in OD and UD1+OD2 mice. Conclusions: This study suggests that single and mixed extracts of Ulmus davidiana and Oldenlandia diffusa have anti-neoplastic and anti-inflammatory effects on AOM/DSS-induced colonic neoplasms in mice. Therefore, we conclude that UD, OD, and a mixture of UD and OD are potential therapeutic agents for colitis-associated colonic neoplasms.
Intracellular ions which have a major role in cellular function have been reported to affect repair of radiation damage. Recently it has been reported that ouabain sensitizes A549 tumor cellls but not CCL-120 normal cells to radiation. Ouabain inhibits the $Na^+-K^+$-pump rapidly thus it increases intracellular Na concentration, Vanadate which is distributed extensively in almost all living organisms is known to be a $Na^+-K^+$-ATPase inhibitors, This study was performed to see any change in radiosensitivity of tumor cell by vanadate and any role of $Na^+-K^+$ATPase in radiosensitization. Experiments have been carried out by pretreatment with vanadate in human cell line(A549, JMG) and mouse cell line(L1210, spleen). For the cell survival MTT assay was performed for A549 and JMC cells and frypan blue dye exclusion test for L120, and spleen cells. Measurements of $Na^+-K^+$-ATPase activity in control, vanadate treated cell, radiation treated cell (9 Gy for A549 and JMG, 2 Gy for L1201, spleen), and combined $10^{-6}M$ vanadate and radiation treated cells were done. The results were summerized as fellows. 1. L1210 cell was most radiosensitive, and spleen cell and JMG cell were intermediate, and A549 cell was least radiosensitive. 2. Mininum or no cytotoxicity was seen with vanadate below concentration of $10^{-6}M$. 3. In A549 cells there was a little change in radiosensitivity with treatment of vanadate. However radiation sensitization was shown in low dose level of radiation i. e. 2- Gy. In JMG cells no change in radiosensitivity was noted. Both L1210 and spleen cell had radiosensitization but change was greater in tumor cell. 4. $Na^+-K^+$-ATPase activity was inhibited significantly in tumor cell by treatment of vanadate. 5. Radiaiton itself inhibited $Na^+-K^+$-ATPase activity of tumor cell with high $Na^+-K^+$-ATPase concention. Increase in radiosensitivity by vanadate was closely associated with orginal $Na^+-K^+$-ATPase contents. From the above results vanadate had little cytotoxicity and it sensitized tumor cells to radiation. Inhibitory effect of vanadate on $Na^+-K^+$-ATPase activity might be one of the contributing factors for radiosensitization to tumor cells which has greater enzyme activity than that of normal cell. It was suggested vanadate could be used as a potential radiosensitizer for tumor cells.
Human pancreatic lipase is a digestive enzyme which is synthesized in pancreas, secreted into small intestine, and there hydrolyze the fat in food. Pancreatic lipase protein composes of catalytic domain and colipase-binding domain. In this research, the gene segments corresponding to total protein, catalytic domain, and co lipase-binding domain were cloned by PCR method, inserted into an expression vector, and then used to transform Escherichia coli BL21 (DE3). The recombinant proteins produced were purified and injected intramuscularly three times into laying hens. The egg yolk antibodies (IgY) were obtained from the egg yolks and tested for their antibody titer. Among three IgY, the IgY against colipase-binding domain showed the highest antibody titer. All three IgY had inhibitory effects on the porcine pancreatic lipase. Among them, the IgY against colipase-binding domain showed the highest inhibition effects. The fat diet with corn oil and IgY was administrated to the experimental rats and their blood compositions were examined with time course. The triglyceride concentration of treated rats was decrease meaningfully when compared with those of control rats. This suggested that the IgY against colipase-binding domain antigen inhibited pancreatic lipase in vivo.
