• The Korean Journal of Pharmacology
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• v.26 no.1
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• pp.35-49
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• 1990

A highly purified $(Na^+,\;K^+)-ATPase$ from the rectal gland of Squalus acanthias and from the electric organ of Electrophorus electricus has been used to raise antibodies in rabbits. The 97,000 dalton catalytic subunit and glycoprotein derived from the rectal gland of spiny shark were also used as antigens. The two $(Na^+,\;K^+)-ATPase$ holoenzymes and the two shark subunits were antigenic. In Ouchterlony double diffusion experiments, these antibodies formed precipitation bands with their antigens. Antibodies prepared against the two subunits of shark holoenzyme also formed precipitation bands with their antigens and shark holoenzyme, but not with eel holoenzyme. These observations are in good agreement with inhibitory effect of these antibodies on the catalytic activity of $(Na^+,\;K^+)-ATPase$ both from the shark and the eel, since there is very little cross-reaction between the shark anticatalytic subunit antibodies and the eel holoenzyme. The maximum antibodies titer of the anticatalytic subunit antibodies is found to be 6 weeks after the initial single exposure to this antigen. Multiple injections of the antigen increased the antibody titer. However, the time required to produce the maximum antibody titer was approximately the same. These antibodies also inhibit catalytic activity of $(Na^+,\;K^+)-ATPase$ vesicles reconstituted by a slow dialysis of cholate after solubilization of the enzyme in a presonicated mixture of cholate and phospholipid. In these reconstituted $(Na^+,\;K^+)-ATPase$ vesicles, effects of these antibodies on the fluxes of $Na^+$, $Rb^+$, and $K^+$ were investigated. Control or preimmune serum had no effect on the influx of $^{22}Na^+$ or the efflux of $^{86}Rb^+$. Immunized sera against the shark $(Na^+,\;K^+)-ATPase$ holoenzyme, its glycoprotein or catalytic subunit did inhibit the influx of $^{22}Na^+$ and the efflux of $^{86}Rb^+$. It was also demonstrated that these antibodies inhibit the coupled counter-transport of $Na^+$ and $K^+$ as studied by means of dual labeling experiments. However, this inhibitory effect of the antibodies on transport of ions in the $(Na^+,\;K^+)-ATPase$ vesicles is manifested only on the portion of energy and temperature dependent alkali metal fluxes, not on the portion of ATP and ouabain insensitive ion movement. Simultaneous determination of effects of the antibodies on ion fluxes and vesicular catalytic activity indicates that an inhibition of active ion transport in reconstituted $(Na^+,\;K^+)-ATPase$ vesicles appears to be due to the inhibitory action of the antibodies on the enzymatic activity of $(Na^+,\;K^+)-ATPase$ molecules incorporated in the vesicles. These findings that the inhibitory effects of the antibodies specific to $(Na^+,\;K^+)-ATPase$ or to its subunits on ATP and temperature sensitive monovalent cation transport in parallel with the inhibitory effect of vesicular catalytic activity by these antibodies provide direct evidence that $(Na^+,\;K^+)-ATPase$ is the molecular machinery of active cation transport in this reconstituted $(Na^+,\;K^+)-ATPase$ vesicular system.

• Transactions of the Korean hydrogen and new energy society
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• v.11 no.3
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• pp.119-125
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• 2000

To improve the photochemical energy conversion efficiency and the stability of CdS particulate film electrode which is used to produce hydrogen from the aqueous $H_2S$ solution photoelectrochemically, surface treatment of this film was carried out using $TiCl_4$ solution. CdS particles for preparation of the films were synthesized by precipitation reaction of $Cd({NO_3})_2{\cdot}9H_2O$ and $Na_2S{\cdot}4H_2O$. Then, the CdS sol was hydrothermally treated for 12hr in an autoclave with the variation of treatment temperature to control the crystalline phase of particles. CdS film electrode was thus prepared by annealing at $400^{\circ}C$ for 12hr of the wet-film cast at room temperature, and subsequently surface treated with $TiCl_4$ solution. The electrodes were characterized using XRD, SEM, and the photocurrent meter. The photocurrents of Cds film electrodes prepared with surface treatment were up to two times higher than the electrodes without surface treatment, indicating about $4.0mA/cm^2$. Hydrogen production rate in a continuous flow system using photoelectrochemical or photochemical cells prepared with surface treatment also increased in proportion to the increase of photocurrents.

