Background: Ginsenoside Rd (GSRd), one of the most abundant ingredients of Panax ginseng, protects the heart via multiple mechanisms including the inhibition of $Ca^{2+}$ influx.We intended to explore the effects of GSRd on L-type $Ca^{2+}$ current ($I_{Ca,L}$) and define the mechanism of the suppression of $I_{Ca,L}$ by GSRd. Methods: Perforated-patch recording and whole-cell voltage clamp techniques were applied in isolated rat ventricular myocytes. Results: (1) GSRd reduced $I_{Ca,L}$ peak amplitude in a concentration-dependent manner [half-maximal inhibitory concentration $(IC_{50})=32.4{\pm}7.1{\mu}mol/L$] and up-shifted the current-voltage (I-V) curve. (2) GSRd ($30{\mu}mol/L$) significantly changed the steady-state activation curve of $I_{Ca,L}$ ($V_{0.5}:-19.12{\pm}0.68$ vs. $-6.26{\pm}0.38mV$; n = 5, p < 0.05) and slowed down the recovery of $I_{Ca,L}$ from inactivation [the time content (${\zeta}$) from 91 ms to 136 ms, n = 5, p < 0.01]. (3) A more significant inhibitive effect of GSRd ($100{\mu}mol/L$) was identified in perforated-patch recording when compared with whole-cell recording [$65.7{\pm}3.2%$ (n = 10) vs. $31.4{\pm}5.2%$ (n = 5), p < 0.01]. (4) Pertussis toxin ($G_i$ protein inhibitor) completely abolished the $I_{Ca,L}$ inhibition induced by GSRd. There was a significant difference in inhibition potency between the two cyclic adenosine monophosphate elevating agents (isoprenaline and forskolin) prestimulation [$55{\pm}7.8%$ (n = 5) vs. $17.2{\pm}3.5%$ (n = 5), p < 0.01]. (5) 1H-[1,2,4]Oxadiazolo[4,3-a]-quinoxalin-1-one (a guanylate cyclase inhibitor) and N-acetyl-$\small{L}$-cysteine (a nitric oxide scavenger) partly recovered the $I_{Ca,L}$ inhibition induced by GSRd. (6) Phorbol-12-myristate-13-acetate (a protein kinase C activator) and GF109203X (a protein kinase C inhibitor) did not contribute to the inhibition of GSRd. Conclusion: These findings suggest that GSRd could inhibit $I_{Ca,L}$ through pertussis toxin-sensitive G protein ($G_i$) and a nitric oxide-cyclic guanosine monophosphate-dependent mechanism.
Kim, Byong-Wan;Lin, Guang-Zhe;Park, Byung-Ki;Kim, Jong-Duk;Shin, Jong-Suh
Journal of The Korean Society of Grassland and Forage Science
/
v.27
no.4
/
pp.323-334
/
2007
An in vitro study was conducted to examine the influence of alcohol-fermented feedstuff formulated with byproducts on the fermentation characteristics and NDF disappearance in the rumen. Dietary treatments were either a soybean curd-based alcohol-fermented feedstuff (AFS) or brewery grain-based alcohol-fermented feedstuff (AFB). The AFS and AFB are composed of 50% commercial beef cattle feed, 50% soybean curd dreg, 5% molasses and 0.5% yeast and 25% commercial beef cattle feed, 25% brewery grain, 25% soybean curd dreg, 25% corn grit, 5% molasses and 0.5% yeast, respectively. The change of ammonia, pH alcohol, volatile fatty acids, and NDF disappearance were measured at 0, 2, 4, 8 and 12 hr after in vitro incubation in the rumen. After 2 hr incubation, higher ammonia concentrations were resulted in AFS (12.47 mg/dl) and AFB (12.85 mg/dl) compared to control (11.84 mg/dl) (p<0.05). Ruminal pH of AFS and AFB were significantly higher than control during 1 to 6 hr fermentation, but the pH of AFS and AFB were decreased after 6 hr. At 12 hr fermentation, the alcohol concentration of AFS and AFB was significantly increased by 43.9% and 48.0%, respectively. The acetate concentration was rapidly decreased in control, while the concentration was slowly decreased in AFS and AFB with increasing the fermentation time. Lower concentrations propionate and butyrate were observed in AFS and AFB during every fermentation time (p<0.05). The NDF disappearance was significantly lower in AFS and AFB after 4 hr fermentation. These results suggest that alcohol fermented feedstuff can control the fermentation pattern in the rumen.
