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Volume 66, Issue 6
Cotribution of Analytical Chemistry to Medical Field
Displaying 1-9 of 9 articles from this issue
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Accounts
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Kazuhide MIYAMOTO, Mayumi SUNAGAWA, Kazuki SAITOArticle type: Accounts
2017 Volume 66 Issue 6 Pages 393-402
Published: June 05, 2017
Released on J-STAGE: July 12, 2017
JOURNAL FREE ACCESSProtein ubiquitination induces diverse effects, such as protein degradation, protein interaction, and subcellular localization, on the substrate. E2 enzymes in the ubiquitination are associated with various diseases, such as leukemia, breast cancer, and colorectal cancer. Assessing E2 activity during the ubiquitination reaction leads to the monitoring of tumor development for diagnostic purposes. We engineered an artificial RING finger (ARF) using our ‘Alpha-helix region substitution’ method, and then developed the ARF detection system for simply measuring E2 activities in the ubiquitination. ARFs was created by inserting the helical regions (Active sites) of E3s into the amino-acid sequences of –50-mer peptide. The insertion of only Active sites successfully downsized E3s to the small molecule ARFs, which possess specific E2-binding capabilities, and are ubiquitinated upon themselves without a substrate and a tag. Thus, identification of the target substrate is not needed in the ARF detection system for E2 activities. In this review, the authors mention the design method of ARFs and their structural and functional features. Moreover, the monitoring method of E2 activities was explained using human acute promyelocytic leukemia cells (NB4) following treatment with the anticancer drug bortezomib. This strategy is extremely simple and convenient for the assessment of pathological conditions.View full abstractDownload PDF (2278K) -
Kazuma OGAWA, Ryohei MASUDA, Kazuhiro SHIBAArticle type: Accounts
2017 Volume 66 Issue 6 Pages 403-411
Published: June 05, 2017
Released on J-STAGE: July 12, 2017
JOURNAL FREE ACCESSIt has been reported that sigma-1 receptors regulate the release of signaling substances in the central nervous systems and are related to various diseases, such as schizophrenia, stress disorders, dementia, amyotrophic lateral sclerosis (ALS), and cancer. If the quantification of the sigma-1 receptors is possible, the pathophysiology, the stage, and the early detection of the diseases could be understandable. Molecular imaging using Positron Emission Tomography (PET) or Single Photon Emission Computed Tomography (SPECT) and radioactive probes makes noninvasive quantification of the in vivo metabolism and function possible. Currently, only nuclear medicine diagnosis using PET or SPECT can quantify the sigma-1 receptors. Therefore, there is great expectation for the development of molecular probes to image the sigma-1 receptors specifically. In this paper, we introduce our research on the development of radiohalogen-labeled molecular probes directed against the sigma-1 receptors.View full abstractDownload PDF (898K) -
Masamitsu MAEKAWA, Hiroaki YAMAGUCHI, Nariyasu MANOArticle type: Accounts
2017 Volume 66 Issue 6 Pages 413-419
Published: June 05, 2017
Released on J-STAGE: July 12, 2017
JOURNAL FREE ACCESSAnalytical technologies are used at various clinical sites. In particular, recent developments of analytical chemistry potentiate the usefulness of analytical chemistry in the medical front. The department of pharmaceutical sciences in our hospital has a lot of advanced analytical technologies. Many clinical chemical studies are being developed by analytical technologies and the many analytical methods are used in routine medical care. A new chemical diagnostic method has been developed by a highly accurate and precision LC/MS/MS quantification method and metabolomics technologies. The LC/MS/MS technique is also used for the analysis of the poison substances in patients with the intoxication of some drug or poison. In addition, LC/MS/MS analyses play important roles for the highly precision therapeutic drug monitoring; they are expected to contribute to the advancement of precision- or personalized medicine.View full abstractDownload PDF (774K) -
Shuhei MURAYAMA, Masaru KATOArticle type: Accounts
2017 Volume 66 Issue 6 Pages 421-430
Published: June 05, 2017
Released on J-STAGE: July 12, 2017
