• Asian Pacific Journal of Cancer Prevention
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• v.14 no.10
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• pp.5741-5745
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• 2013

Background: Heat-shock protein70 (HSP70) are intracellular protein chaperones, with emerging evidence of their association with various diseases. We have previously reported significantly elevated plasma-HSP70 (pHSP70) in pancreatic cancer. Current methods of pHSP70 isolation are ELISA-based which lack specificity due to cross-reactivity by similarities in the amino-acid sequence in regions of the protein backbone resulting in overestimated HSP70 value. Materials and Methods: This study was undertaken to develop a methodology to capture all isoforms of pHSP70, while further defining their tyrosine and serine phosphorylation status. Results: The methodology included gel electrophoresis on centrifuged supernatant obtained from plasma incubated with HSP70 antibody-coupled beads. After blocking non-specific binding sites, blots were immunostained with monoclonal-antibody specific for human-HSP70, phosphoserine and phosphotyrosine. Conclusions: Our novel immunocapture approach has distinct advantages over the commercially available methods of pHSP70 quantification by allowing isolation of molecular aggregates of HSP70 with additional ability to precisely distinguish phosphorylation state of HSP70 molecules at serine and tyrosine residues.

• BMB Reports
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• v.31 no.1
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• pp.64-69
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• 1998

Recent studies have suggested that integrin (CR3) participates in the signal transduction pathways of certain GPI-anchored phagocytic receptors including $Fc{\gamma}RIIIB$. One consequence of this functional linkage is an inducible association between CR3 and cortical microfilaments that is triggered by $Fc{\gamma}RIIIB$ binding to immobilized immune complexes (IC). That this signaling event requires the co-expression of $Fc{\gamma}RIIIB$ with CR3 was documented by the use of NIH 3T3 transfectants expressing both CR3 and $Fc{\gamma}RIIIB$ (clone 3-23), CR3 alone (clone 3-19), and $Fc{\gamma}RIIIB$ alone (clone 3-15). Pretreatment of 3-23 cells with protein kinase inhibitors such as staurosporine and methyl 2,5-dihydroxycinnamate (MDHC) blocked IC-stimulated CR3 microfilament proximity without affecting the extent to which $Fc{\gamma}RIIIB$ constrains the lateral membrane mobility of a subset of CR3 on the cell surface (as measured in fluorescence recovery after photobleaching experiments). These data support that CR3 and $Fc{\gamma}RIIIB$ molecules are physically and functionally associated and that ligation of FcgRIIIB triggers CR3-dependent signal transduction.

• THE INTERNATIONAL COMMERCE & LAW REVIEW
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• v.12
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• pp.405-427
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• 1999

Although Uniform Customs and practice for Documentary(UCP) is not a law, it applies to most documentary credits and is binding on all parties unless otherwise expressly stipulated. Besides, Uniform Commercial Code(UCC) Article 5 was codified by the United States and was adopted by every state. Moreover, the New York version of the UCC Article 5-102(4) specifically providing that the UCC does not apply to letters of credit where the parties agree to be governed by the UCP. Identical nonuniform Articles were latter added in Alabama, Arizona, and Missouri. The fact that courts in forty-six of the fifty states are bound by Article 5. Until now, Article 5 of the UCC has probably had an impact on the decisions in New York and the New York common law. Therefore, I examined a few issues on application between Article 5 of the UCC and the UCP. First, although the UCP attempt to introduce a new for examination of document by incorporating "standard practice of financial institutions" and "international banking practice", the standards for documentary compliance are not clear. The UCC attempt to rely on the matter of interpretation for the court, but the UCP would probably be interested in examining in about bank's internal practices as reflected in UCP Articles. Second, the rule for nondocumentary conditions is a useful for stand-by credit transactions under the UCC, but these conditions would probably put the bank in an even worse position in case of documentary credit transactions under the UCP. Third, the UCP does not contain any provision governing the fraud exception, but the UCC codified the fraud and forgery rules developed through American case law. Fourth, the UCP treats the issue of transfer in much more detail than the UCC does. In contract, the UCP's treatment of assignment of proceeds is brief. Finally, I suggest that the fraud exception rules should be prescribed in the UCP in order to protect the issuing bank and the applicant when an unscrupulous party attempts to defraud.

• Journal of Nutrition and Health
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• v.27 no.1
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• pp.12-22
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• 1994

