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

• KSBB Journal
• /
• v.23 no.3
• /
• pp.205-212
• /
• 2008

The efficient soluble expression of human epidermal growth factor (hEGF) was achieved by using functional fusion partners in cytoplasm and periplasm of Escherichia coli (E. coli). hEGF was over-expressed in inactive inclusion body form in cytoplasm of E. coli due to improper disulfide bond formation and hydrophobic interaction, yielding about 5.9 mg/L in flask culture. Six functional fusion partners were introduced by linking to N-terminal part of hEGF gene for the high-level expression of soluble and active hEGF in cytoplasm and peri plasm region. Three fusion partners for cytoplasmic expression such as acidic tail of synuclein (ATS), thioredoxin (Trx) and lipase, and three fusion partners for periplasmic expression such as periplasmic cystein oxidoreductases (DsbA and DsbC) and maltose binding protein (MBP) were investigated. hEGF fused with ATS and DsbA showed over 90% of solubility in cytoplasm and periplasm, respectively. Especially DsbA was found to be an efficient fusion partner for soluble and high-level expression of hEGF, yielding about 18.1 mg/L and three-fold higher level compared to that of insoluble non-fusion hEGF in cytoplasm. Thus, heterologous proteins containing complex disulfide bond and many hydrophobic amino acids can effectively be produced as an active form in E. coli by introducing a suitable peptide or protein.

• Journal of Hospice and Palliative Care
• /
• v.17 no.3
• /
• pp.170-178
• /
• 2014

Purpose: Delirium is a common and serious neuropsychiatric complication among terminally ill cancer patients. We investigated risk factors related to the development of delirium among hospice care patients. Methods: Between May 2011 and September 2012, we included patients who were mentally alert and had no psychiatric disease or drug addiction at the hospice ward of two local hospitals. Among them, participants who had been diagnosed with delirium by two doctors according to the DSM-IV (Diagnostic and Statistical Manual of Mental Disorders-4th edition) criteria were grouped as Delirium Group. We analyzed results of psychometric and other laboratory tests performed at the time of patient's admission - psychometric tests included cognitive function (mini-mental status examination, MMSE), depression (Beck Depression Inventory, BDI), anxiety, and insomnia (Insomnia Severity Index, ISI). Logistic regression analysis was used to compare delirium and the related factors. Cox's proportional hazard model was performed using significant factors of logistic regression analysis. Results: Of the 96 patients who met the inclusion criteria, 41 (42.7%) developed delirium. According to the logistic regression analysis, primary cancer site, cognitive impairment (MMSE < 24), depression ($BDI{\geq}16$), and insomnia ($ISI{\geq}15$) were significant factors related to delirium. Among the four factors, depression (OR 5.130; 95% CI, 2.009~13.097) and cognitive impairment (OR 5.130; 95% CI, 2.009~13.097) were found significant using Cox's proportional hazard model. Conclusion: The development of delirium was significantly related to depression and cognitive impairment among patients receiving hospice care. It is necessary to carefully monitor depression and cognitive function in hospice care.

• Horticultural Science & Technology
• /
• v.33 no.2
• /
• pp.186-195
• /
• 2015

This research was conducted to investigate effect the plant growth regulators (PGRs) on the characteristics of fruit and flesh softening, using GA3 and thidiazuron (TDZ) treatments in 'Heukboseok' grapes. The total yield obtained under PGR treatment was 88.3% lower than the target production when a single treatment with $GA_3$ of low concentrations was used, but the expected yield was recovered by combined treatment with TDZ and $GA_3$. When harvested on the basis of color, the harvest rate up to 100 days after full bloom (DAFB) was low with the $GA_3$ single treatment, but was increased by the addition of TDZ, with the second TDZ mixed treatment being particularly effective. The soluble solids content in the PGR-treated samples demonstrated no significant changes after 90 DAFB, while the acidity content decreased rapidly starting from 90 DAFB. Measured on the basis of sugar and acidity content, maturity was reached much earlier in treated fruit than in the non-treated fruit. Firmness was maintained at a higher level until the final harvest time after PGR treatment compared to untreated grapes according to epidermis thickness and flesh density increase to activity cell division and expansion by $GA_3$ and TDZ. In particular, the fruit quality was improved based on the delay of softening in primary and secondary treatments of $25mg{\cdot}L^{-1}\;GA_3+2.5mg{\cdot}L^{-1}$ TDZ. The production of seedless fruit was difficult, even with the inclusion of TDZ, reaching the highest seedless rate of only 65.5%. Fruit cracking was rare, occurring at a rate of about 0.0~0.9% in all treatments. Accordingly, 'Heukboseok' grapes should be harvested within 100 days after full bloom (DBFB) before a rapid decrease of firmness, conferred by primary and secondary treatments with $25mg{\cdot}L^{-1}\;GA_3+2.5mg{\cdot}L^{-1}$ TDZ.

