• Development and Reproduction
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• v.5 no.1
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• pp.23-33
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• 2001

When mammalian oocytes ovulate into the oviduct, associating follicular fluid components are exposed to the oviductal environment, possibly resulting in the mutual interaction between fillicu1ar and oviductal fluids. In the Present study, we have demonstrated for the first time that components of fallicular fluid could be modified by the oviductal fluid. Gelatin zymographic analyses of human follicular fluid (hFF) obtained from IVF patients showed consistently the presence of 110 kDa gelatinase (GA110) in addition to many bands among which 62 kDa gelatinase was predominant. Addition of EDTA or phenanfhroline to the gelatinase substrate buffer during gel incubation abolished GA110 band whereas phenylmethylsulffnyl fluoride (PMSF) did not. In contrast, bovine oviductal fluid(bOF) exhibited only 62 kDa gelatinase. Surprisingly, when bOF was added to hFF in 1:1 ratio and then the mixture was incubated for 3 h at 37$^{\circ}$C, GA110 of hFF disappeared. Disappearance of GA110 by bOF was observed even within 30 min after mixing with hFF. Addition of aminophenylmercuric acetate (APMA) to hFF also abolished enzymatic activity of GA110 but increased the activityof 62 kDa gelatinase. However, APMA abolished many other gelatinases as well unlike bOF. Interestingly, treatment of hFF with EDTA for 3 h remarkably increased the enzymatic activity of GA110 but not that of other gelatinases. Addition of phenanthroline, PMSF or soybean trypsin inhibitor (SBTI) did not affect overall gelatinase activities. Again, addition of bOF to the hFF pretreated with any of the above proteinase inhibitors abolished the appearance of GA110. Human serum also showed GAI 10 of which activity was greatlyenhanced by EDTA treatment. Similar to hFF, serum GA110 also disappeared by the addition of bOF. Human granulosa cell homogenate did not reveal any appreciable gelatinase activity except 92 kDa gelatinase. Anti-human gelatinase A antibody reacted with 62 kDa gelatinase of hFF. Based upon these results, it is concluded that bOF could selectively degrade an isoform of gelatinase A present in hFF and human serum.

• Analytical Science and Technology
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• v.23 no.6
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• pp.531-536
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• 2010

A quantitative analytical method has been established for the measurement of inosine 5'-monophosphate dehydrogenase (IMPDH) activity in human peripheral blood mononuclear cells (PBMCs) by ion-pair reversed-phase high performance liquid chromatography equipped with ultraviolet detection (HPLC/UV). IMPDH is a ${\beta}$-nicotinamide adenine dinucleotide hydrate (NAD+)-dependent dehydrogenase in which the enzyme converts inosine 5'-monophosphate (IMP) into xanthosine 5'-monophosphate (XMP). Its activity was measured by quantifying a HPLC chromatogram corresponding to XMP produced during the incubation of lysed PBMCs with IMP as a substrate and $NAD^+$ as a coenzyme. XMP produced was detected at a wavelength of 260 nm. The mobile phase was composed of a mixture of 37 mM potassium dihydrogen phosphate containing 7 mM tetra-n-butylammonium hydrogen sulfate adjusted to pH 5.5 and methanol (85:15, v/v) with a flow rate of 1 mL/min. The calibration curve was linear ($r^2$=0.999999) in the range of $0.2-50.0\;{\mu}M$ and the limit of quantification (LOQ) was $0.2\;{\mu}M$. The intra- and inter-day precisions were between 0.88-1.47% and 0.85-5.24%, respectively. The intra- and inter-day accuracies were between 98.74-99.99% and 99.95-101.65%, respectively. IMPDH activity in 11 Korean healthy volunteers ranged from 18.29 to 36.60 nmol/h/mg protein (mean = $27.70{\pm}6.28\;nmol/h/mg$ protein).

