• Korean Journal of Microbiology
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• v.45 no.3
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• pp.239-245
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• 2009

Translation initiation factor eIF1A performs essential functions in various initiation steps including 43S preinitiation complex formation in eukaryotes, and contains a highly conserved oligonucleotide-binding (OB) fold. In our previous study, we discovered that eIF1A possesses RNA annealing activity and forms a stable complex with double-stranded RNA. In this study, we initiated site-directed mutations in eIF1A to find the active sites for these biochemical activities and to investigate whether they are essential functions for yeast cell growth. A truncated protein, eIF1A($\Delta$T), devoid of both N- and C-terminal domains but containing an intact OB-fold exhibited RNA annealing activity. In contrast, all point mutations in OB-fold domain, except R57D, impaired both RNA annealing and dsRNA binding activities, indicating that the intact OB-fold domain is required for both activities. Viabilities of the mutant yeast cells were not correlated with RNA annealing activity but with the in vivo protein stabilities in the case of R57D and K94D.

• Journal of the Korean Wood Science and Technology
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• v.34 no.2
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• pp.68-77
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• 2006

The heartwood of cengal (Neobalanocarpus heimii) is known to have a high degree of decay resistance by virtue of its high extractive content. After 30 years in ground contact an utility pole of this tropical hardwood was found to be degraded only in the surface layers by cavity-forming soft rot fungi. The present work was undertaken 1) to characterize the degradation of cengal heartwood from the aspect of ultrastructure and chemistry and 2) to investigate the correlation between soft rot decay and its extractive microdistribution in wood tissues. The chemical analysis of cengal heartwood revealed the presence of a high amount of extractives as well as lignin. The wood contained a relatively high amount of condensed lignin and the guaiacyl units. Microscopic observations revealed that vessels, fibers and parenchyma cells (both ray and axial parenchyma) all contained extractives in their lumina, but in variable amounts. The lumina of fibers and most axial parenchyma were completely or almost completely filled with the extractives. TEM micrographs showed that cell walls were also impregnated with extractives and that pit membranes connecting parenchyma cells were well coated and impregnated with extractives. However, fungal hyphae were present in the extractive masses localized in cell lumina, and indications were that the extractives did not completely inhibit fungal growth. The extent of cell wall degradation varied with tissue types. The fibers appeared to be more susceptible to decay than vessels and parenchyma. Middle lamella was the only cell wall region which remained intact in all cell types which were severely degraded. The microscopic observations suggested a close correlation between extractive microdistribution and the pattern and extent of cell wall degradation. In addition to the toxicity to fungi, the physical constraint of the extractive material present in cengal heartwood cells is likely to have a profound effect on the growth and path of invasion of colonizing fungi, thus conferring protection to wood by restricting fungal entry into cell walls. The presence of relatively high amount of condensed lignin is also likely to be a factor in the resistance of cengal heartwood to soft rot decay.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.30 no.1
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• pp.22-29
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• 2008

Purpose: The purpose of this study is to test the efficacy of various methods of fat harvesting in animal model by viability comparison with assay including cell counting, MTT assay, and histologic evaluation. Materials and methods: New Zealand white rabbits experiments were used. Groin fat pads were subjected to different harvest method varying ingredients of solution(Experiment 1: T1 solution= lidocaine 1000mg/L, epinephrine 1mg/L, sodium bicarbonate 10mgEq/L, Triamcinolone 10mgEq/L; T2 solution=lidocaine 1000mg/L, epinephrine 1mg/L, sodium bicarbonate 0mgEq/L, Triamcinolone 0mgEq/L) and pressure exerted on harvesting with Luer-Lock syringe connected to suction cannula.(Experiment 2: P1 group=3cc intermittent pressure; P2 group=10cc sustained pressure) Fat cell viability was assessed with cell counting with a hemocytometer, MTT assay, and histologic evaluation. Results: Experiment 1 Cell count: T1=2.4/3.4/4.2, T2=9.6/8.4/7.2($\times10^5$ per mL); MTT assay: T1=0.516/0.41/0.453/0.412/0.421, T2=0.925/0.765/0.54/0.634/0.614 in 21 days(absorbance); Histology: T1 showed elongated and, different in size and shape, and ruptured adipocytes with only a few normal adipocytes whereas T2 showed central core of fat with almost intact fat cells Experiment 2 Cell count: P1=1.2/3.2/4.2, P2=1.2/2.4/3.8($\times10^5$ per mL); MTT assay:P1=0.256/0.245/0.258/0.21/0.264, P2=0.12/0.231/0.245/0.313/0.281 in 21 days(absorbance); Histology: P1 showed somewhat evenly distributed normal-looking fat cells and P2 showed relatively irregular shape of fat cells with small blood vessel amongst adiopocytes. Conclusion: Viability was higher in ‘modified tumescent solution’without sodium bicarbonate and triamcinolone and we also found no significantly different viability between using intermittent pressure and using sustained pressure. But in terms of initial viability of fat cell, we can assume that lower intermittent pressure would make better clinical results.

