• Food Science and Biotechnology
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• v.14 no.6
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• pp.732-737
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• 2005

The combinational effect of gamma irradiation and moist heating on the trypsin inhibitor activity (TIA) in soaked and dried soybeans was evaluated by measuring the inhibition using N-benzoyl-DL-arginine-p-nitroanilide (BAPNA) as substrate. Gamma irradiation significantly decreased the TIA level in soybean at doses above 5 kGy, and the $ID_{50}$ (the gamma irradiation dose required to reach 50% inhibition) value for TIA was 13.53 kGy. Soaking prior to gamma irradiation significantly lowered the $ID_{50}$ to 8.44 kGy, and the soaking process enhanced the efficiency to inactivate TIA by as much as 48%. When soaking prior to gamma irradiation was followed by subsequent mild heating ($60^{\circ}C$) process, the $IT_{50}$ (heating time required to reach the 50% inhibition of TIA) value at even 1 kGy (5.28 min) was greatly reduced by over 50% compared to the level for the no-soaking process. In addition, the activation energy of soaking prior to gamma irradiation at 1 kGy was 2.45 kcal/mole, which was also about 50% lower than the 5.10 kcal/mole of dried soybean gamma-irradiated. Based on these results, soaking prior to gamma irradiation is an effective method for TIA inhibition. Furthermore, a combination of two or more processing methods such as soaking, heating and gamma irradiation is much more effective than any single processing method.

• Korean Journal of Veterinary Research
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• v.34 no.2
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• pp.407-415
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• 1994

Present experiments were undertaken in order to find out the most available diagnostic method for acute pancreatitis. Experimental dogs were divided into experimental group and control group. The experimental dogs were laparotomized and their pancreatic ducts were ligated for the induction of pancreatitis. The control dogs were laparotomized only for comparison. In addition to the complete blood count, serum amylase and lipase activities, serum glucose, total protein and albumin contents were measured daily for 11 days after the operation. Fecal fat droplet count by Sudan III staining and fecal trypsin activity examination by x-ray film digetion test were also undertaken daily. The results obtained were summarized as follows; 1. Serum amylase activities of the experimental group increased to peak on the third day and returned to the preoperative values on the eighth day. 2. Serum lipase activities of the experimental group increased to peak on the first day and returned to the preopertive values within six to eight days. 3. Serum glucose contents of the experimental group showed significant increse only on the first day. 4. Serum albumin contents of the experimental group decreased significantly during the experimental period. Whereas those of the control group significantly decreased only on the first day. 5. The experimental group showed marked leukocytosis, neutrophilia, and lymphopenia for the first 5 or 8 days. Whereas the control group showed only neutrophilia for the first 3 days. 6. The results of fecal fat droplet count showed some diagnostic value on the third and fourth day in only one experimental dog(No 1). No significant changes in the fecal trypsin activity were noticed in all dogs. 7. Histopathologically. all dogs of experimental group showed changes of pancreatitis. However the degree of the pancreatic lesion was not pararell to the degree of the serum amylase or lipase activity changes.

• Journal of Life Science
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• v.18 no.3
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• pp.350-356
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• 2008

Biochemical characterization including hemagglutination of erythrocytes, molecular weight, optimum temperature, thermal stability, optimum pH, carbohydrate specificity, and inhibitory effect of metal ion were studied in lectin of eggplant (Solanum melongena L.) fruit prepared by ammonium sulfate fractionation and affinity chromatography. This lectin was agglutinated by trypsin-treated rat blood erythrocyte. The molecular weight of this lectin by SDS-PAGE was estimated to be approximately 19.3 kDa of a single band. This lectin has no activity by 7 carbohydrates containing D-glucose. The optimum range of temperature and pH were $10-20^{\circ}C$ and pH 6.2-7.2, respectively. This lectin was relatively stable at $20-70^{\circ}C$. And the activity of this lectin was not inhibited by $Ca^{2+},\;Co^{2+},\;Cu^{2+},\;Fe^{2+},\;Mg^{2+}$, and $Mn^{2+}$.