Effects of very low level of lead in diet and exposure time on the accumulation and distribution in organs and tissues was investigated with growing rats. 21 days old Sprague-Dawley rats were exposed to lead for 7, 14 and 21 days by feeding of 0.03, 0.42, 0.92 and 1.46mg/kg Pb as Pb-acetate containing diet, respectively. Lead concentrations in blood, liver, kidney and bone exhibit a linear relationship with lead levels in diet. After 7 days of exposure, the greatest dose dependent accumulation of lead was found in kidney and followed in bone. However, after 14 and 21 days, the dose dependent accumlation of lead in bone was about two fold greater than that in kidney. The accumulation of lead in liver and blood was relatively low. As continuous exposure to lead, the concentrations of lead in liver, kidney, blood and intestinal tracts were rather not increased with exposure time. However, bone lead concentration was increased with exposure time by feeding of 0.92 and 1.46mg/kg Pb in diet, but not 0.42mg/kg. The lead concentration in gastrointestinal tracts tends also to increasing with lead levels in diet after 7 and 14 days of exposure. However, by 21 days of exposure the lead concentration revealed relatively constant value regardless of the dietary lead levels. It is concluded that the binding capacity of the lead in blood, liver, kidney and bone seems to be increased with increasing lead levels in diet. The lead concentration in these organs, with the exception of the lead in bone, seems, however, to be standing under steady state regulation by continued exposure with the same dietary lead level. Therefore, by chronic exposure condition with environmental relevant lead level bone might be a principle targe organ for lead and blood lead repesents better the current lead exposure than the lead body burden.

• Asian-Australasian Journal of Animal Sciences
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• v.28 no.4
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• pp.573-583
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• 2015

B-32 is one of a panel of monoclonal anti-idiotypic antibodies to growth hormone (GH) that we developed. To characterize and identify its potential role as a novel growth hormone receptor (GHR) agonist, we determined that B-32 behaved as a typical $Ab2{\beta}$ based on a series of enzyme-linked immunosorbent assay assays. The results of fluorescence-activated cell sorting, indirect immunofluorescence and competitive receptor binding assays demonstrated that B-32 specifically binds to the GHR expressed on target cells. Next, we examined the resulting signal transduction pathways triggered by this antibody in primary porcine hepatocytes. We found that B-32 can activate the GHR and Janus kinase (2)/signal transducers and activators of transcription (JAK2/STAT5) signalling pathways. The phosphorylation kinetics of JAK2/STAT5 induced by either GH or B-32 were analysed in dose-response and time course experiments. In addition, B32 could also stimulate porcine hepatocytes to secrete insulin-like growth factors-1. Our work indicates that a monoclonal anti-idiotypic antibody to GH (B-32) can serve as a GHR agonist or GH mimic and has application potential in domestic animal (pig) production.

• The Korean Journal of Physiology
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• v.25 no.1
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• pp.17-26
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• 1991

Single smooth muscle cells of the rabbit pulmonary artery were isolated by treatment with collagenase and elastase. Using the patch clamp technique, potassium channel activity was recorded from the inside-out membrane patch. The channel had a sin히e channel conductance of about 360 pS in symmetrical concentration of K on both sides of the patch, 150 mM, and had a linear current-voltage relationship. During the application of 10 mM tetraethylammonium (TEA) to the intracellular membrane surface, the amplitude of single channel current was reduced and very rapid flickering appeared. The open probability $(P_0)$ of this channel was increased by increasing positivity of the potential across the patch membrane, with e-fold increase by 20 mV depolarization, and by increasing the internal $Ca^{2+}$ concentration. These findings are consistent with those of large conductance Ca-activated K channels reported in other tissues. But the shortening of the mean open time by increasing $[Ca^{2+}]_i$, was an unexpected result and one additional closed state which might be arisen from a block of the open channel by Ca binding was suggested. The $P_0-membrane$ potential relationship was modulated by internal pH. Decreasing pH reduced $P_0$. Increasing pH not only increased $P_0$ but also weakened the voltage dependency of the channel opening. The modulation of Ca-activated K channel by pH was thought to be related to the mechanism of regulation of vascular tone by the pH change.

• Journal of Microbiology and Biotechnology
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• v.20 no.3
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• pp.532-541
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• 2010

Peroxidase-like activity of Vitreoscilla hemoglobin (VHb) has been recently disclosed. To maximize such activity, two catalytically conserved residues (histidine and arginine) found in the distal pocket of peroxidases have successfully been introduced into that of the VHb. A 15-fold increase in catalytic constant ($k_{cat}$) was obtained in P54R variant,which was presumably attributable to the lower rigidity and higher hydrophilicity of the distal cavity arising from substitution of proline to arginine. None of the modifications altered the affinity towards either $H_2O_2$ or ABTS substrate. Spectroscopic studies revealed that VHb variants harboring the T29H mutation apparently demonstrated a spectral shift in both ferric and ferrous forms (406-408 to 411 nm, and 432 to 424-425 nm, respectively). All VHb proteins in the ferrous state had a $\lambda_{soret}$ peak at ~419 nm following the carbon monoxide (CO) binding. Expression of the P54R mutant mediated the downregulation of iron superoxide dismutase (FeSOD) as identified by two-dimensional gel electrophoresis (2-DE) and peptide mass fingerprinting (PMF). According to the high peroxidase activity of P54R, it could effectively eliminate autoxidation-derived $H_2O_2$, which is a cause of heme degradation and iron release. This decreased the iron availability and consequently reduced the formation of the $Fe^{2+}$-ferric uptake regulator protein ($Fe^{2+}$-Fur), an inducer of FeSOD expression.