• The Korean Journal of Physiology
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• v.29 no.2
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• pp.269-277
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• 1995

Membrane vesicles were prepared by differential centrifugation from epithelial cells of porcine trachea. Total activity of microsomal ATPases was measured spectrophotometrically by a coupled enzyme assay. The steady-state activity of the enzyme was $329{\pm}10$ nmol/min mg protein. Thapsigargin, a specific antagonist of intracellular $Ca^{2+}-ATPase$, inhibited about 50% of the activity, leaving $178{\pm}18\;nmol/min .mg$ protein (n=6), indicating that the $Ca^{2+}-ATPase$ is one of the major microsomal ATPases. The microsomes used in this study appeared to be tight-sealed vesicles since they showed saturation in $^{45}Ca^{2+}$ uptake experiments. Inositol 1,4,5-trisphosphate $InsP_{3}, 4\;{\mu}M$, an agonist of $InsP_{3}$-sensitive $Ca^{2+}$ release channel ($InsP_{3}$, receptor), and Ca-ionophore A23187 $(10\;{\mu}M)$ induced $^{45}Ca^{2+}$ releases of 20% and 50% of stored $^{45}Ca^{2+}$, respectively. The addition of $(10\;{\mu}M\;InsP_{3}$ also increased the microsomal ATPase activity from $282{\pm}8$ nmol/min mg protein to $334{\pm}21$ nmol/min . mg protein in the intact vesicles. Similar increase in the activity was observed by making microsomes leaky (uncoupling) using the Ca-ionophore A23187. ;$InsP_{3}-induced$ effects were blocked by either thapsigargin or heparin suggesting that: 1) the $InsP_{3}-induced$ increase in ATPase activity is mediated by microsomal $Ca^{2+}-ATPase$, and 2) dissipation of $Ca^{2+}$ gradient across the microsomal membrane is responsible for the $InsP_{3}-induced$ effect. In order to test the dependence of the $Ca^{2+}-ATPase$ activity on the activity of $InsP_{3}-induced$ the activity of ATPases was monitored in various concentrations of free $Ca^{2+}$ using $EGTA-Ca^{2+}$ buffers. The $Ca^{2+}$-dependent biphasic change is the well-known character of $InsP_{3} receptor but not of microsomal $Ca^{2+}-ATPase$ in non-excitable cells; however, the activity of microsomal ATPase appeared biphasic and a maxim진 activity of $397{\pm}36nmol/min\;.mg$ protein was obtained in the solution containing 100 nM free $Ca^{2+}$. Below or above this concentration, the activity of ATPases was lower. These results strongly support a positive correlation of microsomal $Ca^{2+}-ATPase$ to the $InsP_{3}$ receptors in epithelial microsomes.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.10
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• pp.1510-1516
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• 2007

