This study was conducted to investigate the genetic analysis of esterase isozymes in maize with tillers. The materials used for the study were stele tissue for five day old seedlings of IK inbred line with tiller and A-type inbred line with no tiller, their Fl and F,. The methods employed for the study were same as previous report by Lee and Choe. A total of thirteen isoesterase enzyme bands were identified and five zones were distinguised according to both migration distance and genetic segregation patterns. The E$\sub$0.3/, E$\sub$0.4/ and E$\sub$0.5/ loci appeared from orgin to 0.5cm migration distance were controlled by the two alleles in (IK/A-type)F$_2$ and the E$\sub$0.3/+E$\sub$0.4/ of variants was controlled by codominant alleles. The E$\sub$1.0/, E$\sub$1.2/, E$\sub$1.5/ and E$\sub$1.8/ loci appeared from 1.0cm to 1.8cm were also controlled by the two alleles. However, the null band was functioned alleles. The E$\sub$2.8/, E$\sub$3.0/ and E$\sub$3.5/ loci appeared from 2.8cm to 3.5cm migration distance were very active and near location. A total of individuals with two paried bands of these loci were more than those of three paired bands(x$^2$=0.327$\^$**/). The activity of bands appeared over 3.8cm were very low and these were controlled by the two alleles. In above results, genetic segregations of stele tissue of maize with tillers were suggested to be controlled by Mendelian genetic laws.
This research was conducted to determine the amount of water absorbed by a polyacrylamide hydrogel such as Stocksorb C (STSB), effect of salts on inhibition in hydration of STSB, and the hydrogel effects on changes of nutrient concentration in external solution. Absorption of deionized water by STSB reached a maximum of 180 $mL{\cdot}g^{-1}$. Monovalent soluble salts such as $KH_2PO_4,\;KNO_3$, and $(NH_4)_2SO_4$ reduced absorption of the hydrogel, but the degrees of inhibition in absorption were similar in three kinds of salts. Twenty milliequivalents per liter of $Ca_{2+}\;or\;Mg_{2+}$ reduced water absorption of STSB to $14\%$ compared to those of deionized water. Solution absorption was consistently lower in the presence of divalent cations than in the presence of the monovalent cations. But the absorption was unaffected by the uncharged salt such as urea in all concentrations tested. The final $K^+\;and\;NH_4^+-N$ concentrations of the solution remaining after absorption by STSB was higher than those of the initial solution. The soaking of STSB to full strength of Hoagland solution resulted in increase of $NO_3^--N,\;H_2PO_4^-\;and\;SO_4^{2-}$ concentrations in external solution compared to initial solution, reaching 5,300, 250 and 1,500 $mL{\cdot}g^{-1}$, respectively, at 24 hrs after soaking.
Three-dimensional microscopic approaches in histopathology display multiplex properties that present puzzling questions for specimens as related to their comprehensive volumetric information. This information includes spatial distribution of molecules, three-dimensional co-localization, structural formation and whole data set that cannot be determined by two-dimensional section slides due to the inevitable loss of spatial information. Advancement of optical instruments such as two-photon microscopy and high performance objectives with motorized correction collars have narrowed the gap between optical theories and the actual reality of deep tissue imaging. However, the benefits gained by a prolonged working distance, two-photon laser and optimized beam alignment are inevitably diminished because of the light scattering phenomenon that is deeply related to the refractive index mismatch between each cellular component and the surrounding medium. From the first approaches with simple crude refractive index matching techniques to the recent cutting-edge integrated tissue clearing methods, an achievement of transparency without morphological denaturation and eradication of natural and fixation-induced nonspecific autofluorescence out of real signal are key factors to determine the perfection of tissue clearing and the immunofluorescent staining for high contrast images. When performing integrated laboratory workflow of tissue for processing frozen and formalin-fixed tissues, clear lipid-exchanged acrylamide-hybridized rigid imaging/immunostaining/in situ hybridization-compatible tissue hydrogel (CLARITY), an equipment-based tissue clearing method, is compatible with routine procedures in a histopathology laboratory.