JOURNAL FREE ACCESSThe maintenance of biological functions depends on the balance between the amounts and function of the many kinds of molecules in the cells, tissues and body fluids. Most of the biochemical and physiological processes (e.g., proliferation, differentiation, migration, and death) are controlled by concentration changes of intracellular compounds. The induction and real-time detection of these compounds gives information concerning the role molecules. The described methods for controlling the molecular activity are usually based on external signals, such as temperature, magnetism, pH, etc. However, these signals do not allow hard local irradiation, as well as most of the techniques are invasive. Among them, light (in particular, gamma rays) is one of the most preferred signals to control the molecular function, due to its low invasiveness and deep-tissue penetration. We developed a simple preparation procedure for protein-encapsulated nanoparticle (nanogels with mesh structure) and used them for spatiotemporal activity control of various proteins and other molecules. The method was named "PARCEL". The encapsulated molecule was kept by the gel matrix. The change of the mesh size allows a control of many kinds of molecules, as: proteins, enzymes, nucleic acids, and small compounds. The nano-gels were delivered into the cells by endocytosis and were retained into the cells for at least 4 days. We can change the molecular release signal by changing reactive unit. The nano-gels with protein were excreted rapidly through the urine without collapse. On the other hand, the nano-gels with quantum dots were characterized by long-term circulation into the bloodstream and have accumulated in solid tumors. We expect the "PARCEL" method to be useful in the preparation of photo-activated biosensors, theranostic drug delivery systems, and catalysis.View full abstractDownload PDF (1413K)
Research Papers
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Setsuko YAJIMA, Masato KISHI, Keiichi KIMURAArticle type: Research Papers
2017 Volume 66 Issue 6 Pages 431-436
Published: June 05, 2017
Released on J-STAGE: July 12, 2017
JOURNAL FREE ACCESSWe used polysilsesquioxanes (PSQ), which are inorganic-organic composites, as ion-sensing membrane materials of ion sensors. In this study, PSQ-bearing methyl and vinyl groups at a ratio of 4 : 1 were modified chemically with crown ether derivatives. The membranes were formed from solutions of these compounds on the gates of ion-sensitive field-effect transistors, and used as ion-sensing membranes. The ion-sensor properties of these membranes were investigated at 25 °C. Ion-sensing membranes based on these PSQ responded to K+. In the case of PSQ chemically immobilized with 18-crown-6, the Nernstian response for K+ activity changes was obtained, and the K+ selectivity of the membrane was similar to that of the conventional 18-crown-6 derivative based ion-sensing membranes. In addition, the adsorption of fibrinogen on PSQ derivatives were highly suppressed; it was therefore suggested that the membranes based on PSQ showed good biocompatibility.View full abstractDownload PDF (793K) -
Shinobu SATO, Kazuya HARAGUCHI, Mana HAYAKAWA, Kazuhiro TOMINAGA, Shig ...Article type: Research Papers
2017 Volume 66 Issue 6 Pages 437-443
Published: June 05, 2017
Released on J-STAGE: July 12, 2017
JOURNAL FREE ACCESSThe human telomerase reverse transcriptase (hTERT) gene is encoded as a catalytic unit of telomerase. Since the methylation pattern of the CpG islands in the promoter region of hTERT gene is related with cancer development, this should serve as a suitable cancer marker. Electrochemical hybridization assay was applied to this detection using ferrocenylnaphthalene diimide (FND) as a hybrid indicator. Ten different 24-meric DNA probes were designed and immobilized on the individual electrodes of multi-electrode chip, which can hybridize with sample DNAs carrying different methylation patterns: M0, M1, M1’, M2, M2’, M3, M3’, M4, M4’, or M5, where number refers to the number of methylated cytosine of CpG islands in the promoter region of hTERT gene. Aiming for simple diagnosis of oral cancer, the total exfoliated oral cells as clinical samples were obtained from oral cancer patients (oral squamous cell carcinoma, OSCC), oral leukoplakia as precancerous lesion, and healthy volunteers. The clinical samples were amplified by a methylation-specific polymerase chain reaction (MSP) with bisulfite treated genome DNA from the total exfoliated oral cells and hybridized with 10-probe DNA immobilized multi-electrode chip. The current increase ratio of M4, M4’, M5 probe for detecting methylation was increased with cancer progress according to the following order: healthy volunteers, oral leukoplakia, and OSCC. Receiver operatorating characteristic curve of the obtained data gave the threshold values in the case of the detection of OSCC from all samples to obtain 91 % of the accuracy rate for total exfoliated oral cells using 96.6 % of the threshold values.View full abstractDownload PDF (857K)
