Benign Specimens Research Articles

Human urothelial bladder cancer generates a clonal immune response: The results of T-cell receptor sequencing

BackgroundHigh T-cell receptor (TCR) repertoire clonality is associated with clinical response to immune checkpoint blockade in bladder cancer. ObjectiveTo determine if TCR repertoire is more clonal in tumors than in benign inflammation. MethodsWe prospectively identified 12 patients with bladder lesions undergoing transurethral resection. Specimens were collected at time of transurethral resection and stored at −80C. DNA was extracted and high throughput DNA sequencing of the CDR3 region of the TCR beta chain using the immunoSEQ assay (Adaptive Biotechnologies) was performed. T-cell fraction, clonal dominance, and maximum frequency of TCR clone were assessed. ResultsOf the 12 bladder lesions resected, 3 of 12 were cT0, 3 of 12 were cTa, 3 of 12 were cT1, and 3 of 12 were cT2 or greater. The median number of T cells in urothelial carcinoma specimens (UC+) and benign (UC−) specimens was 5,569 and 25,872, respectively. The number of unique TCRs sequenced in UC+ and UC− specimens was 3,069 and 9,680, respectively. The median tumor infiltrating lymphocyte percentage in UC+ and UC− specimens was 2% and 12%, respectively. The UC+ specimens demonstrated clonality as evidenced by identification of a specific T-cell clone being present in up to 17% of the total tumor infiltrating lymphocyte pool, in contrast to 2% among UC− specimens. ConclusionsPrimary urothelial tumors contain clonally expanded T-cell populations. These data support the hypothesis that bladder tumors induce an antigen-driven immunogenic host response, in contrast to the benign inflammatory response, which does not appear to demonstrate any T-cell clonal dominance.

Probe-based confocal laser endomicroscopy for rapid on-site evaluation of transbronchial biopsy specimens.

BackgroundProbe‐based confocal laser endomicroscopy (pCLE) is a novel, noninvasive technology that provides real‐time lung imaging during bronchoscopy. pCLE shows the elastic fiber network without the use of a fluorescent dye. Elastic fibers produce argon laser‐induced autofluorescence at a wavelength of 488 nm, but tumor cells do not produce autofluorescence at this wavelength. As a result, the tumor cells cannot be observed directly. Therefore, we stained transbronchial biopsy (TBB) specimens with acriflavine to evaluate the benign and malignant structures using pCLE of ex vivo samples and to determine whether rapid histopathological diagnosis of TBB specimens could be made via pCLE.MethodsAfter bronchoscopy, 36 TBB specimens were stained with acriflavine and observed using pCLE. Benign and malignant lesions were classified by cell density and nuclear magnitude disparity.ResultsWe defined the confocal laser endomicroscopic atypia classification from the findings of the cells. The sensitivity for malignancy was 91.3%, and the specificity was 76.9%. Both inter‐observer (κ = 0.48) and intra‐observer (κ = 0.57) agreement confirmed moderate agreement.ConclusionpCLE with acriflavine staining was useful to differentiate malignant from benign TBB specimens, and might be useful as a substitute for rapid on‐site evaluation of histopathological diagnosis.

Analysis of microRNA expression in brush cytology specimens improves the diagnosis of pancreatobiliary cancer

