Publications by authors named "Yasushi Nakano"

35 Publications

Potential biomarker enhancing the activity of tuberculosis, hsa-miR-346.

Tuberculosis (Edinb) 2021 Jul 10;129:102101. Epub 2021 Jun 10.

Department of Infectious Diseases, Keio University School of Medicine, Japan.

Objectives: To determine the usefulness of hsa-miR-346, a potential biomarker enhancing the activity of non-tuberculous mycobacterial diseases, as a biomarker of tuberculosis activity.

Methods: We investigated whether hsa-miR-346 is secreted by human macrophages infected with Mycobacterium tuberculosis (M. tuberculosis) in an in vitro study. In addition, a cross-sectional study was conducted first to evaluate whether serum hsa-miR-346 is elevated in patients with tuberculosis compared with that in healthy individuals. Second, we conducted a retrospective study to evaluate whether anti-tuberculosis treatment reduces serum hsa-miR-346 levels.

Results: Log hsa-miR-346 levels were significantly elevated in the supernatant of human macrophages infected with M. tuberculosis in a dose-dependent manner. The mean serum log hsa-miR-346 levels were -15.48 (-15.76 to -15.21) in patients with tuberculosis and -16.12 (-16.29 to -15.95) in healthy volunteers, which significantly differed. In addition, hsa-miR-346 significantly decreased at 2 months from starting an anti-tuberculosis treatment.

Conclusions: We consider hsa-miR-346 as a potential biomarker enhancing the tuberculosis activity.
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http://dx.doi.org/10.1016/j.tube.2021.102101DOI Listing
July 2021

Development of X-ray contrast agents using single nanometer-sized gold nanoparticles and lactoferrin complex and their application in vascular imaging.

Colloids Surf B Biointerfaces 2021 Jul 1;203:111732. Epub 2021 Apr 1.

Department of Medical Physics, Graduate School of Medicine, Tohoku University, 2-1 Seiryo-machi, Aoba-ku, Sendai, Miyagi, 980-8575, Japan; International Center for Synchrotron Radiation InnovationSmart (SRIS), Tohoku University, 2-1-1, Katahira, Aoba-ku, Sendai 980-8577, Japan. Electronic address:

The technology to accurately image the morphology of tumor vessels with X-ray contrast agents is important to clarify mechanisms underlying tumor progression and evaluate the efficacy of chemotherapy. However, in clinical practice, iodine-based contrast agents present problems such as short blood retention owing to a high clearance ability and insufficient X-ray absorption capacity when compared with other high atomic number elements. To resolve these issues, gold nanoparticles (AuNPs), with a high atomic number, have attracted a great deal of attention as contrast agents for angiography, and have been employed in small animal models. Herein, we developed novel contrast agents using AuNPs and captured changes in tumor vessel morphology with time using X-ray computed tomography (CT). First, glutathione-supported single nanometer-sized AuNPs (sAu/GSH) (diameter, 2.2 nm) were fabricated using tetrakis(hydroxymethyl)phosphonium chloride as a reducing agent. The sAu/GSH particles were intravenously injected into mice, remained in vessels for a few minutes, and were then excreted by the kidneys after 24 h, similar to the commercial contrast agent iopamidol. Next, the Au/GSH and lactoferrin (sAu/GSH-LF) (long axis size, 17.3 nm) complex was produced by adding lactoferrin to the sAu/GSH solution under the influence of a condensing agent. On intravenously administering sAu/GSH-LF to mice, the blood retention time was 1-3 h, which was considerably longer than that observed with iopamidol and sAu/GSH. Moreover, we succeeded in imaging morphological changes in identical tumor vessels for several days using X-ray CT with sAu/GSH-LF.
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http://dx.doi.org/10.1016/j.colsurfb.2021.111732DOI Listing
July 2021

Clinical Features and Prognosis of Nontuberculous Mycobacterial Pleuritis: A Multicenter Retrospective Study.

Ann Am Thorac Soc 2021 Apr 8. Epub 2021 Apr 8.

Keio University School of Medicine, Department of Infectious Diseases, Shinjuku-ku, Tokyo, Japan;

Rationale: The clinical features and prognosis of nontuberculous mycobacterial (NTM) pleuritis and pleural effusion combined with NTM lung disease (NTM-LD) remain unclear.

Objectives: To investigate the clinical features and prognosis of NTM pleuritis.

Methods: This retrospective observational study included patients with NTM pleuritis from January 2001 to June 2018 across eight hospitals in Japan. NTM pleuritis was defined by a positive culture of NTM from pleural effusion. We matched patients with Mycobacterium complex lung disease (MAC-LD) without pleuritis by sex and age to obtain comparative data and investigated the association between clinical parameters and prognosis.

Results: We identified 64 patients with NTM pleuritis (median age, 73 years; 37 female). The median follow-up duration was 11 months, and 27 patients died. Patients with MAC pleuritis had a significantly worse survival compared to matched patients with MAC-LD without pleuritis. Multivariate analysis revealed that pleuritis (adjusted hazard ratio [aHR], 6.99; 95% confidence interval [CI], 2.58-19.00) and underlying pulmonary diseases (aHR, 3.01; 95% CI, 1.44-6.28) were independently associated with all-cause mortality in MAC-LD patients.

Conclusions: The prognosis of MAC pleuritis is poorer than that of MAC-LD without pleuritis. Pleuritis is an independent prognostic factor in patients with MAC-LD.
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http://dx.doi.org/10.1513/AnnalsATS.202008-938OCDOI Listing
April 2021

[Supporting Technology for Development of Antibody Drug and Diagnostic Method to Predict Response to the Drug, by Using Fluorescence Imaging].

Gan To Kagaku Ryoho 2021 Feb;48(2):170-175

Dept. of Medical Physics, Graduate School of Medicine, Tohoku University.

