• Archives of Pharmacal Research
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• v.20 no.6
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• pp.550-554
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• 1997

Chlorambucil is known to alkylate primarily N7 of guanine and N3 of adenine to induce DNA monofunctional adducts and interstrand cross-links (ISC). We have investigated the sequence specificity for DNA ISC induced by chlorambucil using duplex oligomers containing a defined cross-linkable sequences $ 5^{I}-A*TT, 5^{I}-G*TTor5^{I}-G*CC$ under bar which asterisk indicates the potential cross-linking site and underlined base indicates the potential cross-linking site on the opposite strand. An analysis of 20% denaturing polyacrylamide gel electrophoresis showed that chlorambucil was albe to induce DNA ISC in the duplex oligomers containing a sequence $5^I-GCC$. The formation of DNA ISC was not observed in the duplex oligomers containing sequences $5^I-ATT$. or $5^I-GTT$. These results indicate that chlorambucil induces guanine-guanine DNA ISC but not guanine-adenine or adenine-adenine DNA ISC. In addition, we have tested the ability of chlorambucil to induce DNA ISC within $5^I-GNNC$ or $5^I-GNNC$sequences using duplex oligomers containing the sequence$5^I-G^4G^3G^2^C$. The result of DNA strand cleavage assay showed that DNA ISC was formed at the $5^I-GGC$ sequence (an 1,3 cross-link, $G^1-G^3$) but not at $5^I-GGGC$ (an 1,4 cross-link, $G^1-G^4$) or $5^I-GC$ sequence (an 1,2 cross-link, $G^1-G^2$).

• Archives of Pharmacal Research
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• v.19 no.3
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• pp.191-196
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• 1996

Bizelesin is a promising novel anticancer agent which is known to alkylate N3 of adenine to induce DNA interstrand cross-links (ISC) with in $5^I-TAATTA\; and\; 5^I-TAAAAAA$. We have investigated the base specificity for DNA ISC induced by bizelesin using oligomers containing the cross-linkable sequence $5^I-TAATTA\; and\; 5^I-TAAAAAA$. in which "N" was either A, C, G, or T. An analysis of denaturing polyacrylamide gel showed that bizelesin is able to induce DNA ISC in the duplex oligomer containing sequences $5^I-TAATTA\; and\; 5^I-TAAAAAA$. The formation of interstrand crosslinking did not occur in the sequences $5^I-TAATTA\; and\; 5^I-TAAAAAA$. DNA strand cleavage assay to determine the cross-linking site within $5^I-TAATTA$sequence showed that bizelesin alkylates guanine. These results demonstrate that bizelesin is able to induce DNA ISC at guanine but not at cytosine or thymine. In addition, guanine adducts have been found to be susceptible to DNA strand cleavage by exposure to hot piperidine. The extent of DNA strand cleavage, however, was not 100% efficient in either neutral pH buffer or hot piperidine.

• Archives of Pharmacal Research
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• v.21 no.2
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• pp.135-139
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• 1998

High titer rabbit polyclonal antibodies (pAbs) which show a specificity for saikosaponin a (SSA), have been generated. The immunogen used was a conjugate of SSA linked through its glucose moiety to bovine serum albumin by periodate oxidation method. The antibody titers obtained from two rabbits, innoculated with the immunogen, reached a plateau after the fourth and third booster injection, respectively. The specificity of the pAbs was determined by hapten inhibition assays using several SSA-like structures. SSA competitively inhibited the binding of the rabbit anti-SSA pAbs to SSA-ovalbumin on solid phase, a coated antigen on the well. The antibodies showed high specificity to SSA, exhibiting no significant cross-reactivity with any of SSA analogues tested.

• Biomedical Science Letters
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• v.7 no.4
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• pp.161-166
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• 2001

Most mammalian cells take up glucose by passive transport proteins in the plasma membranes. The best known of these proteins is the human erythrocyte glucose transporter, GLUT1. High levels of heterologous expression far the transporter are necessary for the investigation of its three-dimensional structure by crystallization. To achieve this, the baculovirus expression system has become popular choice. However, Spodoptera frugiperda Clone 9 (Sf9) cells, which are commonly employed as the host permissive cell line to support baculovirus replication and protein synthesis, grow well on TC-100 medium that contains 0.1% D-glucose as the major carbon source, suggesting the presence of endogenous glucose transporters. Furthermore, very little is known of the endogenous transporters properties of Sf9 cells. Therefore, human GLUT1 antibodies would play an important role for characterization of the GLUT1 expressed in insect cell. However, the successful use of such antibodies for characterization of GLUT1 expression m insect cells relies upon their specificity for the human protein and lack of cross-reaction with endogenous transporters. It is therefore important to determine the potential cross-reactivity of the antibodies with the endogenous insect cell glucose transporters. In the present study, the potential cross-reactivity of the human GLUT1 antibodies with the endogenous insect cell glucose transporters was examined by Western blotting. Neither the antibodies against intact GLUT1 nor those against the C-terminus labelled any band migrating in the region expected fur a protein of M$_r$ comparable to GLUT1, whereas these antibodies specifically recognized the human GLUT1. Specificity of the human GLUT1 antibodies tested was also shown by cross-reaction with the GLUT1 expressed in insect cells. In addition, the insect cell glucose transporter was found to have very low affinity for cytochalasin B, a potent inhibitor of human erythrocyte glucose transporter.

