• Animal cells and systems
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• 제1권3호
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• pp.483-489
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• 1997

This study was performed to investigate the differentiation and distribution of choline acetyltransferase (ChAT}-immunoreactive cells in the magnacellular preoptic nucleus (MCPO) of the postnatal and adult rat forebrains, utilizing techniques of immunocytochemistry. According to the cell shape and the ratio of long axis versus short axis of cell soma, the ChATimmunoreactive nerve cells in the MCPO were classified into six types: 1) round, 2) oval, 3) elongated, 4) fusiform, 5) triangular, and 6) polygonal types. Frequency distributions of the oval and round nerve cells on the postnatal day (PND) 0 were observed to be high. But in the adult, frequency distributions of the same cells were shown to decrease. Compared to those of the postnatal rats, frequency distributions of elongated, fusiform, triangular, and polygonal nerve cells in the adult were increased. The total mean volumes of ChAT-immunoreactive cell somata in the MCPO of PND 0 rat were the lowest, while those in the PND 17 rat were shown to be the highest and decreased in the adult. The soma volumes of the immunoreactive cells at the PND 17 were evenly distributed, but those in the other developmental stages (e.g. PND 7 and adult) appeared to exhibit unimodal distributions. On the electron micrography, the free ribosomes, polysomes, and rough endoplasmic reticula (RER) of the nerve cells in the MCPO of PND 21 rat forebrains were immunoreactive to ChAT in the tissues untreated with triton X-100. According to the observations in the present study, it is considered that the ChAT-immunoreactive nerve cells in the MCPO of the rat forebrains are differentiated throughout the following processes during the postnatal development: 1) increase in cell soma volumes, 2) development of neurites, 3) increase in the frequency of differentiated cell types, and 4) decrease in cell soma volumes. The ribosomes, polysomes, and RER are considered to be closely related to the intracellular localization and biosynthesis of the ChAT but not Golgi complex.

• Asian-Australasian Journal of Animal Sciences
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• 제13권1호
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• pp.10-18
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• 2000

This paper describes the magnetic orientation of the intact and demembranated bull sperm treated by DTT or heparin in a 5,400 G static field. Semen samples collected from four bulls (Japanese Black) were mixed to the same sperm density. One percentage triton X-100 was used to extract the plasma membrane. The intact and demembranated sperm suspensions were treated with 20, 200, 2,000 mM DTT, 100, 1,000 or 10,000 units heparin solutions at $4{^{\circ}C}$ for 6 days. The decondensation of the sperm nuclei treated by DTT or heparin was examined by measuring the sperm head area at 1, 3, and 6 days. After measuring the area, each sperm sample was exposed to a 5,400 G static magnetic field generated by Nd-Fe-B permanent magnets for 24 hours at room temperature. Results showed that the decondensation of bull sperm nuclei was not induced by the heparin treatment, however, incomplete decondensation was induced by the DTT treatment. During the magnetic orientation, bull sperms treated by DTT or heparin had low percentages of long axis perpendicular to the magnetic lines of force. However, different aspects were obtained for long axis perpendicular orientations following treatment of DTT or heparin. Through the DTT treatment, the decline of long axis perpendicularly oriented percentages was due to the increase of long axis parallel orientation with the head of the flat plane perpendicular to the magnetic lines of force, whereas, using the heparin treatment, the decline of long axis perpendicular orientation was due to the increment of long axis parallel orientation with the head of the flat plane parallel to the magnetic lines of force. Also, percentages of the head of the flat plane perpendicular were decreased by the heparin treatment. These findings suggest that maintaining the structure of protamine in the chromatin is necessary for the sperm head to orient with its flat plane perpendicular, and maintaining the disulfide bond in the chromatin is necessary for the long axis of sperm to orient perpendicularly.

• 한국분말재료학회지
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• 제25권3호
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• pp.257-262
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• 2018

The coupling of two semiconducting materials is an efficient method to improve photocatalytic activity via the suppression of recombination of electron-hole pairs. In particular, the coupling between two different phases of $TiO_2$, i.e., anatase and rutile, is particularly attractive for photocatalytic activity improvement of rutile $TiO_2$ because these coupled $TiO_2$ powders can retain the benefits of $TiO_2$, one of the best photocatalysts. In this study, anatase $TiO_2$ nanoparticles are synthesized and coupled on the surface of rutile $TiO_2$ powders using a microemulsion method and heat treatment. Triton X-100, as a surfactant, is used to suppress the aggregation of anatase $TiO_2$ nanoparticles and disperse anatase $TiO_2$ nanoparticles uniformly on the surface of rutile $TiO_2$ powders. Rutile $TiO_2$ powders coupled with anatase $TiO_2$ nanoparticles are successfully prepared. Additionally, we compare the photocatalytic activity of these rutile-anatase coupled $TiO_2$ powders under ultraviolet (UV) light and demonstrate that the reason for the improvement of photocatalytic activity is microstructural.