• Journal of Life Science
• /
• v.27 no.8
• /
• pp.937-944
• /
• 2017

To investigate the beneficial effects of fiber derived from agar on the lipid and glucose metabolism of obese model mice, changes in the fat accumulation, lipid content, and insulin concentration were measured in C57BL/6N mice fed a high-fat diet (HFD) and treated with a beverage containing agar (BCA) for 9 weeks. The feed efficiency ratio was significantly decreased in the HFD+BCA treatment group, although the body weight and food intake were maintained a constant level. Also, the fat accumulation in the fatty tissue and liver were lower in the HFD+BCA treatment group than in the HFD+Vehicle treatment group. Furthermore, a significant decrease was detected in the levels of total cholesterol (TC) and LDL after the BCA treatment, while a constant HDL level was maintained in the same group. Moreover, the serum insulin concentration was significantly decreased in the HFD+BCALo (Low concentration) and HFD+BCAMi (Middle concentration) groups compared to the HFD+Vehicle group. Therefore, the results suggest that the long-term administration of dietary fiber from agar can improve fat accumulation in the fatty tissue and liver, the serum lipid profile, and the insulin concentration in the blood. Also, the results provide evidence that the inclusion of agar fiber in beverages results in significant hypolipidemic and hypoinsulinemic effects.

• Journal of Genetic Medicine
• /
• v.4 no.2
• /
• pp.186-189
• /
• 2007

More than 6,000 rare disorders including genetic diseases have been reported. Of them, 1,500 diseases (1,211 for clinical diagnosis and 289 for research only) are technically possible for genetic testing. In Korea, since 2005, only 63 genetic diseases is permitted for prenatal genetic testing by the "Bioethics and Biosafety Law". The article 25 in the law prescribes 63 genetic diseases without clear indication for its selection and inclusion criteria. In EU, USA, and other foreign countries, however, there is no provision in the statute on prenatal genetic testing; it is not restricted by a law. Recently, a woman (Mrs. L, 38y) who is a carrier for Menkes disease made an appeal to a government for an amendment of the "Bioethics and Biosafety Law" prohibiting the prenatal diagnosis of her pregnancy at risk for Menkes disease. Menkes disease (MNK) is an X-linked recessive disorder characterized by neurodegeneration, connective tissue defects and hair abnormalities, and no effective treatment is available yet. The prevalence rate of MNK is one in about 250,000 live births. Menkes syndrome patients fail to absorb copper from the gastrointestinal tract in quantities adequate for meeting nutritional needs. These needs seem particularly acute during the initial 12 month of life, when the velocity of brain growth and motor neurodevelopment. Most of pts. die around 3yrs. of age. Mrs. L had a boy with Menkes disease who died at 2y.o. in 2001. Subsequent pregnancy in 2003, she was able to have prenatal genetic testing for mutation of the Menkes (ATP7A) gene and delivered a healthy baby boy. Now, She is pregnant again and wants to have prenatal diagnosis. however, this time, she was not allowed to have any more because Menkes disease is not included in 63 genetic diseases permitted by the law for prenatal genetic testing, in spite of the fact that she is a Menkes disease carrier and her pregnancy is at risk to have an affected baby. This case shows the practical problem of the legal restriction for prenatal genetic testing in Korea. In this study, we report a arguable case and discuss the controversial issues in the legal restriction for prenatal genetic testing in Korea.

