Serum-contain is commoly used for the production of in vitro-derived bovine embryos. However, were biological activity of serum varies from lot to lot, time consuming to choose better serum with good quality and risks of virus, bacteria and mycoplasma infection. This study established serum-free culture systems of in vitro embryo development to efficiently obtain a large number of blastocysts from ovaries of Hanwoo and oocytes maturation, cell number, tlerance of cryopreservation. Secondly, serum-contain medium is suspected of contributing to the large calf size, dystocia, cersarean sections, calf mortality and confirmed these blastocysts are high quality in terms of cyotolerance, high rates of pregancy and normal birth. For these reasons, Culture media (IVMD101 and IVD101) designed specifically for the preimplantation bovine embryo are rather simplistic, being based on salt solutions with additional energy substrates and growth factors. An improved serum-free medium (IVMD101) was developed for bovine oocytes maturation in vitro. Proportions of embryos developing to the blastocyst stage cultured in both IVD101(32.4%) and IVD101(34.5%) serum-free media were higher than in TCM199+10% FBS(12.4%) serumcontaining medium. Futhermore, the cell numbers per blastosyst obtained in the serum-free media were superior to those of blastocysts developed in serum-supplemented medium. Also, cell numbers of blastocysts obtained in the serum-free media were similar with blastocysts derived in vivo. Survival rate blastocysts after 24 hr incubation after thawing, the blastocysts cultured in both IVD101(94.5%) and IVD101(95.8%) serum-free media were higher than in TCM199+10% FBS (52.5%) serum-containing medium. After 72 hr incubation after thawing, hatching rates of blastocysts developed in IVD101(78.4%) and IVMD101(83.7%) were sighnificantly higher than that developed in the serum-supplemented medium(32.0%). The pregnancy rates almost not different between fresh blastocysts(38.2%) and frozen blastocysts(34.9%). The results suggested that the improved serum-free media(IVMD101 and IVD101) offer several advantages over culture in serum-cotaining medium, including increased rates of blastocyst formation and high cel numbers. Additionally, the survival and hatching rates of embryos product in serum-free media after post-thaw culture were superior to those of embryos produced in the serum-containg medium and useful for the production of high quality bovine embryos for cryo-preservation. These improved serum-free media are beneficial not only for the study of the mechanisms of early embryogenesis but also for mass production of good quality embryos for embryo transfer, cloning and transgenesis.
The purpose of this study was to characterize equine heat-shock protein (Hsp) genes and analyze their expression pattern in various horse tissues and blood leukocytes after exercise. In a previous study, RNA sequencing of blood and skeletal muscles of thoroughbreds before and after exercise was performed using differently expressed gene (DEG) analysis. Three Hsp genes (HspH1, Hsp90${\alpha}$ and Hsp70) were selected by DEG analysis and were found to be differentially expressed in either blood or muscle. To validate and extend previous observations on these genes, we performed RT-PCR analyses of horse tissue as well as real-time qPCR analyses of blood leukocytes after exercise. mRNA expression of these Hsp genes was found to be ubiquitous in the analyzed tissues (including thyroid, colon, skeletal muscle, cecum, kidney, spinal cord, heart, and lung). In addition, Hsp mRNA expression of these genes in extracted whole blood increased after 120 minutes of exercise compared to the baseline condition. These results are in agreement with the results of human and other experimental animals, suggesting that regulatory mechanisms that are responsible for upregulation of Hsp gene transcription may be conserved among species. Further investigations to correlate Hsp gene expression patterns with athletic performance or recovery processes after exercise are warranted.
Moon, Yea-Hwang;Kim, Su Cheol;Cho, Woong Gi;Lee, Sung Sill;Cho, Soo Jeong
Journal of Mushroom
/
v.12
no.2
/
pp.138-143
/
2014
This study was conducted to know the optimal supplementary level of spent mushroom substrates (Flammulina velutipes) as an energy source and fermentation period in manufacturing of rye silage. Whole crop rye was harvested at full bloom stage and ensiled with spent mushroom substrates of 20%(S-20), 40%(S-40) and 60%(S-60) as fresh matter basis. Each silage was prepared in plastic buckets included with vinyl bag by three replications and fermented for 3, 6 and 9 weeks, respectively. Moisture contents of whole crop rye at full bloom stage and spent mushroom substrates were 62.3% and 54.3%, respectively, and those of silages was ranged from 58% to 64%. Ether extracts content of silages was significantly (P<0.05) increased in 6 weeks of fermentation period. The pH of silages ranged from 4.46 to 5.05, and fluctuated in the changes by fermentation period beside of decreased with elapsing the period in the S-60. The organic acid content of silages was higher in the order of lactic acid, butyric acid, acetic acid and propionic acid. Lactic acid content was higher when fermented for 6 weeks compared to the other fermentation period. Flieg's score for estimation of silage quality ranged from 63 to 80, and was relatively high level in the S-20 and the S-40 fermented for 6 weeks, and was high level in S-20, and was relatively low level in the silages fermented for 3 week. Hence, we suggest that supplementation of 20% spent mushroom substrates in fresh matter basis and fermentation for 6 weeks are resonable for a good quality of rye silage.