This study was conducted to determine the optimum condition of extraction and antioxidant activity of ethanol extracts of Epimedium koreanum Nakai based on the icariin quantity. Also, further organic solvent fractions of hexane, chloroform, ethyl acetate and butanol were obtained from the ethanol extract of Epimedium koreanum Nakai at different temperatures. Total ethanol extraction yield of wild grape seed ranged from 11.8% to 39.3% depending on the concentration of icariin as well as different ethanol concentration, extraction temperature and time condition. Among different extraction temperatures and time, the highest extraction yield of 39.3% was obtained at 70% ethanol for 3 hour at $50^{\circ}C$ in the sample containing the 0.596% icariin (KE9412). In the meantime, the strongest free radical scavenging effect $(RC_{50})$ with $18.9{\mu}g/mL$ in the KE9412 sample was observed in 70% ethanol extract of Epimedium koreanum Nakai extracted for 7 hour at $70^{\circ}C$, while $RC_{50}$ with $35.2{\mu}g/mL$ was observed in the KE9405 (0.20% icariin content) sample at same condition. Also, antioxidant activity of ethanol extracts of Epimedium koreanum Nakai increased as icariin concentration increased. Among each fraction obtained from organic solvents, butanol fraction was found to have the strongest $RC_{50}\;(39.2{\mu}g/mL)$ and followed by ethylacetate $(49.0{\mu}g/mL)$, water $(118.8{\mu}g/mL)$, ethylacetate $(119.8{\mu}g/mL)$, and chloroform fraction $(138.5{\mu}g/mL)$ respectively.
Sorafenib is a multikinase inhibitor and an oral anticancer drug approved for the treatment of patients with advanced renal cell carcinoma and those with unresectable hepatocellular carcinoma. The purpose of this study was to develop an efficient method of the determination of sorafenib in human plasma using tandem mass spectrometry coupled with liquid chromatography (LC/MS/MS) and validate the method by the guidelines of the Korean Food and Drug Administration (KFDA). Plasma samples ($100{\mu}l$) were added with chlorantraniliprole as an internal standard and then mixed with the 0.1% formic acid-containing extraction solution composed of isopropyl alcohol and ethyl acetate (1:4, v/v). After centrifugation, the supernatant was concentrated at $45^{\circ}C$ under negative pressure and centrifugal force. The residue was reconstituted with a mobile phase and injected into the HPLC instrument using a reverse phase Waters XTerra$^{TM}$ C18 column (particle size $3.5{\mu}m$). Liquid chromatography was carried out within the run time of 5 min using a mobile phase composed of buffer (0.1% formic acid and 10 mM ammonium formate), methanol, and acetonitrile (1:6:3, v/v/v). The analytes were monitored by tandem mass spectrometry in the multiple reaction monitoring method programmed to detect sorafenib at 'm/z 465.2 ${\rightarrow}$ 252.5' and chlorantraniliprole at 'm/z 484.4 ${\rightarrow}$ 286.2' with positive electrospray ionization mode ($ES^+$). The result showed the proper linearity ($r^2$ > 0.99) over the range of 2,000-5,000 ng/ml with good accuracy (90.7-103.9%) and precision (less than 10%). The newly developed method using LC/MS/MS was validated by the guideline of KFDA and identified as more sensitive compared to the previous methods.
