• Applied Microscopy
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• v.19 no.1
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• pp.34-48
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• 1989

The oval shaped pericardial cells are clustered along the lateral sides of the heart and irregularly connected with the heart. The cells are bounded by a basement membrane. The basement membranes of the connected two peicardial cells are irregularly linked each other there-fore funnels are formed. The multiple invaginations of the cell membrane are observed and septate junctions develope at the part of enterance of the cell membrane. The coated pits are appeared in the inner side of the invaginated cell membrane. The coated vesicles, tubular and spherical shaped vesicle, Golgi complex containing high electron densed material in the cisternae and mitochondria are observed in the cytoplasm and lysosomes are remarkably well developed. The whirled membrane structures in the multiformed complex bounded by single membrane are linked with low electron densed granules and spherical shaped small granules having high electron density with $0.03{\mu}m$ in diameter are located between the whirled membrane in a row and gradually secretes the granules and then they produced the multilamellar body. The lysosomal regions of cytoplasm of pericardial cell are appeared negative reaction to the acid phosphatase and according to the results of the electrophoresis, lipoproteins having acid phosphatase activity are contained. The axon is contacted with the pericardial cells.

• YAKHAK HOEJI
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• v.41 no.1
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• pp.81-85
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• 1997

Ceramide has been suggested as an important mediator of the effects of extracellular agonists on cell growth inhibition, differentiation, apoptosis. However the biochemical sign aling mechanism involved in transducing the effects of ceramide on leukemia cell differentiation is still unclear. In these respects, we examined the regulatory effects of ceramide on c-jun gene expression during differentiation. In U-937 cells. ceramide increased c-jun mRNA levels in a time-dependent manner. The half life, of c-jun mRNA was 30 min. In contrast, inhibition of protein synthesis with cycloheximide in the absence, of transcription with actinomycin D increased the half-life of c-jun mRNA in ceramide-treated U-937 cells to more than 90 min. In order to examine whether ceramide-inhibited c-jun gene expression is regulated through ceramide-activated protein phosphatase (CAPP), a direct target for the action of ceramide, okadaic acid were treated to the cells. Okadaic acid inhibited enhancement of c-jun mRNA induced by C2-ceramide in a dose-dependent manner. These results suggested that ceramide increases c-jun mRNA level during differentiation in U-937 cells and regulates the gene expression on posttranscriptional level. In addition, we provide the evidence that CAPP is involved in ceramide-induced c-jun gene expression in U-937 cells.

• Journal of Microbiology
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• v.37 no.1
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• pp.21-26
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• 1999

Two novel calcium-binding proteins, named CAB-I and CAB-II, have been isolated from Streptomyces coelicolor. Purification of the calcium-binding proteins involved heat treatment, fractionation with ammonium sulfate, acid treatment, anion exchange and hydrophobic interaction column chromatography, FPLC gel filtration, and preparative isoelectric focusing. A chelex competitive assay and 45Ca autoradiography verified the calcium-binding ability of the proteins. The major band CAB-II has an apparent molecular weight of 26,000 determined by SDS-polyacrylamide gel electrophoresis and 340,000 determined by gel filtration. The isoelectric point of this molecule showed the acidic nature of the molecule. N-terminal amino acid sequence analysis shows homology to rat Ca2+/calmodulin-dependent protein kinase-II (CAB-II) and yeast phosphoprotein phosphatase (CAB-I).

• BMB Reports
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• v.56 no.7
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• pp.392-397
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• 2023

In this study, recombinant Fc-fused Prostate acid phosphatase (PAP) proteins were produced in transgenic plants. PAP was fused to immunoglobulin (Ig) A and M Fc domain (PAP-IgA Fc and PAP-IgM Fc), which were tagged to the ER retention sequence KDEL to generate PAP-IgA FcK and PAP-IgM FcK. Agrobacterium-mediated transformation was performed to produce transgenic tobacco plants expressing four recombinant proteins. Genomic PCR and RT-PCR analyses confirmed the transgene insertion and mRNA transcription of PAP-IgA Fc, PAP-IgM Fc, PAP-IgA FcK, and PAP-IgM FcK in tobacco plant leaves. Western blot confirmed the expression of PAP-IgA Fc, PAP-IgM Fc, PAP-IgA FcK, and PAP-IgM FcK proteins. SEC-HPLC and Bio-TEM analyses were performed to confirm the size and shape of the plant-derived recombinant PAP-Fc fusion proteins. In mice experiments, the plant-derived IgA and IgM Fc fused proteins induced production of total IgGs including IgG1 against PAP. This result suggests that IgA and IgM Fc fusion can be applied to produce recombinant PAP proteins as a prostate cancer vaccine in plant expression system.