• Clinical and Experimental Pediatrics
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• v.62 no.6
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• pp.217-223
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• 2019

Purpose: To determine the diagnostic value of eosinopenia and the neutrophil-to-lymphocyte ratio (NLR) in the diagnosis of early onset neonatal sepsis (EONS). Methods: This cross-sectional study was conducted in the Neonatology Ward of R.D. Kandou General Hospital Manado between July and October 2017. Samples were obtained from all neonates meeting the inclusion criteria for EONS. Data were encoded using logistic regression analysis, the point-biserial correlation coefficient, chi-square test, and receiver operating characteristic curve analysis, with a P value <0.05 considered significant. Results: Of 120 neonates who met the inclusion criteria, 73 (60.8%) were males and 47 (39.2%) were females. Ninety (75%) were included in the sepsis group and 30 (25%) in the nonsepsis group. The mean eosinophil count in EONS and non-EONS groups was $169.8{\pm}197.1cells/mm^3$ and $405.7{\pm}288.9cells/mm^3$, respectively, with statistically significant difference (P<0.001). The diagnostic value of eosinopenia in the EONS group (cutoff point: $140cells/mm^3$) showed 60.0% sensitivity and 90.0% specificity. The mean NLR in EONS and non-EONS groups was $2.82{\pm}2.29$ and $0.82{\pm}0.32$, respectively, with statistically significant difference (P<0.001). The diagnostic value of NLR in the EONS group (cutoff point, 1.24) showed 83.3% sensitivity and 93.3% specificity. Conclusion: Eosinopenia has high specificity as a diagnostic marker for EONS and an increased NLR has high sensitivity and specificity as a diagnostic marker for EONS.

• Parasites, Hosts and Diseases
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• v.56 no.1
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• pp.61-70
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• 2018

We developed a Rapid Diagnostic Test (RDT) kit for detecting IgG/IgM antibodies against Zika virus (ZIKV) using monoclonal antibodies to the envelope (E) and non-structural protein 1 (NS1) of ZIKV. These proteins were produced using baculovirus expression vector with Sf9 cells. Monoclonal antibodies J2G7 to NS1 and J5E1 to E protein were selected and conjugated with colloidal gold to produce the Zika IgG/IgM RDT kit (Zika RDT). Comparisons with ELISA, plaque reduction neutralization test (PRNT), and PCR were done to investigate the analytical sensitivity of Zika RDT, which resulted in 100% identical results. Sensitivity and specificity of Zika RDT in a field test was determined using positive and negative samples from Brazil and Korea. The diagnostic accuracy of Zika RDT was fairly high; sensitivity and specificity for IgG was 99.0 and 99.3%, respectively, while for IgM it was 96.7 and 98.7%, respectively. Cross reaction with dengue virus was evaluated using anti-Dengue Mixed Titer Performance Panel (PVD201), in which the Zika RDT showed cross-reactions with DENV in 16.7% and 5.6% in IgG and IgM, respectively. Cross reactions were not observed with West Nile, yellow fever, and hepatitis C virus infected sera. Zika RDT kit is very simple to use, rapid to assay, and very sensitive, and highly specific. Therefore, it would serve as a choice of method for point-of-care diagnosis and large scale surveys of ZIKV infection under clinical or field conditions worldwide in endemic areas.