The effects of seedtime and maturity stage on nutritive value of five maize stover varieties, including conventional maize (Kexiangyu 11, CM), fodder maize (Huqing 1, FM), high oil maize (Gaoyou 115, HOM), sweet maize (Kexiangtianyu 1, SM) and waxy maize (Kexiangluoyu 1, WM), were examined based on chemical composition, in vitro gas production and in situ incubation techniques. Maize stover was sampled at d 17 and d 30 after tasseling, and designated as maturity stage 1 and stage 2, respectively. The average dry matter (DM) organic matter (OM), crude protein (CP) and fiber contents were the greatest for HOM, SM and FM, respectively. CM had the highest in vitro organic matter disappearance (IVOMD) and volatile fatty acid (VFA) concentration. The highest ammonia nitrogen ($NH_3$-N) concentration in the incubation solution, and effective degradability of DM ($ED_{DM}$) and neutral detergent fiber ($ED_{NDF}$) were observed in SM. Advanced maturity stage increased (p<0.05) DM content, $ED_{DM}$ and $ED_{NDF}$, but decreased (p<0.05) OM and CP contents, and decreased (p<0.05) b and a+b values, IVOMD and molar proportion of valerate in the incubation solution for maize stover. Maize sown in summer had greater (p<0.05) OM content, but lower DM, CP, neutral detergent fiber (NDF) and acid detergent fiber (ADF) content compared with maize sown in spring. Maize sown in summer had greater (p<0.001) IVOMD, $NH_3$-N concentration in the incubation solution and $ED_{NDF}$, but lower (p<0.01) ratio of acetate to propionate compared to maize sown in spring. The interaction effect of variety${\times}$seedtime was observed running through almost all chemical composition, in vitro gas production parameters and in situ DM and NDF degradability. The overall results suggested that SM had the highest nutrient quality, and also indicated the possibility of selecting maize variety and seedtime for the utilization of maize stover in ruminants.
Cho, Sangbuem;Mbiriri, David Tinotenda;Shim, Kwanseob;Lee, A-Leum;Oh, Seong-Jin;Yang, Jinho;Ryu, Chaehwa;Kim, Young-Hoon;Seo, Kang-Seok;Chae, Jung-Il;Oh, Young Kyoon;Choi, Nag-Jin
Asian-Australasian Journal of Animal Sciences
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v.27
no.11
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pp.1652-1662
/
2014
The present study investigated the optimum blending condition of protected fat, choline and yeast culture for lowering of rumen temperature. The Box Benken experimental design, a fractional factorial arrangement, and response surface methodology were employed. The optimum blending condition was determined using the rumen simulated in vitro fermentation. An additive formulated on the optimum condition contained 50% of protected fat, 25% of yeast culture, 5% of choline, 7% of organic zinc, 6.5% of cinnamon, and 6.5% of stevioside. The feed additive was supplemented at a rate of 0.1% of diet (orchard grass:concentrate, 3:7) and compared with a control which had no additive. The treatment resulted in lower volatile fatty acid (VFA) concentration and biogas than the control. To investigate the effect of the optimized additive and feed energy levels on rumen and rectal temperatures, four rumen cannulated Hanwoo (Korean native beef breed) steers were in a $4{\times}4$ Latin square design. Energy levels were varied to low and high by altering the ratio of forage to concentrate in diet: low energy (6:4) and high energy (4:6). The additive was added at a rate of 0.1% of the diet. The following parameters were measured; feed intake, rumen and rectal temperatures, ruminal pH and VFA concentration. This study was conducted in an environmentally controlled house with temperature set at $30^{\circ}C$ and relative humidity levels of 70%. Steers were housed individually in raised crates to facilitate collection of urine and feces. The adaptation period was for 14 days, 2 days for sampling and 7 days for resting the animals. The additive significantly reduced both rumen (p<0.01) and rectal temperatures (p<0.001) without depressed feed intake. There were interactions (p<0.01) between energy level and additive on ruminal temperature. Neither additive nor energy level had an effect on total VFA concentration. The additive however, significantly increased (p<0.01) propionate and subsequently had lower acetate:propionate (A/P) ratios than non-additive supplementation. High concentrate diets had significantly lower pH. Interactions between energy and additive were observed (p<0.01) in ammonia nitrogen production. Supplementation of diets with the additive resulted in lower rumen and rectal temperatures, hence the additive showed promise in alleviating undesirable effects of heat stress in cattle.
