• KSBB Journal
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• v.23 no.3
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• pp.231-238
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• 2008

The second family member of tissue inhibitors of matrix metalloproteinases, TIMP-2, is a 21kDa protein which inhibits matrix metalloproteinases 2 (MMP-2). Expression of mammalian proteins in E. coli often forms inclusion bodies that are made up of mis-folded or insoluble protein aggregates. The requirement for the formation of 6 disulfide bonds in the process of the TIMP-2 folding is likely to be incompatible with the reducing environment of E. coli. However, this incompatibility can be often overcome by introducing a mutagenesis that could lead to enhancement of the protein solubility. In this reason, we have attempted to express the soluble TIMP-2 in E. coli by applying a modified staggered extension process (StEP), one of the in vitro PCR-based recombinant mutagenesis methods, and error-prone PCR. C-terminally located CAT fusion protein with respect to mutated TIMP-2 proteins enables us to differentiate the soluble TIMP-2 from the insoluble in E. coli by virtue of chloramphenicol resistance. According to this scheme, E. coli harboring properly-folded CAT fused to TIMP-2 protein was selected, and some of the resulting colonies exhibited an enhanced, soluble expression of TIMP-2 compared to the wild type, implying (i) the StEP technique is successfully employed to enhance the proper folding thereby increasing the solubility of TIMP-2, and (ii) the CAT dependent screening may be a simple and effective method to differentiate the soluble protein expression in E. coli.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.4
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• pp.508-514
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• 2010

This study was conducted to evaluate the effects of dietary arachidonic acid (AA, 20:4n-6) levels on growth performance and body composition in juvenile eel, Anguilla japonica. Six semi-purified experimental diets were formulated to be isonitrogenous and iso-caloric containing 55.0% crude protein and 15% crude lipid (18.3 kJ of available energy $g^{-1}$). Six different levels of AA were added to the basal diet, with 0, 0.2, 0.4, 0.6, 0.8 or 1.2% on a dry matter (DM) basis, respectively ($AA_{0.07},\;AA_{0.22},\;AA_{0.43},\;AA_{0.57},\;AA_{0.78}\;or\;AA_{1.23}$). After a conditioning period, fish initially averaging 27${\pm}$0.5 g (mean${\pm}$SD) were randomly distributed into each aquarium as triplicate groups of 20 fish each. One of six experimental diets was fed on a DM basis to fish in three randomly selected aquaria at a rate of 2-3% of total body weight twice a day. At the end of the 12-week feeding trial, weight gain (WG) and feed efficiency (FE) of fish fed $AA_{0.78}$ and $AA_{1.23}$ diets were significantly higher than of fish fed $AA_{0.07},\;AA_{0.22},\;AA_{0.43}$ diets (p<0.05). Specific growth rate (SGR) of fish fed the $AA_{0.78}$ diet was significantly higher than of fish fed $AA_{0.07},\;AA_{0.22},\;AA_{0.43}$ diets (p<0.05). However, there were no significant differences in WG, SGR and FE among fish fed $AA_{0.57},\;AA_{0.78}\;or\;AA_{1.23}$ diets (p>0.05). Whole body AA deposition of fish fed the $AA_{1.23}$ diet was significantly higher than for the other diets (p<0.05). Broken-line model analysis on the basis of WG and SGR indicated that the dietary AA requirement could be greater than 0.69% but less than 0.71% of the diet in juvenile eel. The growth-promoting activity of AA observed in the present study provides strong support for the contention that dietary AA is essential for juvenile eel.

• BMB Reports
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• v.40 no.2
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• pp.196-204
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• 2007

Our previous study has shown that sodium selenite can cause apoptosis in acute promyelocytic leukemia-derived NB4 cells in a caspase-dependent manner, but the detailed mechanism is unknown. Here we demonstrate a requirement for extracellular signal-regulated protein kinase (ERK) in mediating sodium selenite -induced apoptosis in NB4 cell. Though no apparent elevation of ERK activity was observed during the apoptosis in NB4 cells caused by 20 μM sodium selenite treatment, PD98059 and U0126, specific chemical inhibitors of the MEK/ERK signaling pathway, were shown to strongly prevent the apoptosis process, while ERK activator TPA enhanced the process. It is also known that p38 MAPK inhibitor SB203580 and JNK inhibitor SP600125 had slight effects on apoptosis. Further study indicated that ERK exerted its proapoptotic effect only at the early stage of apoptosis and played an antiapoptotic role at the later stages. Taken together, our findings suggest that ERK plays an active role in mediating sodium seleniteinduced apoptosis in NB4 cells .