Hydrogeochemical study in Samseung area (Boeun, Chungbuk) and waterrock interaction experiment using rock samples from the area were performed to elucidate the fluoride source in groundwater and explaining geochemical behavior of fluoride ion. Fluoride concentration of public water supply mostly using groundwater in Boeun area was significantly higher in South Korea. The maximum fluoride concentration of the study area was 3.9 mg/L, and 23% of samples exceeded the Korean Drinking Water Standard of fluoride (1.5 mg/L). The average concentration of fluoride was 1.0 mg/L and median was 0.5 mg/L. Because of high skewness (1.3), median value is more appropriate to represent fluoride level of this area. The relationships between fluoride ion and geochemical parameters ($Na^+$, $HCO_3$, pH, etc.) indicated that the degree of waterrock interaction was not significant. However, high fluoride samples were observed in $NaHCO_3$ type on Piper's diagram. The negative relationship between fluoride and $NO_3$ ion which might originate from surface contaminants was obvious. These results indicate that fluoride ion in groundwater is geogenic origin. The source of fluoride was proved by waterrock interaction batch test. Fluoride concentration increased up to 1.2 mg/L after 96 hours of reaction between water and biotite granite. However, the relationship between well depth and fluoride ion, and groundwater age and fluoride ion was not clear. This indicates that fluoride ion is not correlated with degree of waterrock interaction in this area but local heterogeneity of fluoriderich minerals in granite terrain. High fluoride concentration in Boeun area seems to be correlated with distribution of permeable structures in hard rocks such as lineaments and faults of this area. This entails that the deep bedrock groundwater discharges through the permeable structures and mixed with shallow groundwater.
Geochemical characteristics of the Guryongsan (Ogcheon) uraniferous black slate show that this is an analogue to the conventional Chattanooga and Alum shales in occurrences. Whereas, its highest enrichment ratio in metals including uranium, among others, is explained by the cyclic sedimentation of the black muds and quartz-rich silts, and the uniform depositional condition with some what higher pH condition compared to the conditions of the known occurrences. The cyclic sedimentation, caused by the periodic open and close of the silled basin, has brought about the flush-out) of the uranium depleted water and the recharge with the new metal-rich sea water, which consequently contributed to the high concentration of metals in mud. The metal-rich marine black muds, which mostly occur in the early to middle Palaeozoic times, is attributed by the geologic conditions which related to the atmospheric oxygen contents, and these are scarcely met in the late Precambrian and/or with the onset of Palaeozoic era in the geologic evolution of the earth.
Park, Sung-ae;Kim, Taekyu;Shim, Kyuyoung;Kong, Hak-Yang;Yang, Byeong-Gug;Suh, Sanguk;Lee, Chang Seok
Korean Journal of Ecology and Environment
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v.52
no.3
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pp.210-220
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2019
Population growth and the increase of energy consumption due to civilization caused global warming. Temperature on the Earth rose about $0.7^{\circ}C$ for the last 100 years, the rate is accelerated since 2000. Temperature is a factor, which determines physiological action, growth and development, survival, etc. of the plant together with light intensity and precipitation. Therefore, it is expected that global warming would affect broadly geographic distribution of the plant as well as structure and function ecosystem. In order to understand the effect of global warming on the ecosystem, a study about the effect of temperature rise on germination and growth in the plant is required necessarily. This study was carried out to investigate the effects of experimental warming on the germination and growth of two oak species(Quercus mongolica and Q. serrata) in temperature gradient chamber(TGC). This study was conducted in control, medium warming treatment($+1.7^{\circ}C$; Tm), and high warming treatment ($+3.2^{\circ}C$; Th) conditions. The final germination percentage, mean germination time and germination rate of two oak species increased by the warming treatment, and the increase in Q. serrata was higher than that in Q. mongolica. Root collar diameter, seedling height, leaf dry weight, stem dry weight, root dry weight, and total biomass were the highest in Tm treatment. Butthey were not significantly different in the Th treatment. In the Th treatment, Q. serrata had significantly higher H/D ratio, S/R ratio, and low root mass ratio (RMR) compared with control plot. Q. mongolica had lower RMR and higher S/R ratio in the Tm and Th treatments compared with control plot. Therefore, growth of Q. mongolica are expected to be more vulnerable to warming than that of Q. serrata. The main findings of this study, species-specific responses to experimental warming, could be applied to predict ecosystem changes from global warming. From the result of this study, we could deduce that temperature rise would increase germination of Q. serrata and Q. mongolica and consequently contribute to increase establishment rate in the early growth stage of the plants. But we have to consider diverse variables to understand properly the effects that global warming influences germination in natural condition. Treatment of global warming in the medium level increased the growth and the biomass of both Q. serrata and Q. mongolica. But the result of treatment in the high level showed different aspects. In particular, Q. mongolica, which grows in cooler zones of higher elevation on mountains or northward in latitude, responded more sensitively. Synthesized the results mentioned above, continuous global warming would function in stable establishment of both plants unfavorably. Compared the responses of both sample plants on temperature rise, Q. serrata increased germination rate more than Q. mongolica and Q. mongolica responded more sensitively than Q. serrata in biomass allocation with the increase of temperature. It was estimated that these results would due to a difference of microclimate originated from the spatial distribution of both plants.
