Koyu Suzuki

One-step Nucleic Acid Amplification for Intraoperative Detection of Lymph Node Metastasis in Breast Cancer Patients

Purpose: Detection of sentinel lymph node (SLN) metastasis in breast cancer patients has conventionally been determined by intraoperative histopathologic examination of frozen sections followed by definitive postoperative examination of permanent sections. The purpose of this study is to develop a more efficient method for intraoperative detection of lymph node metastasis. Experimental Design: Cutoff values to distinguish macrometastasis, micrometastasis, and nonmetastasis were determined by measuring cytokeratin 19 (CK19) mRNA in histopathologically positive and negative lymph nodes using one-step nucleic acid amplification (OSNA). In an intraoperative clinical study involving six facilities, 325 lymph nodes (101 patients), including 81 SLNs, were divided into four blocks. Alternate blocks were used for the OSNA assay with CK19 mRNA, and the remaining blocks were used for H&E and CK19 immunohistochemistry–based three-level histopathologic examination. The results from the two methods were then compared. Results: We established CK19 mRNA cutoff values of 2.5 × 102 and 5 × 103 copies/μL. In the clinical study, an overall concordance rate between the OSNA assay and the three-level histopathology was 98.2%. Similar results were obtained with 81 SLNs. The OSNA assay discriminated macrometastasis from micrometastasis. No false positive was observed in the OSNA assay of 144 histopathologically negative lymph nodes from pN0 patients, indicating an extremely low false positive for the OSNA assay. Conclusion: The OSNA assay of half of a lymph node provided results similar to those of three-level histopathology. Clinical results indicate that the OSNA assay provides a useful intraoperative detection method of lymph node metastasis in breast cancer patients.

Molecular Detection of Lymph Node Metastases in Breast Cancer Patients: Results of a Multicenter Trial Using the One-Step Nucleic Acid Amplification Assay

Purpose: Accurate assessment of metastasis in sentinel lymph nodes (SLN) of breast cancer is important but involves a heavy workload for the pathologist. We conducted a multicenter clinical trial in Japan to evaluate a new automated assay system for cytokeratin 19 mRNA, the one-step nucleic acid amplification (OSNA) assay (Sysmex), to detect lymph node metastasis of breast cancer. Experimental Design: Surgically obtained axillary lymph nodes were sectioned into four pieces, two of which were examined with the OSNA assay. The other two adjacent pieces were examined with H&E and immunohistochemical staining for cytokeratin 19. Serial sections at 0.2-mm intervals were used in trial 1 to determine the specificity of the OSNA assay, and three pairs of sections cut from the sliced surfaces of the pieces were used in trial 2 to compare the accuracy of the OSNA assay with that of a routine pathologic examination for SLNs in Japan. Results: In trial 1, the sensitivity and specificity were 95.0% [95% confidence interval (95% CI), 75.1-99.9%] and 97.1% (95% CI, 91.8-99.4%), respectively, for 124 axillary lymph nodes obtained from 34 patients. In trial 2, the agreement between findings of the assay and of the pathologic examination was 92.9% (95% CI, 90.1-95.1%) for 450 axillary lymph nodes obtained from 164 patients. Conclusion: The OSNA assay can detect lymph node metastasis as accurately as can conventional pathology and thus can be an effective addition to or alternative for rapid intraoperative examination of SLNs.

Anatomic Properties of Myocardial Bridge Predisposing to Myocardial Infarction

Background— A myocardial bridge (MB) that partially covers the course of the left anterior descending coronary artery (LAD) sometimes causes myocardial ischemia, primarily because of hemodynamic deterioration, but without atherosclerosis. However, the mechanism of occurrence of myocardial infarction (MI) as a result of an MB in patients with spontaneously developing atherosclerosis is unclear. Methods and Results— One hundred consecutive autopsied MI hearts either with MBs [MI(+)MB(+) group; n=46] or without MBs (n=54) were obtained, as were 200 normal hearts, 100 with MBs [MI(−)MB(+) group] and 100 without MBs. By microscopy on LADs that were consecutively cross-sectioned at 5-mm intervals, the extent and distribution of LAD atherosclerosis were investigated histomorphometrically in conjunction with the anatomic properties of the MB, such as its thickness, length, and location and the MB muscle index (MB thickness multiplied by MB length), according to MI and MB status. In the MI(+)MB(+) group, the MB showed a significantly greater thickness and greater MB muscle index (P<0.05) than in the MI(−)MB(+) group. The intima-media ratio (intimal area/medial area) within 1.0 cm of the left coronary ostium was also greater (P<0.05) in the MI(+)MB(+) group than in the other groups. In addition, in the MI(+)MB(+) group, the location of the segment that exhibited the greatest intima-media ratio in the LAD proximal to the MB correlated significantly (P<0.001) with the location of the MB entrance, and furthermore, atherosclerosis progression in the LAD proximal to the MB was largest at 2.0 cm from the MB entrance. Conclusions— In the proximal LAD with an MB, MB muscle index is associated with a shift of coronary disease more proximally, an effect that may increase the risk of MI.

