• Reproductive and Developmental Biology
• /
• v.33 no.2
• /
• pp.77-83
• /
• 2009

Erythropoietin (EPO), a glycoprotein hormone produced from primarily cells of the peritubular capillary endothelium of the kidney, is responsible for the regulation of red blood cell production. We have been investigating the roles of glycosylation site added in the biosynthesis and function of recombinant protein. We constructed three EPO mutants ($\Delta$69, $\Delta$105 and $\Delta$69,105), containing an additional oligosaccharide chains. EPOWT and EPO$\Delta$69 were effectively expressed in transient and stably transfected CHO-K1 cell lines. But, it wasn't detected any protein in the culture medium of EPO$\Delta$105 and EPO$\Delta$69,105 mutants. The growth and differentiation of EPO-dependent human leukemic cell line (F36E) were used to measure the cytokine dependency and in vitro bioactivity of rec-hEPO. MTT assay values were increased by survival of F36E cells at 24h. To analysis biological activity in vivo, two groups of ICR-mice (7 weeks old) were injected subcutaneously with 10 IU per mice of rec-hEPO molecules on days 0 and 2. Red blood cell and hematocrit values were measured on 6 days after the first injection. The hematocrit values were remarkably increased in all treatment groups. The pharmacokinetic analysis was also affected in the mice injected with rec-hEPO molecules 2.5 IU by tail intravenous. Protein samples were detected by Western blotting. An EPO$\Delta$69 protein migrated as a broad band with an average apparent molecular and detected slightly high band. Enzymatic N-deglycosylation resulted in narrow band and was the same molecular size. The biological activity of EPO$\Delta$69 was enhanced to compare with wt-hEPO. The half-life was longer than wt-hEPO. The results suggest that hyperglycosyalted recombinant human erythropoietin (EPO$\Delta$69) may have important biological and therapeutic good points.

• The Korean Journal of Food And Nutrition
• /
• v.31 no.5
• /
• pp.689-695
• /
• 2018

This study was carried out to select apple cultivars suited for the fresh cutting or minimal processing as investigating correlation between total phenol and ascorbic acid as browning factors and the enzymatic browning degree. In soluble solid-acid ratio, 'Gamhong' and 'Arisoo' were highest value of 71.69 and 71.52 compared with other cultivars. And the change of lightness (${\Delta}L$) and color (${\Delta}E$) in 'Gamhong' and 'Arisoo' showed lower than other cultivars while 'Arkansas black' was highest change of lightness and color as ${\Delta}L$ 8.27, ${\Delta}E$ 13.86 respectively. Total phenol and ascorbic acid contents in 'Arkansas black' were highest such as 334.3 mg GAE/100 g and 9.22 mg/100 g, respectively. And polyphenol oxidase (PPO) activity also was the highest such as 14.43unit in 'Arkansas black'. The correlation test showed browning of apple was significantly dependent on ${\Delta}L$, ${\Delta}E$, total phenol, ascorbic acid contents and PPO activity (p<0.01). On the contrary, no significant correlation had been determined between the browning and ascorbic acid oxidase (AAO) activity. Thus, it seemed that the browning of apple was not only due to oxidation of ascorbic acid but also total phenol. It was suggested that 'Gamhong' and 'Arisoo' were suitable for fresh cutting or minimal processing.

• Journal of the Korean Chemical Society
• /
• v.21 no.6
• /
• pp.395-403
• /
• 1977

Significant structure theory was applied to some liquid mixture systems ranging from simple monatomic molecule systems to polyatomic molecule systems, and the activity coefficients ${\gamma}$ of the liquid mixture systems were calculated over whole mole fractions using the following thermodynamic relation $RTln_{{\gamma}i} = (\frac{{\partial A}^E}{{\partial N}_i})_{T,V,N_i} $ where $A^E$ represents the excess Helmholtz free energy, and $N_i$ is the number of molecules of component i. The activity coefficients of the solutions such as monatomic molecule systems (Ar-Kr, Kr-Xe) and diatomic molecule systems $(Ar-O_2,\;N_2-CO)$ and $CH_4-Kr$ systems whose components have similar shapes for intermolecular potential curves were calculated successfully only with the ${\delta}E_s$, correction parameter for energy $E_s$, for mixture systems. For other systems such as $Ar-N_2,\;O_2-N_2\;and\;CH_4-C_3H_8$ whose components have dissimilar intermolecular potential curve shapes an additional correction parameters ${\delta}$V and even one more parameter ${\delta}$n were necessary [see Eqs.(10)∼(12)].