• Fisheries and Aquatic Sciences
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• v.9 no.1
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• pp.7-13
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• 2006

Using biochemical methods, we determined the potential of local female shrimp populations as breeding stock to select the best adult prawns for improving larval production. As condition indexes, we selected total RNA, DNA, their ratio, and trypsin activity. The DNA content in the pleopods of each local population was similar, i.e., between $0.90{\pm}0.06\;and\;1.02{\pm}0.04(SE){\mu}g/mg$. In comparison, the RNA contents differed markedly between $2.00{\pm}0.09$ and $0.96{\pm}0.08\;{\mu}g/mg$. Therefore, the RNA/DNA (R/D) ratio in the pleopod could be used as a condition index because it represents a biochemical characteristic of the population. The mean pleopodal R/D ratio of the Goheung population was the highest at $2.52{\pm}0.19$, which indicated the best condition. Trypsin activity was influenced little by shrimp condition and more by the amount of food ingested. The gonadosomatic index (GSI) and R/D ratio in the gonads provided offsetting information about the instantaneous gonad maturity. The Goheung population had the highest instantaneous GSI, despite some spawning. Based on the condition indexes and time of gonad maturation, the Goheung shrimp population is suitable for use as breeding stock.

• The Korean Journal of Zoology
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• v.39 no.1
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• pp.65-74
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• 1996

Treatment of regenerating amphibian limbs with retinoic acid (RA) is known to induce paftern duplication, which is closely related to the extent of dedifferentiation. In the present study, the activities of trypsin- and chymotrypsin-like proteases are examined to delineate a possible role in the process of dedifferentiation in the regenerating limbs of urodeles, the Korean salamander (Hynobius leechii) and the Mexican axolod (Ambystoma mexicanum). Specifically, we were interested to know if there is any correlation between trypsin- and chymotrypsin-like protease activities and the state of dedifferentiation which is augmented by RA treatment. We were also interested in expoloring if there is any species-specific difference in the profile of enzyme activities during limb regeneration. The results showed that the activities of these two enzymes reached a peak level at dedifferentiation stage, and RA treatment caused elevation of their activities, especially in the case of trypsin-like protease. The increase of trypsin-like protease activity after RA treatment was pronounced in the Korean salamander, which might reflect a species-specific responsiveness to RA. The present results imply that trypsin and chymotrypsin or similar proteases may play an active role in the process of dedifferentiation in regenerating limbs, and that trypsin or trypsin-like eryrymes might be involved in the RA-evoked enhancement of dedifferentiation which precedes overt pattern duplication.

• Microbiology and Biotechnology Letters
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• v.17 no.5
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• pp.412-420
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• 1989

To know how the ribosomes involved in secretory protein synthesis were attached to the cytoplasmic membrane in Bacillus amyloliquefaciens, the cells were treated with puromycin combinated with magnesium at the logarithmic phase, and the variation of cell-bound and extracellular $\alpha$-amylase activity was assayed for determining the $\alpha$-amylase translocation blocking through the cytoplasmic membrane. In the abnormal $\alpha$-amylase producing mutant in which the C-terminal of the $\alpha$-amylase structure was deleted, B. umytotiquefaciens CH10-2, the $\alpha$-amylase was translocated normally through the cytoplasmic membranes, and the translocation blocking by puromycin was revealed to have a similar pattern as that in the wild type. This means that the C-terminal part of the enzyme structure may not have a signal for secretion. The cell death of the logarithmic phase cells in both strains was not affected much under 20$\mu\textrm{g}$/$m\ell$ of puromycin, however, the $\alpha$-amylase translocation was blocked markedly under less than 10$\mu\textrm{g}$/$m\ell$ of the puromycin concentration. The blocking of the enzyme secretion by puromycin may be due to the detachment of the ribosomes from cytoplasmic membranes by disturbing the nascent polypeptide synthesis. Further evidence for confirming this was that the detachment was increased in 50 mM of magnesium ion because the extracellular $\alpha$-amylase activity was decreased more under this condition. If the cells were treated with trypsin combinated with Iysozyme, the extracellular $\alpha$-amylase activity from the cultured medium was reduced markedly, however, the activity from the cells treated with trypsin only was not reduced. This means that the nascent polypeptides protruding from the cytoplasmic membrane were sensitive to the trypsin digestion, whereas the matured ones were not. Therefore, the protruding polypeptides from the cytoplasmic membranes may be truncated by trypsin before forming their final tertiary structures by folding in the cell wall layer.

