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Improved method of plasma 8-Isoprostane measurement and association analyses with habitual drinking and smoking

World Journal of Gastroenterology 2006年第36期12卷 5846-5852页

作者： Soichi Kitano Hisashi Hisatomi Nozomu Hibi Katsumi Kawano Shoji Harada Technology Development Department SRL Inc. Hachioji Tokyo Japan Center for Molecular Biology and Cytogenetics SRL Inc. Hino Tokyo Japan

AIM: To develop a simple and accurate method for quantifying 8-isoprostane in plasma by employing a combination of two-step solid-phase extraction of samples and a commercially available ELISA kit, and by this method to examine the effects of drinking and smoking habits against the levels of plasma 8-isoprostane in healthy Japanese ***: Plasma 8-isoprostane was extracted with ODS gel suspension followed by NH2 Sep-Pak column. The 8-isoprostane fractions were assayed using a commercially available ELISA kit. We measured plasma 8-isoprostane levels in 157 healthy Japanese volunteers divided into three groups (64 non-habitual drinkers, 56 moderate drinkers and 37 habitual drinkers) according to their alcohol consumption per week. Genotypes of aldehyde dehydrogenase 2 (ALDH2) were also determined to investigate the plasma 8-isoprostane levels with reference to drinking habits. In addition, the plasma 8-isoprostane levels of 96 non-smokers and 61 smokers from the same subjects were ***: Our method fulfilled all the requirements for use in routine clinical assays with respect to sensitivity, intra- and inter-assay reproducibility, accuracy and dynamic assay range. Significant inc.eases of plasma 8-isoprostane levels were observed in female habitual drinkers when compared with those of non-habitual drinkers (t = 5.494, P inc.ease oxidization stress, especially in females. On the contrary, no significant difference of the plasma 8-isoprostane levels was observed between non-smokers and ***: Our present method was proved to be a simple and accurate tool for measuring plasma 8-isoprostane. However, the clinical utility of plasma 8-isoprostane for drinking and smoking habits was

关键词： 8-Isoprostane ELISA Lipid peroxidation Drinking Smoking

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Decrease of reactive-oxygen-producing granulocytes and release of IL-10 into the peripheral blood following leukocytapheresis in patients with active ulcerative colitis

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World Journal of Gastroenterology 2005年第20期11卷 3085-3090页

作者： Hiroyuki Hanai Takayuki Iida Ken Takeuchi Fumitoshi Watanabe Yasuhiko Maruyama Masataka Kikuyama Tatsuo Tanaka Kenji Kondo Kou Tanaka Kenji Takai Department of Endoscopic and Photodynamic Medicine Hamamatsu University School of Medicine HamamatsuJapan Department of Gastroenterology Fujieda Municipal General Hospital FujiedaJapan Department of Gastroenterology Hamamatsu Rosai Hospital Hamamatsu Japan Tanaka Clinic Shizuoka Japan srl inc. Tokyo Japan

AIM: To investigate the clinical efficacy of leukocytapheresis (LCAP) in patients with active ulcerative colitis (UC), and to elucidate the mechanisms by determining the changes in the cytokine levels in the peripheral blood and of the functions of the peripheral blood leukocytes in these patients. METHODS: The subjects were 19 patients with active UC, with a mean clinical activity index (CAI) of 9.2. The LCAP was conducted using Cellsorba E. In each session of LCAP, 2-3 L of blood at the flow rate of 30-50 mL/min was processed. The treatment was carried out in approximately 1-h sessions, once a week, for 5-10 wk. Blood samples for determination of the cytokine levels were collected from the inflow side of the column (site of dehematization; at the start of LCAP) and outflow side of the column (at the end of LCAP). Blood samples for the determination of reactive-oxygen-producing cells were collected from the peripheral blood before and after LCAP. RESULTS: LCAP resulted in clinical improvement in all the 19 patients of UC recruited for this study. Remission (CAI: ≤4) was noted in 15 (79%) of the 19 patients. The blood level of the pro-inflammatory cytokine IL-6 was found to be decreased following treatment by LCAP, and the level of the anti-inflammatory cytokine IL-10 at the outflow side of the LCAP column was found to be significantly elevated as compared to that at the inflow side of the column. The reactive-oxygen-producing granulocytes in the peripheral blood of UC patients was inc.eased as compared to that in healthy persons and the inc.ease was found to be decreased following treatment by LCAP. CONCLUSION: LCAP exerted a high therapeutic efficacy in patients with active UC. Our findings suggest that LCAP is associated with enhanced production of the inhibitory cytokine IL-10 to indirectly inhibit the functions of the inflammatory leukocytes, and that inflammation is also considerably attenuated by the direct removal of reactive-oxygen-producing neutrophils fro

