The cloning and expression of $\beta-glucosidase$ II, encoded by the gene ${\beta}glu2$, from thermotolerant yeast Pichia etchellsii into Escherichia coli is described. Cloning of the 7.3 kb BamHI/SalI yeast insert containing ${\beta}glu2$ in pUC18, which allowed for reverse orientation of the insert, resulted in better enzyme expression. Transformation of this plasmid into E. coli JM109 resulted in accumulation of the enzyme in periplasmic space. At $50^{\circ}C$, the highest hydrolytic activity of 1686 IU/g protein was obtained on sophorose. Batch and fed-batch techniques were employed for enzyme production in a 14 L bioreactor. Exponential feeding rates were determined from mass balance equations and these were employed to control specific growth rate and in turn maximize cell growth and enzyme production. Media optimization coupled with this strategy resulted in increased enzyme units of 1.2 kU/L at a stabilized growth rate of $0.14\;h^{-l}$. Increased enzyme production in bioreactor was accompanied by formation of inclusion bodies.
Objective : Spondylitis is often chemotherapy resistant and requires long-term treatment. Without adequate chemotherapy, the outcome can be fatal or result in severe neurologic damage. Therefore, differentiating the etiology of spondylitis is very important, particularly in spontaneous cases. As the prevalence of tuberculosis in Korea has decreased in recent years, updated clinical research about spondylitis is warranted. Methods : From April 2010 to March 2016, data from spondylitis patients were collected retrospectively. In total, 69 patients (51 with pyogenic spondylitis and 18 with tuberculous spondylitis) were included. Clinical data, laboratory findings including erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) level, measurements of Cobb angles at the initial and final follow-up, and radiologic features on magnetic resonance imaging (MRI) scans were evaluated. To test differences between the pyogenic and tuberculous groups, numerical data were compared using the student's t-test and Mann-Whitney U test, and categorical data were compared using the chi-square test and Fisher's exact test. Results : The patients' mean age was 60.0 years. Male sex was slightly predominant (56.5%). There was no difference in mean age and sex between the two groups. The pyogenic group had a relatively higher proportion of immunocompromised patients. The peak CRP value was higher in the pyogenic group than in the tuberculous group (14.08 mg/dL and 8.50 mg/dL, respectively, p=0.009), whereas the ESR was not significantly different between the groups (81.5 mm/h and 75.6 mm/h, respectively, p=0.442). Radiologically, the presence of disc space sparing and vertebral body collapse differed between the groups. In the tuberculous group, the disc was more commonly preserved on contrast-enhanced MRI (50% and 23.5%, respectively, p=0.044), and vertebral body collapse was more common (66.6% and 15.7%, respectively, p<0.001). The mean length of hospitalization was longer in the pyogenic group (56.5 days and 41.2 days, respectively, p=0.001). Four mortality cases were observed only in the pyogenic group. The most commonly isolated microorganism in the pyogenic group was Staphylococcus aureus(S. aureus) (methicillin susceptible S. aureus and methicillin resistant S. aureus [MRSA] in 8 and 4 cases, respectively). Conclusion : The clinical and radiological manifestations of spontaneous spondylitis differ based on the causative organism. Pyogenic spondylitis patients tend to have a higher CRP level and a more severe clinical course, whereas tuberculous spondylitis patients present with destruction of the vertebral body with disc sparing more frequently. The presence of MRSA is increasing in community-acquired spondylitis cases.