Endoscopic band ligation for colonic diverticular hemorrhage.

BACKGROUND The number of sample cases of colonic diverticular hemorrhage treated with endoscopic band ligation (EBL) has been small to date. OBJECTIVE To elucidate the safety and efficacy of EBL for colonic diverticular hemorrhage. DESIGN Retrospective study. SETTING General hospital. PATIENTS A total of 29 patients with 31 colonic diverticula with stigmata of recent hemorrhage (SRH). INTERVENTIONS Urgent colonoscopy was performed after bowel preparation. When diverticula with SRH were identified, marking with hemoclips was done near the diverticula. The endoscope was removed and reinserted after a band-ligator device was attached to the tip of endoscope. At first, EBL was attempted. In patients who could not be treated with EBL, epinephrine injection or endoscopic clipping was performed. MAIN OUTCOME MEASUREMENTS Procedure time, rate of hemostasis and rebleeding, complications. RESULTS The mean procedure time was 47 ± 19 minutes. EBL was successfully completed in 27 colonic diverticula (87%); except in 3 diverticula with a small orifice and large dome and 1 diverticula in which the orifice was too large. Early rebleeding after EBL occurred in 3 of 27 cases (11%). Although 2 cases of sigmoid rebleeding could be managed by repeat EBL or conservatively, right hemicolectomy was performed in 1 ascending diverticulum, in which the bleeding source was not identified on repeat colonoscopy. Scar formation at previously banded diverticula was identified in 7 of 11 patients who underwent follow-up colonoscopy. There were no complications after EBL in any of the patients. LIMITATIONS Retrospective study. CONCLUSIONS EBL is a safe and effective treatment for colonic diverticular hemorrhage, and colonic diverticula resolve after EBL.

Histopathological predictors of regional lymph node metastasis at the invasive front in early colorectal cancer

Akishima‐Fukasawa Y, Ishikawa Y, Akasaka Y, Uzuki M, Inomata N, Yokoo T, Ishii R, Shimokawa R, Mukai K, Kiguchi H, Suzuki K, Fujiwara M, Ogata K, Niino H, Sugiura H, Ichinose A, Kuroda Y, Kuroda D & Ishii T
(2011) Histopathology59, 470–481

Efficacy of 3D-MR mammography for breast conserving surgery after neoadjuvant chemotherapy.

BackgroundOne of the main roles of neoadjuvant chemotherapy for breast cancer is to shrink large tumors to increase patient eligibility for breast conserving surgery. Three dimensional MR Mammography (3D-MRM) can detect tumor extension more accurately compared with mammography and Ultrasonography (US). Therefore, the shrinkage pattern observed on 3D-MRM was analyzed with regard to several pathological factors.MethodsA total of 27 breast cancer cases were examined by 3D-MRM before and after neoadjuvant chemotherapy. The volume reduction and shrinkage patterns were assessed and compared with the pathological diagnosis.ResultsThere were two shrinkage patterns. Twelve of 25 evaluable breast cancers (48%) showed a concentric shrinkage pattern while 13 cases (52%) showed a dendritic shrinkage pattern. The cases with concentric shrinkage were good candidates for breast coserving surgery, But tumors showing dendritic shrinkage often had positive margins necessitating mastectomy. Pathologically, tumors with a papillotubular pattern, Estrogen receptor (ER) positivity, low nuclear grade and c-erbB 2 negativity tended to show dendritic shrinkage.Conclusions3D-MRM is a useful modality for evalumting whether breast conserving surgery can be safely done in the neoadjuvant setting.

Severe steatosis resulted from anorexia nervosa leading to fatal hepatic failure

1. Coussens LM, Werb Z. Inflammation and cancer. Nature 2002;420:860– 7. 2. Hussain SP, Hofseth LJ, Harris CC. Radical causes of cancer. Nat Rev Cancer 2003;3:276–85. 3. Horiike S, Kawanishi S, Kaito M, Ma N, Tanaka H, Fujita N, et al. Accumulation of 8-nitroguanine in the liver of patients with chronic hepatitis C. J Hepatol 2005;43:403–10. 4. Nishiguchi S, Kuroki T, Nakatani S, Morimoto H, Takeda T, Nakajima S, et al. Randomized trial of effects of interferon-alpha on incidence of hepatocellular carcinoma in chronic active hepatitis C with cirrhosis. Lancet 1995;346:1051–5. 5. Kasahara A, Hayashi N, Mochizuki K, Takayanagi M, Yoshioka K, Kakumu S, et al. Risk factors for hepatocellular carcinoma and its incidence after interferon treatment in patients with chronic hepatitis C. Osaka Liver Disease Study Group. Hepatology. 1998;27:1394–402. 6. Ikeda K, Saitoh S, Arase Y, Chayama K, Suzuki Y, Kobayashi M, et al. Effect of interferon therapy on hepatocellular carcinogenesis in patients with chronic hepatitis type C: a long-term observation study of 1,643 patients using statistical bias correction with proportional hazard analysis. Hepatology 1999;29:1124–30. 7. Arase Y, Ikeda K, Tsubota A, Suzuki F, Suzuki Y, Saitoh S, et al. Interferon therapy for 2 years or longer reduces the incidence of hepatocarcinogenesis in patients with chronic hepatitis C viral infection. Intervirology 2004;47:355–61. 8. Ikeda M, Fujiyama S, Tanaka M, Sata M, Ide T, Yatsuhashi H, et al. Risk factors for development of hepatocellular carcinoma in patients with chronic hepatitis C after sustained response to interferon. J Gastroenterol 2005;40:148–56.

