• Biomolecules & Therapeutics
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• v.17 no.1
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• pp.104-110
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• 2009

Phenylketonuria (PKU) is an inherited metabolic disorder caused by mutations in the phenylalanine catabolic enzyme, phenylalanine hydroxylase (PAH). The use of phenylalanine ammonia-lase (PAL) by oral and parenteral routes as a therapeutic drug for PKU has been severely limited due to inactivation by intestinal proteolysis and immune reactions. PEGylation was applied to PAL to reduce the degrees of antigenicity and proteolytic inactivation. Kinetic experiments with native PAL and pegylated PALs were performed, and pH stability, temperature stability, and protease susceptibility were evaluated. Enzyme linked immunosorbent assay (ELISA) was carried out to measure the immune complex between pegylated PALs and antiserum that had been extracted from a PAL-immunized mouse. Pegylated PAL, especially branched pegylated PAL (10 kDa, 1:32), was more active for phenylalanine and more stable in pancreatic proteases than native PAL. Native PAL was optimal at pH 8.5, corresponding to the average pH range of the small intestine; the same finding was noted for pegylated PALs. All linear and branched pegylated PALs had low reactivity with mouse antiserum, especially the 1:16 formulation with linear 5-kDa PEG and the 1:32 formulation with branched 10-kDa PEG. Therefore, we suggest the 1:32 formulation with branched 10-kDa PEG as the most promising formulation for enzyme replacement therapy.

• Journal of Microbiology
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• v.37 no.4
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• pp.229-233
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• 1999

The mitochondrial plasmid pMLP1 from a white-rot fungus, Pleurotus ostreatus, is a double-stranded DNA containing 381 bp terminal inverted repeat (TIR) whose 5'-ends are covalently bound by terminal proteins. The plasmid contains two major open reading frames (ORFs), encoding putative DNA and RNA polymerases, and a minor ORF encoding a small, highly basic protein. To identify the DNA binding activity that recognizes the TIR region of pMLP1, gel retardation assays were performed with mitochondrial extracts. A specific protein binding to a region between 123 and 248 nt within TIR was observed. We examined whether the gene product of mORF1 bindes to this region specifically. E. coli cell extract which contains an overproduced mORF1 protein formed a complex specific to the region between 123 and 248 nt. Inclusion of mORF1 protein in the specific complex formed between P. ostreatus mitochondrial extract and TIR was confirmed by a supershift assay using polyclonal antibodies against the mORF1 protein. Our result suggest that the product of mORF1 may function as a terminal region recognition factor (TRF), recognizing an internal region in TIR.

• The Korean Journal of Nuclear Medicine
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• v.34 no.1
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• pp.74-81
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• 2000

Purpose: The purpose of this study was to develop in vivo dosimetries using both chromosomal aberrations and micronuclei in mice to assess biological effects of radiations. Materials and Methods: Five each mice were irradiated with 0, 1, 2, 3, 4, 5, 10 Gy of Cs-137 gamma-rays. We scored numbers of chromosomal aberrations in metaphase spreads and numbers of micronuclei in bone marrow smears under light microscope, and obtained the dose-response relationships. We also examined the relationship between the two dose-response curves. Results: The frequency of both chromosomal aberrations and micronuclei increased with dose, in a linear-quadratic manner The delta, beta, and alpha coefficients were 0.0176, 0.0324, and 0.0567 for metaphase analysis (r=1.0, p<0.001) and 0.0019, 0.0073, and 0.0506 for micronuclei assay (r=1.0, p<0.001). The frequency of chromosomal aberrations and micronuclei in different radiation doses was significantly correlated (r=0.99, p<0.01). Conclusion: In vivo dosimetry using either metaphase analysis or micronucleus assay was feasible in mice. These methods could be useful to evaluate biological effects of radiation.