When the cattail pollen was identified by using fibrinolytic agents, we found that the fibrinolytic activity was controlled by an enzyme. Therefore, for determining the fibrinolytic activity of cattail pollen, the fibrinolytic enzyme in cattail pollen was purified by gel filtration using DEAE-cellulose, Sephadex G-150 and HPLC. Also, its purity was certified by polyacrylamide gel electrophoresis, and its physico-chemical properties, such as pH and temperature stabilities and effects of metal, inhibitors and substrates, were examined. The specific activity, purification fold, and molecular weight of the enzyme were 38U/mg, 86.4,and 75kDa, respectively. The optimum pH for the purified enzyme was at 4.0 and it was stable at pH 4.0-6.0. The optimum temperature was $55^{\circ}C$ and it was stable at $30-60^{\circ}C$. But the enzyme began to be inactivated at $70^{\circ}C$ and its activity was totally lost at temperatures above $80^{\circ}C$. As for substrate specificity, the enzyme was most effective in dissolving fibrin, followed by whole casein, ${\kappa}$-casein, ${\alpha}$-casein, ${\beta}$-casein, and BSA. With casein as the substrate, Km value was found to be 0.44mM and the enzyme showed a high affinity for casein. As for the metal ions affecting enzyme activity, $K^+$, $Na^+$, and $Mg^{2+}$ had no effect on enzyme reaction while $Zn^{2+}$ and $Fe^{2+}$ showed potent inhibitory activity. Judging from the fact that the purified enzyme was also strongly inhibited by PMSF, iodoacetic acid, and SDA, it assumed to be a serine protease.
Bead type super-absorbent resins to be used for release-control were prepared by modification of the inverse suspension polymerization, and their physical properties were characterized. Acrylic acid and acrylamide were used as monomers, and N,N-methylenebisacrylamide was used as crosslinker, controlling the viscosity of monomer solution by adding hydroxyethylcellulose (HEC). SEM studies of the synthesized beads verified that the bead surfaces had many pores with their diameters of several tens nm. The bead sizes were in the range of $500{\sim}3000{\mu}m$, depending on the viscosity of the monomer solution. Both absorbent amount and absorbent rate of the beads were inversely proportional to the bead size, and the maximum water absorbent amount of 1 g beads was determined to be ca. 170~200 g for 5 hrs. The absorbent rate was also dependent on pH change of the aqueous solution, exhibiting the maximum rate in pH ranging from 5 to 11. The absorbent rate decreased as the concentration of salt (NaCl and $MgCl_2$) or ethanol and ethylene glycol increased. Release time of the water absorbed into the bead resins was 700 hrs, confirming the usefulness of the resin for the good release-control materials.
Park, Mijung;Lee, Keum Hee;Lee, Eun Kyung;Park, Sang Hee;Kim, So Ra;Lee, Heum Sook
Journal of Korean Ophthalmic Optics Society
/
v.13
no.4
/
pp.43-49
/
2008
Purpose: The current study was conducted to evaluate the compatibility of UV-A blocking contact lens on eye protection with regular contact lens. Methods: The protective activity of regular contact lens (UV-A blocking: 20%) and UV-A blocking contact lens (UV-A blocking: 85%) on the denaturation of RNase A, catalase, and superoxide dismutase (SOD) induced UV-A irradiation were compared by acrylamide gel electrophoresis. The enzyme solutions were irradiated with UV-A for 1, 3, 6, 24 and 96 hours at the wavelength of 365 nm. Covering area with contact lenses were varied as 50%, 70% and 100% according to the calculation of blocking areas of anterior eye that could be covered with RGP lens, soft contact lens, and eye glasses, respectively. Results: Denaturations of RNase, catalase and SOD were exaggerated when they were exposed to UV-A for a longer period. The denaturation was effectively prevented by UV-A blocking contact lens compared to regular contact lens. The capability of UV-A blocking contact lens was considerably reduced when the covering area with contact lens decreased and exposure time to UV-A extended. Conclusion: Therefore, it would be suggested that wearing contact lens for a long time under sunlight is carefully considered since the activity of UV-A blocking contact lens against UV-A irradiation may not be enough to protect enzymes presented in eyes when exposure time to UV-A increased.
This experiment was conducted to investigate the combined effects of radiation and antiseptics on the keeping qualities of pork sausage, which was treated with potassium sorbate and AF-2(2-(-2-furyl)-3-(5-nitro-2-furyl)-acrylamide), and then followed by gamma radiation of 0.25, 0.5, and 0.75 Mrad. Amounts of treated antiseptics were a quarter, half, and full levels of their maximum permissible concentration. Irradiated and unirradiated sausages were stored for 50 days at $5^{\circ}C\;and\;25^{\circ}C$, and their changes in rancidity, volatile basic nitrogen, bacterial counts, pH, and sensory analysis were examined during the storage period. The results obtained are as follows: 1) Preservative effects of antiseptics were manifested at cold storage; antiseptics treatment of a quarter-level and unirradiation following low-temperature storage showed the same good keeping qualities as the combined treatment of full-level antiseptics and radiation of 0.25 Mrad following high-temperature storage. 2) There did not appear to recognize irradiation-odor, while color and odor were deteriorated intensively by storage temperature. Sausage irradiated with 0.75 Mrad has shown slightly noticeable off-odor at the end of storage at $25^{\circ}C$. 3) The most suitable radiation dose was considered to be 0.5 Mrad, which could extend the storage life about $2{\sim}3$ times longer than untreated.