Objective: Two experiments were performed to evaluate the effects of coated slow-release urea on nutrient digestion, ruminal fermentation, nitrogen utilization, blood glucose and urea concentration (Exp 1), and average daily gain (ADG; Exp 2) of steers. Methods: Exp 1: Eight ruminally fistulated steers [$503{\pm}28.5kg$ body weight (BW)] were distributed into a d $4{\times}4$ Latin square design and assigned to treatments: control (CON), feed grade urea (U2), polymer-coated slow-release urea A (SRA2), and polymer-coated slow-release urea B (SRB2). Dietary urea sources were set at 20 g/kg DM. Exp 2: 84 steers ($350.5{\pm}26.5kg$ initial BW) were distributed to treatments: CON, FGU at 10 or 20 g/kg diet DM (U1 and U2, respectively), coated SRA2 at 10 or 20 g/kg diet DM (SRA1 and SRA2, respectively), and coated SRB at 10 or 20 g/kg diet DM (SRB1 and SRB2, respectively). Results: Exp 1: Urea treatments (U2+SRA2+SRB2) decreased (7.4%, p = 0.03) the DM intake and increased (11.4%, p<0.01) crude protein digestibility. Coated slow-release urea (SRA2+-SRB2) showed similar nutrient digestibility compwared to feed grade urea (FGU). However, steers fed SRB2 had higher (p = 0.02) DM digestibility compared to those fed SRA2. Urea sources did not affect ruminal fermentation when compared to CON. Although, coated slow-release urea showed lower (p = 0.01) concentration of $NH_3-N$ (-10.4%) and acetate to propionate ratio than U2. Coated slow-release urea showed lower (p = 0.02) urinary N and blood urea concentration compared to FGU. Exp 2: Urea sources decreased (p = 0.01) the ADG in relation to CON. Animals fed urea sources at 10 g/kg DM showed higher (12.33%, p = 0.01) ADG compared to those fed urea at 20 g/kg DM. Conclusion: Feeding urea decreased the nutrient intake without largely affected the nutrient digestibility. In addition, polymer-coated slow-release urea sources decreased ruminal ammonia concentration and increased ruminal propionate production. Urea at 20 g/kg DM, regardless of source, decreased ADG compared both to CON and diets with urea at 10 g/kg DM.