• Korean Journal of Veterinary Research
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• v.29 no.3
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• pp.393-405
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• 1989

This study was carried out to treatment test for bovine mastitis by the determination of adenosine triphosphate (ATP) based on firefly bioluminescence. The results obtained are followed; 1. In the susceptibility test, cephalothin which looks the most effective were sensitive to Staphylococcus sp. (72.3%), Micrococcus sp. (84.2%), Streptococcus sp. (72.7%) and Gram positive bacilli (72.7%), Gram negative bacilli were sensitive to gentamicin (92.3%) and Yeast-like-fungi was the most sensitive to clotrimazole, and nystatin in order. 2. When the number of bacteria, such as Staphylococcus aureus, Candida tropicalis isolated from the mastitis milk were counted by conventional agar plating technique, and compared with the concentration of bacterial ATP, it gave a good linear relationship. The content of ATP per Staphylococcus aureus, cell was 3.1fM and Candida tropicalis showed the high level of A TP (90fM). 3. The ATP assay was applied to the determination of minimal inhibitory concentration (MIC) of various antibiotics. When Staphylococcus aureus was incubated in the presence of different concentration of tetracycline, erythromycin, kanamycin and streptomycin sulfate and the growth was monitored by the conventional agar plating technique and ATP assay, both methods shown the same results that they were 1mcg/ml, 2mcg/ml, 6.25mcg/ml and 8mcg/ml, respectively. 4. For the determination of susceptibility of sensitive and resistant Staphylococcus au reus isolated for the milk with mastitis to tetracycline, erythromycin kanamycin and streptomycin sulfate, the minimum time required for the test was determined by the assay of ATP every 30 minutes during incubation of 3 hours at $37^{\circ}C$. ATP concentration time curve calculated on both resistant and sensitive strains incubated 3 hours as the optimum time for the determination of susceptibilities of various antibiotics exemed. The ATP concentration of each test broth (antibiotic containing), expressed as a percentage of its own control brith (antibioticfree) indicated values of 30% to be indicative of each antibiotic sensitivity. Single time point ATP assay carried out on the various sensitive and resistant of Staphylococcus aureus to antibiotics examined after 3 hours at $37^{\circ}C$ correlated exactly with disc diffusion and MIC. 5. In the cure of intramammary treatment of bovine mastitis in lactating quarters, the cure rate of Staphylococcal mastitis showed to cephalexin (80%), cloxacillin and gentamicin (70%), ampicillin and oxytetracycline (60%), and Streptococcal mastitis showed to cephalexin (85%), penicillin (80%), cloxacillin and oxytetracycline (75%), and ampicillin (70%), but intramammary antimycotic drug (clotrimazol) were only a little effect about fungal mastitis.

• Korean Journal of Clinical Laboratory Science
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• v.52 no.1
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• pp.53-61
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• 2020

Cell migration is a central process for recovering from wounds triggered by physical distress besides embryogenesis and cancer metastasis. Wound healing assay is widely used as a fundamental research technique for investigation of two-dimensional cell migration in vitro. The most common approach for imitating physical wound in vitro is mechanical scratching on the surface of the confluent monolayer by using sharp materials. The iron metal pin with a suspension spring for fine adjustment of the orthogonal contact surface between the scratching point and the individual bottom of multi-well plate with planar curvatures were adopted for the creative invention of a 96-well plate wound maker. While classic tips drew diverse and zigzag scratching patterns on the confluent monolayer, our wound maker displayed synchronized linear wounds in the middle of each well of a 96-well plate that was seeded with several cell lines. Given that several types of multi-well plates commercially available are compatible with our lab-made wound maker for creating uniform scratches on the confluent monolayer for the collective cell migration in wound healing assay, it is certain that the application of this wound maker to the real-time wound healing assay in high content screening (HCS) is superior than utilization of typical polypropylene pipette tips.

• Food Science of Animal Resources
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• v.25 no.3
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• pp.316-321
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• 2005

A total of ninety six pigs ($L{\times}Y{\times}D$, 20.92(2.13kg average initial body weight) were used in a 16-week performance growth assay to determine the effect of supplemental medicinal plane (Artemisia, Acanthopanax and Garlic) on growth performance, IGF-1 of serum and carcass characteristics in finishing pigs. The dietary treatments were included 1) CON (basal diet; Control), 2) MP1 (basal diet added $0.02\%$ of medicinal plant mixtures), 3) MP2 (basal diet added $0.04\%$ of medicinal plant mixtures) and 4) MP3 (basal diet added $0.06\%$ of medicinal plant mixtures). Through entire experimental period, as medicinal plants mixture (MP) increased, there was a decrease (linear, P<0.08) in average daily feed intake and an increase (linear, P<0.02; quadratic, P<0.08) in gain/feed. The backfat thickness tended to decrease in pigs fed MP diet compared to pigs fed CON diet (linear, P<0.09; quadratic, P<0.01). Increasing medicinal plane mixture tended to increase in IGF-1 content in serum (linear, P<0.09). The hunter $a^{*}$ (redness) (linear, P<0.01) and $b^{*}$ (yellowness) (linear, P<0.02) values of longissimus muscle were affected by the dietary MP treatments. The color of longissimus muscle was higher in the dietary MP treatments than that of the muscle in the control diet (linear, P<0.03). In conclusion, the result obtained from this feeding triad suggest that the medicinal plants mixture supplementation below $0.06\%$ in diets for growing-finishing pigs can be improved growth performance, IGF-1 and meat quality.