A new concept of "photo" -antisense method has been evaluated, where the inhibition of gene expression by the conventional antisense method is enhanced by photochemical binding between antisense oligonucleotides conjugated with photo-reactive compound and target mRNA or DNA. Fluorescein labeled oligodeoxyribonucleotides (F-DNA) was delivered to cell nuclei in the encapsulated form in multilamellar lecithin liposomes with neutral charge. F-DNA was previously shown to photo-bind to the complementary stranded DNA, and the delivery system using neutral liposome to be effective in normal human keratinocytes. In the present study, we used human kidney cancer G401.2/6TG.1 cell line to be advantageous in reproducible experiments. p53 was adopted as a target gene since antisense sequence information has been accumulated. The nuclear localization ofF-DNA was identified by comparing the fluorescence ofF-DNA with that of Hoechst 33258 under fluorescence microscope. After 7hr incubation to accumulate p53 protein induced by UV -B, p53 protein was quantified by Western blot. After 2hrs from F-DNA application, about 30% of cell population incorporated F-DNA in their nuclei with some morphological change possibly due to liposomal toxicity. Irradiation of visible light longer than 400nm from solar simulator at this time enhanced the inhibitory action of antisense F-DNA. The present results suggest that photo-antisense method is promising to control gene expression in time and space dependent manner. Further improvement of F-DNA delivery to cancer cells in the stability and toxicity is in progress. progress.
Objectives : The purpose of this study was to investigate the anti-caner effect of cobrotoxin on the prostatic cancer cell line(PC-3). Methods : After the treatment of PC-3 cells with cobrotoxin, we performed fluorescence microscope, MTT assay, Western blotting, Flow cytometry, PAGE electrophoresis and Surface plasmon resonance analysis to identify $NF-{\kappa}B$ the change of calcium and NO. Results : 1. The expression of $NF-{\kappa}B$ was decreased at 1nM and It·as decreased significantly at 2, 4, 8nM. 2. $I{\kappa}B,\;NF-{\kappa}B$ inhibitor, was decreased significantly at 8nM and $p-l{\kappa}Ba$, phosphrylation of $I{\kappa}B$, was decreased significantly at all concentrations of cobrotoxin. 3. The expressions of p50 and p65 were decreased significantly and dose-dependently at 1, 2, 4, 8nM. 4. The expression of p53 was increased significantly at 1, 2, 4, 8nM. 5. The calcium concentration in cell wasn't changed at 1, 2, 4, 8nM, but was increased dose-dependently at 30, 70, 130, 250nM comparing with lower dose of cobrotoxin. 6. The NO concentration in cell was increased significantly at 1, 2, 4, 8nM. 7. In immunochemical staining, we found that cobrotoxin-immunochemical complex move into intracellular space dose-dependently. Conclusion : These results indicate that cobrotoxin has anti-cancer effects by inducing apoptosis.
PURPOSE. This in vitro study evaluated the effects of four different cements on the color attributes of a zirconia ceramic. MATERIALS AND METHODS. 40 zirconia ceramic disk specimens (0.5 mm thickness, 10 mm diameter, 0.1 mm cement space) were fabricated by a computer-aided design and computer-aided manufacturing system. The specimens were divided into 4 groups of 10 specimens and cemented to composite substrates using four different cements including: Glass Ionomer, Panavia F2.0, Zinc Phosphate, and TempBond. The $L^*$, $a^*$, and $b^*$ color attributes of the specimens were measured before and after cementation by a spectrophotometer. Additionally, ${\Delta}E$ values were measured to determine color changes for the groups and then compared with the perceptional threshold of ${\Delta}E=3.3$. Repeated Measures ANOVA, Tukey Post Hoc, Bonferroni, One-way ANOVA, and One-sample t-test tests were used to analyze the data. All tests were carried out at the 0.05 level of significance. RESULTS. Statistically significant differences were detected in the ${\Delta}E$ values for Zinc Phosphate (P<.0001) and TempBond (P<.0001) groups. However, there were no statistically significant differences in this respect for Glass Ionomer (P=.99) and Panavia F2.0 (P=1) groups. The means and standard deviations of the ${\Delta}E$ values for Glass Ionomer, Panavia F2.0, Zinc Phosphate, and Tempbond groups were $2.11{\pm}0.66$, $0.94{\pm}0.39$, $5.77{\pm}0.83$, and $7.50{\pm}1.16$ Unit, respectively. CONCLUSION. Within the limitations of this study, it was concluded that Zinc Phosphate and Tempbond cements affected the color attributes of the tested zirconia ceramic beyond the perceptional threshold. However, Glass Ionomer and Panavia F2.0 cements created acceptable color changes.