关键词： Ulcerative colitis Reactive-oxygen Leukocyta-pheresis Interleukin-10

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New combination test for hepatitis C virus genotype and viral load determination using Amplicor GT HCV MONITOR test v2.0

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World Journal of Gastroenterology 2005年第4期11卷 469-475页

作者： Motokazu Mukaide Yasuhito Tanaka Hirokazu Kakuda Kei Fujiwara Fuat Kurbanov Eturo Orito Kentaro Yoshioka Kiyotaka Fujise Shoji Harada Takazumi Kozaki Kazuo Takemura Kazumasa Hikiji Masashi Mizokami srl inc. 5-6-50ShinmachiHino-shiTokyo 191-0002Japan Department of Clinical Molecular Informative Medicine Nagoya City University Graduate School of Medical Sciences KawasumiMizuhoNagoya 467-8601Japan Department of Internal Medicine and Molecular Science Nagoya City University Graduate School of Medical Sciences KawasumiMizuhoNagoya 467-8601Japan Department of Clinical Molecular Informative Medicine Nagoya City University Graduate School of Medical Sciences KawasumiMizuhoNagoya 467-8601 Japan Third Department of Internal Medicine Nagoya University School of Medicine65 TsurumaShowaNagoya 466-8550Japan Division of Gastroenterology and Hepatology Department of Internal Medicine (Kashiwa) The Jikei University School of Medicine163-1KashiwashitaKashiwaChiba 277-8567Japan srl inc. 5-6-50ShinmachiHmo-shiTokyo 191-0002Japan

AIM: To develop a new sensitive and inexpensive hepatitis C virus (HCV) combination test (HCV Guideline test) that enables the determination of HCV genotypes 1, 2 and 3, and simultaneous determination of HCV viral load using commercial Amplicor GT HOV MONITOR test v2.0 (microwell version). METHODS: The HCV Guideline test used the PCR product generated in commercial Amplicor GT HCV Monitor test v2.0 for viral load measurement using microwell plate version of Amplicor HCV Monitor and also captured on separate plates containing capture probes and competitive oligonucleotide probes specific for HCV genotypes 1, 2 and 3, The HCV genotype was subsequently determined using the biotin-labeled PCR product and five biotin-labeled HCV-specific probes. RESULTS: The sensitivity of the HCV Guideline test was 0.5 KIU/mL. Specificity of the HCV Guideline test was confirmed by direct sequencing of HCV core region and molecular evolutionary analyses based on a panel of 31 samples. The comparison of the HCV Guideline test and an in-house HCV core genotyping assay using 252 samples from chronic hepatitis C patients indicated concordant results for 97.2% of samples (59.5% genotype 1, 33.7% genotype 2, 6.0% genotype 3, and 0.8% mixed genotypes). Similarly, the HCV Guideline test showed concordance with a serological test, and the serological test failed to assign any serotype in 12.7% of the samples, indicating a better sensitivity of the HCV Guideline test. CONCLUSION: Clinically, both viral load and genotypes (1, 2 and 3) have been found to be major predictors of antiviral therapy outcome regarding chronic hepatitis C based on guidelines and they are, in normal circumstances, performed as separate stand-alone assays. The HCV Guideline test is a useful method for screening large cohorts in a routine clinical setting for determining the treatment regimen and for predicting the outcome of antiviral therapy of chronic hepatitis C.