Background/ObjectivesMalignant pancreatobiliary strictures are in many cases clinically indistinguishable and present a major problem to endoscopy specialists. Intraductal sampling procedures such as brush cytology are commonly used for diagnosis with a sensitivity that is low for a diagnostic test used in daily clinical practice. MicroRNA (miR) alterations detected in many cancers are disease-specific, which can be utilized in clinical applications. The aim of the present study was to analyze whether determination of miR expression levels in intraductal brush cytology specimens is a feasible approach to improve the diagnosis of pancreatobiliary cancer. MethodsBrush cytology specimens have been collected during endoscopic retrograde cholangio-pancreatography (ERCP) and analyzed by routine cytology and ancillary miR assays. Total RNA was extracted using the miRNeasy Mini Kit and the expression of miRs frequently dysregulated in pancreatobiliary cancer (miR-16, miR-21, miR-196a, miR-221) were analyzed by quantitative real-time PCR using RNU6B as internal control. ResultsRoutine cytology resulted in no false positive diagnoses, however, the combined sensitivity remained at 53.8%. Expression (ΔCt values) of miR-16 (p = 0.0039), miR-196a (p = 0.0003) and miR-221 (p = 0.0049) showed a clear statistical significance between malignant and benign pancreatobiliary specimens (n = 35). Malignancy could be detected combining routine cytology and the miR-196a single marker expression levels with a sensitivity of 84.6% (92.9% in biliary strictures) with no false positives. ConclusionsThe results offer the first direct demonstration that microRNAs are readily detectable in brush cytology specimens obtained during ERCP, and have the potential to help the cytological diagnosis of pancreatobiliary malignancy.

Expression and clinical significance of SLP-2 in ovarian tumors.

The expression and clinical significance of stomatin-like protein 2 (SLP-2) in ovarian tumors were investigated. A total of 280 cases of ovarian tissue specimens preserved from inpatients after surgical treatments in the Department of Oncology of Yidu Central Hospital of Weifang from April 2013 to May 2016 were collected, including 130 cases of malignant ovarian tumor tissue specimens (malignant tumor group), 75 cases of benign ovarian tumor tissue specimens (benign tumor group) and 75 cases of normal ovarian tissue specimens from bilateral ovariectomy for unilateral ovarian lesions (control group). Immunohistochemistry was used to detect the expression of SLP-2 protein in the three groups. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was utilized to detect the relative expression of SLP-2 mRNA in the three groups, and the relationship between SLP-2 and clinicopathological parameters of the ovarian cancer patients was analyzed. The patients with ovarian cancer were divided into the SLP-2 high-expression group and the SLP-2 low-expression group according to the median of SLP-2 relative expression. The survival of patients was analyzed using the Kaplan-Meier and Cox regression model. The results of immunohistochemistry showed that the positive expression rate of SLP-2 protein in the malignant tumor group was significantly higher than that in the benign tumor and control groups (P<0.001). The results of RT-qPCR showed that compared with the control group, the relative expression of SLP-2 mRNA in the ovarian tissues in the benign tumor group and the malignant tumor group was increased (P<0.001). The relative expression of SLP-2 mRNA in the malignant tumor group was higher than that in the benign tumor group (P<0.001). The relative expression of SLP-2 mRNA correlated with clinical stage, pathological differentiation and lymph node metastasis of the patients with ovarian cancer (P<0.05). The 5-year overall survival (OS) in the SLP-2 mRNA high expression group was significantly lower than that in the SLP-2 mRNA low expression group at 5 years (P<0.05). SLP-2 mRNA was an independent prognostic factor influencing OS of the patients (P<0.05). SLP-2 may be involved in the occurrence and development of ovarian cancer and related to the clinical stage, pathological differentiation and lymph node metastasis of the patients with ovarian cancer, which may also play a role in promoting the invasion and metastasis processes of ovarian cancer. Therefore, SLP-2 is expected to be an effective biomarker for targeted treatment and prognosis of ovarian tumor.

Targeting mediator subunits CDK8/CDK19 for treatment of advanced prostate cancer.