• Journal of The Korean Society of Grassland and Forage Science
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• v.31 no.2
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• pp.177-190
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• 2011

This study was conducted to determine effects of dietary herbaceous peat on in vitro fermentation and milk production in dairy cows. Ruminal pH, gas production, VFA (volatile fatty acid), Ammonia-N, and rumen degradability were examined by the addition of three times over 0, 1, and 5% herbaceous peat with substrate of timothy hay, and the change of rumen fermentation characteristics were evaluated. In 0, 3, 12 and 24 hours cultivation, all treatments did not show a significant difference but the control at 6 hours appeared significantly lower pH compared to 1 and 5% treatments (p<0.05). The gas production of the treatments significantly increased until 12 hours of cultivation compared to control (p<0.05), the rumen ammonia concentration showed a tendency to increase until 24 hours in all treatment groups, and there was no significant difference between treatments. About the rumen degradability, 5% treatment showed higher rumen degradability in all hours than control and 1% treatment (p<0.05). Meanwhile, for in vivo trial, 16 heads of Holstein lactation dairy cows were selected for experiment for four weeks in order to research the change of milk yield, milk compositions and change of somatic cell counts of lactation dairy cows by herbaceous peat feeding. The milk yield of vitamin C and herbaceous peat treatments (T3) was 25.0 kg but the control was 23.2 kg, herbaceous peat treatment (T1) was 23.1 kg, and vitamin C treatment (T2) was 23.4 kg, so there was linear increase effect of milk yield by T3. The partial significance of the milk (fat, milk protein, lactose, MUN and SNF) and change of somatic cell count before and after experiment by the control and treatments about change of milk and somatic cell counts (p<0.05) were recognized. About change of milk in the first half (1~2 weeks) and latter half (3~4 weeks) during four weeks of experiments period, the herbaceous peat supplement treatments showed a tendency of significant decrease of quality of milk protein and SNF. The control and treatments did not show significant change of blood nutrients (total protein, cholesterol, NEFA, BUN), liver function component (AST, GGT) and minerals (Ca, P, Mg) before and after experiment. In summary, it is judged that herbaceous peat feeding for lactation dairy cows would be recommendable based on the results of milk, somatic cell count physiologically.

• Horticultural Science & Technology
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• v.29 no.3
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• pp.187-194
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• 2011

The objective of this study was to determine the effects of light intensity and electrical conductivity (EC) of nutrient solution on the growth and nutrient uptake of potted kalanchoe plants (Kalanchoe blossfeldiana 'Marlene') with growth stage in ebb and flow subirrigation systems. The plants were grown at four ECs of 0.5, 1.0, 1.5, and 2.0 $dS{\cdot}m^{-1}$ for seedling stage and four ECs of 1.0, 1.5, 2.0, and 3.0 $dS{\cdot}m^{-1}$ for short day stage under three daily photosynthetic photon flux (PPF) of 6.5, 10.3, 18.2 $mol{\cdot}m^{-2}{\cdot}d^{-1}$. At seedling stage, plant height was the longest under the lowest light intensity, and particularly dry weights and leaf areas were the highest at PPF 10.3 $mol{\cdot}m^{-2}{\cdot}d^{-1}$. Dry weights and leaf areas were the highest at EC 1.5 $dS{\cdot}m^{-1}$ regardless of light intensity. At short day exposure, plant height was the longest under the lowest light intensity. Dry weights, leaf areas, and number of pedicels of the plants significantly increased as light intensity increased. Under all light intensity conditions, dry weights, leaf areas, and number of pedicles increased until EC becomes to 1.0 - 2.0 $dS{\cdot}m^{-1}$. And after reached the highest at EC 2.0 $dS{\cdot}m^{-1}$, they decreased at EC 3.0 $dS{\cdot}m^{-1}$. By comparing the ion uptakes at EC 1.5 $dS{\cdot}m^{-1}$ of seedling stage and EC 2.0 $dS{\cdot}m^{-1}$ of short day stage in which the plants grew better, we confirmed that ion balance of nutrient solution among $NO_3{^-}$-N, $H_2PO_4{^-}$, $K^+$, $Ca^{2+}$, and $Mg^{2+}$ were significantly changed at short day stage compared to seedling stage. For better growth of the plants, both ion balance and EC of nutrient solution should be considered under different light intensities at short day stage while control of EC is enough at seedling stage.

• Applied Chemistry for Engineering
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• v.5 no.4
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• pp.681-697
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• 1994