Fluorescence imaging is a very useful method for visualizing molecules and cells, but when tissues are measured", decrease in resolution due to increased scattering and absorption of light in proportion to tissue thickness (problem 1)" and "decrease in signal to noise(S/N)ratio of positive signal due to tissue autofluorescence(problem 2)"are problems to be solved. In this paper, to develop a technology to improve the analysis accuracy of drug efficacy mechanisms in preclinical trial of drug discovery, we performed development of a supporting technology for drug discovery of antibody drug conjugates by imaging living tumor tissues, while solving problem 1. This technology is expected to lead to an improvement in the success rate of clinical trials. Next, to develop a diagnostic method to predict the response to neoadjuvant chemotherapy with antibody drugs for breast cancer, we performed development of fluorescence imaging of pathological tissues using fluorescent nanoparticles with ultra-high brightness, while solving problem 2. This diagnostic technology makes it possible to evaluate the expression level of the target protein of antibody drug with high quantitative and wide range sensitivity. This improved the accuracy of drug efficacy prediction. Therefore, patients who are expected to have a low drug efficacy will be able to select anticancer drugs with different mechanisms of action. These results of this study showed the reduction of drug discovery costs and improvement of individualized medicine. Thus, this study will greatly contribute to the development of precision medicine.
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February 2021

Prolonged viral shedding of SARS-CoV-2 in an immunocompromised patient.

J Infect Chemother 2021 Feb 4;27(2):387-389. Epub 2020 Dec 4.

Kawasaki City Institute for Public Health, 210-0821 3-25-13 Tono-machi, Kawasaki-ku, Kawasaki City, Kanagawa prefecture, Japan.

The duration of viral shedding of SARS-CoV-2 is usually less than 10 days. We experienced a COVID-19 case with prolonged viral shedding for 2 months. His cell mediated immunity has been depressed (CD4T cell <100/μl) due to advanced malignant lymphoma and chemotherapy which had been completed 4 months prior to the onset of symptoms of COVID-19. We administered several treatments against COVID-19, however the results of Polymerase Chain Reaction (PCR) from nasopharyngeal specimens remained positive to SARS-CoV-2 for 2 months. Moreover, virus isolation assays performed on Day 59 also remained positive. He was finally discharged on Day 69 with two consecutive negative PCR results for SARS-CoV-2. Immunocompromised status may prolong viral shedding and it is therefore important for the clinician to take into account this when assessing such patients.
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http://dx.doi.org/10.1016/j.jiac.2020.12.001DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7836222PMC
February 2021

Heterogeneous Drug Efficacy of an Antibody-Drug Conjugate Visualized Using Simultaneous Imaging of Its Delivery and Intracellular Damage in Living Tumor Tissues.

Transl Oncol 2020 Jun 11;13(6):100764. Epub 2020 May 11.

Department of Breast and Endocrine Surgical Oncology, Graduate School of Medicine, Tohoku University, Sendai, Miyagi 980-8574, Japan.

Anticancer drug efficacy varies because the delivery of drugs within tumors and tumor responses are heterogeneous; however, these features are often more homogenous in vitro. This difference makes it difficult to accurately determine drug efficacy. Therefore, it is important to use living tumor tissues in preclinical trials to observe the heterogeneity in drug distribution and cell characteristics in tumors. In the present study, to accurately evaluate the efficacy of an antibody-drug conjugate (ADC) containing a microtubule inhibitor, we established a cell line that expresses a fusion of end-binding protein 1 and enhanced green fluorescent protein that serves as a microtubule plus-end-tracking protein allowing the visualization of microtubule dynamics. This cell line was xenografted into mice to create a model of living tumor tissue. The tumor cells possessed a greater number of microtubules with plus-ends, a greater number of meandering microtubules, and a slower rate of microtubule polymerization than the in vitro cells. In tumor tissues treated with fluorescent dye-labeled ADCs, heterogeneity was observed in the delivery of the drug to tumor cells, and microtubule dynamics were inhibited in a concentration-dependent manner. Moreover, a difference in drug sensitivity was observed between in vitro cells and tumor cells; compared with in vitro cells, tumor cells were more sensitive to changes in the concentration of the ADC. This study is the first to simultaneously evaluate the delivery and intracellular efficacy of ADCs in living tumor tissue. Accurate evaluation of the efficacy of ADCs is important for the development of effective anticancer drugs.
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http://dx.doi.org/10.1016/j.tranon.2020.100764DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7218300PMC
June 2020

Automated Quantification of Extranuclear ERα using Phosphor-integrated Dots for Predicting Endocrine Therapy Resistance in HR/HER2 Breast Cancer.

Cancers (Basel) 2019 Apr 12;11(4). Epub 2019 Apr 12.

Department of Medical Physics, Graduate School of Medicine, Tohoku University, Sendai, Miyagi 980-8574, Japan.

In addition to genomic signaling, Estrogen receptor alpha (ERα) is associated with cell proliferation and survival through extranuclear signaling contributing to endocrine therapy (ET) resistance. However, the relationship between extranuclear ERα and ET resistance has not been extensively studied. We sought to measure extranuclear ERα expression by immunohistochemistry using phosphor-integrated dots (IHC-PIDs) and to assess its predictive value for ET resistance. After quantitative detection of ERα by IHC-PIDs in vitro, we developed "the nearest-neighbor method" to calculate the extranuclear ERα. Furthermore, tissue sections from 65 patients with HR+/HER2- BC were examined by IHC-PIDs, and the total ERα, nuclear ERα, extranuclear ERα PIDs score, and ratio of extranuclear-to-nuclear ERα (ENR) were measured using the novel method. We demonstrate that quantification of ERα using IHC-PIDs exhibited strong correlations to real-time qRT-PCR ( = 0.94) and flow cytometry ( = 0.98). High ERα ENR was significantly associated with poor overall survival ( = 0.048) and disease-free survival (DFS) ( = 0.007). Multivariate analysis revealed that the ERα ENR was an independent prognostic factor for DFS [hazard ratio, 3.8; 95% CI, 1.4-11.8; = 0.006]. Our automated measurement has high accuracy to localize and assess extranuclear ERα. A high ERα ENR in HR/HER2 BC indicates decreased likelihood of benefiting from ET.
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http://dx.doi.org/10.3390/cancers11040526DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6520781PMC
April 2019

Quantitative immunohistochemical assay with novel digital immunostaining for comparisons of PD-L1 antibodies.

