This study was conducted to evaluate effects of plant extracts on methanogenesis and rumen microbial diversity in in vitro. Plant extracts (Artemisia princeps var. Orientalis; Wormwood, Allium sativum for. Pekinense; Garlic, Allium cepa; Onion, Zingiber officinale; Ginger, Citrus unshiu; Mandarin orange, Lonicera japonica; Honeysuckle) were obtained from the Plant Extract Bank at Korea Research Institute of Bioscience and Biotechnology. The rumen fluid was collected before morning feeding from a fistulated Holstein cow fed timothy and commercial concentrate (TDN; 73.5%, crude protein; 19%, crude fat; 3%, crude fiber; 12%, crude ash; 10%, Ca; 0.8%, P; 1.2%) in the ratio of 3 to 2. The 30 ml of mixture, comprising McDougall buffer and rumen liquor in the ratio of 4 to 1, was dispensed anaerobically into serum bottles containing 0.3 g of timothy substrate and plant extracts (1% of total volume, respectively) filled with $O_2$-free $N_2$ gas and capped with a rubber stopper. The serum bottles were held in a shaking incubator at $39^{\circ}C$ for 24 h. Total gas production in all plant extracts was higher (p<0.05) than that of the control, and total gas production of ginger extract was highest (p<0.05). The methane emission was highest (p<0.05) at control, but lowest (p<0.05) at garlic extract which was reduced to about 20% of methane emission (40.2 vs 32.5 ml/g DM). Other plant extracts also resulted in a decrease in methane emissions (wormwood; 8%, onion; 16%, ginger; 16.7%, mandarin orange; 12%, honeysuckle; 12.2%). Total VFAs concentration and pH were not influenced by the addition of plant extracts. Acetate to propionate ratios from garlic and ginger extracts addition samples were lower (p<0.05, 3.36 and 3.38 vs 3.53) than that of the control. Real-time PCR indicted that the ciliate-associated methanogen population in all added plant extracts decreased more than that of the control, while the fibrolytic bacteria population increased. In particular, the F. succinogens community in added wormwood, garlic, mandarin orange and honeysuckle extracts increased more than that of the others. The addition of onion extract increased R. albus diversity, while other extracts did not influence the R. albus community. The R. flavefaciens population in added wormwood and garlic extracts decreased, while other extracts increased its abundance compared to the control. In conclusion, the results indicated that the plant extracts used in the experiment could be promising feed additives to decrease methane gas emission from ruminant animals while improving ruminal fermentation.
Objectives : Oxidative stress due to excessive accumulation of reactive oxygen species (ROS) is one of the risk factors for the development of several chronic diseases, including neurodegenerative diseases. Methods : In the present study, we investigated the protective effects of cheongnoemyeongsin-hwan (CNMSH) against oxidative stress‑induced cellular damage and elucidated the underlying mechanisms in neuronal-derived SH-SY5Y cells. Results : Our results revealed that treatment with CNMSH prior to hydrogen peroxide (H2O2) exposure significantly increased the SH-SY5Y cell viability, indicating that the exposure of the SH-SY5Y cells to CNMSH conferred a protective effect against oxidative stress. CNMSH also effectively attenuated H2O2‑induced comet tail formation, and decreased the phosphorylation levels of the histone ${\gamma}H2AX$, as well as the number of apoptotic bodies and Annexin V‑positive cells. In addition, CNMSH exhibited scavenging activity against intracellular ROS generation and restored the mitochondria membrane potential (MMP) loss that were induced by H2O2, suggesting that CNMSH prevents H2O2‑induced DNA damage and cell apoptosis. Moreover, H2O2 enhanced the cleavage of caspase-3 and degradation of poly (ADP-ribose)-polymerase, a typical substrate protein of activated caspase-3, as well as DNA fragmentation; however, these events were almost totally reversed by pretreatment with CNMSH. Furthermore, CNMSH increased the levels of heme oxygenase-1 (HO-1), which is a potent antioxidant enzyme, associated with the induction of nuclear factor-erythroid 2-related factor 2 (Nrf2). According to our data, CNMSH is able to protect SH-SY5Y cells from H2O2-induced apoptosis throughout blocking cellular damage related to oxidative stress through a mechanism that would affect ROS elimination and activating Nrf2/HO-1 signaling pathway. Conclusions : Therefore, we believed that CNMSH may potentially serve as an agent for the treatment and prevention of neurodegenerative diseases caused by oxidative stress.