• Korean Journal of Soil Science and Fertilizer
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• v.44 no.5
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• pp.801-807
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• 2011

This study was carried out to evaluate effect of consecutive application of organic matter on soil chemical properties and dehydrogenase, acid phosphatase activity in non-volcanic ash soil during three cropping season. Organic matter mixture and organic fertilizer (MOF, $2,000kg\;10a^{-1}$), food waste compost (FWC, $2,000kg\;10a^{-1}$), and pig manure compost (PMC, 2,000, 4,000, and $6,000kg\;10a^{-1}$) were applied for each cropping season. Soil pH values were increased after three cropping season in all treatment. In the soils of the increased application of PMC, soil pH, total-nitrogen, available phosphate, exchangeable cations (K, Ca, and Mg), and heavy metal (Zn and Cu) contents were increased. In addition, Soil dehydrogenase activity was significantly increased in proportions to PMC application rate and cropping season during potato cultivation period. The activity was two times higher in PMC ($4,000kg\;10a^{-1}$) than control after the third cropping season. Soil dehydrogenase activity was in order of PMC>FWC>NPK+PMC>MOF. Acid phosphatase activity was higher in PMC ($6,000kg\;10a^{-1}$) than other treatment. Soil Zn content and dehydrogenase activity showed linearly correlation, which were MOF ($R^2$=0.427), FWC ($R^2$=0.427) and PMC ($R^2$=0.411, p<0.01), respectively. This study demonstrated that soil chemical properties and enzyme activity could be affected greatly by consecutive application of different organic matter in the potato cultivation field.

• Applied Microscopy
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• v.12 no.1
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• pp.11-21
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• 1982

A number of recent ultrastructural studies have shown marked differences between the two lining cell types in adult liver sinusoids, endothelial cells and Kupffer cells. In the present study, the ultrastructural features and electron microscopic cytochemistry of sinusoidal lining cells in the fetal liver were studied through fetal period to neonate in the rat. At fetal period, the sinusoid, which contains various blood component, in lined by the endothelial cells, the Kupffer cells and the fat storing cells that located in the space of Disse. As gestation proceeded, these eel's are arranged as adult liver sinusoids. The sinusoidal wall appears to be discontinuous with open fenestration between endothelial cells, but no basal lamina can observed. It seems to be morphologically and functionally distinct at the early gestation between the endothelial cells and the Kupffer cells, the latter showing marked phagocytized activity. The fat storing cells, which contain several fat droplets, are located in the space of Disse. Ultrastructural localization of the acid and alkaline phosphatase activity were noted on the sinusoidal lining cells.

• BMB Reports
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• v.30 no.3
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• pp.222-228
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• 1997

Protein phosphatase 2C (PP2C) is one of the four major serine/threonine phosphatases which is dependent on $Mg^{2+}$ for its activity. PP2C was purified from rat liver cytosol and its characteristics were investigated. The substrate employed for routine assay was $[^{32}P]casein$ phosphorylated by PKA. The purification process involved DEAE chromatography, ammonium sulfate fractionation, phenyl sepharose chromatography, sephacryl 5-200 gel filtration, and histone agarose chromatography. The SDS-PAGE of PP2C showed one major single protein band at a position corresponding to a molecular mass of 43 kd and the purification fold was 637. The enzyme showed a pH optimum of 8 and $K_M$ value was $1.9\;{\mu}M$. However, when the substrate was changed to $[^{32}P]histone$, the pH optimum was shifted to 7 and $K_M$ value was $2.3\;{\mu}M.\;Mg^{2+}$ was essential to the enzyme activity and okadaic acid did not exert any inhibitory effect on the enzyme. To examine residue in the active site of PP2C effects of some protein-modifying reagents were tested.