• Journal of Microbiology and Biotechnology
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• v.20 no.3
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• pp.574-578
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• 2010

Archaebacteria Sulfolobus acidocaldarius contains the highly thermophilic cytochrome P450 enzyme (CYP119). CYP119 possesses stable enzymatic activity at up to $85^{\circ}C$. However, this enzyme is still considered as an orphan P450 without known physiological function with endogenous or xenobiotic substrates. We characterized the regioselectivity of lauric acid by CYP119 using the auxiliary redox partner proteins putidaredoxin (Pd) and putidaredoxin reductase (PdR). Purified CYP119 protein showed a tight binding affinity to lauric acid ($K_d=1.1{\pm}0.1{\mu}M$) and dominantly hydroxylated (${\omega}-1$) position of lauric acid. We determined the steady-state kinetic parameters; $k_{cat}$ was 10.8 $min^{-1}$ and $K_m$, was 12 ${\mu}M$. The increased ratio to $\omega$-hydroxylated production of lauric acid catalyzed by CYP119 was observed with increase in the reaction temperature. These studies suggested that the regioselectivity of CYP119 provide the critical clue for the physiological enzyme function in this thermophilic archaebacteria. In addition, regioselectivity control of CYP119 without altering its thermostability can lead to the development of novel CYP119-based catalysts through protein engineering.

• THE INTERNATIONAL COMMERCE & LAW REVIEW
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• v.17
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• pp.7-35
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• 2002

In many international financing transactions Korean companies are required to issue a guarantee. Thus far, however, legal issues on international guarantees have not been fully discussed in Korea. This is partly because most of the international guarantees are governed by a foreign law such as English law or the laws of the State of New York. In this articles the author examines major concepts or terms and conditions of a typical international guarantee, e.g., language on consideration, primary obligor, joint and several guarantee, unconditional and irrevocable guarantee, continuing guarantee, right of subrogation, representations and warranties, covenant or undertaking, currency indemnity, assignment, participation, governing law and jurisdiction clause, etc. For reference, standard forms of a guarantee and a standby letter of credit are attached to the article. In examining the terms and conditions, the author compares them with similar or equivalent concepts under Korean law. The author further discusses some Korean law issues that may arise under international guarantees governed by a foreign law. These issues include the application of the ultra vires doctrine under Article 34 of the Civil Code of Korea, the validity of an international guarantee which a Korean company has issued in violation of the guarantee ceiling set under Article 10 of the Law on Monopoly Regulation and Fair Trade of Korea and the validity of an international guarantee which a Korean party has issued in violation of the Foreign Exchange Transaction Law. In addition, the author discusses some issues under a so-called independent guarantee and a standby letter of credit. In this regard, reference is made to the Uniform Rules for Demand Guarantee (URDG), International Standby Practices (ISP98) and the Convention on Independent Guarantees and Stand-by Letters of Credit adopted by the United Nations in 1995. Finally, the author examines major terms and conditions of typical comfort letters and discusses some legal issues, such as the binding force of the comfort letter. In dealing with the issues the author underscores that to the extent the issues are not properly dealt with by an international norm such as Uniform Customs and Practice for Documentary Credits or ISP 98, the issues must be analyzed by reference to the governing law of the relevant instrument.

• The Korean Journal of Physiology and Pharmacology
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• v.22 no.5
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• pp.585-595
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• 2018

Amitriptyline, a tricyclic antidepressant, is commonly used to treat depression and neuropathic pain, but its mechanism is still unclear. We tested the effect of amitriptyline on 5-hydroxytryptamine 3 ($5-HT_3$) receptor currents and studied its blocking mechanism because the clinical applications of amitriptyline overlapped with $5-HT_3$ receptor therapeutic potentials. Using a whole-cell voltage clamp method, we recorded the currents of the $5-HT_3$ receptor when 5-HT was applied alone or co-applied with amitriptyline in cultured NCB-20 neuroblastoma cells known to express $5-HT_3$ receptors. To elucidate the mechanism of amitriptyline, we simulated the $5-HT_3$ receptor currents using Berkeley $Madonna^{(R)}$ software and calculated the rate constants of the agonist binding and receptor transition steps. The $5-HT_3$ receptor currents were inhibited by amitriptyline in a concentration-dependent, voltage-independent manner, and a competitive mode. Amitriptyline accelerated the desensitization of the $5-HT_3$ receptor. When amitriptyline was applied before 5-HT treatment, the currents rose slowly until the end of 5-HT treatment. When amitriptyline was co-applied with 5-HT, currents rose and decayed rapidly. Peak current amplitudes were decreased in both applications. All macroscopic currents recorded in whole cell voltage clamping experiments were reproduced by simulation and the changes of rate constants by amitriptyline were correlated with macroscopic current recording data. These results suggest that amitriptyline blocks the $5-HT_3$ receptor by close and open state blocking mechanisms, in a competitive manner. We could expand an understanding of pharmacological mechanisms of amitriptyline related to the modulation of a $5-HT_3$ receptor, a potential target of neurologic and psychiatric diseases through this study.