The deposition characteristics of MOCVO Cu using the (hfac)Cu(I) (1,5-DMCOD)(1,1,1,5,5,5-hexafluoro-2,4-pentanedionato Cu(I) 1,5-dimethyl-cyclooctadine) as a precursor have been investigated in terms of the effects of hydrogen and H(hfac) ligand addition with He carrier gas. MOCVD Cu using a Helium carrier gas showed a low deposition rate (20~$125{\AA}/min$) at the substrate temperature range of 180~$230^{\circ}C$. Moreover, the Cu film deposited at 19$0^{\circ}C$ was very thin (~$700{\AA}$) and showed the lowest resistivity value of $2.8{\mu}{\Omega}-cm$. The deposition rate of MOCVD Cu using $H_2$or H(hfac) addition was significantly enhanced especially at the low temperature region (180~$190^{\circ}C$). Furthermore, thinner Cu films (~$500{\AA}$) provided low resistivity (3.6~$2.86{\mu}{\Omega}-cm$). From surface reflectance measurement, very thin films deposited by using different gas system revealed good surface morphology comparable with sputtered Cu film ($300^{\circ}C$, vacuum-anneal). Hence, Cu film using (hfac)Cu(1,5-DMCOD) as a precursor is expected as a good seed layer in the electrochemical deposition process for Cu metallization.
Sung, Ha Guyn;Min, Dong Myung;Kim, Dong Kyun;Li, De Yun;Kim, Hyun Jin;Upadhaya, Santi Devi;Ha, J.K.
Asian-Australasian Journal of Animal Sciences
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v.19
no.12
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pp.1761-1768
/
2006
In this study, the comparative effects of transgenic corn (Mon 810 and Event 176) and isogenic corn (DK729) were investigated for their influence on in vitro rumen fermentation. This study consisted of three treatments with 0.25 g rice straw, 0.25 g of corn (Mon810/Event176/DK 729) mixed with 30 ml rumen fluid-basal medium in a serum bottle. They were prepared in oxygen free conditions and incubated at $39^{\circ}C$ in a shaking incubator. The influence of transgenic corn on the number of bacterial population, F. succinogenes (cellulolytic) and S. bovis (amylolytic), was quantified using RT-PCR. Fermentative parameters were measured at 0, 2, 4, 8, 12 and 24 h and substrate digestibility was measured at 12 and 24 h. No significant differences were observed in digestibility of dry matter, NDF, ADF at 12 and 24 h for both transgenic and isogenic form of corns (p>0.05) as well as in fermentative parameters. Fluid pH remained unaffected by hybrid trait and decreased with VFA accumulation as incubation time progressed. No influence of corn trait itself was seen on concentration of total VFA, acetic, propionic, butyric and valeric acids. There were no significant differences (p<0.05) in total gas production, composition of gas (methane and hydrogen) at all times of sampling, as well as in NH3-N production. Bacterial quantification using RT-PCR showed that the population number was not affected by transgenic corn. From this study it is concluded that transgenic corn (Mon810 and Event 176) had no adverse effects on rumen fermentation and digestibility compared to isogenic corn. However, regular monitoring of these transgenic feeds is needed by present day researchers to enable consumers with the option to select their preferred food source for animal or human consumption.
Temperature increase and precipitation changes caused by change alter aquatic environments including water quantity and quality that eventually affects the habitat of aquatic organisms. Such changes in habitat lead to changes in habitat suitability of the organisms, which eventually determines species distribution. Therefore, conventional habitat suitability models were investigated to evaluate habitat suitability changes of freshwater fish cause by change. Habitat suitability models can be divided into habitat-hydraulic (PHABSIM, CCHE2D, CASiMiR, RHABSIM, RHYHABSIM, and River2D) and habitat-physiologic (CLIMEX) models. Habitat-hydraulic models use hydraulic variables (velocity, depth, substrate) to assess habitat suitability, but lack the ability to evaluate the effect of water quality, including temperature. On the contrary, CLIMEX evaluates the physiological response against climatic variables, but lacks the ability to interpret the effects of physical habitat (hydraulic variables). A new concept of ecological habitat suitability modeling (EHSM) is proposed to overcome such limitations by combining the habitat-hydraulic model (PHABSIM) and the habitat-physiologic model (CLIMEX), which is able to evaluate the effect of more environmental variables than each conventional model. This model is expected to predict fish habitat suitability according to climate change more accurately.