The enzymatic hydrolysate of gelatin extracted from fish skin was fractionated and recycled through the membrane reactor according to the molecular weight for the purpose of using as functional material. In addition, the enzymatic hydrolysis conditions of gelatin, enzyme stability by membrane and mechanical shear, and effect on the long-term operational stability of the recycle membrane reactor were investigated. Using the pH-drop technique, Alcalase, pronase E and collagenase were identified as the most suitable enzymes for the hydrolysis of fish skin gelatin. The optimum hydrolysis conditions in the 1st-step membrane reactor(1st-SMR) by Alcalase were enzyme concentration 0.2mg/ml, substrate-to-enzyme ratio(S/E) 50(w/w), $50^{\circ}C$, pH 8.0, reaction volume 600ml and flow rate 6.14ml/min. In the 2nd-SMR by pronase E were enzyme concentration 0.3mg/ml, S/E 33(w/w), $50^{\circ}C$, pH 8.0, reaction volume 600ml and flow rate 6.14ml/min. In the case of 3rd-SMR, enzyme concentration 0.1mg/ml, S/E 100(w/w), $37^{\circ}C$, pH 7.5, reaction volume 600ml and flow rate 10ml/min. Decreased enzyme activities by mechanical shear and membrane were 30% and 15% in the 1st-SMR, were 14% and 5% in the 2nd-SMR, and 18% and 8% in the 3rd-SMR, respectively. Under the optimum conditions, the degree of hydrolysis in the 1st, 2nd and 3rd-SMR were 3.5%(Kjeldahl method, 87%), 3.1%(77%) and 2.7%(70%), respectively. The productivity of hydrolysate in the continuous three-step membrane reactor was 430mg per enzyme(mg) for 10 times of volume replacements.

• Korean Journal of Ecology and Environment
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• v.39 no.2 s.116
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• pp.167-177
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• 2006

Both fish community and water quality in Lake Doam were investigated from September 2004 to August 2005. The turbidity of Lake Doam located in the upper region of the Songchun River in the South River system, Korea was high whole year due to the effects of distributed non point source pollutions in the watersheds. During the experimental periods, mean concentration of chlorophyll-a in epilimnetic layer (0 ${\sim}$ 5 m) was 18.5 ${\mu}g\;L^{-1}$ and transparency ranged from 0.3 m to 2.4 m. Average TP and TN concentrations were 111 ${\mu}g\;L^{-1}$ and 4.1 $mg\;L^{-1}$, respectively. Lake was classified as eutrophic state based on the nutrient concentrations suggested by U.S.EPA (1976). Total number of fish collected in Lake Doam was 9,600 individuals in 26 species of 6 family. Both dominant and subdominant species in the lake were P. herzi (34.9%) and Z. platypus (22.5%), respectively. Occurrence of water column species was high at upper region of the lake, whereas benthic type of species highly ,appeared in downstream area. The different fish assemblage between upper and lower area would be considered as the difference of bottom substrate and concentrations of suspended solids. In addition high appearance of Comat type of fish that is hybrid between gold fish (C, auratus) and C. auratus was found in the lake. It was unclear the reasons that high proportion of mutant species appeared in the lake. More researches are required in this area in future.

• Korean Journal of Ecology and Environment
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• v.43 no.2
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• pp.288-297
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• 2010

This study was to evaluate the ecological stream health through the Multimetric Fish Assessment Index (MFAI) along with fish fauna analysis based on the tolerance and trophic guilds at Miho stream in 2008 and 2009. Also, we analysed total mercury concentration in fish tissues to examine heavy metal contamination. Total sampled fish were 40 species and 2,557 individuals and Zacco platypus was the most dominant with 35% relative abundance. It was sampled with 11.4% RA for Korean endemic species (10 species 291 individuals) less than average RA 39.3% for the Geum river watersheds. According to the tolarance guild analysis, tolerant species was more dominant with 58.9% RA (15 species, 1,507 individuals) than sensitive species with 6.6% RA. Trophic guild analysis also suggested that omnivores were more dominant (60.5% RA) than insectivores (31.5% RA). Riffle-benthic species was also sampled with 7.7% RA. Ecological stream health based on the MFAI were averaged 25.3 (n=3) with fair-poor condition in 2008 and also 26.3 (n=3) with fair condition in 2009, just slightly increased than 2008. Qualitative habitat evaluation index was averaged 134 (n=3) with fair condition but most of sites had sediment accumulation that reflected substrate degradations proceeding. From the result of total mercury accumulation in fish tissues, kidney and liver tissues showed the highest but the lowest for gill tissues. Overall mercury concentration were not exceed the national standards by Korean Federation of Drug and Administration (KFDA). Consequently, our result could correspond with the characteristics of Miho stream where point sources such industrial complexes and wastewater treatment plant widely distributed around the stream along the gradient of up and downstream.