• 대한본초학회지
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• 제30권5호
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• pp.15-21
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• 2015

Objectives : Methicillin-resistantStaphylococcus aureus(MRSA) is a human pathogen. New antibacterial agents are needed to treat MRSA-related infections. This study investigated the antibacterial activity of EtOH 70% extracts ofTonghyeonipal-dan(THD) which prescription is composed of oriental medicine against MRSA.Methods : The antibacterial activity of THD was evaluated against MRSA strains by using the Disc diffusion method, broth microdilution method, Checkerboard dilution test, and Time-kill test; its mechanism of action was investigated by bacteriolysis, detergent or ATPase inhibitors were used.Results : The minimum inhibitory concentration (MIC) of THD is 1,000~2,000 μg/mL against MRSA. In the checkerboard dilution test, fractional inhibitory concentration index (FICI) of THD in combination with antibiotics indicated synergy or partial synergism againstS. aureus. Furthermore, a time-kill assay showed that the growth of the tasted bacteria was considerably inhibited after 24 h of treatment with the combination of THD with selected antibiotics. For measurement of cell membrane permeability, THD 500 μg/mL along with concentration of Triton X-100 (TX) and Tris-(hydroxymethyl) aminomethane (TRIS) were used. In the other hand, N,N-dicyclohexylcarbodimide (DCCD) and Sodium azide (NaN3) were used as an inhibitor of ATPase. TX, TRIS, DCCD and NaN3 cooperation againstS. aureusshowed synergistic action.Conclusions : Accordingly, antimicrobial activity of THD was affected by cell membrane and inhibitor of ATPase were assessed. These results suggest that THD has antibacterial activity, and that THD extract offers great potential as a natural antibiotic against MRSA.

• KSBB Journal
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• 제13권5호
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• pp.606-614
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• 1998

In this study, biodegradation rate of Arabian light crude oil by mixed cultures of selected petroleum-degraders was determined. Their modes of hydrocarbon uptake were then observed to determine whether there are differences in biodegradation rate by the mixed cultures. By the mixed cultures of petroleum-degraders having same modes of hydrocarbon uptake, such as strain US1 and K1 (using pseudo-solubilized hydrocarbons by a biosurfactants), K2-2 and P1(using hydrocarbons by direct contact), CL 180 and IC-10 (mixed type of uptake modes), the biodegradation rates of aliphatics in the crude oil were increased more than those by their pure cultures, about 40%, 25% and 20%, respectively. Biodegradation rate of strain KH3-2 (using only water- dissolved hydrocarbons) was increased by mixed cultures with strain K1, CL180 or IC-10 possessing high emulsifying activity. However, the biodegradation rate of the crude oil was decreased about 20%-40% by the mixed cultures of petroleum-degraders having different mode of hydrocarbon uptake, such as addition of strain US1 or K1 in the cultures of K2-2 or P1. Biosurfactants produced by US1 or K1 seems to enhance the emulsification of crude oil in aqueous phase but inhibit the attachment of K2-2 or P1 to crude oil. As same phenomena, the addition to Triton X-100 into the culture of strain US1, K1, CL180, IC-10 or KH3-2 increased the biodegradation rate, but the addition in the culture of strain K2-2 or P1 decreased the biodegradation rate. The mixed culture made of CL180, IC-10 and KH3-2 degraded 61.5% of aliphatics and 69% of aromatics in 3% (v/v) of Arabian light crude oil added.

• KSBB Journal
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• 제30권6호
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• pp.291-295
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• 2015

The limited productivity of natural shell matrix proteins has hampered the investigation of their biochemical properties and practical applications, although biominerals in nature obtained by organic-inorganic assemblies have attractive mechanical and biological properties. Here, we prepared a vector for the expression of a fusion protein of a shell matrix protein from Pinctada fucata (named as GRP_BA) with the GRGDSP residue. The fusion protein of BA-RGD was simply produced in E. coli and purified through sequential steps including the treatment with $CaCl_2$ and EDTA solution for cell membrane washing, mechanical cell disruption and the application of non-ionic surfactant of Triton X-100 for BA-RGD inclusion body washing. The production yield was approximately 60 mg/L, any other protein band was not observed in SDS-PAGE and it was estimated that above 97% endotoxin was removed compared to the endotoxin level of whole cell. This study showed this simple and easy purification approach could be applied to the purification of BA-RGD fusion protein. It is expected that the protein could be utilized for the preparation of biominerals in practical aspects.

• BMB Reports
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• 제42권7호
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• pp.462-466
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• 2009

Lipopolysaccharide (LPS), found in the outer membrane of Gram negative bacteria, only exerts its toxic effects when in free form. LPS has three major parts, lipid A, the toxic component, along with a core polysaccharide and O-specific polysaccharide. LPS monomers are known to have molecular masses between 10 to 30 kDa. Under physiological conditions, LPS exists in equilibrium between monomer and vesicle forms. LPS removal by 100 kDa ultrafiltration was more efficient (99.6% of LPS removed) with a low concentration of protein (2.0 mg/ml) compared to a high concentration (20.1 mg/ml). In the presence of different detergents (0.5% Tween 20, 1.0% taurodeoxycholate and 1.0% Triton X-100), LPS removal was more efficient at low protein concentrations (2.0 mg/ml) compared to high protein concentrations (20.1 mg/ml).