Piao, X.S.;Chae, B.J.;Kim, J.H.;Jin, J.;Cho, W.T.;Han, In K.
Asian-Australasian Journal of Animal Sciences
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v.12
no.5
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pp.783-787
/
1999
To study the effects of different extrusion conditions of barley on growth performance, nutrient digestibility and nutrient excretion in feces, a total of 150 growing pigs ($Landrace{\times}Duroc{\times}Large$ White; average 24.4 kg body weight) were allotted to five treatments, in a completely randomized block design. The experimental diets were based on corn-soybean and 30% of barley was included in each diet; barley was the only extruded ingredient. The treatments were 1) no extrusion (Control); 2) extrusion at $100^{\circ}C$ without preconditioning (ENLT); 3) extrusion at $150^{\circ}C$ without preconditioning (ENHT); 4) extrusion at $100^{\circ}C$ with preconditioning (ECLT); 5) extrusion at $150^{\circ}C$ with preconditioning (ECHT). Temperature in the barrel was controlled within ${\pm}5^{\circ}C$ by feed rate with the addition of water at the rate of $3{\ell}\;per\;min$. in the extruder for each treatment. For the 6 week experimental period, extrusion of barley improved the average daily gain (ADG) and digestibilities of dry matter, crude protein and gross energy in growing pigs. As compared to control, significant improvements in ADG (p<0.05) were shown in the groups of feeding extruded barley at high temperature (ENHT and ECHT). There were also significant differences in the digestibilities of DM, CP and P between extrusion temperatures. Barley extruded at high temperature gave better digestibilities of DM, CP and GE than barley extruded at low temperature. Extruded barley diet groups showed significantly (p<0.05) lower excretions of DM, nitrogen (N) and P per kg gain as compared to the ground barley group. DM, N and P excretion per kg gain were also significantly lower in pigs fed barley extruded at $150^{\circ}C$ than at $100^{\circ}C$. In conclusion, extrusion considerably improved the nutritive value of barley and it appeared that temperature is the most important variable.
Park, Chang-Hoon;Kwak, Jin-Won;Park, Bong-Soo;Kim, Yong-Ho;Kim, Yong-Deok;Yoon, Ji-Uk;Yoon, Ji-Young;Kim, Cheul-Hong
Journal of The Korean Dental Society of Anesthesiology
/
v.14
no.1
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pp.41-47
/
2014
Background: Autophagy is a self-eating process that is important for balancing sources of energy at critical times in development and in response stress. Autophagy also plays a protective role in removing clearing damaged intracellular organelles and aggregated proteins as well as eliminating intracellular pathogens. The purpose of the present study was to examine the protective effect of propofol against hypoxic damage using keratinocytes. Methods: Human keratinocytes (HaCaT cells) were obtained from the American Type Culture Collection. Propofol which were made by dissolving them in DMSO were kept frozen at $-4^{\circ}C$ until use. The stock was diluted to their concentration with DMEM when needed. Prior to propofol treatment cells were grown to about 80% confluence and then exposed to propofol at different concentrations (0, 25, 50, 75, $100{\mu}M$) for 2 h pretreatment. Cell viability was measured using a quantitative colorimetric assay with thiazolyl blue tetrazolium bromide (MTT assay), and fluorescence microscopy and western blot analysis were used for evaluation of autophagy processes. Results: The viability of propofol-treated HaCaT cells was increased in a dose-dependent manner. Propofol did not show any significant toxic effect on the HaCaT cells. The autophagy inhibitor, 3-methyladenine, reduced cell viability of hypoxia-injured HaCat cells. Fluorescence microscopy and western blot analysis showed propofol induce autophagy pathway signals. Conclusions: Propofol enhanced viability of hypoxia-injured HaCaT cells and we suggest propofol has cellular protective effects by autophagy signal pathway activation.
