Underestimated Amoebic Appendicitis among HIV-1-Infected Individuals in Japan.

Entamoeba histolytica is not a common causative agent of acute appendicitis. However, amoebic appendicitis can sometimes be severe and life threatening, mainly due to a lack of awareness. Also, its frequency, clinical features, and pathogenesis remain unclear. The study subjects were HIV-1-infected individuals who presented with acute appendicitis and later underwent appendectomy at our hospital between 1996 and 2014. Formalin-fixed paraffin-embedded preserved appendix specimens were reexamined by periodic acid-Schiff (PAS) staining and PCR to identify undiagnosed amoebic appendicitis. Appendectomies were performed in 57 patients with acute appendicitis. The seroprevalence of E. histolytica was 33% (14/43) from the available stored sera. Based on the medical records, only 3 cases were clinically diagnosed as amoebic appendicitis, including 2 diagnosed at the time of appendectomy and 1 case diagnosed by rereview of the appendix after the development of postoperative complications. Retrospective analyses using PAS staining and PCR identified 3 and 3 more cases, respectively. Thus, E. histolytica infection was confirmed in 9 cases (15.8%) in the present study. Apart from a significantly higher leukocyte count in E. histolytica-positive patients than in negative patients (median, 13,760 versus 10,385 cells/µl, respectively, P = 0.02), there were no other differences in the clinical features of the PCR-positive and -negative groups. In conclusion, E. histolytica infection was confirmed in 9 (15.8%) of the appendicitis cases. However, only 3, including one diagnosed after intestinal perforation, were diagnosed before the present analyses. These results strongly suggest there is frequently a failure to detect trophozoites in routine examination, resulting in an underestimation of the incidence of amoebic appendicitis.

High expression level of geminin predicts a poor clinical outcome in salivary gland carcinomas.

AIMS: To examine the prognostic impact of geminin expression, a negative regulator of DNA replication, in salivary gland carcinomas. METHODS AND RESULTS: Tissues from 170 patients with salivary gland carcinoma were assembled in a tissue microarray format, and immunohistochemistry staining for geminin and Ki67 was performed. Patients were categorized into three groups using the tertile values of the labelling index (LI) for each marker as the cut-off points for constructing Kaplan-Meier survival curves. The LI for geminin was relatively low in acinic cell carcinoma (mean 1.55%) and mucoepidermoid carcinoma (mean 2.43%), intermediate in adenoid cystic carcinoma (mean 4.09%), and relatively high in salivary duct carcinoma (mean 15.22%). The geminin LI was significantly correlated with the Ki67 LI and histopathological factors, including pathological T factor, lymphatic and blood vessel infiltration, and lymph node metastasis. The increased expression of geminin was more strongly associated with reduced overall and relapse-free survival rates than with Ki67 expression and was a significant and independent prognostic factor in patient survival and tumour relapse. CONCLUSIONS: Geminin expression is potentially a useful marker for predicting tumour aggressiveness and clinical outcome in salivary gland carcinomas.

Immunohistochemical differential diagnosis between thymic carcinoma and type B3 thymoma: diagnostic utility of hypoxic marker, GLUT-1, in thymic epithelial neoplasms.

There are only a few immunohistochemical markers that are useful for differentiating thymic carcinomas from type B3 thymomas. The purpose of this study is to examine the additional markers that would be useful for differentiating between thymic carcinoma and thymoma type B3. We performed a tissue microarray analysis of surgically resected thymic tumor specimens from 12 cases of thymic carcinoma, 7 cases of type B3 thymoma, and 68 cases of other types of thymoma. Immunostaining using 49 antibodies was scored based on staining intensity and the percentage of cells that stained positive. Seven proteins that were selected by the staining scores, namely, GLUT-1 (167 vs 4), CA-IX (110 vs 15), c-kit (162 vs 44), CD5 (33 vs 0), MUC-1 (54 vs 0), CEA (42 vs 0), and CK18 (110 vs 42), were significantly higher in the thymic carcinomas than in the type B3 thymomas. The staining sensitivity and specificity of the antibodies for thymic carcinoma were GLUT-1, sensitivity 72% and specificity 100%; CA-IX, 58 and 71%; c-kit, 72 and 85%; CD5, 33 and 100%; CK18, 58 and 71%; MUC-1, 25 and 100%; and CEA, 33 and 100%. Glucose transporter 1 (GLUT-1) is the best marker for thymic carcinoma because it had the highest sensitivity and specificity. Positive immunostaining for a combination of three markers, namely, GLUT-1, CD5, and CEA, enabled differentiation of thymic carcinoma with 91.6% sensitivity and 100% specificity. In conclusion, we identified GLUT-1 as an additional marker that will be useful for differentiating thymic carcinoma from type B3 thymoma, especially in biopsy specimens that have been crushed or are otherwise difficult to examine morphologically in thymic tumors.

