Light- and Electron- microscopic and Immunohistochemical Studies of Human Rhabdomyosarcomas. Comparisons Among Primary Tumors, Heterotransplants in Nude Mice, and Cultured Cells from 13 Patients

Eighteen human rhabdomyosarcomas (RMS) were transplanted into the sub-cutaneous space on the back of nude mice. Thirteen of the tumors gave rise to transplantable tumors that were further examined morphologically and immuno-histochemically. The morphology of the transplanted tumors was similar to that of the primary tumors. Immunohistochemically, five primary tumors and six transplanted tumors were reacted with both desmin and myoglobin. However, in eleven cases cultured cells derived from the transplanted tumors, which showed elongated to strap-spindle-shaped cytoplasm resembling myotubules, reacted more intensely with both myoglobin and desmin. On ultrastructural examination, six primary tumors and seven transplanted tumors were found to have myofilaments or Z-bands. However, cultured cells showed myofilaments or Z-bands in their cytoplasm in all cases examined. We concluded that, on xeno-grafting, the histologic characteristics of the primary tumor are essentially con-served, and that tumor cells under culture conditions undergo an increased differentiation of skeletal muscle. These human RMS strains in nude mice and in cell lines will provide an excellent model system for investigating the biology of RMS and for further study of the molecular events underlying the genesis of this tumor

Persistent homology index as a robust quantitative measure of immunohistochemical scoring

Immunohistochemical data (IHC) plays an important role in clinical practice, and is typically gathered in a semi-quantitative fashion that relies on some degree of visual scoring. However, visual scoring by a pathologist is inherently subjective and manifests both intra-observer and inter-observer variability. In this study, we introduce a novel computer-aided quantification methodology for immunohistochemical scoring that uses the algebraic concept of persistent homology. Using 8 bit grayscale image data derived from 90 specimens of invasive ductal carcinoma of the breast, stained for the replicative marker Ki-67, we computed homology classes. These were then compared to nuclear grades and the Ki-67 labeling indices obtained by visual scoring. Three metrics for IHC staining were newly defined: Persistent Homology Index (PHI), center coordinates of positive and negative groups, and the sum of squares within groups (WSS). This study demonstrates that PHI, a novel index for immunohistochemical labeling using persistent homology, can produce highly similar data to that generated by a pathologist using visual evaluation. The potential benefits associated with our novel technology include both improved quantification and reproducibility. Since our method reflects cellularity and nuclear atypia, it carries a greater quantity of biologic data compared to conventional evaluation using Ki-67

Isolated tumor cells and micrometastases in regional lymph nodes in stage I to II endometrial cancer

Objective: The aim of this study was to clarify the clinical significance of isolated tumor cells (ITCs) or micrometastasis (MM) in regional lymph nodes in patients with International Federation of Gynecology and Obstetrics (FIGO) stage Ito II endometrial cancer. Methods: In this study, a series of 63 patients with FIGO stage Ito II were included, who had at least one of the following risk factors for recurrence: G3 endometrioid/serous/clear cell adenocarcinomas, deep myometrial invasion, cervical involvement, lympho-vascular space invasion, and positive peritoneal cytology. These cases were classified as intermediate-risk endometrial cancer. Ultrastaging by multiple slicing, staining with hematoxylin and eosin and cytokeratin, and microscopic examination was performed on regional lymph nodes that had been diagnosed as negative for metastases. Results: Among 61 patients in whom paraffin-embedded block was available, ITC/MM was identified in nine patients (14.8%). Deep myometrial invasion was significantly associated with ITC/MM (p=0.028). ITC/MM was an independent risk factor for extrapelvic recurrence (hazard ratio, 17.9; 95% confidence interval [CI], 1.4 to 232.2). The 8-year overall survival (OS) and recurrence-free survival (RFS) rates were more than 20% lower in the ITC/MM group than in the node-negative group (OS, 71.4% vs. 91.9%; RFS, 55.6% vs. 84.0%), which were statistically not significant (OS, p=0.074; RFS, p=0.066). Time to recurrence tended to be longer in the ITC/MM group than in the node-negative group (median, 49 months vs. 16.5 months; p=0.080). Conclusions: It remains unclear whether ITC/MM have an adverse influence on prognosis of intermediate-risk endometrial cancer. A multicenter cooperative study is needed to clarify the clinical significance of ITC/MM

Volume-based parameters on FDG PET may predict the proliferative potential of soft-tissue sarcomas