• Korean Journal of Clinical Laboratory Science
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• v.43 no.1
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• pp.6-11
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• 2011

In clinical chemistry tests, the interfering substances such as hemoglobin, lipid, bilirubin, and drugs, etc. can cause the changes of test results performed by spectrophotometrical methods. We evaluated the effects of interfering substances on the test results by adding interfering substances on the samples in the 19 kinds of clinical chemistry tests such as aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, lactate dehydrogenase, gamma-glutamyltransferase, total protein, albumin, glucose, total cholesterol, total bilirubin, triglyceride, uric acid, calcium, inorganic phosphours, high density lipoprotein cholesterol, low density lipoprotein cholesterol, creatinine, blood urea nitrogen, and C-reactive protein using newly implemented automatic chemical analyzer Toshiba TBA-C8000 under the direction of CLSI EP07-A guideline. Hemolytic samples show increased concentration of total protein, aspartate aminotransferase, alanine aminotransferase, and lactate dehydrogenase and reduced concentration of total bilirubin, alkaline phosphatase by interfering effect. Hyperlipemic samples show increased concentration of total protein and alkaline phosphatase and reduced concentration of low density lipoprotein cholesterol. The samples with conjugated bilirubinemia show increased concentration of inorganic phosphours, otherwise the samples with unconjugated bilirubinemia show no interference or allowable range in the test result.

• Applied Biological Chemistry
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• v.51 no.3
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• pp.171-176
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• 2008

The comparative molecular similarity indices analysis (CoMSIA) models between 3${\beta}$-Hydroxy-12-oleanen-28-oic acid (1-30) analogues as substrate molecule and their inhibitory activities ($pI_{50}$) against protein tyrosine phosphatase (PTP)-1B were derived and discussed quantitatively. Listing in order, the CoMFA>CoMSIA${\geq}$HQSAR>2D-QSAR model, these QSAR models had the better statistical values. The optimized CoMSIA F1 model at grid 3.0${\AA}$ had the best predictability and fitness ($q^2$=0.754 and $r^2$=0.976) by field fit alignment. The order of contribution ratio (%) of CoMSIA fields concerning the inhibitory activities was a H-bond acceptor (48.9%), steric field (25.8%) and hydrophobic field (25.4%), respectively. Therefore, the inhibitory activities of substrate molecules against PTP-1B were dependent upon H-bond acceptor field (A) of $R_4$-group. From the analytical results of CoMSIA contour maps, oleanolic acid derivatives will have better inhibition activities if $R_1$ group has H-bond acceptor disfavor, $R_3$group has steric disfavor and $R_4$ group has steric, hydrophobic, H-bond favor.

• Journal of Microbiology and Biotechnology
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• v.16 no.2
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• pp.272-279
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• 2006

The gene encoding Thermus sp. T351 alkaline phosphatase (T351 APase) was cloned and sequenced. The gene consisted of 1,503 bp coding for a protein with 500 amino acid residues including a signal peptide. The deduced amino acid sequence of T351 APase showed relatively low similarity to other Thermus APases. The T351 APase gene was expressed under the control of the T7lac promoter on the expression vector pET-22b(+) in Escherichia coli BL21 (DE3). The expressed enzyme was purified by heat treatment, and $UNO^{TM}$ Q and $HiTrap^{TM}$ Heparin HP column chromatographies. The purified enzyme exhibited high activity at extremely alkaline pHs, reaching a maximum at pH 12.0. The optimum temperature of the enzyme was $80^{\circ}C$, and the half-life at $85^{\circ}C$ was approximately 103 min. The enzyme activity was found to be dependent on metal ions: the addition of $Mg^{2+}$ and $CO^{2+}$ increased the activity, whereas EDTA inhibited it. With p-nitrophenyl phosphate as the substrate, T351 APase had a Michaelis constant ($K_{m}$) of $3.9{\times}10^{-5}M$. The enzyme catalyzed the hydrolysis of a wide variety of phosphorylated compounds.