Low podoplanin expression of tumor cells predicts poor prognosis in pathological stage IB squamous cell carcinoma of the lung, tissue microarray analysis of 136 patients using 24 antibodies.

The aim of this study was to identify clinicopathological and biological prognostic markers for patients who had undergone complete resection of pathological stage IB squamous cell carcinoma (SqCC) of the lung. A total of 136 consecutive stage IB SqCC patients fulfilled eligibility criteria, and their clinicopathological factors were evaluated. Tissue microarrays were also constracted, and immunohistochemical staining with 24 antibodies was performed. Correlations between clinicopathological factors, antibody immunohistochemical reactions, the patients' overall survival (OS) and relapse-free survival (RFS) were evaluated. The univariate analysis showed that 70-year-old and over elderly group had a shorter OS time and RFS time than the younger group (p=0.0086 and p=0.0091, respectively). The univariate analysis for immunohistochemical staining showed that the podoplanin-negative group had a shorter OS time and RFS time than the podoplanin-positive group (p=0.0106 and p=0.0308, respectively). Multivariate analysis revealed a significant correlation between both the 70-year-old and over elderly group and the podoplanin-negative group and poor outcome (OS, p=0.007 and p=0.008, respectively; RFS, p=0.008 and p=0.024, respectively). The results showed that patient age and a novel biological prognostic marker, podoplanin, are useful for predicting a poor outcome of patients after complete resection of stage IB SqCC of the lung.

Immunohistochemical expression of BCRP and ERCC1 in biopsy specimen predicts survival in advanced non-small-cell lung cancer treated with cisplatin-based chemotherapy.

PURPOSE: The aim of this study was to determine the prognostic value of expression of ATP binding cassette (ABC) transporter proteins and DNA repair gene proteins by immunohistochemically staining tumor biopsy specimens from patients with advanced non-small-cell lung cancer (NSCLC) being treated with platinum-based chemotherapy. EXPERIMENTAL DESIGN: Expression of ABC transporter proteins, including BCRP (breast cancer resistance protein) and MRP2 (multidrug resistance proteins 2), and the DNA-repair-related proteins, ERCC1 (excision repair cross-complementation group 1) and BRCA1 (breast cancer type 1 susceptibility protein) was assessed immunohistochemically in 156 tumor samples from untreated stage IV NSCLC patients. All of the patients had received platinum-based chemotherapy. Response to chemotherapy, progression-free survival (PFS), and overall survival were compared in relation to expression of each of the proteins and to clinicopathological factors. RESULTS: High ERCC1 expression was associated with short survival (237 days vs. 453 days, log-rank P = 0.03), but not with response to chemotherapy or PFS. And high BCRP expression was associated with short survival (214 days vs. 412 days, log-rank P = 0.02) but not with response to chemotherapy or PFS. Multivariate analysis confirmed that negativity for the expression of BCRP tends to be an independent variable related to overall survival (P = 0.06). CONCLUSIONS: This study examined ERCC1 and BCRP expression in biopsy specimens as candidates for predictors of the survival of patients with advanced NSCLC treated with platinum-based chemotherapy.

Podoplanin expression by cancer associated fibroblasts predicts poor prognosis of lung adenocarcinoma.