Introduction: Soft-tissue sarcomas (STS) are rare types of tumors that have variable levels of tumor differentiation. F-18 fluorodeoxyglucose positron emission tomography (FDG PET) has been established as an useful tool for STS patients, and the metabolic tumor volume (MTV) and total lesion glycolysis (TLG) are reported to be useful in various cancers. We compared the diagnostic value of four PET parameters (maximum standardized uptake value [SUVmax], SUVmean, MTV, and TLG) from two acquisition timings for predicting the expression of the pathological marker of cell proliferation Ki-67, based on pathological investigation. Materials and methods: In this retrospective study, we investigated 20 patients (59±19 years old, 18-87 years old) with pathologically confirmed STS who underwent FDG PET before surgical intervention. The patients fasted ≥6 h before the intravenous injection of FDG. The whole body was scanned twice; at an early phase (61.5±2.6 min) and at a delayed phase (118.0±2.1 min) post-injection. The SUVmax, SUVmean, MTV, and TLG of the primary lesion were measured with a tumor boundary determined by SUV ≥2.0. Ki-67 was measured using MIB-1 immunohistochemistry. We used Pearson's correlation coefficient to analyze the relationships between the PET parameters and Ki-67 expressions. The Kaplan-Meier analysis with the log-rank test was performed to compare overall survival between high-group and low-group at each of the four PET parameters and Ki-67 expression. Results: All four PET parameters at each phase showed significant correlations with Ki-67. Among them, the Pearson's correlation coefficient (r) was largest for TLG (r=0.76 and 0.77 at the early and delayed phases, respectively), followed by MTV (0.70 and 0.72), SUVmax (r=0.65 and 0.66), and SUVmean (r=0.62 and r=0.64). From early to delayed phases, the SUVmax and SUVmean both increased in all 20 patients, whereas the MTV and TLG increased in 13/20 (65%) and 16/20 (80%) patients, respectively. None of the %increases of the PET parameters were significantly correlated with Ki-67. The overall survival was shorter for high-SUVmax, high-SUVmean, high-TLG, and high-Ki-67 groups than the other groups, although the difference did not reach statistical significance. Conclusion: The SUVmax, SUVmean, MTV, and TLG acquired at both 1 and 2 h after injection showed significant correlations with Ki-67. Among them, correlation coefficient with Ki-67 expression was highest for TLG, although the best parameter should be determined in a larger population. The delayed-phase FDG PET was equally useful as that of early-phase to predict tumor aggressiveness in STS

Strong cytoplasmic expression of NF-κB/p65 correlates with a good prognosis in patients with triple-negative breast cancer

Purpose: Recent studies have indicated that constitutive NF-κB activity could be involved in the proliferation of triple-negative breast cancer. Methods: NF-κB/p65 expression and the effects of the NF-κB inhibitor, (-)-DHMEQ were examined in triple-negative MDA-MB-231 breast cancer cells. Women with triple-negative breast cancer treated with neoadjuvant chemotherapy between 2002 and 2012 were retrospectively analyzed. Expression of NF-κB/p65, Bcl2 and Ki67 was examined by immunohistochemistry in pre- and post-treatment specimens, and predictive factors for neoadjuvant chemotherapy and prognosis were analyzed. Results: NF-κB/p65 was dominantly expressed in the cytoplasm in MDA-MB-231 cells. Of 34 triple-negative breast cancer patients, positive staining for NF-κB/p65 expression was detected in the nuclei of a few cells in 7 tumors before neoadjuvant chemotherapy, although expression of NF-κB/p65 in the cytoplasm was detected in almost all tumor cells of 33 tumors. Expression levels of NF-κB/p65 were not associated with response to neoaduvant chemotherapy, although expression levels of cytoplasmic NF-κB/p65 intensity were significantly decreased in post-treatment tumor samples compared with those in pretreatment samples. All patients whose tumors showed strong cytoplasmic NF-κB/p65 expression before neoadjuvant chemotherapy are currently disease-free. Conclusion: Our results suggest that strong cytoplasmic NF-κB/p65 expression could be a prognostic marker in triple-negative breast cancer

A feasibility study of sentinel lymph node mapping by cervical injection of a tracer in Japanese women with early stage endometrial cancer

Objective: The aim of this study was to investigate the feasibility of sentinel lymph node mapping characterized by a cervical tracer injection in endometrial cancer. Materials and methods: This retrospective study was carried out using data for 57 patients with endometrial carcinoma who had undergone intraoperative sentinel lymph node mapping and subsequent surgical staging. Technetium colloid and/or indocyanine green was injected into the uterine cervix and a gamma-detecting probe and/or photodynamic eye camera system was used intraoperatively to locate hot spots. Results: Of the 57 patients, 52 (91.2%) had FIGO Stage I disease. Successful unilateral or bilateral mapping occurred in 54 patients (94.7%) and 46 (80.7%), respectively. The median number of sentinel lymph nodes detected was two (range, 0–5). Following sentinel lymph node mapping, 41 patients (71.9%) underwent pelvic lymphadenectomy alone and 16 (28.1%) full lymphadenectomy. The median number of lymph nodes resected was 17 (range, 8–110). Sentinel lymph nodes were involved in four patients (7.0%), two with macrometastases and two with low-volume metastases. The sensitivity and negative predictive value for detecting lymph node metastasis were both 100%. Conclusion: Sentinel lymph node mapping with the use of cervical tracer injection is highly feasible in Japanese women with early stage endometrial cancer. Keywords: Endometrial cancer, Uterine cancer, Sentinel lymph node, Cervical injection, Sensitivit