Journal of the korean academy of Pediatric Dentistry
/
v.34
no.4
/
pp.639-645
/
2007
Removable or fixed space maintainer could be needed if one or some anterior primary teeth were missing, where resin pontics well-matched to natural primary teeth should be demanded to get an esthetic satisfaction. Resin Natural $Teeth^{TM}$(Nissin dental, Japan) is available currently in Korea, which consists of two shades of colors; type A1 and A2. The purpose of this study is to elucidate the colors of the anterior primary resin teeth and to establish the data to compare those with the natural anterior primary teeth. CIE $L^*a^*b^*s$ were measured each three times labially from 17 sets of maxillary four anterior teeth for type A1 and A2 Resin Natural $Teeth^{TM}$ using ShadeEye $NCC^{TM}$(Shofu, Japan) which is one of spectrophotometers. The data were analysed statistically using Kruskall-Wallis Test and Mann-Whitney U Test. The results were as follows : 1. There were smaller teeth color differences in group A1 than in group A2 when it comes to distributions of ${\Delta}E$, $L^*$ and $b^*$. There were no statistically significant differences of $a^*$ between teeth in the same group(P>0.05). 2. ${\Delta}E$ in group A1 and A2 (maximum ${\Delta}E{\le}1.23$) were very small, which could not be discerned by eyesight. ${\Delta}E$ between mean CIE $L^*a^*b^*s$ of group A1 and A2 was 3.97, which could be discernible by eyesight. 3. Mean measurements of group A1 were $L^*=73.8$, $a^*=-1.8$, $b^*=-4.7$, and those of group A2 were $L^*=75.8$, $a^*=-2.7$, $b^*=-1.4$. It would be recommended that resin teeth compatible to the colors of the natural primary ones needed to be developed by investigating in vivo study.
Nitrogen dioxide ($NO_2$) gas damage on vegetable crops commonly occurs in plastic film houses where relatively large amounts of $NO_3{^-}$ are applied in acid soils. In acid soils, $HNO_2$ can be formed from the $NO_2{^-}$ accumulated during denitrification, and $NO_2$ can be evolved from the chemical self-decomposition of $HNO_2$. In this study, $NO_2$ gas production and its detrimental effects on plants were investigated in soils of various conditions to elucidate the mechanisms involved in the gas production. A silty loam soil was amended with $NO_3{^-}$ (500 mg N $kg^{-1}$) and glucose, and pH and moisture of the soil were adjusted respectively to 5.0 and 34.6% water holding capacity (WHC) with 0.01 M phosphate buffer. The soil was placed in a 0.5-L glass jar with strawberry leaf or $NO_2$ gas absorption badge in air space of the jar, and the jar was incubated at $30^{\circ}C$. After 4-5 days of incubation, dark burning was observed along the outside edge of strawberry leaf and $NO_2$ production was confirmed in the air space of jar. However, when the soil was sterilized, $NO_2$ emission was minimal and any visible damage was not found in strawberry leaf. In the soil where water or $NO_3{^-}$ content was reduced to 17.3% WHC or 250 mg N $kg^{-1}$, $NO_2$ production was greatly reduced and toxicity symptom was not found in strawberry leaf. Also in the soil where glucose was not amended, $NO_2$ production was significantly reduced. In soil with pH of 6.5, $NO_2$ was evolved to the level causing damage to strawberry leaf when the soil conditions were favorable for denitrification. However, compared to the soil of pH 5.0, the $NO_2$ production and its damage to plants were much less serious in pH 6.5. Therefore, the production of $NO_2$ damaging plants might be occurred in acid soils when the conditions are favorable for denitrification.