Recent studies have reported increased podoplanin expression by cancer cells and stromal cells, but little is known about its expression and biological significance in adenocarcinoma of the lung. We examined podoplanin expression by both cancer cells and stromal cells in 177 consecutive lung adenocarcinoma cases and analyzed relations between podoplanin expression and both clinicopathological factors and outcome. Podoplanin expression was observed on the apical membrane of the cancer cells in only 9 of the 177 (5.1%) cases. By contrast, cancer-associated fibroblasts (CAFs) were found to express podoplanin in 54 cases (30.5%). Podoplanin (+) CAFs were found only in invasive adenocarcinoma and none were found in noninvasive adenocarcinoma. Conventional prognostic factors were significantly correlated with podoplanin expression by CAFs. The univariate analyses and log-rank test showed that podoplanin expression was significantly associated with shorter survival time (p < 0.001 and p < 0.001, respectively). We divided the cases into 3 groups according grade based on the proportion of CAFs expressing podoplanin [a grade 0 group (n = 123), a grade 1 group (n = 36) and a grade 2 group (n = 18)]. The result showed that conventional prognostic factors were significantly correlated with the grade of podoplanin expression by CAFs. Furthermore, the grade 2 group tended to have a shorter survival time than the grade 1 group (p = 0.092). The results of this study highlight the importance of podoplanin expression by CAFs and provide new insights into the biology of the cancer microenvironment in adenocarcinoma of the lung.

Pleomorphic carcinoma of the lung expressing podoplanin and calretinin.

Pleomorphic carcinoma (PC) of the lung is classified as a subtype of sarcomatoid carcinoma of the lung, and peripheral PC is sometimes difficult to differentiate from the sarcomatoid component of mesothelioma. An 80-year-old man was referred to National Cancer Center Hospital East because a chest X-ray showed an abnormal shadow. CT scans of the chest indicated two solid masses located in the right lower lobe, and CT-guided needle biopsy yielded spindle-shaped tumor cells that were immunoreactive for both podoplanin and calretinin. Mesothelioma could not be ruled out, and the tumors were surgically resected to facilitate definitive pathological diagnosis. Both tumors were composed of undifferentiated carcinoma, bronchioloalveolar carcinoma and spindle cell carcinoma, and spindle cell component was immunoreactive for podoplanin and calretinin. Ten other tumors diagnosed as peripheral PC were also tested for podoplanin and calretinin expression. The sarcomatoid component in four of the 11 cases (36%) was immunoreactive with podoplanin, and it was calretinin positive in nine of the 11 cases (82%). When making the differential diagnosis between PC and the sarcomatoid component of mesothelioma, care is required in diagnosing biopsy specimens of peripheral lung spindle-cell tumors that are positive for both podoplanin and calretinin.

The lymphatic infiltration identified by D2-40 monoclonal antibody predicts lymph node metastasis in submucosal invasive colorectal cancer.

BACKGROUND AND STUDY AIMS: Lymphatic infiltration has been recognized as a significant risk factor for lymph node metastasis of submucosal invasive colorectal cancer (SICC), but it is difficult to detect microscopically on hematoxylin and eosin (H&E)-stained slides. We therefore identified lymphatic infiltration of tumor cells with D2-40 monoclonal antibody, which reacts specifically against the endothelium of lymphatic vessels, to make an objective and precise diagnosis. PATIENTS AND METHODS: The surgical specimens of 122 consecutive patients with nonpedunculated SICC were examined for lymphatic infiltration by immunohistochemical staining with D2-40 monoclonal antibody (LI-D) and for venous infiltration by Elastica van Gieson staining (VI-E). RESULTS: Lymph node metastasis was found in 20 patients. Multivariate analysis showed that LI-D (p = 0.0415) and VI-E (p = 0.0119) were significant risk factors for lymph node metastasis. Regardless of the presence of risk factors including at least either lymphatic infiltration or venous infiltration, no lymph node metastasis-positive patients were found (0%) among the 25 patients whose colorectal cancer had a submucosal invasive depth of less than 1,500 microm. No lymph node metastasis was found in any of the patients with a depth of submucosal invasion of less than 3,000 microm, who had no risk factors, including LI-D or VI-E. CONCLUSIONS: Correct evaluation of lymphatic infiltration by immunohistochemical staining with D2-40 monoclonal antibody may play a crucial role in determining whether there are indications for additional treatment in the management of endoscopically resected SICC.

Carbonic anhydrase IX expression is associated with tumor progression and a poor prognosis of lung adenocarcinoma.