关键词： Hepatitis CVirus HCV Guideline test Viral Load Genotype

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Standardized Assessment of Ki-67 in Breast Cancer Patients Using Virtual Slides and an Automated Analyzer in Comparison to Central/Local Pathological Assessments

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Journal of Cancer Therapy 2014年第2期5卷 141-146页

作者： Yoshio Mizuno Hiromi Fuchikami Tsuneo Natori Naoko Takeda Yuko Inoue Junichi Yamada Hiroaki Abe Hiroshi Seto Kazuhiko Sato Clinical Testing Business Testing Division SRL Inc. Tokyo Japan Department of Breast Oncology Tokyo-West Tokushukai Hospital Tokyo Japan Department of Clinical Pathology Tokyo-West Tokushukai Hospital Tokyo Japan Inoue Ladies Clinic Tokyo Japan Seto Hospital Tokorozawa Japan

Purpose: To standardize the methods to measure Ki-67, there is an interest in automating the assessment of Ki-67. Therefore, we reviewed the possibility of introducing an automated analyzer to standardize the Ki-67 evaluation method. Methods: We retrospectively reviewed a clinical database of patients who underwent surgery for early breast cancer at Tokyo-West Tokushukai Hospital. Among them, those who underwent preoperative core needle biopsy (CNB) were enrolled. The concordance rates of estrogen receptor (ER), progesterone receptor (PgR), human epidermal growth factor receptor 2 (HER2), and Ki-67 by local pathologists were reviewed (valuations made by local pathologists), and nonmatching cases (from August 2008 to October 2011) were reassessed both by central review and using an automated analyzer with virtual slides. The results were compared with the evaluations made by local pathologists, and we reexamined the concordance rate by using central review and the automated analyzer. Results: The concordance rate of Ki-67 evaluations made by local pathologists in the preoperative CNB and surgical specimens was 78.7% in 287 cases pathologically assessed from October 2008 to March 2013. This rate was significantly lower (p < 0.01) than that of ER (95.6%), PgR (88.5%), and HER2 (91.6%). Reassessment of the 37 cases of nonmatching Ki-67 values from 2008 to October 2011 using central review and an automated analyzer resulted in clear improvement in matching of 22 (92.1%) and 24 (93.1%) of 37 cases, respectively. Conclusion: The concordance rate of Ki-67 in preoperative CNB and surgical specimens was lower than that of other biological markers;however, they were nearly equal by reassessment using central review and an automated analyzer.

关键词： Ki-67 Core Needle Biopsy Concordance Rate Virtual Slides Automated Analyzer Standardized Assessment

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An Autoantibody Based Protein Microarray Blood Test to Enhance the Specificity of a Negative Screening Mammogram

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Advances in Breast Cancer Research 2015年第1期4卷 22-38页

作者： T. M. Allweis L. Strauss Z. Malyutin A. Bassein Kapov-Kagan I. Novikov T. B. Bevers S. Iacobelli M. T. Sandri A. Bitterman P. Engelman B. Piura M. Rosenberg G. Yahalom Kaplan Medical Center Rehovot Israel Eventus Diagnostics (Israel) LTD Ora Israel University of Texas MD Anderson Cancer Center Houston USA Media Pharma srl Chieti Italy Laboratory Medicine Division Istituto Europeo di Oncologia Milan Italy Carmel Medical Center Surgery A Haifa Israel Zvulun Breast Center Clalit Organization Kiryat Bialik Israel Eventus Diagnostics inc. Miami USA

Background: Current screening mammography for breast cancer is associated with misdiagnosis in as many as 30% of cases. Objectives: To develop and clinically evaluate a unique autoantibody based protein microarray blood test to improve the accuracy of breast cancer screening. Materials and Methods: A microarray was constructed from commercial antigens and antigens selected from screened cDNA libraries of breast cancer tissue samples. A training set containing 439 healthy controls and 276 biopsy proven breast cancer cases was used to establish a set of separating models between the two groups. These models were used to assign a diagnosis to 285 blind samples from 120 breast cancer patients and 165 healthy controls. Results: The test identified 82 of the 120 breast cancer patients and 160 of the 165 healthy controls. These results can be translated into a sensitivity of 68.3% [CI: 59% -77%] and a specificity of 97% [CI: 93% -99%], with a PPV for this validation set of 94.3% (CI: 87.10% -98.11%), NPV of 80.81% [CI: 74.62% -86.05%] and an AUC of 89.2% [CI: 78% -87%]. Conclusions: The protein microarray can be utilized to reduce the false negative rate of routine screening mammography. Women with a negative mammography and a negative blood test can be reassured and encouraged to continue routine breast cancer screening. A positive test should alert the physician about the possible presence of a breast cancer not detected by routine screening mammography and drive to perform additional investigation, such as breast ultrasound and MRI.

关键词： Autoantibodies Breast Cancer Diagnostic Screening

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