It has been reported that the glycoprotein extracted from Aloe has strong anti-inflammatory response. However, there has been no research report yet about the effect of Aloe on allergic hypersensitivity reactivity. By using guinea pig lung mast cells, this study aimed to examine the effects of Aloe glycoprotein (NY945) on the mediator releases caused by mast cell activation, and also aimed to assess the effects of NY945 on the mechanism of mediator releases in the mast cell activation. We partially purified mast cell from guinea pig lung tissues by using the enzyme digestion, the rough and the discontinuous density percoll gradient method. Mast cells were sensitized with IgG1 (anti-OA) and challenged with ovalbumin. Histamine was assayed by fluorometric analyzer, leukotrienes by radioimmunoassay. The phospholipase D activity was assessed by the production of labeled phosphatidylalcohol. The amount of mass 1, 2-diacylglycerol (DAG) was measured by the $[^3H]DAG$ produced when prelabeled with $[^3H]myristic$ acid. The phospholipid methylation was assessed by measuring the incorporation of the $[^3H]methyl$ moiety into phospholipids of cellular membranes. Pretreatment of NY945 (10 ${\mu}g$) significantly decreased histamine and leukotrienes releases during mast cell activation. The decrease of histamine release was stronger than that of leukotriene during mast cell activation. The phospholipase D activity increased by the mast cell activation was decreased by the dose-dependent manner in the pretreatment of NY945. The amount of DAG produced by PLC activity was decreased by NY945 pretreatment. The amount of mass 1, 2-diacylglycerol produced by activation of mast cells was decreased in the pretreatment of NY945. NY945 pretreatment strongly inhibited the incorporation of the $[^3H]methyl$ moiety into phospholipids. The data suggest that NY945 purified from Aloe inhibits in part an increase of 1, 2-diacylglycerol which is produced by activating mast cells with antigen-antibody reactions, which is mediated via phosphatidylcholine-phospholipase D and phosphatidylinositol-phospholipase C systems, and then followed by the inhibition of histamine release. Furthermore, NY945 reduces the production of phosphatidylcholine by inhibiting the methyltransferase I and II, which decreases the conversion of phosphatidylcholine into arachidonic acid and inhibits the production of leukotrienes.
While ornithine decarboxylase (ODC) is considered a key enzyme in the biosynthesis of polyamines, difluoromethylornithine(DFMO) acts as an inhibitor of polyamine synthesis. Cycling crossbred gilts were randomly assigned to one of two (treatment and control) groups (6/group). An indwelling silicone catheter was surgically implanted in the jugular vein of each animal. DFMO was dissolved in saline(200 mg/ml) and adminstered by i. m. injection at a dose of 80 mg/kg/day. The control group received an equivalent volume saline injection. DFMO was injected 3 times daily(08:00. 16:00. 24:00h) from day 16 of estrous cycle to 21 or until estrus. Once daily blood samples (10ml) were taken from day 14 until two days after the last DFMO treatment. Window blood samples were collected every 15 min for 8 h (from 08:00 to 16:00h) starting on day 16 and continuing until day 21 from one gilt per day. Serum progesterone (P$_4$), estradiol (E$_2$), LH and FSH were measured. Typical concentration profiles for P$_4$ and E$_2$ were seen during the follicular phase regardless of DFMO treatment. Injection of DFMO suppressed the preovulatory LH concentration in the serum(p<0.01) while having no effect on FSH profile. The present results indicate that DFMO had an inhibitory effect on LH secretion in the pig, but did not affect PI, E2 or FSH release.