Mol Clin Oncol 2019 Mar 17;10(3):391-396. Epub 2019 Jan 17.

Department of Otolaryngology, Head and Neck Surgery, Kansai Medical University, Osaka 573-1010, Japan.

One obstacle in diagnostic pathology is the harmonization of one drug-one diagnostic tests for programmed death ligand-1 (PD-L1). There are many challenges in accurate comparisons of diagnostic tests, such as differences in the titer of each antibody, detection system and dynamic range of visualization. Our previously developed digital immunostaining technique is highly sensitive and quantitative with the ability to quantify particles that bind in a one-to-one fashion with antibody in each cell. Determining the differences in the titer of each antibody with digital immunostaining may be beneficial for future harmonized analysis. To demonstrate the accuracy of digital immunostaining, the present study compared the number of particles with ELISA and nCounter data from five cell lines. NCI-H460 exhib-ited the highest level of PD-L1 protein, followed by A549, PC-3, NCI-H1299, and NCI-H446 cells. In addition, the PD-L1 mRNA values determined by nCounter corresponded with the order of the protein levels determined by ELISA. The present study revealed that digital immunostaining for PD-L1 was highly associated with ELISA and nCounter data. Among the four antibodies tested, the titer of all but SP142 coincided with ELISA and nCounter data. These results indicated that our digital immunostaining technique may be beneficial for future harmonized analysis.
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http://dx.doi.org/10.3892/mco.2019.1801DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6388504PMC
March 2019

Preventability of Early Versus Late Hospital Readmissions.

Ann Intern Med 2019 02;170(3):217-218

Sendai City Hospital, Sendai, Japan (J.H.).

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http://dx.doi.org/10.7326/L18-0610DOI Listing
February 2019

A novel detection methodology for HER2 protein quantitation in formalin-fixed, paraffin embedded clinical samples using fluorescent nanoparticles: an analytical and clinical validation study.

BMC Cancer 2018 Dec 18;18(1):1266. Epub 2018 Dec 18.

Konica Minolta INC., Bio Health Care Business Development Division, Corporate R&D Headquarters, No. 1 Sakura-machi, Hino-shi Tokyo, 191-8511, Japan.

Background: Clinical assays for the assessment of the human epidermal growth factor receptor-2 (HER2) status in breast cancer include immunohistochemistry (IHC) and in situ hybridization (ISH), both of which have limitations. Recent studies have suggested that a more quantitative approach to the measurement of HER2 protein expression may improve specificity in selecting patients for HER-2 targeted therapy. In the current study, we have used HER2 expression in breast cancer cell lines and clinical samples as a model to explore the potential utility of a novel immunodetection technique, using streptavidin coated Phosphor Integrated Dot fluorescent nanoparticles (PID), which can be quantitatively measured using computer analysis.

Methods: The expression of HER2 protein in cell lines was evaluated with antibody-binding capacity using fluorescence-activated cell sorting (FACS) for comparison with PID measurements to test for correlations with existing quantitative protein analysis methodologies. Various other analytic validation tests were also performed, including accuracy, precision, sensitivity, robustness and reproducibility. A methods comparison study investigated correlations between PID versus IHC and ISH in clinical samples. Lastly, we measured HER2 protein expression using PID in the pretreatment biopsies from 34 HER2-positive carcinomas that had undergone neoadjuvant trastuzumab-based chemotherapy.

Results: In the analytic validation, PID HER2 measurements showed a strong linear correlation with FACS analysis in breast cell lines, and demonstrated significant correlations with all aspects of precision, sensitivity, robustness and reproducibility. PID also showed strong correlations with conventional HER2 testing methodologies (IHC and ISH). In the neoadjuvant study, patients with a pathologic complete response (pCR) had a significantly higher PID score compared with patients who did not achieve a pCR (p = 0.011), and was significantly correlated to residual cancer burden (RCB) class (p = 0.026, R = 0.9975).

Conclusions: Analytic testing of PID showed that it may be a viable testing methodology that could offer advantages over other experimental or conventional biomarker diagnostic methodologies. Our data also suggests that PID quantitation of HER2 protein may offer an improvement over conventional HER2 testing in the selection of patients who will be the most likely to benefit from HER2-targeted therapy. Further studies with a larger cohort are warranted.
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http://dx.doi.org/10.1186/s12885-018-5172-1DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6299600PMC
December 2018

Concerns About the Effect of Transfusion in Critically Ill Septic Patients.

Crit Care Med 2018 07;46(7):e723-e724

Department of Nephrology, Saiseikai Utsunomiya Hospital, Utsunomiya, Japan; Department of Palliative Care Medicine, Asoka Vihara Hospital, Kyoto, Japan; Department of Pulmonary Medicine, Kawasaki Municipal Ida Hospital, Kawasaki, Japan; Department of Pharmacoepidemiology, Graduate School of Medicine and Public Health, Kyoto University, Kyoto, Japan.

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http://dx.doi.org/10.1097/CCM.0000000000003064DOI Listing
July 2018

Aspergillus precipitating antibody in patients with Mycobacterium avium complex lung disease: A cross-sectional study.

Respir Med 2018 05 12;138:1-6. Epub 2018 Mar 12.

Center for Infectious Diseases and Infection Control, Keio University School of Medicine, Tokyo, Japan. Electronic address:

Rationale: Little is known about the role of Aspergillus precipitating antibody (APAb) in patients with Mycobacterium avium complex lung disease (MAC-LD).

Objectives: We investigated the clinical characteristics of patients with MAC-LD positive for APAb.