To produce ${\beta}-cyclodextrin({\beta}-CD)$, a cyclodextrin glucanotransferase(CGTase) producing Aspergillus sp. CC-2-1 was isolated from soil. The enzyme was purified and its enzymological characteristics were investigated. It was found that production of CGTase reached to the maximum when the wheat bran medium containing 0.1% albumin, 2% $(NH_4)_2S_2O_8$, 2% soluble starch and 0.2% $KH_2PO_4$ was cultured for 5 days at $37^{\circ}C$. The purity of CGTase was increased by 13.14 folds after DEAE-cellulose ion exchange chromatography and Sephadex G-100, G-150 gel filtration and the specific activity was 172.14 unit/mg. Purified enzyme was confirmed as a single band by the polyacrylamide gel electrophoresis. The molecular weight of CGTase was estimated to be 27,800 by Sephadex G-100 gel filtration and SDS-polyacrylamide gel electrophoresis. The optimum pH and temperature for the CGTase activity were 9.0 and $80^{\circ}C$, respectively. The enzyme was stable in pH $8.0{\sim}11.0$ at $60{\sim}80^{\circ}C$. The activity of purified enzyme was activated by $K^+,\;Cu^{2+}$ and $Zn^{2+}$. The activity of the CGTase was inhibited by the treatment with 2,4-dinitrophenol and iodine. The result suggests that the purified enzyme has phenolic hydroxyl group of tyrosine, histidine imidazole group and terminal amino group at active site. The reaction of this enzyme followed typical Michaelis-Menten kinetics with the $K_m$ value of 18.182 g/L with the $V_{max}$ of 188.68 ${\mu}mole/min$. The activation energy for the CGTase was calculated by Arrhenius equation was 1.548 kcal/mol.
This study was conducted to elucidate phylogenetic relationships and genetic characterization of silkworms that might be recognized as the Korean native strains. Genetic characterization in isozymes and the proteins of larval hemolymph of 17 silkworms were observed by acrylamide gel eletrophoresis, on 12 genes; Bph, Bes, les, Amy-hc, Ict-A, -B, -D,-E,-H, Pfl, Pst, Lp. Gene frequencies in each locus were compared other geographic strains. Korean native strains were remarkably different from others considered as the genetic characterization of Korean native strains. Phylogenetic relationships in Korean native strains were analysed using RAPD-PCR markers. A total of 40 primers were used and 346 bands of amplified DNA were generated from geographic strains. Genetic similarity based on the RAPD bands was used to construct phylogenetic dendrogram based on analysis of bard sharing data of amplified markers. Genetic similarity ranged from 0.595 to 0.860. In the genetic relationship based on dendrogram, they were classified into Bombyx mori group (including 16 domesticated silkworm strains) and B. mandarina group. The Bombyx mori group was separated into three sub-groups at the genetic similarity of 0.6930, including Korean, Japanese and Chinese groups. According to this result, the Korean native variety can be considered as a clearly different variety from other geographic strains. It may be concluded that the Korean native strains are also one of original geographic variety such as Japanese, Chinese, etc.
Aegilops genus is known to include the donor species of the Band D genome of the bread wheat(ABD). An effort to establish a better strategy for phylogenetic relationships about Aegilops polyploids by AFLPs(Amplified Fragment Length Polymorphisms) was conducted using the 19 Aegilops sPP. and T. aestivum. The 207 polymorphic bands from the amplified products on the 6% acrylamide denaturing sequencing gels were obtained with the 7 AFLP primer combinations, and used to account for the genetic similarities and cluster analysis using NTSYS program. According to the genome analysis, the $M^h$-genome of Ae. heldreichii was estimated as an intermediate genome between the M-genome of Ae. comosa and N-genome of Ae. uniaristata and supposed to be incorporated in the establishing process of UM-genome as a possible diploid donor. And Ae. ventricosa(DN) was more close to Ae. umbellulata(U) than Ae. squarrosa(D). The close relationship between Ae. squarrosa and T. aestivum was perceived as a diploid donor of D-genome. As for the polyploid species, hexaploid Ae. triaristata was more closely related to Ae. columnaris rather than tetraploid Ae. triaristata. The clustered groups were, basically same to the previous Gihara's sections based on phenotypes and pairing analysis of interspecific hybrids. AFLP was evaluated as an efficient and powerful method in the genome evaluation of closely related species.
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