Volatile organic compounds (VOCs) are increasingly been recognized as the chemical mediators of mold interactions, shaping their community dynamics, growth, and metabolism. Herein, we selectively examined the time-correlated (0 D-11 D, where D = incubation days) effects of intraspecies VOC-mediated interactions (VMI) on Aspergillus oryzae KCCM 60345 (S1), following co-cultivation with partner strain A. oryzae KACC 44967 (S2), in a specially designed twin plate assembly. The comparative evaluation of $S1_{VMI}$ (S1 subjected to VMI with S2) and its control ($S1_{Con}$) showed a notable disparity in their radial growth ($S1_{VMI}$ < $S1_{Con}$) at 5 D, protease activity ($S1_{VMI}$ > $S1_{Con}$) at 3-5 D, amylase activity ($S1_{VMI}$ < $S1_{Con}$) at 3-5 D, and antioxidant levels ($S1_{VMI}$ > $S1_{Con}$) at 3 D. Furthermore, we observed a distinct clustering pattern for gas chromatography-time of flight-mass spectrometry datasets from 5 D extracts of $S1_{VMI}$ and $S1_{Con}$ in principle component analysis (PC1: 30.85%; PC2: 10.31%) and partial least squares discriminant analysis (PLS-DA) (PLS1: 30.77; PLS2: 10.15%). Overall, 43 significantly discriminant metabolites were determined for engendering the metabolic variance based on the PLS-DA model (VIP > 0.7, p < 0.05). In general, a marked disparity in the relative abundance of amino acids ($S1_{VMI}$ > $S1_{Con}$) at 5 D, organic acids ($S1_{VMI}$ > $S1_{Con}$) at 5 D, and kojic acid ($S1_{VMI}$ < $S1_{Con}$) at 5-7 D were observed. Examining the headspace VOCs shared between S1 and S2 in the twin plate for 5 D incubated samples, we observed the relatively higher abundance of C-8 VOCs (1-octen-3-ol, (5Z)-octa-1,5-dien-3-ol, 3-octanone, 1-octen-3-ol acetate) having known semiochemical functions. The present study potentially illuminates the effects of VMI on commercially important A. oryzae's growth and biochemical phenotypes with subtle details of altered metabolomes.
The effects of organic supplement (acetate) and dissolved oxygen concentration on the nitrification rate of wastewater were investigated in the 27.7 L pilot-scale airlift biofilm reactor with the granular activated carbon media of 0.613 mm diameter. The ammonium oxidation rate increased stepwise up to 5 kg N/㎥$.$d at the riser air velocity of 0.063 m/s, when the air velocity and the ammonium loading rate were raised alternately. The nitrite build-up was observed during the early stage of the biofilm formation, which disappeared after the reactor operation of 128 days. As increasing the organic loading rate, the organic oxidation rate increased up to 25.0 kg COD/㎥$.$d with the removal efficiency of 94% but the oxidation rates of ammonium and nitrite decreased. The oxidation rates of ammonium and nitrite increased with the dissolved oxygen concentrations. When the pure oxygen was sparged, the ammonium oxidation rate was almost five times higher than that with air at the same velocity.
The effects of organic acids mix (0.4%) and modified atmosphere packaging (MAP) on the storage quality of sliced bacon were investigated. Pork bellies were treated with or without organic acids at the curing stage. The organic acids mix comprised 35% sodium acetate, 25% salt, 15% calcium lactate, 11% trisodium citrate, 7% ascorbate, and 7% citric acid. The cured pork bellies were smoked and packaged with 50% $CO_2$ + 50% $N_2$ (50% $CO_2$-MAP) and 100% $N_2$ (100% $N_2$-MAP), and stored at $5^{\circ}C$ for 14 d. The 50% $CO_2$-MAP showed a higher pH value (p<0.05) up to 10 d, a lower protein deterioration (p<0.05) as measured by volatile basic nitrogen (VBN) from 6 to 14 d, and a higher color value of lightness (CIE $L^*$) compared to 100% $N_2$-MAP. The development of lipid oxidation measured by thiobarbituric acid reactive substance (TBARS) values seemed to be effectively controlled throughout the storage period in both 50% $CO_2$-MAP and 100% $N_2$-MAP regardless of the application of organic acids. The 50% $CO_2$-MAP inhibited the growth of aerobic and anaerobic bacteria (p<0.05) both in non-added and bacon added with organic acids mix. The 50% $CO_2$-MAP alone seemed to be effective in delaying the growth of bacteria since the use of organic acids mix gave no additional effects. The addition of organic acids mix lowered the pH value (p<0.05), effectively retarded the protein deterioration (p<0.05), and showed a higher color value of lightness (CIE $L^*$) value (p<0.05) and lower color value of redness (CIE $a^*$) value (p<0.05). In conclusion, 50% $CO^2$-MAP showed better quality and self-life of sliced bacon during storage. However, the beneficial effect of organic acids mix was not noticed in the concentration used in this experiment.
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