• Journal of Yeungnam Medical Science
• /
• v.9 no.2
• /
• pp.288-301
• /
• 1992

The objective of the present study was to identify the characteristics of phospholipase C (PLC) isozymes purified from bovine brain and to investigate their interrelationship with G protein. The purified PLC isozymes ${\beta}$, ${\gamma}$ and ${\delta}$ were obtained and the characteristics of PLC activity on various concentrations of free $Ca^{2+}$ were observed. The activity of PLC was increased with increasing $Ca^{2+}$ concentration and the activity PLC ${\delta}$ was increased higher in the presence of phosphatidyl choline(PC) than in the abscence of PC. For vesicle formation as the structure of cell membrane, cholic acid and deoxycholic acid as detergent on phosphatidylinositol bisphosphate($PIP_2$) substrate containing PC were used, and then the activity of PLC isozymes were increased with increasing concentration of cholate, from 0.2% to 1% and were increased slightly in deoxycholate. In the $PIP_2$ containing phospholipid and glycolipid as brain extract, the activity of PLC isozymes were checked in 0.2%-1% cholic acid. The activities of PLC isoyzmes were continuously increased up to 1% cholic acid. The quantitation of PLC isozymes from several bovine organs by radioimmunoassay was made. Brain was the most sufficient organ in terms of amount of PLC ${\beta}$and ${\delta}$. A large amount of PLC ${\delta}$ was existed in adrenal gland. The binding capacity of GTPrS and G protein was observed and other observations of the binding effect of GTPrS-G protein and PLC monoclonal Ab-Protein A from tissue homogenate with PLC were made. From the observation the binding capacity was revealed the range of 0.11%-1.49%. The effects of each type of G protein on the percent activity of purified PLC isozymes were observed. From the observation, activities of isozymes were increased in $Go{\alpha}$ & Gmix, and the activities of PLC ${\beta}$ and ${\delta}$ were increased in $G{\beta}{\gamma}$ and $Gi{\alpha}$. Activities of PLC ${\beta}$ and ${\gamma}$ were decreased in $Gt{\alpha}$ but PLC ${\delta}$ increased.

• Horticultural Science & Technology
• /
• v.28 no.5
• /
• pp.850-856
• /
• 2010

An in vivo oral glucose tolerance test (OGTT) was performed on hyperglycemic male Sprague-Dawley rats to assess the effect of fruits and vegetables ($1g{\cdot}kg^{-1}$ body weight) on blood glucose levels (${\Delta}BGLs$) at different time intervals of 0, 5, 15, 30, 60, 90 and 120 min. The areas under glucose curve (${\Delta}AUCs$) were calculated at 120 min of OGTT by trapezoid method. Total phenolic content (TPC) and anti-oxidant activity (AOA) of fruits and vegetables were assayed in vitro by Folin Ciocalteu and DPPH (2, 2-diphenyl-1-picrylhydrazyl) methods, respectively. At the end of the experiment the correlations among the parameters TPC, AOA and ${\Delta}AUC$ was estimated by Pearson's correlations. Among fruit crops, tangerine, plum, grape and pear and among vegetables, blue leaf mustard, cabbage, chicory, broccoli and others exhibited significant hypoglycemic effects by reducing ${\Delta}BGLs$ with significant ${\Delta}AUC$. The effective ${\Delta}AUC$ ranged from $5548.2{\pm}462.1$ to $3823.3{\pm}282.0mg-min{\cdot}dL^{-1}$. The TPC and AOA ranged from $0.063{\pm}0.00$ to $0.913{\pm}0.14mg{\cdot}g^{-1}$ GAE and $01.05{\pm}0.08$ to $75.46{\pm}0.06%$, respectively. Overall, six fruits and fifteen vegetables exhibited higher TPC and one fruit and four vegetables exhibited higher AOA. There was a better correlation among TPC, AOA and ${\Delta}AUC$ of fruits and TPC & AOA of vegetables. We report that hypoglycemically significant fruits and vegetables investigated in this study have pharmacological importance which reduced ${\Delta}BGLs$ through insulin like activity and AOA in prevention of type-2 diabetes.