• BMB Reports
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• v.32 no.6
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• pp.567-572
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• 1999

We have previously shown that a DNA fragment is responsible for the anticoagulatory effect of an earthworm, Lumbricus rubellus. The anticoagluant increased the activated partial thromboplastin time (APTT) and also inhibited the thrombin activity observed with either N-${\alpha}$-p-tosyl-L-arginine methyl ester (TAME) or H-D-phenyl-alanyl-L-pipecoil-L-arginine-p-nitroanilide (S-2238). Since trypsin digestion of the anticoagulant further increased the APTT, the possible presence of a regulatory protein for the anticoagulatory DNA was investigated by digesting the anticoagulant with trypsin and isolating the DNA fragment with C4-reversed phase HPLC. The DNA fragment lacking a regulatory protein was eluted in the flow-through fraction, and analyzed with thrombin and activated factor X. Activated factor X activity was more strongly inhibited than thrombin activity. For DNA digestion, we treated the anticoagulant with DNase and purified the DNA-binding protein with a FPLC Resource-S cation exchange column. The regulatory protein, with an $M_r$ of 55.0 kDa, reduced the anticoagulatory effect of the DNA fragment.

• BMB Reports
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• v.29 no.1
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• pp.58-62
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• 1996

The structural changes in the electron donor side of the PSII reaction center have been monitored since heat treatment ($45^{\circ}C$ for 5 min) of thylakoids is known to decrease the oxygen evolving activity. In heat-treated spinach chloroplast thylakoids, the inhibitory effect of 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) on the electron transport activity of the PSII reaction center from diphenyl carbazide to dichlorophenolindophenol became reduced approximately 3.8 times and [$^{14}C$]-labeled DCMU binding on the D1 polypeptide decreased to 25~30% that of intact thylakoid membranes, implying that the conformational changes of the DCMU binding pocket, residing on the D1 polypeptide, occur by heat treatment. The accessibility of trypsin to the $NH_2$-terminus of the cytochrome b-559 ${\alpha}$-subunit, assayed with Western blot using an antibody generated against the synthetic peptide (Arg-68 to Arg-80) of the COOH-terminal domain, was also increased, indicating that heat-treatment caused changes in the structural environments near the stromal side of the cytochrome b-559 ${\alpha}$-subunit, allowing trypsin more easily to cleave the $NH_2$-terminal domain. Therefore, the structural changes in the electron donor side of the PSII reaction center complexes could be one of the reasons why the oxygen evolving activity of the heat-treated thylakoid membranes decreased.

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.4
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• pp.500-508
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• 2011

Forty-four Gansu Alpine Fine-wool lambs were used to study changes in the activities of three gastric and five pancreatic enzymes under grazing conditions between 0 and 56 days of age. The lambs were slaughtered on days 0, 3, 7, 14, 21, 28, 42 and 56, the abomasal contents, mucosa and pancreas were immediately removed and placed into liquid nitrogen and enzyme activities were determined. Gastric enzyme (chymosin, pepsin and pregastrc esterase) activities were relatively high at birth, especially chymosin, but decreased quickly between day 0 and 21. The activity of pepsin changed insignificantly with increasing age. There was no significant change in the pancreatic enzyme activities (trypsin, chymotrypsin, ${\alpha}$-amylase, lipase and lactase). The activity of trypsin was relatively higher than that of the other pancreatic enzymes, and lactase activity was low. These ontogenic patterns might be under the control of many gut regulatory peptides, the plasma concentrations of which changed simultaneously. Some gastric and pancreatic enzymes were correlated with plasma concentrations of these gut regulatory peptides.

• Macromolecular Research
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• v.16 no.7
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• pp.637-643
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• 2008

A combination of magnetic and temperature-responsive properties in the same polymer composites is expected to increase their potential applications in the biomedical field. Accordingly, micron-sized magnetite/polystyrene/poly(2-dimethylaminoethyl methacrylate-ethyleneglycol dimethacrylate), which are abbreviated as $Fe_3O_4$/PS/P (DM-EGDM) composite polymer particles, were prepared by the seeded copolymerization of DM and EGDM in the presence of magnetite/polystyrene ($Fe_3O_4$/PS) particles. $Fe_3O_4$/PS/P(DM-EGDM) composite particles with magnetic properties showed a temperature-sensitive phase transition at approximately $31^{\circ}C$. The adsorption behavior of the low molecular weight emulsifiers and trypsin (TR) as biomolecules were examined on $Fe_3O_4$/PS/P(DM-EGDM) composite polymer particles at different temperatures. The native conformation of TR was followed by measuring the specific activity under various adsorption conditions. The activity of the adsorbed TR on composite polymer particles was higher than those of the tree TR and TR adsorbed on $Fe_3O_4$/PS particles.