• Journal of fish pathology
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• v.6 no.2
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• pp.133-142
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• 1993

Vitamin B6(pyridoxine, pyridoxamine. and pyridoxal) is a dietary requirement in relatively small quantities for growth, health, and function in animals and fish. The metabolically active B6 is pyridoxal-5-phosphate(PLP). It does function as a coenzyme in number of enzymes(PLP-dependent enzymes) in which amino acids are metabolized, including decarboxylases, aminotransferases, sulfhydrases, tryptophanase, and hydroxylases. Vitamin B6 requirement is higher for fish because fish are fed much higher protein diet than land animals. B6 is also involved in metabolism of carbohydrates and lipids and essential for the synthesis of heme and serotonin. Deficiency signs in fish develop quickly, in cluding nervous disorders, convulsions, poor swimming coordination, skin lesions, edema, exophthalmos, and tetany. The conversion of vitamin B6 to metabolically active form(PLP) is catalyzed by pyridoxal kinase and pridoxine(pyridoxamine) oxidase. In this review, we summarized in detail the enzymatic studies on vitamin B6 metabolism and about the mechanisms and properties of a PLP-dependent enzyme.

• Journal of Nutrition and Health
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• v.39 no.5
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• pp.451-459
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• 2006

The purpose of our study was to evaluate an association between milk consumption and bone mineral density (BMD) of female college students in Korea. The dietary data of 1,410 Korean college students living in the vicinity of Seoul area were collected through the Food Frequency Questionnaire. The mean age, height, weight and 8MI of the subjects were 20.4 years, 162.1 cm, 52.7 kg, and $20.0\;kg/m^2$ respectively. The mean intakes of energy, protein, and phosphorus were 2,398.7 kcal, 95.2 g, and 1,302.9 mg respectively, which were above their EER or Recommended Intake (RI), but the mean intake of calcium was 657.8 mg which was below its RI, and 44.0% of the subjects did not meet Estimated Average Requirement (EAR) of calcium. The daily milk consumption was 131.1 g, and the total consumption of milk and dairy product was 228.7 g. Forty five percent of the subjects consumed $50{\sim}200\;g$ of milk or dairy products. After the analysis of their dietary intakes, the subjects were divided into quintiles of total consumption of milk and dairy product, which were considered as major source of dietary calcium. Ninety two subjects from the upper quintile (UQ) and 85 subjects from the lower quintile (LQ) were selected and their bone mineral densities (BMD) of lumbar spine (L1-L4) and the femur were examined by the dual energy X-ray absorptiometry. The BMD $(g/cm^2)$of L1, L2, L3, and average of femur were 1.065, 1.150, 1.187, and 0.988 for the UQ subjects, which were significantly higher than those of the LQ subjects, 1.010, 1.090, 1.146, and 0.942 respectively. Moreover, the T-scores of the UQ were significantly higher than those of the LQ. The prevalence rates of osteopenia classified according to WHO's criterion were 12.0% in the UQ. and 20.0% in the LQ group which showed non significant. Blood levels of calcium, and phosphorus of UQ and LQ group were both within the normal range, and were not significantly different. Our results showed that UQ group presented the better status of bone mineral density and bone health. Therfore our results suggest that continuous consumption of more than one and a half cups of milk per day, which corresponds to the least amount of UQ group's consumption, would affect positively on the bone health of Korean adults.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.29 no.5
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• pp.643-650
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• 1996