• The Korean Journal of Pain
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• v.33 no.1
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• pp.60-65
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• 2020

Background: Anatomic changes in the acromion have been considered a main cause of shoulder impingement syndrome (SIS). To evaluate the relationship between SIS and the acromion process, we devised a new morphological parameter called the acromion process cross-sectional area (APA). We hypothesized that the APA could be an important morphologic diagnostic parameter in SIS. Methods: We collected APA data from 95 patients with SIS and 126 control subjects who underwent shoulder magnetic resonance imaging (MRI). Then we measured the maximal cross-sectional area of the bone margin of the acromion process on MRI scans. Results: The mean of APAs were 136.50 ± 21.75 ㎟ in the male control group and 202.91 ± 31.78 ㎟ in the male SIS group; SIS patients had significantly greater APAs (P < 0.001). The average of APAs were 105.38 ± 19.07 ㎟ in the female control group and 147.62 ± 22.90 ㎟ in the female SIS group, and the SIS patients had significantly greater APAs (P < 0.001). The optimal APA cut-off in the male group was 165.14 ㎟ with 90.2% sensitivity, 91.4% specificity, and an area under the curve (AUC) of 0.968. In the female group, the optimal cut-off was 122.50 ㎟ with 85.2% sensitivity, 84.9% specificity, and an AUC of 0.928. Conclusions: The newly devised APA is a sensitive parameter for assessing SIS; greater APA is associated with a higher possibility of SIS. We think that this result will be helpful for the diagnosis of SIS.

• YAKHAK HOEJI
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• v.47 no.5
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• pp.266-270
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• 2003

We evaluated four commercially available methamphetamine immunoassays for their relative cross-reactivities of amphetamine analogues in human urine: Abbott TDx, Vitalab Selectra and on-site test kits (Accusign MET, SD bioline MET). High cross-reactivities were shown at designer's drugs such as methylenedioxyamphetamine (MDA), methylenedioxymethamphetamine (MDMA) and methylenedioxyethylamphetamine (MDEA) in all of the tested immunoassays. Methoxyphenamine, fenfluramine and phentermine were positive in TDx and Selectra, but were not positive in on-site test kits. Pseudoephedrine, norpseudoephedrine, ephedrine, norephedrine, MDMA, MDA, fenfluramine and phentermine were detected by gas chromatography/mass spectrometry(GC/MS) in false positive urines. Since the overall specificity of any of the devices was not 100％, we found it is important to confirm any positive screening test result, so we developed simultaneous determination of amphetamine analogues in urines. After alkalinization of the urine samples with 6-N NaOH, the analytes were extracted using ethyl acetate, derivatized with pentafluoropropyl anhydride (PFPA) prior at GC/MS analysis.

• Journal of Periodontal and Implant Science
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• v.41 no.2
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• pp.54-59
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• 2011

Purpose: The aim of this study was to define the immunoreactive specificity of Porphyromonas gingivalis (P. gingivalis) heat shock protein (HSP) 60 in periodontitis and atherosclerosis. Methods: In an attempt to define the cross-reactive bacterial heat-shock protein with human self-antigen at molecular level, we have introduced a novel strategy for cloning hybridoma producing anti-P. gingivalis HSP 60 which is polyreactive to bacterial HSPs or to the human homolog. Results: Five cross-reactive clones were obtained which recognized the #19 peptide (TLVVNRLRGSLKICAVKAPG) among 37 synthetic peptides (20-mer, 5 amino acids overlapping) spanning the whole molecule of P. gingivalis HSP 60. We have also established three anti-P. gingivalis HSP 60 monoclonal antibodies demonstrating mono-specificity. These clones recognized the #29 peptide (TVPGGGTTYIRAIAALEGLK). Conclusions: Peptide #19 and #29 of P. gingivalis HSP 60 might be important immunoreactive epitopes in the immuno-pathogenic mechanism of bacterial antigen-triggered autoimmune diseases.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.5
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• pp.2001-2006
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• 2014

The objective of this study was to investigate the diagnostic significance of EBV antibody combined detection for nasopharyngeal carcinoma (NPC) in a high incidence region of southern China. Two hundred and eleven untreated NPC patients, 203 non-NPC ENT patients, and 210 healthy controls were recruited for the study. The titers of VCA/IgA and EA/IgA were assessed by immunoenzyme assay, and the levels of Rta/IgG and EBNA1/IgA were determined by enzyme-linked immunosorbent assay. The levels of VCA/IgA, EA/IgA, Rta/IgG and EBNA1/IgA demonstrated no association with gender or age (p>0.05). The receiver operating characteristic curve and the area under the curve were used to evaluate the diagnostic value. The sensitivity of VCA/IgA (98.1%) and the specificity of EA/IgA (98.5%) were the highest. When a logistic regression model was used to combine the results from multiple antibodies to increase the accuracy, the combination of VCA/IgA+Rta/IgG, whose area under the curve was 0.99, had the highest diagnostic efficiency, and its sensitivity, specificity and Youden index were 94.8%, 98.0% and 0.93 respectively. The data suggest that the combination of VCA/IgA+Rta/IgG may be most suitable for NPC serodiagnosis.