Carbonic anhydrase (CA) IX catalyzes the hydration of carbon dioxide into carbonic acid and participates in a variety of physiological and biological processes. The aim of this study was to evaluate the prognostic significance of CA IX expression in patients with lung adenocarcinoma. Standard immunohistochemical techniques were used to study CA IX expression in 134 patients who underwent curative resection for adenocarcinoma of the lung at our hospital between January 1995 and December 1996. We evaluated the correlations between CA IX expression levels on cancer cells and clinicopathological factors. CA IX expression was not observed in normal lung tissue or specimens from non-invasive adenocarcinomas. CA IX immunostaining was detected in 33 (24.6%) invasive adenocarcinoma cases. Poor differentiated histological phenotype (p=0.0015), pathological stage (p=0.0400), vascular invasion (p=0.0009) and lymphatic permeation (p=0.0050) were significantly related to CA IX expression. On univariate analysis, CA IX positive cases showed significantly shorter overall survival (p=0.0083) and disease-free survival (p=0.0122). In particular, the overall and disease-free survivals in stages I+II were significantly shorter in the CA IX positive than in the CA IX negative cases (p=0.0269 and 0.0011, respectively). Our results suggest that CA IX expression is strongly associated with tumor progression and indicates a poor prognosis for patients with stages I+II lung adenocarcinoma.

Immunohistochemical differential diagnosis between large cell neuroendocrine carcinoma and small cell carcinoma by tissue microarray analysis with a large antibody panel.

To elucidate additional phenotypic differences between large cell neuroendocrine carcinoma (LCNEC) and small cell lung carcinoma (SCLC), we performed tissue microarray (TMA) analysis of surgically resected LCNEC and SCLC specimens. Immunostaining with 48 antibodies was scored based on staining intensity and the percentage of cells that stained positively. Four proteins were identified as significantly expressed in LCNEC as compared with SCLC: cytokeratin (CK)7, 113 vs 49 (P < .0301); CK18, 171 vs 60 (P < .0008); E-cadherin, 77 vs 9 (P < .0073); and beta-catenin, 191 vs 120 (P < .0286). Immunostaining of cross-sections containing LCNEC and SCLC components revealed significant expression of CK7, CK18 and beta-catenin in the LCNEC component compared with the SCLC component in 2 of 3 cases. Our results indicate that significant expression of CK7, CK18, E-cadherin, and beta-catenin is more characteristic of LCNEC than of SCLC, and these findings provide further support that these tumor types are separate entities morphologically and immunophenotypically, if not biologically.

Telomere Visualization in Tissue Sections using Pyrrole-Imidazole Polyamide Probes.

Pyrrole-Imidazole (PI) polyamides bind to specific DNA sequences in the minor groove with high affinity. Specific DNA labeling by PI polyamides does not require DNA denaturation with harsh treatments of heat and formamide and has the advantages of rapid and less disruptive processing. Previously, we developed tandem hairpin PI polyamide probes (TH59 series), which label telomeres in cultured cell lines more efficiently than conventional methods, such as fluorescence in situ hybridization (FISH). Here, we demonstrate that a TH59 derivative, HPTH59-b, along with immunostaining for specifying cell types in the tissues, visualizes telomeres in mouse and human tissue sections. Quantitative measurements of telomere length with single-cell resolution suggested shorter telomeres in the proliferating cell fractions of tumor than in non-tumor tissues. Thus, PI polyamides are a promising alternative for telomere labeling in clinical research, as well as in cell biology.

Feasibility study on radiofrequency ablation followed by partial mastectomy for stage I breast cancer patients.

To evaluate the safety and reliability of thermal ablation therapy instead of breast-conserving surgery (BCS), we performed radiofrequency ablation (RFA) for clinical stage I breast cancer patients. Subjects were T1N0 breast cancer patients with no extensive intraductal components. Under general anesthesia, sentinel node biopsy was performed, followed by RFA and BCS. Resected specimens were examined at 5-mm intervals by hematoxylin-eosin (H&E) staining and nicotinamide adenine dinucleotide (NADH) diaphorase staining. Thirty of the 34 eligible patients were enrolled. RFA-related adverse events were observed in nine patients: two with skin burn and seven with muscle burn. Twenty-six patients (87%) showed pathological degenerative changes in tumor specimens with H&E staining. In 24 of the 26 cases (92%) examined by NADH diaphorase staining, tumor cell viability was diagnosed as negative. RFA proved to be reliable and feasible in clinical stage I breast cancer, with no extensive intraductal components. Randomized clinical trials are needed to compare RFA with BCS.