A long term feeding trial was conducted to study the effects of different dietary vitamin C levels on growth and its tissue distributions in juvenile Korean rockfish, Sebastes schlegeli. Prior to the start of feeding trial, fish were fed the basal diet supplementing no L-ascorbic acid for four weeks to minimize their body reserves of vitamin C. Then fish were divided into six groups with triplicates and given one of the laboratory semipurified diets supplementing either 0, 25, 50, 75, 150, or 1500 mg L-ascovbic acid (AA)/kg diet $(C_0,\;C_{25},\;C_{50},\;C_{75},\;C_{150},\;&\;C_{1500})$. Fish fed the $C_0$ diet had lower percent weight gain, feed conversion ratio, specific growth rate, and protein efficiency ratio than did fish fed the other diets (P<0.05). After 28 weeks of feeding trial, tissue AA concentrations of fish fed $C_0$ diet were lower than those of fish fed $C_{1500}$ diet (P<0.05). A large amount of total tissue Ah may be reserved in muscle, but the unit AA concentration seemed to be higher in brain than did the other tissues. The growth performances of fish fed $C_{25}$ diet were not different compared to those of fish fed $C_{50}-C_{1500}$ diets (P>0.05), and diet analysis of vitamin C showed that the $C_{25}$ diet had 65 mg AA/kg diet. Therefore, the present long-term study may suggest that the dietary vitamin C requirement is approximately 65 mg AA/kg diet in juvenile Korean rockfish.

• Animal Bioscience
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• v.35 no.5
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• pp.690-697
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• 2022

Objective: This study aimed to evaluate the effect of the ad libitum and restricted feeding regimen on fasting heat production (FHP) and body composition. Methods: Twelve Hubbard broilers breeders were selected with the same body weight and submitted in two feeding regimes: Restricted (T1) with feed intake of 150 g/bird/d and ad libitum (T2). The birds were randomly distributed on the treatments in two runs with three replications per treatment (per run). The birds were adapted to the feed regimens for ten days. After that, they were allocated in the open-circuit chambers and kept for three days for adaptation. On the last day, oxygen consumption (VO₂) and carbon dioxide production (VCO₂) were measured by 30 h under fasting. The respiratory quotient (RQ) was calculated as the VCO₂/VO₂ ratio, and the heat production (HP) was obtained using the Brower equation (1985). The FHP was estimated throughout the plateau of HP 12 hours after the feed deprivation. The body composition was analyzed by dual-energy X-ray absorptiometry scanning at the end of each period. Data were analyzed for one-way analysis of variance using the Minitab software. Results: The daily feed intake was 30 g higher to T2 (p<0.01) than the T1. Also, the birds of the T2 had significatively (p<0.05) more oxygen consumption (+3.1 L/kg^0.75/d) and CO₂ production (+2.2 L/kg^0.75/d). That resulted in a higher FHP 359±14 kJ/kg^0.75/d for T2 than T1 296±17.23 kJ/kg^0.75/d. In contrast, the RQ was not different between treatments, with an average of 0.77 for the fasting condition. In addition, protein and fat composition were not affected by the treatment, while a tendency (p<0.1) was shown to higher bone mineral content on the T1. Conclusion: The birds under ad libitum feeding had a higher maintenance energy requirement but their body composition was not affected compared to restricted feeding.

• Journal of Animal Science and Technology
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• v.45 no.4
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• pp.593-606
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• 2003

This study was conducted to develop effective manufacturing methods of a total mixed ration(TMR) composed of broiler litter(BL) and bakery by-product(BB) for ruminants. Five experiments included a small-scaled manufacture of TMR using a deepstacking method(Exp. 1), its pelletization(Exp. 2), its field-scaled manufacture(Exp. 3), a field-scaled manufacture using an ensiling method(Exp. 4), and a mixing process of deepstacked BL and BB prior to feeding(Exp. 5). BL and BB were mixed at a ratio which makes total digestible nutrients of the TMR 69%. For each experiment, temperature, appearance and physico-chemical properties were recorded and analyzed. The chemical composition data revealed that the mixture of BL and BB showed nutritionally additive balance which resulted from a considerable increase(P<0.05) of organic matter and a desirable decrease(P<0.05) of protein and fiber up to the requirement level for growing ‘Hanwoo’ steers. Deepstacking of BL and BB in Exp. 1 and 3 resulted in a sufficient increase of stack temperature for pasteurization, little chemical losses, appearance of white fungi on the surface, and partial charring due to excess stack temperature. For Exp. 2, its pelleting, which was successful using a simple, small-scaled pelletizer, resulted in a little loss(P<0.05) of organic matter and an increase(P<0.05) of indigestible protein(ADF-CP). Ensiling the mixture in Exp. 4 made little effect on chemical composition; however, one month of the ensiling period was not enough for favorable silage parameters. Deepstacking BL alone in Exp. 5 tended(P<0.1) to decrease true protein : NPN ratio and hemicellulose content and increase ADF-CP content due to the heat damage occurred. Deepstacking or ensiling of BL-BB mixtures and simple incorporating of BB into deepstacked BL prior to feeding could be practical and nutrients-preservative methods in TMR manufacture for beef cattle, although ensiling needed further hygienic evaluation.