Methods: We conducted a cross-sectional study targeting patients with MAC-LD. APAb was checked in all participants. Clinical variables included laboratory data, pulmonary function, high-resolution computed tomography findings, and health-related quality of life.

Results: We analyzed 109 consecutive patients. Their median age was 68 years, and the median duration of MAC-LD was 4.8 years. Twenty (18.3%) patients tested positive for APAb. APAb-positive patients had significantly longer duration of MAC-LD (9.4 vs. 4.0 years, P = 0.017), more severe bronchiectasis evaluated by modified Reiff score (6.5 vs. 4, P = 0.0049), and lower forced expiratory volume in 1 s (%FEV) (75.1% vs. 86.2%, P = 0.013) than APAb-negative patients. Analysis of covariance adjusted for background factors and underlying pulmonary disease revealed that %FEV was also significantly lower in patients with APAb (P = 0.045). Ten patients were newly diagnosed with chronic pulmonary aspergillosis (N = 5) or allergic bronchopulmonary aspergillosis (N = 5).

Conclusions: APAb is associated with lower pulmonary function, and observed especially in patients with longer duration of MAC-LD and severe bronchiectasis, even in the absence of cavitary lesions.
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http://dx.doi.org/10.1016/j.rmed.2018.03.013DOI Listing
May 2018

hsa-miR-346 is a potential serum biomarker of Mycobacterium avium complex pulmonary disease activity.

J Infect Chemother 2017 Oct 19;23(10):703-708. Epub 2017 Aug 19.

Center for Infectious Diseases and Infection Control, Keio University School of Medicine, Japan. Electronic address:

MicroRNA (miRNA) has been recently recognized as a biomarker of various diseases; however, there are no known miRNAs associated with Mycobacterium avium complex (MAC) pulmonary disease. In addition, there are no known biomarkers to precisely reflect disease activity after the diagnosis of MAC pulmonary disease. Thus, we sought to identify a miRNA which is a candidate biomarker of MAC pulmonary disease activity. Serum hsa-miR-346 concentrations of 16 patients with M. avium pulmonary disease were significantly higher than those of 16 healthy controls (p = 0.047). The secretion of hsa-miR-346 increased in a multiplicity of infection-dependent manner in M. avium-infected macrophages. Serum hsa-miR-346 levels of 5 patients with bacterial conversion at the end of follow-up were significantly lower than those at the beginning of the follow-up (p = 0.043). In addition, the longitudinal change in serum hsa-miR-346 concentration correlated with bacterial load in 2 patients with M. avium pulmonary disease. Based on our results, it is supposed that MAC-infected macrophages in pulmonary lesions produce hsa-miR-346, which is then secreted into the bloodstream. The magnitude of this process could be quantitatively controlled by the bacterial load, suggesting that serum hsa-miR-346 is a potentially useful biomarker of MAC pulmonary disease activity.
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http://dx.doi.org/10.1016/j.jiac.2017.07.015DOI Listing
October 2017

Quantitative diagnostic imaging of cancer tissues by using phosphor-integrated dots with ultra-high brightness.

Sci Rep 2017 08 8;7(1):7509. Epub 2017 Aug 8.

Department of Nano-Medical Science, Graduate School of Medicine, Tohoku University, Seiryo-machi, Aoba-ku, Sendai, 980-8575, Japan.

The quantitative sensitivity and dynamic range of conventional immunohistochemistry (IHC) with 3,3'-diaminobenzidine (IHC-DAB) used in pathological diagnosis in hospitals are poor, because enzyme activity can affect the IHC-DAB chromogenic reaction. Although fluorescent IHC can effectively increase the quantitative sensitivity of conventional IHC, tissue autofluorescence interferes with the sensitivity. Here, we created new fluorescent nanoparticles called phosphor-integrated dots (PIDs). PIDs have 100-fold greater brightness and a more than 300-fold greater dynamic range than those of commercially available fluorescent nanoparticles, quantum dots, whose fluorescence intensity is comparable to tissue autofluorescence. Additionally, a newly developed image-processing method enabled the calculation of the PID particle number in the obtained image. To quantify the sensitivity of IHC using PIDs (IHC-PIDs), the IHC-PIDs method was compared with fluorescence-activated cell sorting (FACS), a method well suited for evaluating total protein amount, and the two values exhibited strong correlation (R = 0.94). We next applied IHC-PIDs to categorize the response to molecular target-based drug therapy in breast cancer patients. The results suggested that the PID particle number estimated by IHC-PIDs of breast cancer tissues obtained from biopsy before chemotherapy can provide a score for predicting the therapeutic effect of the human epidermal growth factor receptor 2-targeted drug trastuzumab.
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http://dx.doi.org/10.1038/s41598-017-06534-zDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5548777PMC
August 2017

Regulation of MUC5B Expression in Idiopathic Pulmonary Fibrosis.

Am J Respir Cell Mol Biol 2017 07;57(1):91-99

1 Department of Medicine, School of Medicine, University of Colorado-Denver, Denver, Colorado.

The gain-of-function mucin 5B (MUC5B) promoter variant, rs35705950, confers the largest risk, genetic or otherwise, for the development of idiopathic pulmonary fibrosis; however, the mechanisms underlying the regulation of MUC5B expression have yet to be elucidated. Here, we identify a critical regulatory domain that contains the MUC5B promoter variant and has a highly conserved forkhead box protein A2 (FOXA2) binding motif. This region is differentially methylated in association with idiopathic pulmonary fibrosis, MUC5B expression, and rs35705950. In addition, we show that this locus binds FOXA2 dynamically, and that binding of FOXA2 is necessary for enhanced expression of MUC5B. In aggregate, our findings identify novel targets to regulate the expression of MUC5B.
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http://dx.doi.org/10.1165/rcmb.2017-0046OCDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5516283PMC
July 2017

X-ray computed tomography imaging of a tumor with high sensitivity using gold nanoparticles conjugated to a cancer-specific antibody via polyethylene glycol chains on their surface.

Sci Technol Adv Mater 2016 26;17(1):387-397. Epub 2016 Jul 26.