• Journal of Plant Biotechnology
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• v.34 no.1
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• pp.75-80
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• 2007

The release of genetically modified organisms ($GMO_{s}$) into the environment has the potential risks regarding the possibility of gene transfer from $GMO_{s}$ to natural organisms and this needs to be evaluated. This study was conducted to monitor the possible horizontal gene transfer from herbicide-resistant zoysiagrass (Zoysia japonica Steud.) to indigenous microorganisms. We have first examined the effect of field-released GM zoysiagrass on the microbial flora in the gut of locust (Locusts mlgratoria). The microbial flora was analyzed through determining the 165 rDHA sequences of microorganisms. The comparison of the microbial flora in the gut of locusts that were captured at the field of GM zoysiagrass and of wild-type revealed that there is no noticeable difference between these two groups. This result indicates that the GM zoysiagrass does not have negative impact on microbial flora in the gut of locust. We then investigated whether the horizontal gene transfer occurred from GM zoysiagrass to microbes in soil, rhizosphere and faecal pellets from locusts by utilizing molecular tools such as Southern hybridization and polymerase chain reaction (PCR). When the total DNAs isolated from microbes in GM zoysiagrass and in wild-type zoysiagrass fields were hybridized with probes for bar or hpt gene, no hybridization signal was detected from both field isolates, while the probes were hybridized with DNA from the positive control. Absence of these genes in the FNAs of soil microorganisms as well as microbes in the gut of locust was further confirmed by PCR. Taken together, our data showed that horizontal gene transfer did not occur in this system. These results further indicate that frequencies of transfer of engineered plant DNA to bacteria are likely to be negligible.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.17
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• pp.7169-7174
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• 2014

Background: NPAS2 is a product of the circadian clock gene. It acts as a putative tumor suppressor by playing an important role in DNA damage responses, cell cycle control and apoptosis. Chronic lymphocytic leukemia (CLL) appears to be an apoptosis related disorder and alteration in the NPAS2 gene might therefore be directly involved in the etiology of CLL. Here, the Ala394Thr polymorphism (rs2305160:G>A) in the NPAS2 gene was genotyped and melatonin concentrations were measured in a total of seventy-four individuals, including thirty-seven CLL cases and an equal number of age- and sex-matched healthy controls in order to examine the effect of NPAS2 polymorphism and melatonin concentrations on CLL risk in a Pakistani population. Materials and Methods: Genotyping of rs2305160:G>A polymorphism at NPAS2 locus was carried out by amplification refractory mutation system-polymerase chain reaction (ARMS-PCR). Melatonin concentrations were determined by enzyme linked immunosorbent assay (ELISA). Statistical analysis was performed using Statistical Package for Social Sciences software. Results: Our results demonstrated no association of the variant Thr genotypes (Ala/Thr and Thr/Thr) with risk of CLL. Similarly, no association of rs2305160 with CLL was observed in either females or males after stratification of study population on a gender basis. Moreover, when the subjects with CLL were further stratified into shift-workers and non-shift-workers, no association of rs2305160 with CLL was seen in either case. However, significantly low serum melatonin levels were observed in CLL patients as compared to healthy subjects (p<0.05). Also, lower melatonin levels were seen in shift-workers as compared to non-shift-workers (p<0.05). There was no significant difference (p>0.05) in the melatonin levels across NPAS2 genotypes in all subjects, subjects with CLL who were either shift workers or non-shift-workers. General Linear Model (GLM) univariate analysis revealed no significant association (p>0.05) of the rs2305160 polymorphism of the NPAS2 gene with melatonin levels in any of the groups. Conclusions: While low melatonin levels and shift-work can be considered as one of the risk factors for CLL, the NPAS2 rs2305160 polymorphism does not appear to have any association with risk of CLL in our Pakistani population.

• Korean Journal of Organic Agriculture
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• v.23 no.4
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• pp.847-858
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• 2015

Bacillus amyloliquefaciens GR4-5 was isolated from the rhizosphere soil of Korean ginseng and displayed broad-spectrum suppression of ginseng root rot pathogens. The survivability of B. amyloliquefaciens GR4-5 in soil was investigated under three different conditions; indoor, outdoor - of which soil was put in 14 mL tube after treatment - and field environments. Soil samples were collected over a four-week period from three experimental designs, and assessed for 16S rRNA gene copy number by quantitative polymerase chain reaction (qPCR). In outdoor condition, the 16S rRNA gene copy number of Bacillus spp. was 8.35 log copies g $soil^{-1}$ immediately after the GR4-5 treatment. Two weeks later, the 16S rRNA gene copy number of Bacillus spp. (6.70 log copies g $soil^{-1}$) was similar to that of the control (6.38 log copies g $soil^{-1}$). In indoor condition, the 16S rRNA gene copy number of Bacillus spp. maintained in a certain level for a longer period than those in outdoor and field. The 16S rRNA gene copy number of Bacillus spp. in field experiment was reduced faster than that of outdoor condition. Our results show that B. amyloliquefaciens GR4-5 can survive in bulk soil for 1 week, indicating its potential use as a biocontrol agent following 7 day application intervals. This study presents that outdoor microcosm system design could be a useful method to assess easily the survivability of beneficial microorganisms.