• Korean Journal of Microbiology
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• v.21 no.2
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• pp.51-60
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• 1983

Cultural characteristics of Strptomyces griseolus isolated from the soil were investigated. This strain was disclosed to utilize D-xylose, and D-glactose in preference order as a carbon source with the formation of glucose isomerase. The addition of sweet potato starch also proved effective promoting the total enzyme activity measured at 29% higher than the control. Corn cob, one of waste agricultural resources, was hydrolyzed in 2~3% $H_2SO_4$ solution at $100^{\circ}C$, 3~5 hours to produce a xylose syrup which gave rise to the recovery of 19.9% in a batch system and 28.2% in a repeated system. By the addition of both 2% of xylose syrup(Be'28) prepared by and us 65% of corn steep liquor (total nitrogen 1.2%), enzyme induction was maximized. The enzyme activity was stimulated by the xylose and the cell growth by the C.S.L. Also, remarkable increase of enzyme activity was noticed by the addition of protein acid hydrolysate 86.2% higher than the control. $QO_2$ of the biomass cultured in 30L capacity jarfermentor recorded low oxygen requirement of 251.2 1/hr. Maximum activity of glucose isomerase was observed noted at the 9th hour after inoculation which is 2 hours faster than the stationery was observed noted at the 9th hour after inoculation which is 2 hours faster than the stationery phase of the biomass growth. Glucose isomerase from the strain was activated by adding the $Co^{++}\;and\;Mg^{++}$ with optimum temperature of $73^{\circ}C$ and pH of 7.2. Conversion ratio of 60% glucose to frutose was 42.5% after 70 hours reaction.

• Asian-Australasian Journal of Animal Sciences
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• v.26 no.2
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• pp.227-232
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• 2013

Phytate is not only an unavailable source of phosphorus (P) for broilers but it also acts as an anti-nutrient, reducing protein and mineral absorption, increasing endogenous losses and reducing broiler performance. The objective of this study was to investigate the anti-nutritional effects of phytate by including high levels of phytase in diets not severely limited in available P. A total of 768 male Arbor Acres broilers were distributed in six treatments of eight replicate pens of 16 birds each consisting of a positive control diet (PC), positive control with 500 FTU/kg phytase, negative control (NC) diet with lower available P and calcium (Ca) levels and the same NC diet with 500, 1,000 or 1,500 FTU/kg phytase. Body weight gain (BWG), feed intake (FI), feed conversion ratio (FCR) and mortality were determined at 21 and 35 d of age while foot ash was determined in four birds per pen at 21 d of age. FI, FCR and foot ash where not affected by the lower mineral diets at 21 d of age nor by the enzyme inclusion but broilers fed lower Ca and available P diets had lower BWG. At 35 d of age no difference was observed between broilers fed the positive or NC diets but broilers fed 500, 1,000 and 1,500 FTU/kg on top of the NC diet had better FCR than broilers fed the positive control diet. When compared to birds fed a diet adequate in P, birds fed the same diet included with 500, 1,000 and 1,500 FTU/kg of phytase in marginally deficient available P and Ca diets had an improvement of performance. These results support the concept that hydrolysing phytate and reducing the anti-nutritional effects of phytate improves bird performance on marginally deficient diets that were not covering the P requirement of birds.