Department of Nano-Medical Science, Graduate School of Medicine, Tohoku University, Sendai, Japan; Department of Surgical Oncology, Graduate School of Medicine, Tohoku University, Sendai, Japan.

Contrast agents are often used to enhance the contrast of X-ray computed tomography (CT) imaging of tumors to improve diagnostic accuracy. However, because the iodine-based contrast agents currently used in hospitals are of low molecular weight, the agent is rapidly excreted from the kidney or moves to extravascular tissues through the capillary vessels, depending on its concentration gradient. This leads to nonspecific enhancement of contrast images for tissues. Here, we created gold (Au) nanoparticles as a new contrast agent to specifically image tumors with CT using an enhanced permeability and retention (EPR) effect. Au has a higher X-ray absorption coefficient than does iodine. Au nanoparticles were supported with polyethylene glycol (PEG) chains on their surface to increase the blood retention and were conjugated with a cancer-specific antibody via terminal PEG chains. The developed Au nanoparticles were injected into tumor-bearing mice, and the distribution of Au was examined with CT imaging, transmission electron microscopy, and elemental analysis using inductively coupled plasma optical emission spectrometry. The results show that specific localization of the developed Au nanoparticles in the tumor is affected by a slight difference in particle size and enhanced by the conjugation of a specific antibody against the tumor.
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http://dx.doi.org/10.1080/14686996.2016.1194167DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5101864PMC
July 2016

[A CASE OF MILIARY TUBERCULOSIS ASSOCIATED WITH HEPATOSPLENIC ABSCESSES APPEARING DURING ANTI-TUBERCULOUS TREATMENT].

Kekkaku 2015 Oct;90(10):671-5

A 27-year-old man with a 4-month history of treatment for miliary tuberculosis at another hospital was admitted to our hospital for continued treatment. Computed tomography showed new lesions in the S8 area of the liver and spleen, despite resolution of chest radiographic findings. Because these new lesions were still present after 8 months of treatment, we performed laparoscopic drainage of the liver abscess. Purulent material drained from the lesion revealed positive polymerase chain reaction results for Mycobacterium tuberculosis, and identification of granuloma with infiltrating lymphocytes and plasma cells confirmed the diagnosis of tubercular liver abscess. Pathological changes in the spleen over the clinical course were also regarded as representing tubercular abscess. Postoperative course was good, and tuberculosis treatment ended after 12 months. Tubercular liver abscess subsequently showed prominent reduction, and the tubercular splenic abscess disappeared on abdominal ultrasonography. Tubercular hepatosplenic abscesses appearing during tubercular treatment are rare. We report this valuable case in which laparoscopic drainage of a liver abscess proved useful for diagnosis and treatment.
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October 2015

Development of a quantitative diagnostic method of estrogen receptor expression levels by immunohistochemistry using organic fluorescent material-assembled nanoparticles.

Biochem Biophys Res Commun 2012 Sep 29;426(3):409-14. Epub 2012 Aug 29.

Department of Nano-Medical Science, Graduate School of Medicine, Tohoku University, Seiryo-machi, Sendai 980-8575, Japan.

The detection of estrogen receptors (ERs) by immunohistochemistry (IHC) using 3,3'-diaminobenzidine (DAB) is slightly weak as a prognostic marker, but it is essential to the application of endocrine therapy, such as antiestrogen tamoxifen-based therapy. IHC using DAB is a poor quantitative method because horseradish peroxidase (HRP) activity depends on reaction time, temperature and substrate concentration. However, IHC using fluorescent material provides an effective method to quantitatively use IHC because the signal intensity is proportional to the intensity of the photon excitation energy. However, the high level of autofluorescence has impeded the development of quantitative IHC using fluorescence. We developed organic fluorescent material (tetramethylrhodamine)-assembled nanoparticles for IHC. Tissue autofluorescence is comparable to the fluorescence intensity of quantum dots, which are the most representative fluorescent nanoparticles. The fluorescent intensity of our novel nanoparticles was 10.2-fold greater than quantum dots, and they did not bind non-specifically to breast cancer tissues due to the polyethylene glycol chain that coated their surfaces. Therefore, the fluorescent intensity of our nanoparticles significantly exceeded autofluorescence, which produced a significantly higher signal-to-noise ratio on IHC-imaged cancer tissues than previous methods. Moreover, immunostaining data from our nanoparticle fluorescent IHC and IHC with DAB were compared in the same region of adjacent tissues sections to quantitatively examine the two methods. The results demonstrated that our nanoparticle staining analyzed a wide range of ER expression levels with higher accuracy and quantitative sensitivity than DAB staining. This enhancement in the diagnostic accuracy and sensitivity for ERs using our immunostaining method will improve the prediction of responses to therapies that target ERs and progesterone receptors that are induced by a downstream ER signal.
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http://dx.doi.org/10.1016/j.bbrc.2012.08.105DOI Listing
September 2012

[Case report: a case of pulmonary infectious disease due to Mycobacterium shinjukuense].

Nihon Naika Gakkai Zasshi 2011 Dec;100(12):3637-9

Department of Internal Medicine, Kawasaki Municipal Ida Hospital, Japan.

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http://dx.doi.org/10.2169/naika.100.3637DOI Listing
December 2011

Carbon black nanoparticles enhance bleomycin-induced lung inflammatory and fibrotic changes in mice.

Exp Biol Med (Maywood) 2011 Mar;236(3):315-24

Division of Pulmonary Medicine, Keio University School of Medicine, Tokyo, Japan.