These experiments were carried out to optimize the parameters of electrical activation, methods of parthenogenetic activation and embryo culture in vitro and meanwhile to isolate embryonic stem cells-like (ESCs) derived from porcine parthenogenetic blastocysts (pPBs). These results showed that, as the electric field strength increased from 1.0 to 2.7 kV/cm, the cleavage rate of parthenogenetic embryos increased gradually but the rate of oocyte lysis was significantly increased when using 2.7 kV/cm field strength. The rate of cleavage in 2.2 and 2.7 kV/cm groups was significantly increased in comparison with that of the 1.0 kV/cm group. A voltage field strength of 2.2 kV/cm DC was used to investigate blastocyst development following activation with a single pulse of 30 or $60-{\mu}sec$ pulse duration. The optimum pulse duration was 30-${\mu}sec$, with a blastocyst rate of 20.7%. Multiple pulses were inferior to a single pulse for blastocyst yield (8.0% vs. 29.9) (p<0.05). For porcine oocyte parthenogenetic activation methods, the rates of cleavage (79.0% vs. 59.8%) and blastocysts (19.4% vs. 3.4%) were significantly increased in electrical activation in contrast to chemical activation with ionomycin/6-DMAP (p<0.05). Rates of cleavage and blastocyst formation in NCSU-23 and PZM-3 embryo media were higher than those of G1.3/G2.3 serial culture media, but there was no significant difference among the three groups. The total cell number of blastocysts in PZM-3 embryo culture media containing $5{\mu}g/ml$ insulin was significantly higher than that of the control (no insulin) ($44.3{\pm}9.1$ vs. $33.9{\pm}11.7$). For isolation of PESCs-like, the rates of porcine blastocysts attached to feeder layers and ICM colony formation in Method B (nude embryo culture) were better than those in Method A (intact embryo culture).

• Journal of Veterinary Clinics
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• v.28 no.1
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• pp.122-127
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• 2011

A 5-year old, intact male, Cocker spaniel dog was referred with paraplegia and loss of deep pain perception. Physical, neurological examinations, radiography, and computed tomography were evaluated. Based on the clinical examinations, the dog was diagnosed with severe disc herniation ($L_2$ to $L_3$ intervertebral disc space). On the next day of presentation (6 days after loss of deep pain perception), hemilaminectomy was performed. After decompression of spinal cord and removal of herniated disc materials, $1{\times}10^6$ canine allogenic adipose tissue-derived mesenchymal stem cells (MSCs) diluted by $50{\mu}l$ saline were directly applied to the injured site of the spinal cord. Ten weeks of follow-up after surgery, full recovery of deep pain perception and motor function were evaluated in both hind limbs. Based on the result, we suggest that the transplantation of allogenic adipose tissue-derived MSCs to dogs with spinal cord injuries could be a considerable method to expect better clinical outcomes in veterinary practice.

• Horticultural Science & Technology
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• v.18 no.3
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• pp.360-367
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• 2000

'Yumyeong' peach has desirable characteristics of long shelf-life and specific non-melting nature with long harvesting period. However, some fruits harvested too late tend to show fruit pithiness symptom or internal breakdown. The study was conducted to investigate the differences between fruit flesh pithiness and internal breakdown symptoms, and to find out inducing factors of flesh pithiness in 'Yumyeong' peach. The incidence of flesh pithiness was higher in the fruits harvested late. Sugar and malic acid contents were not different between normal fruits and flesh pithiness fruits, but the acidity was significantly lower affected by low citric acid content in flesh pithiness fruit. In flesh pithiness fruits, calcium contents were low in both skin and flesh. Occurrence of flesh pithiness fruits was high in the years with low precipitation and high temperature for 2 months before harvest. In observations on morphological characteristics, the parts showing flesh pithiness consisted of smaller cells than the normal parts. Tonoplasts were disintegrated and the number of dead cells was high in internal breakdown fruits, while the tonoplasts were intact with contracted vacuoles in flesh pithiness fruits. Tylosises were observed in vascular tissues around the flesh pithiness, therefore, it was assumed that those tylosises restricts flesh tissue development resulting in flesh pithiness. Other varieties such as 'Fantasia' and 'Wolmi' also showed tylosis and smaller cells were observed in the flesh tissue of these cultivars, indicating abnormal growth of the flesh part. These results suggested the possibility of the occurrence of pithiness like symptoms in other peach varieties.