• Korean Journal of Ecology and Environment
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• v.43 no.2
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• pp.307-318
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• 2010

This study was to analyze the ecological conditions, based on physical habitat, chemical, and biological conditions before (2006, 2007) and after ecological restoration (2009) in five sites of Changwon Stream (CS) and six sites of Nam Stream (NS), respectively, and then to compare ecological health between the two period. The analysis of ecological health was based on the multimetric models of Index of Biological Integrity (IBI) and Qualitative Habitat Evaluation Index (QHEI) along with water chemistry in the streams. For the study, the models of IBI and QHEI were modified as 8 and 11 metric attributes, respectively. For the evaluations, the survey was conducted in the period of 2006~2007 before the restoration and in 2009 after the restoration by the city. Chemical conditions, based on conductivity, in both streams showed a typical longitudinal declines along the axis of the upstream-to-downstream. There were no significant differences (p>0.05) in water quality between the two periods. Values of IBI in the CS and NS averaged 21.6 and 19.7, respectively, indicating a C grade in the criteria of Ministry of Environment, Korea, and there was no significant differences in the IBI between the two periods. Values of QHEI after the restoration averaged 29.2 and 63.2 in the CS and NS, respectively and the values decreased markedly especially, in the NS (35.3) after the restoration. The habitat disturbance was mainly attributed to destructions (i.e., the narrower width of riparian vegetation and higher substrate exposure by the air) of artificial materials by massive flood in 2009. Overall, our results suggest that the restoration was not effective in the two streams between the two periods, even if the budget was used a lot and that such ecological restoration, not considered the natural disaster, may not effect for the stream restoration.

• Korean Journal of Ecology and Environment
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• v.43 no.2
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• pp.190-198
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• 2010

This study was conducted to examine the effects of dams on benthic macroinvertebrate communities in Korean streams and rivers. Four low-head dams and three large dams were studied throughout South Korea. Sampling was taken at immediately upper (impoundment), lower (riffle area), and control (riffle area) sites from the dams during 2004-2007. The upper sites, of which substrate heterogeneity and velocity were relatively low, showed a lower degree of species richness, density, and diversity indices, which is very different from the lower and control sites. Heavily polluted streams showed a lesser degree of community differences between the upper and lower sites. In the large dams, the upper and lower sites showed very low values of species diversity indices and very high values of dominance indices compared to the control sites. In the low-head dams, however, the difference of degree of the values was relatively smaller. Compositions of the functional feeding groups and the habitat orientation groups were relatively simpler at the upper sites than at the lower sites and the degree of difference was greater in the large dams. Species richness and community indices of benthic macroinvertebrates were more significantly affected by habitat characteristics than water quality at the upper sites; however, those were more significantly related with water quality at the lower sites. In conclusion, large and low-head dams could simplify stream habitats particularly at the upper sites (impoundment), and they negatively affected on the benthic macroinvertebrate communities inhabited the habitats. The impact was larger in the large dams than in the low-head dams.

• Journal of Life Science
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• v.25 no.5
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• pp.487-495
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• 2015

We carried out pH stability, chemical inhibition, chemical modification, and pH-dependent kinetic parameter assessments to further characterize protocatechuate 3,4-dioxygenase from Pseudomonas pseudoalcaligenes KF707. Protocatechuate 3,4-dioxygenase was stable in the pH range of 4.5~10.5. L-ascorbate and glutathione were competitive inhibitors with $K_{is}$ values of 0.17 mM and 0.86 mM, respectively. DL-dithiothreitol was a noncompetitive inhibitor with a $K_{is}$ value of 1.57 mM and a $K_{ii}$ value of 8.08 mM. Potassium cyanide, p-hydroxybenzoate, and sodium azide showed a noncompetitive inhibition pattern with $K_{is}$ values of 55.7 mM, 0.22 mM, and 15.64 mM, and $K_{ii}$ values of 94.1 mM, 8.08 mM, and 662.64 mM, respectively. $FeCl_{2}$ was the best competitive inhibitor with a $K_{is}$ value of $29{\mu}M$. $FeCl_{3}$, $MnCl_{2}$, $CoCl_{2}$, and $AlCl_{3}$ were also competitive inhibitors with $K_{is}$ values of 1.21 mM, 0.85 mM, 3.98 mM, and 0.21 mM, respectively. Other metal ions showed noncompetitive inhibition patterns. The pH-dependent kinetic parameter data showed that there may be at least two catalytic groups with pK values of 6.2 and 9.4 and two binding groups with pK values of 5.5 and 9.0. Lysine, cysteine, tyrosine, carboxyl, and histidine were modified by their own specific chemical modifiers, indicating that they are involved in substrate binding and catalysis.