With the recent increasing use of nanoparticles, there is concern that they may become an environmental risk factor as airborne particles. However, the impact of these particles on susceptible subjects with predisposing lung disease have not been sufficiently elucidated. In the present study, we investigated the effects of nanoparticles on pulmonary inflammatory and fibrotic changes induced by intratracheal bleomycin (BLM) challenge in mice. Mice were intratracheally administered either vehicle, 14-nm carbon black nanoparticles (CBNPs), BLM or BLM plus CBNP. First, we assessed lung collagen content, lung compliance and fibrotic changes in histopathology on day 21 after instillation. Then, to elucidate how CBNP contributes to the development of BLM-induced fibrosis, we collected bronchoalveolar lavage (BAL) fluid on days 2, 7, 14 and 21 and determined the total and differential cell counts and concentrations of two proinflammatory cytokines (keratinocyte chemoattractant [KC] and interleukin [IL]-6) and two fibrogenic mediators (CC chemokine ligand 2 [CCL2] and transforming growth factor-β(1) [TGF-β(1)]). Expression of nitrotyrosine, an indicator of oxidant injury, was also evaluated on days 7 and 21. CBNP, when combined with BLM, significantly enhanced BLM-induced increase in lung collagen content, decrease in lung compliance, and fibrotic changes in histopathology. CBNP significantly augmented BLM-induced increase in the numbers of inflammatory cells in BAL fluid on days 2 and 7 and levels of KC and IL-6 on day 2. In addition, CBNP administered in combination with BLM significantly elevated the levels of CCL2 on days 2, 7 and 14, and TGF-β(1) on day 14 in BAL fluid as compared with BLM alone. Nitrotyrosine expression was also increased by BLM plus CBNP compared with BLM alone. In contrast, CBNP did not exert any significant effect on these parameters by itself. These results indicate that CBNP can exaggerate BLM-induced inflammatory and fibrotic changes in the lung, suggesting the potential impact of nanoparticles on lung inflammation and fibrosis.
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http://dx.doi.org/10.1258/ebm.2011.010180DOI Listing
March 2011

[A case having chyliform pleural effusion caused by former tuberculous pleurisy].

Kekkaku 2011 Feb;86(2):57-60

Department of Respiratory Medicine, Kawasaki Municipal Ida Hospital, 2-27-1, Ida, Nakahara-ku, Kawasaki-shi, Kanagawa 211-0035, Japan.

A 49-year-old male who had been treated for pulmonary tuberculosis and tuberculous pleurisy in 2007 was referred to our hospital with the complaint of dyspnea on exertion in Nov. 2009. Chest X-ray showed increased pleural effusion compared with that remaining after the previous treatment of pleurisy in 2008. A chest CT revealed that fluid collection was surrounded by thickened pleura. Thoracocentesis was performed, and yellow milky liquid was obtained. The pleural effusion contained few cells. The triglyceride concentration was 83 mg/dl, and the cholesterol level was very high at 628 mg/dl. Based on these findings we diagnosed this case as chyliform pleural effusion. Both smear of acid-fast bacilli and PCR-TB test of the pleural effusion were positive, but culture was negative for mycobacterium, suggesting that this chyliform pleural effusion was produced by the former episode of tuberculous pleurisy, not by the recent reactivation of tuberculous pleurisy. The ADA concentration in the pleural effusion was high at 91.7 IU/l. No increase in the amount of pleural effusion was observed after thoracocentesis without any anti-tuberculosis therapy.
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February 2011

[A case of tuberculous pleurisy with transient new intra-pulmonary lesions during anti-tuberculosis therapy].

Kekkaku 2010 Aug;85(8):667-71

Department of Pulmonary Medicine, Kawasaki Municipal Ida Hospital, 2-27-1, Ida, Nakahara-ku, Kawasaki-shi, Kanagawa 211-0035, Japan.

A 24-year-old man who had been treated 3 months for tuberculous pleurisy presented with thoracic back pain. Chest CT showed a new lesion abutting the pleura, despite the disappearance of pleural effusion. Two weeks later, the mass abutting the pleura progressed to form a new intrapulmonary infiltrative shadow. A transbronchial lung biopsy was performed and the histopathologic examination of the specimen from this lesion revealed granulomatous inflammation without caseous necrosis or acid-fast bacilli. No acid-fast bacilli were cultured from the bronchoalveolar lavage fluid. Anti-tuberculosis medication was continued without change, and the lesions finally resolved. More than 3 years have passed since the completion of anti-tuberculosis chemotherapy, and no recurrence has been observed. We believe that these lesions were pulmonary tuberculomas and transient intra-pulmonary infiltration due to non-specific inflammation, caused secondarily by an excessive immune response, as in paradoxical worsening.
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August 2010

Production and activation of matrix metalloproteinase 7 (matrilysin 1) in the lungs of patients with idiopathic pulmonary fibrosis.

Arch Pathol Lab Med 2010 Aug;134(8):1136-42

Department of Emergency and Critical Care Medicine, School of Medicine, Keio University, Shinjuku-ku, Tokyo, Japan.

Context: Idiopathic pulmonary fibrosis (IPF) is characterized by diffuse interstitial inflammation and fibroblast proliferation with accelerated remodeling of extracellular matrix, which result in irreversible destruction of the lung's architecture.

Objective: To elucidate the production levels, tissue localization, and activation of matrix metalloproteinase 7 (MMP-7) in the lungs of patients with IPF.

Design: Bronchoalveolar lavage analysis was performed in 17 IPF patients and 6 healthy volunteers. Levels of MMP-7 in blood were assayed in 23 IPF patients and 20 controls. Histologic and immunohistochemical analyses were performed on paraffin sections of the lung tissues from patients with IPF, interstitial pneumonia associated with rheumatoid arthritis, or nonspecific interstitial pneumonia.

Results: The proMMP-7 levels in bronchoalveolar lavage fluids from IPF patients were significantly higher than those from healthy controls, although there was no difference in the serum levels between the 2 groups. By immunohistochemistry, proMMP-7 was localized mainly to the hyperplastic alveolar and metaplastic bronchiolar epithelial cells in the lung tissues from IPF patients. Active MMP-7 was immunolocalized on alveolar macrophages and hyperplastic epithelial cells, which were also immunostained with antibody against CD151, a molecule associated with activation of proMMP-7. Immunoblot analysis indicated the overproduction of proMMP-7 together with a small amount of active MMP-7 in bronchoalveolar lavage fluids from IPF patients. The MMP-7 activity was detected in a cross-linked carboxymethylated transferrin film assay.