Jaspine B is an anhydrophytosphingosine that is isolated from a marine sponge. Because of its structural similarity to sphingosine, it shows anti-cancer effects in human carcinomas. Therefore, this study aims to investigate its anti-proliferative effect on various cancer cells and to correlate its association with the intracellular accumulation of Jaspine B in relevant cancer cells. The anti-proliferative effect of Jaspine B in various cancer cells was determined by a cell viability test, and the intracellular concentration of Jaspine B in relevant cancer cells was determined using mass spectrometry coupled with liquid chromatography. The correlation coefficient and p value between the cytotoxicity and the cell accumulation of Jaspine B were determined using SPSS 16.1. The cytotoxicity of Jaspine B varied depending on the type of cancer cell when compared the $EC_{50}$ values of Jaspine B. Breast and melanoma cancer cells were susceptible to Jaspine B, whereas renal carcinoma cells were resistant. The intracellular concentrations of Jaspine B had a reciprocal correlation with the $EC_{50}$ values in the same cells (r = 0.838). The results suggested that the anti-proliferative effect of Jaspine B was associated with the cellular accumulation of this compound. However, Jaspine B was not a substrate for P-glycoprotein and breast cancer resistance protein, as major efflux pumps caused multidrug resistance. The maintenance of a high intracellular concentration is crucial for the cytotoxic effect of Jaspine B; however, efflux pumps may not be a controlling factor for Jaspine B-related resistance in cancer cells.
Park, In-Seon;Kim, Dong-Kyung;Shon, Dong-Hwa;Cho, Yong-Jin;Kim, Nam-Soo
Korean Journal of Food Science and Technology
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v.31
no.3
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pp.593-599
/
1999
Substrate specificity of a flow-injection-analysis (FIA)-type biogenic amine sensor with enzyme reactor was determined. The enzyme reactor was prepared with a diamine oxidase immobilized on preactivated chitosan porous beads (Chitopearl) by intramolecular cross-linking via glutaraldehyde. The sensor showed a rapid response to putrescine and a quasi-linear calibration curve was obtained up to 15.0 mM. The optimal pH and temperature of the enzyme reactor system were 7.5 and $35^{\circ}C$. Interferences due to ATP-related compounds and trimethylamine, and the effects of NaCl and amino acids were measured. Inhibitory effects owing to these components could be mitigated by sample extraction with perchloric acid. Polyamines except putrescine were determined by a putrescine calibration range within 26.7%. This system was confirmed as rapid and convenient for biogenic amine determination.
In order to prepare digested Protein source for the Weanling Food from soybean, an attempt was made to decompose steamed soybean protein to amino acids and peptides by protease and cellulase produced from Aspergillus niger and Aspergillus sojae. In this paper, the optimum condition for digestion of soybean protein were studied and also investigated the effects of decolorization of it. As the results, followings were obtained; 1. As steaming conditions, a treatment under 15 lb of pressure and 10 minutes of heating shows most effective. 2. The optimum pH of Asp, sojae enzyme for the digestion of soybean protein is 6.0, while that of Asp. niger enzyme is 4.4. In successievly-decomposing with Asp. sojae and Asp. niger, it shows the most effective on ratio of water-soluble-nitrogen to total nitrogen and amino-nitrogen to total nitrogen than any other separate treatments. 3. The suitable amount of the enzyme solution to that of the soybean substrate paste, in volume, is 1 : 2. 4. Digestion ratio of soybean protein indicates the gradual and steady effects of increasing time of digestion, but 8 hour-digestion regarding to putrefaction was suitable. 5. The most effective decolorization was successively passed on culumns of active carbon and anion exchanger (Dowex 2-x-8) at room temperature. In separate treatments, the effective order of decolorization was as follows; (Dowex 2-x-8)>Active carborn>Amberite IR-120 6. The powder type of the soy protein source obtained by concentration below $60^{\circ}C$ contains 12.51% of moisture, 66.31% of protein, 4.25% of fat, 12.75% of carbohydrate, 4.18% of ash.