• Asian Pacific Journal of Cancer Prevention
• /
• v.16 no.18
• /
• pp.8177-8186
• /
• 2016

The present study was aimed at determining the effects of alkylating and oxidative stress inducing agents on a newly identified variant of DNA polymerase beta ($pol{\beta}{\Delta}_{208-304}$) specific for ovarian cancer. $Pol{\beta}{\Delta}_{208-304}$ has a deletion of exons 11-13 which lie in the catalytic part of enzyme. We compared the effect of these chemicals on HeLa cells and HeLa cells stably transfected with this variant cloned into in pcDNAI/neo vector by MTT, colony forming and apoptosis assays. $Pol{\beta}{\Delta}_{208-304}$ cells exhibited greater sensitivity to an alkylating agent and less sensitivity towards $H_2O_2$ and UV when compared with HeLa cells alone. It has been shown that cell death in $Pol{\beta}{\Delta}_{208-304}$ transfected HeLa cells is mediated by the caspase 9 cascade. Exon 11 has nucleotidyl selection activity, while exons 12 and 13 have dNTP selection activity. Hence deletion of this part may affect polymerizing activity although single strand binding and double strand binding activity may remain same. The lack of this part may adversely affect catalytic activity of DNA polymerase beta so that the variant may act as a dominant negative mutant. This would represent clinical significance if translated into a clinical setting because resistance to radiation or chemotherapy during the relapse of the disease could be potentially overcome by this approach.

• Restorative Dentistry and Endodontics
• /
• v.17 no.1
• /
• pp.1-9
• /
• 1992

The intradental nerve activity was recorded from single pulp nerve unit dissected from the inferior alveolar nerve in canine teeth of anesthetized cats. The effects of various test solutions on intradental nerve activity evoked by 4M NaCl applied to the deep dentinal cavities were investigated. 1. Total 15 single pulp nerve units were recorded. Of these 9 were Mi - fibers and 6 were C -fibers. The mean conduction velocity and electrical threshold of $A{\delta}$ - fiber were $6.3{\pm}3.7m/sec$, $1.2{\pm}0.7V$ and those of C - fiber were $1.0{\pm}0.2m/sec$, $2.3{\pm}1.3V$, respectively. The response to cold stimuli of $A{\delta}$ - fiber began immediately and that of C - fiber began after a latency. 2. When applied to deep dentinal cavity, 4M NaCl induced irregular bursts of action potential in 4 $A{\delta}$ - fibers and 3 C - fibers, which continued until the solution was washed away. 3. In the $A{\delta}$ - fiber, histamine failed to induce any nerve acitivity and did not produce an increase in intradental nerve activity evoked by 4M NaCl. However following the application of 1M KCl, the response to 4M NaCl was eliminated. 4. In the C - fiber, histamine generated some nerve activity and produced a significant increase in intradental nerve activity evoked by 4M NaCl, but 5M $CaCl_2$ did not abolish this enhandced response.