Conclusions: proMMP-7 is excessively produced by hyperplastic alveolar and metaplastic bronchiolar epithelial cells and activated locally in the lungs of IPF patients, suggesting that MMP-7 may contribute to the pathology of IPF.
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http://dx.doi.org/10.5858/2009-0144-OA.1DOI Listing
August 2010

Role of interleukin-6 in elastase-induced lung inflammatory changes in mice.

Exp Lung Res 2010 Aug;36(6):362-72

Division of Pulmonary Medicine, Keio University School of Medicine, Shinjuku-ku, Tokyo, Japan.

Interleukin-6 (IL-6) is known to be involved in the pathogenesis of various inflammatory diseases, but its role in the development of pulmonary emphysema remains unclear. Wild-type (WT) and IL-6-deficient mice received either phosphate-buffered saline (PBS) or porcine pancreatic elastase (PPE) intratracheally. The development of emphysema was determined by measuring the mean linear intercept (Lm). The lung specimens were also subjected to immunohistochemistry for single-stranded DNA to detect apoptotic cells. Lung mechanics and airway responsiveness to inhaled methacholine were analyzed. Bronchoalveolar lavage (BAL) fluid was subjected to evaluation of inflammatory cell accumulation and cytokine measurement. PPE treatment caused significant increases in Lm and lung compliance, which was attenuated by IL-6 deficiency. The increases in apoptotic cells in the lung were attenuated in IL-6 null mice. Airway responsiveness was not affected by PPE challenge or IL-6 deficiency. Intratracheal PPE increased the cell counts in BAL fluid throughout the observation, which was suppressed in IL-6 null mice. In BAL fluid, PPE-induced increases in the levels of macrophage inflammatory protein (MIP)-1alpha and eotaxin were mitigated by IL-6 deficiency. PPE-induced up-regulation of matrix metalloproteinase (MMP)-12 in the lung was attenuated by IL-6 deficiency. These results indicate that IL-6 may play an important role in the development of elastase-induced lung inflammatory changes.
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http://dx.doi.org/10.3109/01902141003678590DOI Listing
August 2010

Effect of Toll-like receptor 4 inhibitor on LPS-induced lung injury.

Inflamm Res 2010 Oct 13;59(10):837-45. Epub 2010 Apr 13.

Department of Anesthesiology, Keio University School of Medicine, Tokyo, Japan.

Objective And Design: Toll-like receptor 4 (TLR4) plays important roles in the recognition of lipopolysaccharide (LPS) and the activation of inflammatory cascade. In this study, we evaluated the effect of TAK-242, a selective TLR4 signal transduction inhibitor, on acute lung injury (ALI).

Materials And Methods: C57BL/6J mice were intravenously treated with TAK-242 15 min before the intratracheal administration of LPS or Pam3CSK4, a synthetic lipopeptide. Six hours after the challenge, bronchoalveolar lavage fluid was obtained for a differential cell count and the measurement of cytokine and myeloperoxidase levels. Lung permeability and nuclear factor-kappaB (NF-kappaB) DNA binding activity were also evaluated.

Results: TAK-242 effectively attenuated the neutrophil accumulation and activation in the lungs, the increase in lung permeability, production of inflammatory mediators, and NF-kappaB DNA-binding activity induced by the LPS challenge. In contrast, TAK-242 did not suppress inflammatory changes induced by Pam3CSK4.

Conclusion: TAK-242 may be a promising therapeutic agent for ALI, especially injuries associated with pneumonia caused by Gram-negative bacteria.
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http://dx.doi.org/10.1007/s00011-010-0195-3DOI Listing
October 2010

Intratracheal synthetic CpG oligodeoxynucleotide causes acute lung injury with systemic inflammatory response.

Respir Res 2009 Sep 23;10:84. Epub 2009 Sep 23.

Division of Pulmonary Medicine, Keio University School of Medicine, Tokyo, Japan.

Bacterial genome is characterized by frequent unmethylated cytosine-phosphate-guanine (CpG) motifs. Deleterious effects can occur when synthetic oligodeoxynucleotides (ODN) with unmethylated CpG dinucleotides (CpG-ODN) are administered in a systemic fashion. We aimed to evaluate the effect of intratracheal CpG-ODN on lung inflammation and systemic inflammatory response. C57BL/6J mice received intratracheal administration of CpG-ODN (0.01, 0.1, 1.0, 10, or 100 microM) or control ODN without CpG motif. Bronchoalveolar lavage (BAL) fluid was obtained 3 or 6 h or 1, 2, 7, or 14 days after the instillation and subjected to a differential cell count and cytokine measurement. Lung permeability was evaluated as the BAL fluid-to-plasma ratio of the concentration of human serum albumin that was injected 1 h before euthanasia. Nuclear factor (NF)-kappaB DNA binding activity was also evaluated in lung homogenates. Intratracheal administration of 10 microM or higher concentration of CpG-ODN induced significant inflammatory cell accumulation into the airspace. The peak accumulation of neutrophils and lymphocytes occurred 1 and 2 days after the CpG-ODN administration, respectively. Lung permeability was increased 1 day after the 10 microM CpG-ODN challenge. CpG-ODN also induced nuclear translocation of NF-kappaB and upregulation of various inflammatory cytokines in BAL fluid and plasma. Histopathology of the lungs and liver revealed acute lung injury and liver damage with necrosis, respectively. Control ODN without CpG motif did not induce any inflammatory change. Since intratracheal CpG-ODN induced acute lung injury as well as systemic inflammatory response, therapeutic strategies to neutralize bacterial DNA that is released after administration of bactericidal agents should be considered.
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http://dx.doi.org/10.1186/1465-9921-10-84DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2761862PMC
September 2009

Effects of TNF-alpha-converting enzyme inhibition on acute lung injury induced by endotoxin in the rat.