Expression of breast cancer resistance protein is associated with a poor clinical outcome in patients with small-cell lung cancer.

BACKGROUND: ATP-binding cassette (ABC) transporter and DNA excision repair proteins play a pivotal role in the mechanisms of drug resistance. The aim of this study was to investigate the expression of ABC transporter and DNA excision repair proteins, and to elucidate the clinical significance of their expression in biopsy specimens from patients with small-cell lung cancer (SCLC). METHODS: We investigated expression of the ABC transporter proteins, P-glycoprotein (Pgp), multidrug resistance associated-protein 1 (MRP1), MRP2, MRP3, and breast cancer resistance protein (BCRP), and the DNA excision repair proteins, excision repair cross-complementation group 1 (ERCC1) protein and breast cancer susceptibility gene 1 (BRCA1) protein, in tumor biopsy specimens obtained before chemotherapy from 130 SCLC patients who later received platinum-based combination chemotherapy, and investigated the relationship between their expression and both response and survival. RESULTS: No significant associations were found between expression of Pgp, MRP1, MRP2, MRP3, ERCC1, or BRCA1 and either response or survival. However, there was a significant association between BCRP expression and both response (p=0.026) and progression-free survival (PFS; p=0.0103). CONCLUSIONS: BCRP expression was significantly predictive of both response and progression-free survival (PFS) in SCLC patients receiving chemotherapy. These findings suggest that BCRP may play a crucial role in drug resistance mechanisms, and that it may serve as an ideal molecular target for the treatment of SCLC.

Predominant infiltration of macrophages and CD8(+) T Cells in cancer nests is a significant predictor of survival in stage IV nonsmall cell lung cancer.

BACKGROUND: The purpose of this study was to investigate whether tumor-infiltrating immune cells in biopsy specimens can be used to predict the clinical outcome of stage IV nonsmall cell lung cancer (NSCLC) patients. METHOD: The authors performed an immunohistochemical study to identify and count the number of CD68(+) macrophages, c-kit(+) mast cells, and CD8(+) T cells in both cancer nests and cancer stroma in pretreatment biopsy specimens obtained from 199 patients with stage IV NSCLC treated by chemotherapy, and then analyzed for correlations between the number of immune cells and clinical outcome, including chemotherapy response and prognosis. RESULTS: There was no correlation between the number of immune cells in either cancer nests or stroma and chemotherapy response. Patients with more tumor-infiltrating macrophages in cancer nests than in cancer stroma (macrophages, nests > stroma) had significantly better survival than nests < stroma cases median survival time (MST 440 days vs 199 days; P < .0001). Patients with more tumor-infiltrating CD8(+) T cells in cancer nests than in cancer stroma (CD8(+) T cells: nests > stroma) showed significantly better survival than in nests < stroma cases (MST 388 days vs 256 days; P = .0070). The proportion of tumor-infiltrating macrophages or CD8(+) T cells between cancer nests and stroma became independent prognostic factors in the multivariate analysis. Neither the number of mast cells in nests nor in stroma correlated with the clinical outcome. CONCLUSIONS: Evaluation of the numbers of macrophages and CD8(+) T cells in cancer nests and stroma are useful biomarkers for predicting the prognosis of stage IV NSCLC patients treated with chemotherapy, but could fail to predict chemotherapy response.

Predictive factors for local recurrence of resected colorectal lung metastases.

BACKGROUND: Wedge resection or segmentectomy are the preferred treatments for pulmonary metastasis from colorectal cancer. However, local recurrence at the surgical margin is a problem with limited resections. This study attempted to identify predictive factors associated with local recurrences at the surgical margin after resection of pulmonary metastases. METHODS: A total of 96 lesions in 61 patients who had undergone a pulmonary wedge resection or segmentectomy for the treatment of pulmonary metastasis from colorectal cancer were investigated. Various clinical and pathologic factors were reviewed, and the risk of a local recurrence at the surgical margin was investigated. RESULTS: After pulmonary resection, 34 of the 61 patients (56%) experienced recurrences in their lungs. Local recurrences at the surgical margin were found in 17 patients (28%), even though 15 of these 17 cases had been histologically confirmed as completely resected cases. No clinical factors associated with local recurrence at the surgical margin were identified. Pathologically, lesions exhibiting 10 or more aerogenous spreads with floating cancer cell clusters around the main tumor (p = 0.02) and a malignant positive surgical margin (p = 0.04) had a significantly higher risk of local recurrence. CONCLUSIONS: The present study indicated that local recurrence may occur even in cases with a pathologically negative surgical margin. In cases with pulmonary metastases from colorectal cancer, lesions with 10 or more aerogenous spreads with floating cancer cell clusters around the main lesion and a malignant positive surgical margin in the resected specimens have a significantly higher risk of local recurrence at the surgical margin than those without.