• Journal of Animal Science and Technology
• /
• v.50 no.6
• /
• pp.819-830
• /
• 2008

The object of this study was to investigate different levels of extruded rice product(Nutri-RS®) supplementation as an alternative to lactose on growth performance, diarrhea score, fecal E.coli and volatile fatty acid(VFA) in weaning pigs. Treatments were 1) Control(basal), L3(basal+3% extruded rice product), L6(basal+6% extruded rice product). The control diet in Phase I and Phase II contained 16 and 9% of lactose, respectively. For the feeding trial, a total of 81 crossbred pigs, averaging 7.56±1.18 kg body weight, were allotted to three treatments in three replicates with nine pigs per pen by RCBD. The experimental period was total 5 weeks. During the whole experimental period, there were no significant differences in growth performance, diarrhea score, fecal volatile fatty acid concentration among all treatments. Although there were no significant differences, pigs fed diet of L6 tended to show higher BW and ADG, ADFI, and fecal volatile fatty acid concentration compared to other treatments. Treatment L6 showed the lowest fecal E. coli count among all treatments(P<0.05). These results suggested that inclusion of extruded rice product has similar performance a with 6% of lactose in diet for weaning pigs and reduce approximately 9% production cost of weaning pig's diet. Therefore, extruded rice product could be utilized as an alternative feed ingredient to lactose in weaning pig’s diet.

• The Korean Journal of Nuclear Medicine Technology
• /
• v.16 no.1
• /
• pp.8-11
• /
• 2012

Purpose : Among quality control items, the radiochemical impurity must be below 10% of total radioactivity. In this regard, as the recently commercialized automatic synthesis module produces a large amount of 18F-FDG, radiolysis of radiopharmaceuticals is very likely to occur. Thus, this study compared the changes in radiochemical purity regarding radiolysis of $^{18}F$-FDG according to automatic synthesis module. Materials and methods : Cyclotron (PETtrace, GE Healthcare) was used to produce $^{18}F$ and automatic synthesis module (FASTlab, Tracerlab MX, GE Healthcare) was used to achieve synthesis into FDG. For radiochemical purity, Radio-TLC Scanner (AR 2000, Bioscan), GC (Gas Chromatograph, Agilent 7890A) was used to measure the content of ethanol included in $^{18}F$-FDG. Glass board applied with silica gel ($1{\times}10cm$) was used for stationary phase while a mixed liquid formed of acetonitrile and water (ratio 19:1) was used for mobile phase. High-concentration and low-concentration $^{18}F$-FDG were produced in each synthesis module and the radiochemical purity was measured every 2 hours. Results : The purity in low-concentration (below 2.59 GBq/mL) was measured as 99.26%, 98.69%, 98.25%, 98.09% in Tracerlab MX and as 99.09%, 97.83%, 96.89%, 96.62% in FASTlab according to 0, 2, 4, 6 hours changes, respectively. The purity in high-concentration (above 3.7 GBq/mL) was measured as 99.54%, 96.08%, 93.77%, 92.54% in Tracerlab MX and as 99.53%, 95.65%, 92.39%, 89.82% in FASTlab according to 0, 2, 4, 6 hours changes, respectively. Also, ethanol was not detected in GC of $^{18}F$-FDG produced in FASTlab, while 100~300 ppm ethanol was detected in Tracerlab MX. Conclusion : Whereas the change of radiochemical purity was only 3% in low-concentration $^{18}F$-FDG, the change was rapidly increased to 10% in high-concentration. Also, higher radiolysis were observed in $^{18}F$-FDG produced in FASTlab than Tracerlab MX. This is because ethanol is included in the synthesis stage of Tracerlab MX but not in the synthesis stage of FASTlab. Thus, radiolysis is influenced by radioactivity concentration than the inclusion of ethanol, which is the radioprotector. Therefore, after producing high-concentration $^{18}F$-FDG, the content must be diluted through saline to lower concentration.