Shock 2009 Nov;32(5):535-40

Division of Pulmonary Medicine, Department of Medicine, Keio University School of Medicine, Shinjuku-ku, Tokyo, Japan.

We studied the effects of TNF-converting enzyme inhibition with Y-41654, which down-regulates the production of soluble TNF-alpha (sTNF-alpha), on acute lung injury induced by intratracheal administration of LPS. We first verified in vitro that pretreatment of isolated alveolar macrophages from Sprague-Dawley male rats with 20 microL of 0.1-mM Y-41654, decreased significantly (P < 0.05) the concentration of sTNF-alpha in cell supernatants induced by 10 microg/mL of LPS. We then studied four groups of rats (each n = 10) including 1) a control group, 2) an LPS group (300 microg /kg, instilled intratracheally), 3) a Y-41654 group, and 4) a treatment group treated with Y-41654 after LPS instillation. Y-41654, 10 mg/kg in 0.7 mL of phosphate-buffered saline, was administered (i.v.), 15 min before and 0.5, 1.5, 2.5, and 3.5 h after saline or LPS instillation. The animals were observed for 4 h. In the animals treated with Y-41654, the concentrations of sTNF-alpha and protein in bronchoalveolar lavage fluid, and the number of neutrophils in lung tissue and bronchoalveolar lavage fluid were significantly lower at 4 h than in the LPS group (P < 0.05). In conclusion, sTNF-alpha plays an important role in the development of acute lung injury induced by intratracheal administration of LPS, in part modulating neutrophil kinetics.
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http://dx.doi.org/10.1097/SHK.0b013e3181a2adb7DOI Listing
November 2009

Elevated CC chemokine level in bronchoalveolar lavage fluid is predictive of a poor outcome of idiopathic pulmonary fibrosis.

Respiration 2009 6;78(3):285-92. Epub 2009 Mar 6.

Division of Pulmonary Medicine, Department of Medicine, Keio University School of Medicine, Tokyo, Japan.

Background: CC chemokines play important roles in the pathogenesis of interstitial lung diseases. Elevated CC chemokine levels have been observed in bronchoalveolar lavage (BAL) fluid of patients with idiopathic pulmonary fibrosis (IPF).

Objectives: We aimed to examine whether the levels of four CC chemokines, i.e. monocyte chemoattractant protein-1 (MCP-1/CCL2), macrophage inflammatory protein-1 alpha (MIP-1 alpha/CCL3), thymus- and activation-regulated chemokine (TARC/CCL17), and macrophage-derived chemokine (MDC/CCL22), in BAL fluid are predictive of the prognosis of IPF patients.

Methods: We compared the chemokine levels of patients alive 5 years after diagnosis and those who had died. Lung function data, CT scores, and serum markers were also compared.

Results: Among 39 patients (29 males, median age, 60 years), 19 patients (48%) died within 5 years after the diagnosis. Whereas percent vital capacity was not different, percent lung diffusion capacity for carbon monoxide was significantly higher in the surviving patients than in the nonsurviving patients (p < 0.01). Median CCL2 levels of surviving and nonsurviving patients were 154.3 (interquartile range, IQR: 67.3-381.8) and 427.2 (IQR: 329.2-1184.1) pg/ml, respectively (p < 0.02). CCL3 levels in BAL fluid did not differ between the surviving and nonsurviving patients. CCL17 was detected in BAL fluid of 7 patients, 6 of whom died within 5 years. CCL22 was detectable in BAL fluid of 10 patients, only 1 of whom survived. Serum levels of KL-6 and lactate dehydrogenase did not differ between the surviving and nonsurviving patients.

Conclusion: Elevated levels of CCL2, CCL17 and CCL22 in BAL fluid might be predictive of a poor outcome in patients with IPF.
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http://dx.doi.org/10.1159/000207617DOI Listing
January 2010

Concentrations of clarithromycin and active metabolite in the epithelial lining fluid of patients with Mycobacterium avium complex pulmonary disease.

Pulm Pharmacol Ther 2009 Jun 21;22(3):190-3. Epub 2008 Nov 21.

Division of Pulmonary Medicine, Department of Medicine, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582, Japan.

Objectives: Clarithromycin (CAM) is widely accepted for the treatment of Mycobacterium avium complex (MAC) pulmonary diseases. This study measured (a) the concentrations of CAM and its active metabolite (14OH-CAM) in bronchial epithelial lining fluid (ELF) obtained by bronchoscopic microsampling (BMS), and (b) the minimal inhibitory concentrations (MIC) of CAM for each MAC isolate.

Methods: We studied eight patients with MAC pulmonary disease treated with oral CAM, 400 (n=4) or 800 (n=4) mg/day. BMS was performed 3h after the last CAM dose, and the concentrations of CAM and 14OH-CAM were measured in ELF collected from the diseased and normal contralateral pulmonary segments, and in serum.

Results: The mean+/-SEM ELF concentrations of CAM (23.85+/-7.64 microg/ml) and CAM+14OH-CAM (28.71+/-8.37 microg/ml) in the 800 mg/day treatment group were significantly higher than in the 400mg/day group (7.48+/-2.58 microg/ml and 9.63+/-2.99 microg/ml, respectively; both p<0.05), while the serum concentrations were similar in both groups. In both treatment groups, the mean ELF concentrations sampled from diseased and normal segments were higher than the MIC against MAC isolates. The mean ELF concentration of CAM in the 400mg/day treatment group was <8 mircog/ml (the breakpoint concentration of CAM against M. avium recommended by the Clinical Laboratories Standards Institute), while the mean concentration in the 800 mg/day treatment group was >8 microg/ml.

Conclusion: These observations suggest that CAM, 800 mg/day, is an appropriate dose to treat MAC pulmonary disease, and prevent its spread from diseased to non-diseased lung segments.
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http://dx.doi.org/10.1016/j.pupt.2008.11.004DOI Listing
June 2009
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