• 한국축산식품학회지
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• 제38권5호
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• pp.1064-1079
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• 2018

In this study, the antimicrobial activity of a bacteriocin produced by Enterococcus faecalis KT11, isolated from traditional Kargı Tulum cheese, was determined, and bacteriocin KT11 was partially characterized. The results showed that bacteriocin KT11 was antagonistically effective against various Gram-positive and Gram-negative test bacteria, including vancomycin- and/or methicillin-resistant bacteria. The activity of bacteriocin KT11 was completely abolished after treatment with proteolytic enzymes (proteinase K, ${\alpha}$-chymotrypsin, protease and trypsin), which demonstrates the proteinaceous nature of this bacteriocin. Additionally, bacteriocin KT11 remained stable at pH values ranging from 2 to 11 and after autoclaving at $121^{\circ}C$ for 30 min. In addition, the activity of bacteriocin KT11 was stable after treatment with several surfactants (EDTA, SDS, Triton X-100, Tween 80 and urea) and organic solvents (chloroform, propanol, methanol, ethyl alcohol, acetone, hexane and ethyl ether). Cell-free supernatant of E. faecalis KT11 was subjected to ammonium sulfate precipitation and then desalted by using a 3.5-kDa cut-off dialysis membrane. The bacteriocin activity was determined to be 711 AU/mL in the dialysate. After tricine-SDS-PAGE analysis, one peptide band, which had a molecular weight of ~3.5 kDa, exhibited antimicrobial activity. Because the bacteriocin KT11, isolated from E. faecalis KT11, exhibits a broad antimicrobial spectrum, heat stability and stability over a wide pH range, this bacteriocin can be used as a potential bio-preservative in foods. Additionally, bacteriocin KT11 alone or in combination with conventional antibiotics may provide a therapeutic option for the treatment of multidrug-resistant clinical pathogens after further in vivo studies.

• Journal of Microbiology and Biotechnology
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• 제10권6호
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• pp.805-810
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• 2000

An antifungal protein antagonistic to the rice blast fungus, Pyricularia oryzae was purified from Paenibacillus macerans PM-1 by ammonium sulfate fractionation, Q Sepharose Fast Flow column chromatography, Phenyl Sepharose CL-4B column chromatography and Superose 12 gen filtration. An apparent molecular mass of the purified antifungal protein was determined as 8 kDa by SDS-PAGE and 9 kDa by analytical gel filtration, respectively, suggesting that the purified protein is a monomer. The antifungal protein was stable at pH range from 7-12 and up to $100^{\circ}C$. The protein was also stable at 0.1-1% Tween 20 and Triton X-100. The N-terminal amino acid sequence of the antifungal protein was Thr-Glu-Leu-Pro-Leu-Gly-Ile-Val-Met-Asp-Lys-Tyr-Thr-Asp-Ala-Phe-Lys-Phe-Asp-Met-Phe. Comparison of the determined sequence with other peptide and DNA sequences did not reveal homology at all. Therefore, the purified antifungal protein was speculated to be a novel protein. The condidial germination in vitro of P. oryzae KJ301:93-39 by the purified protein ($5.9{\mu} g/ml$) was limited to $9{\pm}3.2%$ only, compared with $69{\pm}2.4%$ of the control. Ungerminated conidia were swollen at basa and mid cell by the purified protein. In vivo bioassay for inhibition of conidial germination of P. oryzae KJ 301, one of the most predominating racesin Korea. the purified protein ($5.9{\mu} g/ml$)strongly inhibited the conidial germination. The conidia, even though germinated, could not develop any further to produce appressoria efficiently.

• 한국정밀공학회:학술대회논문집
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• 한국정밀공학회 2005년도 춘계학술대회 논문집
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• pp.1155-1158
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• 2005

Micro-stereolithography is a newly proposed technology as a means that can fabricate a 3D micro structure of free form. It makes a 3D micro-structure by dividing the shape into many slices of relevant thickness along horizontal surfaces, hardening each layer of slice with a focused laser beam, and stacking them up to a desired shape. However, we do not anticipate the electric conductivity of the final product at the existing micro-stereolithography. The reason is that this technology uses polymer to make the product. Thus the new suspension which was mixed conventional photopolymer with metal powder was developed in this study. The developed suspensions were based on SL5180 which is commercialized resin and IMS03 that is made in our laboratory. And Triton X-100 was added at the suspension for getting the scattering effect and stabilizing effect. The layer recoating device was developed to be flat the mixed high viscosity suspension. A 3D micro structure was manufactured by using recoating system and micro-stereolithography system. The fabricated product was sintered to get the electric conductivity. After sintering, a pure copper product was made. In this study, new process was developed by making metal micro structure having an electric conductivity. This technology broadened the realm of the micro-stereolithography technology. And it will be applied to make the 3D micro structure of free form which has a high hardness and an electric conductivity in the near future.