152 Background: The mediator complex plays a pivotal role in the regulation of gene transcription. The subunits CDK8 and CDK19 are overexpressed in advanced prostate cancer (PCa) and promote migration and invasion, as shown previously by our group. The aim of this study was to analyze if CDK8/CDK19 inhibition can impede PCa progression. Methods: Immunohistochemistry for CDK8 and CDK19 was performed on a large PCa cohort (376 radical prostatectomy (RP) specimens, 39 locally advanced tumors, 32 lymph node metastases, 24 distant metastases, 73 benign prostatic specimens). PCa cell lines (DU145, PC3) were treated with CDK8/CDK19 small molecule inhibitors followed by MTT assay, wound healing assay, and Western Blot for epithelial/mesenchymal markers. Results: CDK19 is higher expressed in primary PCa vs. benign specimens and locally advanced PCa/distant metastases vs. primary PCa. CDK8 and CDK19 are higher expressed in castration-resistant specimens vs. hormone-naive specimens. CDK19 significantly correlates with grade group in RP specimens. Biochemical recurrence free survival rates are lower for patients with high vs. medium vs. no/low CDK19 expression in the tumor. CDK19 predicts recurrence free survival independently from grade group and PSA. CDK8/CDK19 inhibition results in decreased migration, while there is no effect on proliferation. Mesenchymal markers are reduced while epithelial markers are induced following CDK8/CDK19 inhibition. Conclusions: CDK19 qualifies as a prognostic marker predicting biochemical recurrence following RP. CDK8/CDK19 inhibition leads to decreased migratory potential and mesenchymal phenotype. CDK8/19 expression increases during progression to castration-resistance. Collectively, CDK8/CDK19 represents a new target for treatment of advanced and/or castration-resistant PCa.

Immunogenomic landscape of neuroendocrine small cell prostate cancer.

217 Background: Neuroendocrine small cell prostate cancer (NEPC) is a lethal variant of prostate cancer (PCa) unresponsive to hormone therapy and associated with poor prognosis. Cisplatin induces short-lived responses and therefore alternative novel therapeutic options are urgently needed. Methods: Prostate specimens from radical prostatectomy or transurethral resection (benign prostate specimens n = 4, primary untreated or neoadjuvant hormone-treated adenocarcinoma n = 30, castrate-resistant prostate cancer-CRPC n = 38 and NEPC n = 16) were evaluated for PD-L1 (SpringBio, M4420), AR, chromogranin A, synaptophysin, NSE and CD56 expression by immunohistochemistry (IHC). Archival tissue from liver, lymph nodes and prostate from 30 additional patients with de novo and treatment emergent NEPC were analyzed for PD-L1 expression by IHC. The intensity was assessed as percentage of positive cells / mm2 of tissue. Targeted and whole exome sequencing of the high-density tumor areas were performed and correlated to the PD-L1 status (PD-L1+ve: &gt; 1% of positive cells). OS was calculated from the date of diagnosis of NEPC to death. We set out to define PD-L1 expression and immunogenomic characteristics of NEPC. Results: PD-L1 was expressed in 0%, 5%, 10% and 41% of benign, adenocarcinoma, CRPC and NEPC specimens, respectively. The PD-L1 expression intensity was significantly higher in patients with NEPC (mean: 40%, range: 5-100%) compared to benign, adenocarcinoma and CRPC samples (mean: 0%, 2% and 8%, respectively, P &lt; 0.0001). There was a higher prevalence of biallelic DNA Repair Defects (DRD) in the PD-L1+ve vs PD-L1-ve patients (65% vs 0%, P = 0.005). The median OS of the NEPC patients was 8.5 months vs 10.5 months in PD-L1+ve vs PD-L1-ve tumors (HR 1.24, 95% CI: 0.59-2.75, p = 0.55). Conclusions: NEPC have greater PD-L1 expression than adenoCa and CRPC. Biallelic DRD was exclusively observed in PD-L1+ve patients. Since PD-L1 expression and DRD have been associated to response to PARP and PD1/PD-L1 inhibitors in prostate and other cancers, further studies evaluating the activity of those agents in NEPC patients are warranted.