• Journal of Plant Biology
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• v.38 no.1
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• pp.87-93
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• 1995

For understanding physiological nature of phototaxis in Synechocystis sp. PCC 6803 PTX(S. 6803 PTX), we examined the effects of some metabolic inhibitors and cation ionophore on the phototactic movement. In the presence of DCMU, which blocks the photosynthetic electron transport just after photosystem II acceptor, there was no inhibitory effect on the phototaxis up to $100\;\mu\textrm{M}$. Instead, the respiratory electron chain inhibitor such as sodium azide dramatically impaired the phototaxis in S. 6803 PTX. These observations indicate that the phototaxis is linked not to photo-phosphorylation, but to respiratory phosphorylation. When the cells were treated with un couplers such as CCCP or DNP, which dissipate the electrochemical gradient of proton($\Delta\mu_{H}+$) across the cytoplasmic membrane, these chemicals did not affect phototaxis. In contrast, when cells were treated with DCCD or NBD which deprive cells of A TP but leave $\Delta\mu_{H}+$ intact across the membrane, the phototactic movement was severly reduced. These results imply that ATP production, not proton motive force, is involved in the phototactic movement in this organism as a driving motive force. The application of specific calcium ionophore A23187 strongly impaired positive phototaxis. Calcium fluxes should be engaged in the sensory trans-duction of phototactic orientation. Finally, when ethionine was supplimented to culture media, the photomovement of this organism was inhibited. This implies that methylation/demethylation mechanism controls the process of phototaxis in S. 6803 PTX like chemotaxis in E. coli and Salmonella typhimurium.murium.

• Journal of the Korean Society of Food Science and Nutrition
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• v.32 no.1
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• pp.29-34
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• 2003

In development of the processed food, it is important not only to make the food delicious but to enhance its storage span and thermal stability without change in color, which greatly affects the tastes. Protopectinase (PPase) from Bacillus subtilis EK11 hydrolyses or dissolves protopectin in the middle lamella of plant tissues with the resultant separation of plant cells from each other, called enzymatic maceration. With the PPase, persimmon was enzymatically macerated to separate cells to primary cell wall without damage. Recovery rates of persimmon treated with PPase and mechanical maceration were 95% and 85%, respectively. Total and reducing sugars, crude protein and fat in the enzymatic maceration were well preserved as in the mechanical maceration. Importantly, over 50% of vitamin C, which is the most unstable component during the mechanical maceration, remained with an intact form for one day after the enzymatic treatment. When the suspensions of persimmon macerated with both treatments were stored at 4$^{\circ}C$ for 9 days, the mechanically macerated persimmon suspension was decolorized, whereas decolorization, was not found in the enzymatically macerated persimmon suspension. Moreover the mechanically macerated persimmon was greatly deteriorated after heat treatment at 10$0^{\circ}C$ for 60 min, whereas cells of the enzymatically separated persimmon suspension appeared to be stable, indicating increased thermal stability Thus, the PPase treatment of persimmon could be a better choice for preparation of highly valuable and functional processed food as well as for increase in preservation period.

• Journal of Life Science
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• v.31 no.8
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• pp.755-760
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• 2021

Swine diarrhea is a livestock disease that causes huge economic losses to pig farms. In general, diarrhea occurs because of the proliferation of pathogenic Escherichia coli (E. coli). The toxins produced by the proliferated E. coli cause edema in pigs. Although the proliferation of these coliforms can be prevented by using a vaccine, the vaccines containing chemically produced dead bacteria are not very effective, making it difficult to control the proliferation of E. coli. Therefore, there is a need to develop new, more effective vaccines. In this study, we prepared killed F4+ and F18ab+ E. coli, which induce diarrhea and edema in pigs, using the extracts of immune-induced silkworms containing antimicrobial peptides and examined their availability as a killed-bacteria vaccine. First, the antimicrobial activity analysis of the prepared immune-induced silkworm extract was conducted using the radial diffusion assay. The results showed high activity against both F4+ and F18ab+ E. coli. The production efficiency of E. coli dead cells was determined using the colony-counting method. The concentration of the E. coli dead cells was the highest (50 mg/ml) when treated at 4℃. In addition, the analysis of the prepared dead cells using a transmission electron microscope confirmed that E. coli leaked out of the cytoplasm and the cell membrane remained intact. Therefore, F4+ and F18ab+ E. coli produced using immune-induced silkworms extract are considered to be highly available as bacterial ghost vaccines that can help prevent swine diarrhea and the resulting edema.