Neuroendocrine ductal carcinoma in situ (NE‐DCIS) of the breast – comparative clinicopathological study of 20 NE‐DCIS cases and 274 non‐NE‐DCIS cases

Aims:  To clarify the clinicopathological significance of breast neuroendocrine ductal carcinoma in situ (NE‐DCIS), i.e. DCIS in which >50% of cells immunohistochemically express NE markers (chromogranin A and/or synaptophysin), 20 NE‐DCIS were studied and the findings compared with those of 274 non‐NE‐DCIS.

An induction of microRNA, miR-7 through estrogen treatment in breast carcinoma

BackgroundEstrogen plays an important role in the development of estrogen-dependent breast carcinoma. Recently, several studies demonstrated a possible involvement of several micro RNAs (miRNAs) in the development of resistance to endocrine therapy in breast cancer patients, but the correlation between estrogen actions and miRNA expression in breast carcinoma still remains largely unknown. Therefore, in this study, we examined the in vitro effects of estrogen upon miRNA expression profiles in breast carcinoma.MethodsWe first screened the miRNA expression profiles induced by 17β-Estradiol (E2) using RT2 miRNA PCR Array in the ER-positive breast carcinoma cell line MCF-7. We identified miR-7 as the important miRNA associated with estrogen actions in these cells and further examined the changes of estrogen-dependent EGFR expression by miR-7 in ER-positive or -negative breast carcinoma cell lines including MCF-7. We also evaluated the correlation between miR-7 and EGFR expression in breast carcinoma cells derived from 21 patients using laser capture microdissection combined with quantitative reverse transcriptase-PCR.ResultsSeventeen miRNAs were significantly induced by E2 treatment in the MCF-7 cell line. Among 17 miRNAs induced by estradiol treatment, only miR-7 expression was significantly decreased by subsequent ICI treatment. The expression of miR-7 was up-regulated 2.94-fold by E2 treatment. miR-7 was reported to suppress epidermal growth factor receptor (EGFR) expression in several human malignancies. Transfection of miR-7 significantly suppressed EGFR mRNA levels in MCF-7 cells. Depletion of E2 from cell culture media also increased the expression level of EGFR mRNA in MCF-7 and T-47D cells but not in ER-negative, MDA-MB-231 and SK-BR-3 cells. We also evaluated the status of miR-7 in breast carcinoma tissues, but the correlation between the status of miR-7 and EGFR in carcinoma cells isolated by laser capture microscopy was not detected.ConclusionsThese results suggest that miR-7 may play a role in the development of resistance to endocrine therapy in breast cancer patients through regulating EGFR expression of carcinoma cells.

Localization of hepatitis B surface and core antigens in human hepatocellular carcinoma by immunoperoxidase methods. Replication of complete virions of carcinoma cells.

The localization of hepatitis B surface antigen (HBsAg) and core antigen (HBcAg) was investigated by an indirect immunoperoxidase method in formalin‐fixed, paraffin‐embedded liver specimens obtained from 95 Japanese patients with hepatocellular carcinomas. Routine and immune electron microscopic examinations were done in one case. The correlation between expression of hepatitis B virus antigens in the tissue and serum hepatitis B e antigen (HBeAg)/antibody to HBeAg (anti‐HBe) status was examined. Hepatitis B surface antigen was detected in the cytoplasm of noncarcinomatous hepatocytes in 28 (29.5%) cases and of carcinoma cells in 11 (11.6%) cases. Hepatitis B core antigen was stained in noncarcinomatous hepatocytes in 13 (13.7%) cases and in carcinoma cells in 4 (4.2%) cases. Hepatitis B core antigen was present mainly in the nuclei, and all HBcAg‐positive cases were positive for HBsAg. The routine electron microscopic examination revealed many round particles, 25 to 30 nm in diameter both in the nuclei and in the cytoplasm, and larger particles, 40 to 45 nm in diameter in the cytoplasm of carcinoma cells. Both types of particles had reaction products of HBcAg by immunoelectron microscopic study. Therefore, it was confirmed that the former were cores and the latter were Dane particles. There was a tendency that HBeAg‐seropositive cases showed localization of HBcAg in the noncarcinomatous tissue. Among four cases with positive HBcAg in carcinoma cells, two were positive for HBeAg, one was positive for anti‐HBe, and the other was negative both for HBeAg and anti‐HBe in the sera. The data suggested occasional production of complete hepatitis B viruses of carcinoma cells in anti‐HBe‐positive as well as in HBeAg‐positive hepatocellular carcinomas.