• Journal of Microbiology and Biotechnology
• /
• v.24 no.3
• /
• pp.401-407
• /
• 2014

Quorum sensing and the stringent response are well-known regulation systems for the expression of virulence genes in enterohemorrhagic Escherichia coli (EHEC). However, how these two systems interact is not well known. E. coli strains with mutations in two regulation systems, ${\Delta}luxS$ (ECM101) and ${\Delta}luxS{\Delta}relA{\Delta}spoT$ (ECM201), and the ${\Delta}luxS$ complement strain to ECM201 (ECM202) were created from EHEC O157:H7 EDL933 to investigate how the regulatory systems interact. The phenotypic changes of the mutant strains were characterized and compared with the wild type. The mutant strains exhibited no obvious growth defects, although acid resistance and cellular cytotoxicity were decreased significantly in all the mutant strains. Phenotypic characterization revealed that mutations in the stringent response system (ECM201 and ECM202) influenced the metabolic (defective utilization of arabinose and L-sorbose) and enzymatic activities (decreased trypsin activity, and increased ${\alpha}$-glucosidase activity). In contrast, the quorum sensing system mutant (ECM101) did not display these phenotypes. The motility of the quorum sensing system mutant (ECM101) was unchanged, but mutation in the stringent response system influenced the motility. Our results suggest that quorum sensing interacts with the stringent response regulation system.

• KOREAN JOURNAL OF CROP SCIENCE
• /
• v.48 no.1
• /
• pp.50-55
• /
• 2003

Herbicidal activity of $\delta$-aminolevulinic acid(ALA), an intermediate for the biosynthesis of tetrapyrroles such as chlorophyll, heme, bacteriochlorophyll, and vitamin $\textrm{B}_{12}$ analogues, was examined to determine the variation in phytotoxic potential against different plant species as affected by different application methods. Seed-soaking treatment, ALA at low concentrations did not affect shoot and root lengths of test plants while at highest concentration reduced them by 20 to 30%. Alfalfa showed the most tolerant response to ALA in both pre- and post-emergence application, and followed by rice. When applied with pre-emergence, cotyledons of Chinese cabbage were severely bleached with 0.5 mM of ALA at 24 hrs after application, and root growth of rice, barnyard grass, and alfalfa was significantly inhibited with increasing of concentration. With post-emergence application, ALA at 2 to 4 mM reduced shoot and root growths of Chinese cabbage and barnyard grass completely. Herbicidal effects of ALA were more enhanced in the treatment combined with 2,2-dipyridyl sthan single application in barnyard grass and Chinese cabbage. The results suggest that alfalfa was the most tolerant to ALA among the tested plants, and that post-emergence application of ALA exhibited greatest photodynamic activity against tested plants.

• Journal of Microbiology and Biotechnology
• /
• v.19 no.10
• /
• pp.1169-1175
• /
• 2009

An antifungal chitinase, ChiCW, produced by Bacillus cereus 28-9 is effective against conidial germination of Botrytis elliptica, the causal agent of lily leaf blight. ChiCW as a modular enzyme consists of a signal peptide, a catalytic domain, a fibronectin type-III-like domain, and a chitin-binding domain. When two C-terminal domains of ChiCW were truncated, $ChiCW{\Delta}FC$ (lacking the chitin-binding domain and fibronectin type III-like domain) lost its antifungal activity. Since $ChiCW{\Delta}C$ (lacking the chitin-binding domain) could not be expressed in Escherichia coli as $ChiCW{\Delta}FC$ did, a different strategy based on protein engineering technology was designed to investigate the involvement of the chitin-binding domain of ChiCW ($ChBD_{ChiCW}$) in antifungal activity in this study. Because ChiA1 of Bacillus circulans WL-12 is a modular enzyme with a higher hydrolytic activity than ChiCW but not inhibitory to conidial germination of Bo. elliptica and the similar domain composition of ChiA1 and ChiCW, the C-terminal truncated derivatives of ChiA1 were generated and used to construct chimeric chitinases with $ChBD_{ChiCW}$. When the chitin-binding domain of ChiA1 was replaced with $ChBD_{ChiCW}$, the chimeric chitinase named ChiAAAW exhibited both high enzyme activity and antifungal activity. The results indicate that $ChBD_{ChiCW}$ may play an important role in the antifungal activity of ChiCW.