• Korean Journal of Applied Biomechanics
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• v.21 no.1
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• pp.31-38
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• 2011

This study aimed to determine the effects of the blockage of visual feedback on joint dynamics of the lower extremity. Fifteen healthy male subjects(age: $24.1{\pm}2.3\;yr$, height: $178.7{\pm}5.2\;cm$, weight: $73.6{\pm}6.6\;kg$) participated in this study. Each subject performed single-legged landing from a 45 cm-platform with the eyes open or closed. During the landing performance, three-dimensional kinematics of the lower extremity and ground reaction force(GRF) were recorded using a 8 infrared camera motion analysis system (Vicon MX-F20, Oxford Metric Ltd, Oxford, UK) with a force platform(ORG-6, AMTI, Watertown, MA). The results showed that at 50 ms prior to foot contact and at the time of foot contact, ankle plantar-flexion angle was smaller(p<.05) but the knee joint valgus and the hip flexion angles were greater with the eyes closed as compared to with the eyes open(p<.05). An increase in anterior GRF was observed during single-legged landing with the eyes closed as compared to with the eyes open(p<.05). Time to peak GRF in the medial, vertical and posterior directions occurred significantly earlier when the eyes were closed as compared to when the eyes were open(p<.05). Landing with the eyes closed resulted in a higher peak vertical loading rate(p<.05). In addition, the shock-absorbing power decreased at the ankle joint(p<.05) but increased at the hip joints when landing with the eyes closed(p<.05). When the eyes were closed, landing could be characterized by a less plantarflexed ankle joint and more flexed hip joint, with a faster time to peak GRF. These results imply that subjects are able to adapt the control of landing to different feedback conditions. Therefore, we suggest that training programs be introduced to reduce these injury risk factors.

• Journal of the Korean Society of Food Science and Nutrition
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• v.32 no.6
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• pp.913-920
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• 2003

Soybean sauce fermented with soybean and wheat, has been a major condiment of Korean diets from centuries ago. Melanoidin, a brown pigment generally found in various food systems, is a final product produced in amino-carbonyl reaction during soybean sauce processing. Antioxidative activities of soybean sauce and melanoidin were investigated in vitro system using linoleic acid emulsion. Soybean sauce and glucose-lysine model melanoidin showed the stronger antioxidative effect than control by ferric thiocyanate and conjugated diene assays. In addition, DPPH radical scavenging effect of soybean sauce was higher than melanoidin, which was ascribed to soluble peptide and low molecular protein existing in soybean sauce. To ascertain antioxidative effect of dietary soybean sauce and melanoidin in vivo, the male Wister rats were fed 10% soybean sauce or 10% glucose-lysine model melanoidin with corn oil or fish oil for 5 weeks. Fatty acid compositions in liver and plasma were influenced by oil source. Therefore, EPA and DHA contents of fish oil group were higher than those of corn oil group. When the inhibitory effect of soybean sauce and melanoidin on lipid peroxidation using TBARS methods was measured, fish oil group (FC) showed higher malondialdehyde (MDA) content than corn oil group (CC). However, supplementation of soybean sauce and melanoidin to fish oil group attenuated MDA formation. In the levels of phosphatidyl choline hydroperoxide (PCOOH) in liver and plasma by CL (chemiluminescence)-HPLC method, PCOOH in FC group was significantly higher than that of CC group both in liver and plasma. Supplementation of soybean sauce to fish oil groups significantly inhibited the formation of PCOOH in plasma and liver, while melanoidin suppressed hepatic PCOOH formation. Based on these results, it can be suggested that soybean sauce possesses stronger antioxidative potential than melanoidin.