• Korean Journal of Poultry Science
• /
• v.37 no.3
• /
• pp.255-263
• /
• 2010

Inclusion body hepatitis-hydropericardium syndrome (IBH-HPS) is an acute viral disease usually found in broilers aged from 3 to 5 weeks and causes up to 75% mortality. Among the 12 serotypes of fowl adenovirus group 1, serotype-4 (FAdV-4) was identified as a primary agent of IBH-HPS and was usually isolated in IBH-HPS cases in Korea since 2007. To prevent these IBH-HPS outbreaks in Korea, we developed the FAdV-4 inactivated vaccine using Korean isolate (ADL070244) and evaluated the efficacy of this vaccine. For the efficacy test, 2-week-old specific-pathogen-free (SPF) chickens intramuscularly inoculated with 1 or 2 dose of inactivated vaccine were used and challenged with FAdV-4 through either intramuscular or oral route at 2 weeks after vaccination. The vaccine induced good seroconversion which was confirmed by agar gel precipitation (AGP) test. In addition, the vaccine could decrease the FAdV-4 detection rate and histological lesion severity such as lymphocyte infiltration and necrosis in the liver comparing with those of non-vaccination group. Based on the current results, the developed FAdV-4 inactivated vaccine in this study was effective in the terms of reduction of virus detection rate and histological lesions severity. However, it was difficult to confirm the efficacy of the vaccine clearly because of no mortality and clinical signs in the non-vaccinated group after challenge. Therefore, we need further study to develop a standard challenged model system which could clearly evaluate the efficacy of the vaccines for FAdV-4.

• Journal of the Korean Wood Science and Technology
• /
• v.15 no.3
• /
• pp.44-55
• /
• 1987

In order to study the effect of sawdustboard combined with plastic chips, 0.5mm($T_1$), 1mm($T_2$), 1.4mm($T_3$) thick nylon fiber. polypropylene rope fiber(RP), and 0.23mm thick moth-proof polypropylene net fiber(NP) were cut into 0.5, 1, 2cm long plastic chips. Thereafter, sawdustboard combined with plastic chips prepared as the above and plastic non-combined sawdustboard(control) were manufactured into 3 types of one-, two-, and three layer with 5 or 10% combination level. By the discussions and results at this study, the significant conclusions of mechanical and physical properties were summarized as follows: 1. The MORs were shown in the order of 3 layer> 2 layer> 1 layer among plastic non-combined boards, and $T_3$ < $T_2$ < $T_1$ < RP (NP(5%) < NP(l0%) among plastic combined boards. In 2cm long plastic chip in 1 layer board, the highest strength through all the composition was recognized. 1 layer board showing the lower strength with 0.5cm plastic chip rendered to the bending strength improvement by 2 or 3 layer board composition. On the other hand, 2 or 3 layer combined with 1, 2cm long polypropylene net fiber chips incurred MOR's conspicuous decrease requiring optimum plastic chip combined level and consideration to combined type. 2. MOE in plastic non-combined 3 layer board exhibited sandwich construction effect by higher resin content application to surface layer in the order of 3layer>1layer>2layer with the highest stiffness of the board combined with polypropylene chip, while nylon chip-combined board had little difference from plastic non-combined board. In relevant to length and layer effect, 3 layer board combined with the 0.5cm long polypropylene net fiber chip in 5% and 10% combined level presented 34-43% and 44-76% stiffness increase against plastic non-combined board(control), respectively. Moreover, in 1 layer board, 30% stiffness increase with 10% against 5% combined level in the 1 and 2cm long polypropylene net fiber chip was obtained. 3. Stress at proportional limit(Spl) showing the fiber relationship (r: 0.81-0.97) between MOR presented in the order of 1 layer<2 layer<3 layer in plastic non-combined board. Correspondingly, combined effect by layer and plastic chip length was similar to MOR's. 4. Differently from previous properties(MOR, MOE, Spl). work to maximum load(Wml) of 2 layer board approached to that of 3 layer board. Conforming the above phenomenon. 2 layer combined with 0.5cm long polypropylene net fiber chip kept the greater work than 1 layer. The polypropylene combined board superior to nylon -and plastic non - combined board seemed to have greater anti - failing capacity. 5. Internal bond strength(IB), in contrast to MOR's tendency. showed in the order of T1