• Archives of Pharmacal Research
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• v.11 no.3
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• pp.250-252
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• 1988

A sensitive and simplified HPLC assay of fluconazol is described. The calibration curve of fluconazol in plasma ranging $0-10\;{\mu}g/ml$ was linear with the correlation coefficients of 0.9900. The limit of detection was $0.3\;{\mu}g/ml$. The average recovery of the drug was $89.1\;{\pm}\;9.05%$. After oral administration of single dose(150mg) of fluconazol in man, $C_{max}\;and\;T_{max$ were $3\;{mu}g/ml$ and 4hr., respectively.

• Food Science and Biotechnology
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• v.16 no.3
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• pp.451-456
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• 2007

The potential antioxidant activities of methanolic extracts from soybean and rice fermented with Monascus sp. were investigated. M. pilosus IFO 480 and M. anka IFO 478 were screened as a suitable strain to promote the antioxidant activities in soybean- and rice- fermentation. The methanol extracts from soybean and rice after fermenting for 20 days at $30^{\circ}C$ resulted in a significant increase in the antioxidant capacities expressed as radical (ABTS and DPPH) scavenging assay and peroxidation inhibition (%) by thiocyanate method and increased (p<0.01) by a 2.6 to 3.1-fold compared with those of the unfermented products. The average antioxidant potentials of Monascus-fermented soybean extracts (MFSE) were significantly (p<0.01) stronger than Monascus-fermented rice extracts (MFRE). A linear correlations between free radical scavenging activity of MFSE and the total phenolics content (r=0.84) and total flavonoids content (r=0.81) were observed. These results indicated that MFSE exhibited stronger (p<0.01) antioxidant activity and contained significantly higher levels (p<0.05) of phenolics than MFRE.

• BMB Reports
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• v.28 no.2
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• pp.143-148
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• 1995

In E. coli, chromosomal DNA associated with proteins is condensed into an organized structure known as nucleoid. Using a nitrocellulose filter binding assay to identify proteins forming nucleoid, a 21 kDa protein was purified from E. coli. The molecular weight of the purified protein was 21 kDa on SDS-polyactylamide gel electrophoresis and 24 kDa on gel permeation chromatography. A molecular weight of 21 kDa on SDS-polyacrylamide gel electrophoresis is unique among known proteins which are believed to be involved in the formation of nucleoid in E. coli. The 21 kDa protein nonspecifically binds to both double-stranded and single-stranded DNA. Sedimentation in a sucrose gradient revealed that the protein induced significant condensation of both supercoiled plasmid DNA and linear bacteriophage $\lambda$ DNA On the basis of quantitative Western-blot analysis, approximately 40,000 molecules of the protein were estimated to exist in an E. coli. The biochemical properties and cellular abundance of the 21 kDa protein suggest that this protein participates in the formation of nucleoid in E. coli.

• YAKHAK HOEJI
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• v.31 no.2
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• pp.64-69
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• 1987

Monoclonal antibodies against human chorionic gonadotropin (hCG) were prepared and characterized by examining isotype, epitope binding, cross reactivity and affinity constants. And a sandwich type enzyme immunoassay for native hCG was developed with solid phase monoclonal antibody against the conformational determinant expressed only on native hCG and horseradish peroxidase conjugated monoclonal antibody against the $\beta$-subunit of hCG. The assay was sensitive to 1 mIU hCG/ml and shown a linear response up to 200 mIU hCG/ml. The cross reactivity for luteinizing hormone and $\beta$-subunit of hCG were 1% and 0.18%, respectively.