• Korean Journal of Veterinary Research
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• v.47 no.2
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• pp.175-185
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• 2007

This study was carried out to investigate the pathogenesis and pathogenicity of the porcine circovirus type 2 (PCV2) Korean isolate from weaned pigs. Twenty four weaned pigs, PCV2, porcine reproductive and respiratory syndrome virus (PRRSV) and porcine parvovirus (PPV) antibodies free, were allocated to 4 groups (n = 6). Six pigs were inoculated intranasally with PCV2 alone, 6 with PCV2 and PRRSV, 6 with the combined PCV2/PRRSV/PPV inoculum, and 6 were remained as a uninoculated negative control. Pigs were killed 3 and 6 weeks after inoculation and tissue samples examined for gross and microscopic lesions and for the presence of PCV2 antigens and nucleic acids. Experimentally inoculated pigs were evaluated for 3 considerations: 1. development of postweaning multisystemic wasting syndrome (PMWS), 2. distribution of viral antigens by immunohistochemistry and polymerase chain reaction (PCR), and 3. cytokine mRNA levels in lymph nodes. Pigs inoculated with PCV2/PRRSV/PPV showed typical clinical signs, gross findings, and histopathologic characteristics of PMWS. In the PCV2/PRRSV/PPV inoculated group, the PCV2 antigen was widely distributed in various parenchymal organs such as brain, spinal cord, tonsil, lymph nodes, lung, heart, liver, kidney, spleen, and peyer's patch. Lymph node mRNA expression of IL-$1{\alpha}$, IL-2R and IL-8 was determined by real-time PCR. The pigs of PCV2/PRRSV and PCV2/PRRSV/PPV inoculation group, the mRNA expression was characterized by a decrease of IL-$1{\alpha}$, IL-2R and IL-8. The decrease of cytokine mRNA represent the state of T cell immuno-suppression in pig, and nicely support the evidence for the impairment of immune system in pigs with PMWS. In conclusion, PCV2 infection and some additional infectious causes such as PRRSV and/or PPV are warranted for the presence of PMWS in weaned pigs in Korea.

• Journal of the Korean Society of Food Culture
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• v.28 no.1
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• pp.78-88
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• 2013

Marinated beef galbi is a traditional Korean dish cooked with soy sauce, pear juice, onion, sesame oil, and sugar. However, there are many differences in beef galbi, including flavor and physicochemical aspects, depending on cooking conditions. Therefore, the physicochemical characteristics of marinated beef galbi prepared through various recipes was evaluated for its effects on pH, texture, aging, proteolysis, heating conditions, cooking time, and flavor compounds (pyrazines, IMPs, or FAAs). There were significant differences in salt concentration (0.8~3.03%), pH (4.89~6.22), and solid soluble contents (1.34-6.31 Brix) between recipes in this study. In the Pearson assay for sensory evaluation, overall preference correlated well with texture (a well-known sensory attribute in meat evaluation). Controlling the pH of meat through soaking in lemon solution, alkali water, phosphate, and baking powder solution, improved water holding capacity as much as 9 to 15% compared with the control. The myofibril index (MFI) of marinated meat stored at $4^{\circ}C$ increased 32% with 24 hours of aging and reached 39% at 48 hours of aging, and its fragmentation was observed through microscopy. SDS-PAGE showed hydrolysis of acid-soluble collagen by the pear juice, possibly related to meat tenderness. On the basis of surface temperature, the cooking time was estimated to be 8 minutes with pan heating at $170^{\circ}C$, 6 minutes at $270{\sim}300^{\circ}C$, and 4 minutes with charcoal at $700{\sim}900^{\circ}C$. Different pyrazine compounds, such as 2-methyl-3-phenylpyrrol(2,3-b) pyrazine (the typical product of the browning reaction) was mainly detected, and IMP (one of the main taste compounds in beef) was in higher amounts with the charcoal treatment, potentially related to its flavor preference among treatments. Our results demonstrate an effective case study and cooking system for beef galbi.

• Korean Journal of Microbiology
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• v.43 no.3
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• pp.159-165
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• 2007

For the development of basic genetic materials for specific and effective therapeutic approach to suppress multiplication of hepatitis C virus (HCV), HCV internal ribosome entry site (IRES)-targeting hammerhead ribozyme which activity is allosterically regulated by HCV regulatory protein, NS5B RNA replicase, was developed. The ribozyme targeted most effectively to +382 nucleotide (nt) site of HCV IRES RNA. The allosteric ribozyme was designed to be composed of sequence of RNA aptamer to HCV NS5B, communication module sequence which can transfer structural transition for inducing ribozyme activity upon binding NS5B to the aptamer, and sequence of ribozyme targeting +382 nt of HCV IRES. Noticeably, we employed in vitro selection technology to identify the most appropriate communication module sequence which can induce ribozyme activity depending on the US5B protein. We demonstrated that the ribozyme was nonfunctional either in the absence of any proteins or in the presence of control bovine serum albumin. In sharp contrast, the allosteric ribozyme can induce activity of cleavage reaction with HCV IRES RNA in the presence of the HCV NS5B protein. This allosteric ribozyme can be used as lead compound for specific and effective anti-HCV agent, tool for highthroughput screening to isolate lead chemicals for HCV therapeutics, and ligand for biosensor system for HCV diagnosis.