This experiment was conducted to study the effect of addition of Lucerne juice (LJ) obtained by mechanical extraction of freshly harvested crop on the nutritive value of rice straw silage. Rice straw (RS) was ensiled with intact, NaOH or $NH_3$ treated LJ at 3:7 ratio on fresh weight basis (LJ RS, LJ NaOH RS and LJ $NH_3$ RS, respectively). Each alkali was mixed with fresh juice at a level of 4% of rice straw dry matter just before ensiling. Rice straw ensiled with water was prepared as the control (W RS). In the digestion trial, goats were allocated in a $4{\times}4$ Latin-square design and fed the diet containing three parts of RS silage and one part of wheat bran (DM basis). For the goats receiving the control silage, urea was supplemented at feeding time so as to adjust the nitrogen intake except for goats on LJ $NH_3$ RS silage. Crude protein content of RS silage was increased from 5.2 to 9.1% (DM basis) by the addition of intact LJ and to about 24% by $NH_3$ treated LJ. The control W RS silage contained only trace amount of lactic acid and was dominated by acetic and butyric acid. The addition of intact LJ reduced butyric acid content and $NH_3-N/TN$ of the silage whereas the addition of alkalized LJ increased those values and shifted to a butyrate type fermentation. Nutrient digestibilities and nitrogen balance of goats were almost the same when they were fed W RS and LJ RS silage indicating the addition of intact LJ did not improve the nutritive value. The addition of alkalized LJ significantly increased the fiber digestibilities of RS silage and $NH_3$ treatment was more effective than NaOH treatment. Postprandial ruminal $NH_3-N$ and blood urea nitrogen (BUN) concentrations were decreased by feeding LJ NaOH RS silage suggesting ruminal protein synthesis was enhanced along with the increase of energy supply for supply for rumen microbes by the alkali treatment. The advantageous fiber digestibilities of LJ $NH_3$ RS silage compared with those of LJ NaOH RS silage might be attributable to a sufficient nitrogen supply for microbial fiber digestion in the rumen.
Wang, Kun;Nan, Xuemei;Zhao, Puyi;Liu, Wei;Drackley, James K.;Liu, Shijie;Zhang, Kaizhan;Bu, Dengpan
Asian-Australasian Journal of Animal Sciences
/
v.31
no.5
/
pp.677-685
/
2018
Objective: The objective of the present study was to determine ammonium chloride tolerance of lactating dairy cows, by examining effects of negative dietary cation anion difference (DCAD) induced by ruminal ammonium chloride infusion on performance, serum and urine minerals, serum metabolites and enzymes of lactating dairy cows. Methods: Four primiparous lactating Chinese Holstein cows fitted with ruminal cannulas were infused with increasing amounts (0, 150, 300, or 450 g/d) of ammonium chloride in a crossover design. The DCAD of the base diet was 279 mEq/kg dry matter (DM) using the DCAD formula (Na + K - Cl - S)/kg of DM. Ammonium chloride infusion added the equivalent of 0, 128, 330, and 536 mEq/kg DM of Cl in treatments. According to the different dry matter intakes (DMI), the resulting actual DCAD of the four treatments was 279, 151, -51, and -257 mEq/kg DM, respectively. Results: DMI decreased linearly as DCAD decreased. Yields of milk, 4% fat-corrected milk, energy-corrected milk, milk fat, and milk protein decreased linearly as DCAD decreased. Concentrations of milk protein and milk urea nitrogen increased linearly with decreasing DCAD. Concentration of Cl- in serum increased linearly and concentration of PO43- in serum increased quadratically as DCAD decreased. Urine pH decreased linearly and calculated urine volume increased linearly with decreasing DCAD. Linear increases in daily urinary excretion of $Cl^-$, $Ca^{2+}$, $PO_4{^{3-}}$, urea N, and ammonium were observed as DCAD decreased. Activities of alanine aminotransferase, aspartate aminotransferase, and ${\gamma}-glutamyl$ transferase in serum and urea N concentration in serum increased linearly as DCAD decreased. Conclusion: In conclusion, negative DCAD induced by ruminal ammonium chloride infusion resulted in a metabolic acidosis, had a negative influence on performance, and increased serum enzymes indicating potential liver and kidney damage in lactating dairy cows. Daily ammonium chloride intake by lactating dairy cows should not exceed 300 g, and 150 g/d per cow may be better.
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