Technical Papers
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Hirotake WAKAMATSU, Hiroshi TERASAKI, Mitsunobu SHIMADZUArticle type: Technical Papers
2017 Volume 66 Issue 6 Pages 445-451
Published: June 05, 2017
Released on J-STAGE: July 12, 2017
JOURNAL FREE ACCESSPatients with the T790M mutation in the epidermal growth facter receptor (EGFR) gene are known to acquire resistance to gefitinib. Early detection of this EGFR polymorphism is important for preventing the side effects of gefitinib as well as for considering alternative treatments. In this study, we developed a highly sensitive technique based on photoreactive probes (PREPs) for the detection of single-nucleotide polymorphisms; we detected 2369C > T (T790M) mutation of the EGFR gene by this technique. Our method has a polymerase chain reaction (PCR) clamping efficiency of –99 % and can be applied to virtually all targets. The proposed optical clamping approach is very useful as a simple and efficient pre-treatment method for samples containing only trace amounts of the mutated genes. This method has a potential to be widely used for highly sensitive point-mutation detection.View full abstractDownload PDF (4131K) -
Emi MORI, Yumiko HOSOYA, Ayumi YOSHIZAKI, Kazuma MAWATARI, Takehiko KI ...Article type: Technical Papers
2017 Volume 66 Issue 6 Pages 453-457
Published: June 05, 2017
Released on J-STAGE: July 12, 2017
JOURNAL FREE ACCESSAll of the process from sampling to detection could be performed in microliter order by using a new device, μELISA, which is equipped with microfluidics and a thermal lens microscope. However, as for handling of a small amount of patient sample in clinical sites, when the sample volume is much smaller, a preservation method should be investigated while considering the influences of the S/V ratio in small area. In this research, preservation stability for a small amount of patient sample was investigated, by analyzing CRP. As a result, the inner wall of the container, liquid volume, and preservation time are should be taken into account and evaluated in advance.View full abstractDownload PDF (1009K)
Analytical Reports
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Akira KOTANI, Mototaka KOHAMA, Taisei SHIRATORI, Fumiyo KUSU, Hideki H ...Article type: Analytical Reports
2017 Volume 66 Issue 6 Pages 459-463
Published: June 05, 2017
Released on J-STAGE: July 12, 2017
JOURNAL FREE ACCESSA fecal collection kit, which consisted of a collection tube containing 10 mL of 3 % phenol in water, collection paper, a spoon to scrape the feces, an instruction, glove, and a plastic bag with a zipper, has been developed to collect human feces for determining fecal short-chain fatty acids (SCFAs). Using the fecal collection kit, human healthy volunteers (n = 15) and patients with ulcerative colitis (n = 10) can easily collect their feces specimen and mail them to our laboratory by themselves. The fecal SCFAs in mailed specimens are determined by high-performance liquid chromatography with electrochemical detection (HPLC-ECD) based on the voltammetric reduction of vitamin K3. Fecal SCFAs are stably remained in the present collection tube for 7 days at 4 °C. Fecal lactic acid content in patients with ulcerative colitis is increased, whereas fecal acetic acid content is decreased in comparison to that in healthy volunteers. The ratio of lactic acid to acetic and propionic acids in feces from patients with ulcerative colitis is significantly increased in comparison to that from healthy volunteers (p < 0.1). In conclusion, the present fecal collection kit would be useful to collect feces specimens from humans living in the country to determine fecal SCFAs by HPLC-ECD.View full abstractDownload PDF (795K)
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