Histopathologic prognostic factors in resected colorectal lung metastases.

BACKGROUND: Pulmonary metastasectomy is a standard method of treatment for selective pulmonary metastases of colorectal cancer. However, the pathologic factors of metastatic lesions that affect survival and tumor recurrence after pulmonary resection are less well defined. The pathologic findings of colorectal pulmonary metastases have not been correlated with clinical outcome. The study was conducted to clarify the characteristics and identify the prognostic factors of the pulmonary metastases of colorectal cancer. METHODS: Between July 1992 and November 2002, 89 patients underwent the complete resection of pulmonary metastases of primary colorectal carcinoma, and we pathologically reviewed the surgical specimens of 136 metastatic lesions from these patients. RESULTS: There were no perioperative deaths. The overall 5-year survival rate was 61.4%. No clinical factors were associated with the outcome. The univariate analysis of pathologic factors revealed aerogenous spread with floating cancer cell clusters (ASFC) (p = 0.004), vascular invasion (p = 0.002), lymphatic invasion (p = 0.032), and pleural invasion (p = 0.037) to be prognostic factors. The multivariate analysis showed vascular invasion (p = 0.02) and ASFC (p = 0.02) to be independent prognostic factors. The 5-year survival rate of patients whose lesions were positive for both ASFC and vascular invasion was 24.7% and much poorer than in the patients with ASFC without vascular invasion (78.6%), vascular invasion but without ASFC (80.2%), and patients negative for both (93.3%) (p = 0.0002). CONCLUSIONS: The morphologic features of ASFC and vascular invasion at metastatic sites are prognostic factors for colorectal cancer patients who have undergone pulmonary metastasectomy.

Cytologic factors associated with prognosis in patients with peripheral adenocarcinoma of the lung measuring 3 cm or less in greatest dimension.

BACKGROUND: Recently, peripheral lung adenocarcinomas (PLA) measuring < or = 3 cm in greatest dimension often have been diagnosed using diagnostic radiology. The objective of the current study was to determine which cytologic factors are associated with a favorable outcome and an unfavorable outcome in patients with PLA. METHODS: Imprint smears from 134 patients with PLA were examined. Sixteen cytologic factors, including necrosis, cellular distribution, overlapping of cell clusters, cluster aggregation, cluster size, cluster thickness, nuclear irregularity, nuclear size, variation in nuclear size, multinucleated cells, intranuclear inclusions, type of intranuclear inclusions, appearance of nucleoli, eosinophilic nucleoli, multinucleoli, and mitosis, were evaluated using univariate and multivariate analyses. A counting method was used to determine the prognosis for individual patients. RESULTS: In the univariate analysis, a cluster size that measured > or = 831 microm in short dimension (P = 0.0011), moderate or severe nuclear irregularity (P = 0.0030), > or = 5 multinucleated cells per 100 tumor cells (P = 0.0047), moderate or severe variation in nuclear size (P = 0.0061), medium or large nuclear size (P = 0.0169), and > or = 1 mitotic cell per 100 tumor cells (P = 0.0412) were associated significantly with a poor outcome. In the multivariate analysis, cluster size in short dimension (P = 0.0018), multinucleated cells (P = 0.0066), and nuclear irregularity (P = 0.0310) were found to be independent prognostic factors. CONCLUSIONS: The combination of cytologic features using intraoperative imprint smears, namely, cluster sizes < or = 830 microm in short dimension, < or = 4 multinucleated cells per 100 tumor cells, and mild nuclear irregularity, may provide favorable predictive information in patients with PLA.