Yoon, In Seong;Lee, Gyoon-Woo;Kang, Sang In;Park, Sun Young;Kim, Jin-Soo;Heu, Min Soo
Korean Journal of Fisheries and Aquatic Sciences
/
v.50
no.5
/
pp.506-519
/
2017
This study evaluated the protein recovery and functional properties and biological activity of boiled and steamed process water (BPW and SPW, respectively) generated from the preparation of concentrated roe of bastard halibut (BH; Paralichthys olivaceus), skipjack tuna (ST; Katsuwonus pelamis), and yellowfin tuna (YT; Thunnus albacares) using the cook-dry process. The protein loss from the water extracts (EXT) of 100 g of roe protein was 15.05-19.71% and was significantly (P<0.05) higher than that of BPW (5.47-10.34%) and SPW (3.88-8.18%). The foam capacity of BPW (166-203%) and SPW (15-194%) was better than that of EXT. The emulsifying activity index of the original samples was lower than those ($15.40-107.86m^2/g$) of diluted protein samples. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity and the reducing power of BPW and SPW were stronger than those of EXT. The 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid ($ABTS^+$) radical scavenging activity of EXT (0.028-0.045mg/mL) was significantly higher those of BPW and SPW. The angiotensin I-converting enzyme (ACE) inhibitory activity of SPW was the highest for BH (1.04 mg/mL), followed by YT and ST. The predominant amino acids in SPW were Glu, Ala, Leu, and His. These results demonstrate that processing water containing diluted organic components, including protein, can be consumed directly by humans as a functional reinforcing material after appropriate concentration processes.
To study effects of cellular FLICE (FADD-like IL-$1{\beta}$-converting enzyme)-inhibitory protein (c-FLIP) inhibition by RNA interference (RNAi) on sensitivity of U2OS cells to tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL)-induced apoptosis, plasmid pSUPER-c-FLIP-siRNA was constructed and then transfected into U2OS cells. A stable transfection cell clone U2OS/pSUPER-c-FLIP-siRNA was screened from the c-FLIP-siRNA transfected cells. RT-PCR and Western blotting were applied to measure the expression of c-FLIP at the levels of mRNA and protein. The results indicated that the expression of c-FLIP was significantly suppressed by the c-FLIP-siRNA in the cloned U2OS/pSUPER-c-FLIP-siRNA as compared with the control cells of U2OS/pSUPER. The cloned cell line of U2OS/pSUPER-c-FLIP-siRNA was further examined for TRAILinduced cell death and apoptosis in the presence of a pan-antagonist of inhibitor of apoptosis proteins (IAPs) AT406, with or without 4 hrs pretreatment with rocaglamide, an inhibitor of c-FLIP biosynthesis, for 24 hrs. Cell death effects and apoptosis were measured by the methods of MTT assay with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and flow cytometry, respectively. The results indicated that TRAIL-induced cell death in U2OS/pSUPER-c-FLIP-siRNA was increased compared with control cells U2OS/pSUPER in the presence or absence of AT406. Flow cytometry indicated that TRAIL-induced cell death effects proceeded through cell apoptosis pathway. However, in the presence of rocaglamide, cell death or apoptotic effects of TRAIL were similar and profound in both cell lines, suggesting that the mechanism of action for both c-FLIP-siRNA and rocaglamide was identical. We conclude that the inhibition of c-FLIP by either c-FLIP-siRNA or rocaglamide can enhance the sensitivity of U2OS to TRAIL-induced apopotosis, suggesting that inhibition of c-FLIP is a good target for anti-cancer therapy.
Solubilization of microsomal ${\Delta}^{5}-3{\beta}$-hydroxy steroid acyl transfearse(${\Delta}^{5}-3{\beta}$-OH-SAT) of rat brain and its reconstitution into liposomes were investigated. Among the detergents utilized for the solubilization, deoxycholic acid was superior to Tween 80 or Triton X-100 for the reconstituted activity of ${\Delta}^{5}-3{\beta}$-OH-SAT. The enzyme activity was shown to be affected by the nature of phospholipids used for the preparation of the liposome. Phosphatidylcholines from egg yolk and soybean showed the highest activity of ${\Delta}^{5}-3{\beta}$-OH-SAT and phosphatidylethanolamine came next. However phosphatidylserine and phosphatidic acid showed a lower activity than those obtained before the reconstitution. This study suggests that the presence of quaternary ammonium salt or amine group in the phospholipids stimulates the activity of ${\Delta}^{5}-3{\beta}$-OH-SAT. However the presence of a carboxylic group or the absence of the amine group may have an inhibitory effect on the ${\Delta}^{5}-3{\beta}$-OH SAT.
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