The purpose of this study is to construct ecological pond using LID technique in order to create naturally comfortable community space in urban idle space. The specification of the ecological pond is $110m^2$ of surface area, $0.45{\pm}0.02m$ of average depth, and bed material is composed of gravel (diameter ${\leq}60mm$), sand (diameter ${\leq}2mm$) and bentonite. Rainfall and water depth monitoring were conducted to determine the annual characteristics of inflow of the water for the ecological pond, result of total rainfall was 1,287 mm and showed a seasonal imbalance that accounted for 71.3% (918 mm) during July to August, but the annual mean water depth was kept constant at $0.45{\pm}0.02m$ due to the secondary water source. Annual trends of basic water quality showed a significant changes according to the season, such as water temperature ($5.2{\sim}28.8^{\circ}C$), DO (5.0 ~ 13.8 mg/L), EC ($113{\sim}265{\mu}S/cm$). BOD, COD, TN, and TP in physicochemical water quality tended to increase after October, but the ion parameters such as $NH_3$ and $PO_4{^{3-}}$ were generally low. Phytoplankton indicators Chl-a and BGA (blue green algae) showed a sharp increase from July to August, and green algae (Selenastrum bibraianum, Pediastrum boryanum etc.) and filamentous blue green algae (Phormidium sp.) emerged as a dominant species. The ion parameters ($F^-$, $Na^+$, $K^+$, $Mg^{2+}$, $Ca^{2+}$) were strongly correlated with the $Cl^-$ as a conservative substance (R=0.70~0.97, p<0.05). Water quality was influenced by the ambient environment such as seasonal changes or rainfall, and it was closely related to fluctuation of the inflow of the water. In the future, it is necessary to consider ecological connections by referring to the characteristics surveyed in this study in order to effectively manage the water quality and biodiversity of the ecological pond in idle space.
X-ray methods have been used to determine the chain conformation and packing of the thermotropic liquid crystalline copolyester prepared from 50% tarephthaloyl chloride(TPA) and 50% (1-phenylethyl) hydroquinone(PEHQ). The x-ray patterns of annealed melt-spun fibers contain a series of annealed melt-Pointing to a well ordered crystalline structure, despite the random sense(2 or 3-) of the 1-phenylethyl substiuttion on the TPA-hydroquinone backbone. The crystalline fiber is monoclinic with space group P2l and the unit cell has dimensions 11=12.77 A, b=10.17 A (upique axis), c=12.58 h (fiber axis). and β=90.1° and contains TPA-PEHO units of to or chains. The random substitution of 1-phenylethyl groups was modelled by placing these groups at both the 2and 3 positions and giving each a weight of one-hal(. T he structure has been refined by linked a rom least square methods(LALS) against 16 observed and 21 unobserved reflections. and had a final R value of 0.20. Packing of the side chains is effected by staggering adjacent chains along the b axis by approximately c/2, so that the side chains are interleaved. The phenyl-COO and COO-phenyl torsion angles are -6.1 and 65.6, respectively, such that the main chain phenyls are mutually inclined at 59.5 (the ester groups are assumed to be planar). These torsion angles compare very well with those for the model compounds, notably phenylbenzoate, and can be used in future analyses of the structures of more complex random sequence copolyesters.
The study ooramed has the meanngs as a goal, to examine a character and changes of function a "Maru" in the literatures historically. Methodically the literatures in the temporal oonsequences were selected, and the "Maru", whidl oomes into it, was interpreted in its historical oonnections. By the investigation I oould oome to the following results. First of all, those terms "Maru" and "Daechll1Q" were cfifferently used after the sizes and p1ares of the spaces concerned. I.e, the "Maru" in the center of the house is called "Daed1ung", With the Maru does not concern actually its p1are, Secondly, the functions of the spare "Maru" are characterized as follows. ·Space for the important ceremonies ·Spare for the passage and the switching of two or several spaces. ·Determination of the hierardly of spares in the whole house. ·Spare for the receipt. ·Spare for sleeping and for living. ·Spare for work, storage etc. Thirdly, the meanings of the "Maru" were dlanged as follows in the time. ·The term "Man! was used from the past to today without interrujjion. Arot.l1d 1930 the tenns "Yangsil" and "Geosil" for the tenn "Maru" were emerged. ·The different dlaraderistics for the space "Maru" are found in the cortemporary literatures only rare. This thing explains itself by the fad that the "Maru" becomes the inner-spare after 1900 by the restructuring of the dwelling spares.hing explains itself by the fad that the "Maru" becomes the inner-spare after 1900 by the restructuring of the dwelling spares.
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