Three conmmercial cellulases prepared from Penicillium notatum(cellulalse[K]), Trichoderma viride(cellulase[J]) and Aspergillus niger(cellulase[A]) were nalyzed with respect to their relative purity, activity and the effects of several metal ions on their activities. The activity of cellulase[K] was the strongest of all and that of cellulase[A] being the weaker. The purity of cellulalse[K] was the highest while that of cellulase[J] was the lowest. Under the assay conditions, the optimum concentrations of $Zn^{++}$ and $Mg^{++}$ ions for the activity of cellulase[K] was the highest while that of cellulase[J] was the lowest. Under the assay conditions, the optimum concentrations of $Zn^{++}$ and $Mg^{++}$ ions for the activity of cellulalse[K] was the highest while that of cellulase [A] being weaker. The purity of cellulase[K] was the highest while that of cellulase[J] was the lowest. Under the assay conditions, the optimum concentrations of $Zn^{++}$ and $MG^{++}$ ions for the activity of cellulase[K] were 2 and 7mM while those of cellulase[A] were 5 and 6 mM respectively and those of cellulase[J] were 3mM for both ions. Cellulase[K] and cellulase[J] were more strongly activated by $Zn^{++}$ than $Mg^{++}$ and cellulase[J] by $Mg^{++}$ than $Zn^{++}$. Both $Cu^{++}$ and $Mn^{++}$ ions inhibited by these metal ions. the inhibitory effects of $Mn^{++}$ ions for enzyme activities were stronger than $Cu^{++}$ ions. The Ki values of $Cu^{++}$ and $Mn^{++}$ for cellulase[K] were found to be 6.1 and 0.7mM, those of cellulase[J] were 2.6 and 0.32 mM, and those of cellulalse[A] were 2.0 and 0.2 mM respectively. Both $Cu^{++}$ and $Mn^{++}$ ions showed a pattrn of competitive inhibition of the enzyme activity. When Na-CMC was used as substrate, the Km and V values of celluase [K] were calculated to be $2.0{\times}10^{-4}mM$ and 3.43mmoles/hour, those of cellulase[J] were $2.4{\times}10^{-4}mM$ and 3.77mmoles/hour, and those of cellulase[A] were $4.0{\times}10^{-4}mM$ and 4.00mmoles/hour respectively.
Jeong, Kyung Hui;Hwang, In Sik;Kim, Ji Eun;Lee, Young Ju;Kwak, Moon Hwa;Lee, Young Hee;Lee, Jae Ho;Hwang, Dae Youn;Jung, Young Jin
Textile Coloration and Finishing
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v.26
no.1
/
pp.45-52
/
2014
Red pine (Pinus densiflora) is widely used traditional medicine, pharmacological and nutritional values from which the phytochemical compounds are derived. The present study was aimed to examine the antibacterial effects in the absence and presence of a immature red pine cone extract against 13 microorganisms. The components in the aqueous extract from immature red pine cone were identified by GC-MS. About 1.4% of total polyphenolic compound was measured in aqueous extract collected from immature red pine cone. Also, the high concentration of ${\beta}$-phellenandrene, ${\alpha}$-pinene, limonene, bornyl acetate and aldehyde was detected in total ion chromatograms. Of total 13 microorganisms, 4 microorganisms including Pseudomonas aeruginosa, Vibrio cholera, Listeria monocytogenes, Klebsiella pneumonia were effectively killed by aqueous extract of immature red pine cone. The highest anti-bacterial effect was detected in P. aeruginosa, followed by V. cholera, L. monocytogenes and K. pneumonia. In case of P. aeruginosa, the largest diameter of inhibition zone was maintained to 1/2 solution treated cells and slightly decreased at 1/4 and 1/8 solution treated cells. Also, in test used V. cholera and L. monocytogenes, the inhibition zone was strongly formed in only 1 and 1/2 solution treated cells, while K. pneumonia showed the very small diameter of inhibition zone in all concentrations. Therefore, these results suggested that the aqueous extracts of immature red pine cone should be considered as a new and potentially important anti-bacterial substrate to effectively prevent the microbial infection and penetration.
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