Abstract P2-14-18: Achiveing rapid intrao-operative diagnosis during breast cancer surgery using high-reloution full-field optical coherence imaging and dynamic cell imaging

Abstract Background: Intraoperative pathological diagnosis such as frozen section and imprint cytology is not routinely recommended in clinical practice because of time and accuracy concerns. Full-field optical coherence tomography (FF-OCT) is a new optical imaging technique that could generate sectioning tomogram from fresh tissue and provide depiction of the morphological structure and pathological changes in minutes without conventional tissue preparation, slicing, and staining, and dynamic cell imaging (DCI) added the viability information of cells/tissue, which could be more important in cancer diagnosis. This study was to evaluate the feasibility and diagnostic value of FF-OCT and DCI in breast lesions and lymph node specimens during breast cancer surgery. Methods: We evaluated normal breast tissue, benign breast lesions, breast cancer and axillary lymph node specimens from 107 patients using FF-OCT and DCI. After the optical assessment, the tissue was paraffin embedded and sent to conventional H&amp;E diagnosis. The similar layer of OCT and H&amp;E images were compared and diagnostic criteria were generated. The diagnostic sensitivity and specificity by two trained surgeons without pathology diagnosis experience were evaluated. Results: A total of 194 specimens were examined, including 143 breast tissue(101 malignant and 42 benign/normal) and 51 lymph nodes(26 metastatic and 25 non-metastatic). On FF-OCT and DCI, normal morphological structures such as adipose, collagen, mammary ducts, and lobules in breast tissue and lymphoid follicle and hilum in lymph nodes were easily recognized. Breast cancer characteristics on H&amp;E imaging correspond to collagen distortion, focal hypointensity, micro-calcification, clustered or linear lively cells et etc on FF-OCT or DCI, which could also be easily distinguished. We included the previously mentioned features to build diagnosis criteria for cancer on FF-OCT and DCI. The average acquisition time is 14±11 minutes. The sensitivity and specificity for breast cancer diagnosis were 92.1% and 94.3% respectively. The sensitivity and specificity for lymph node involvement were 92.3% and 84% respectively. Conclusion: The time- and tissue-saving optical imaging technique yielded high accuracy that was comparable to that of traditional intraoperative and postoperative pathological diagnosis in breast cancer and lymph node metastasis. These results implied the promising application in the intraoperative evaluation and possible decrease of the re-excision rate for breast cancer surgery. Citation Format: Yang H, Zhang S, Guo J, Xie F, Tong F, Cao Y, Liu P, Zhou B, Cheng L, Liu M, Wang S, Peng Y, Wang C, Yang Y, Ma Y, Chen D, Shen D, Wang S. Achiveing rapid intrao-operative diagnosis during breast cancer surgery using high-reloution full-field optical coherence imaging and dynamic cell imaging [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P2-14-18.

A Novel Approach to Detect Programed Death Ligand 1 (PD-L1) Status and Multiple Tumor Mutations Using a Single Non–Small-Cell Lung Cancer (NSCLC) Bronchoscopy Specimen

Multiple biomarkers are under evaluation to guide the use of immune checkpoint inhibitors in non-small-cell lung cancer (NSCLC), including programed death ligand 1 (PD-L1) tumor cell staining. We have developed a new approach that accurately quantifies PD-L1 status and identifies multiple mutations by using a single bronchoscopy specimen. A novel molecular marker was identified to detect the presence of malignant cells in radial endobronchial ultrasound bronchial brushings from NSCLC (n = 15) and benign (n = 13) nodules by quantitative real-time RT-PCR (RT-qPCR). The MMP9:TIMP3 transcript ratio was significantly increased in NSCLC and using receiver operating characteristic curve analysis accurately discriminated malignant and benign bronchoscopy specimens (area under the curve=0.98; 95% CI, 0.93-1; P<0.0001). Utilizing the same specimens, PD-L1 expression and multiple oncogenic mutations were detected by RT-qPCR and next-generation sequencing. A second archive of snap-frozen squamous cell carcinoma (n = 40) and control (n = 20) biopsies with matching formalin-fixed, paraffin-embedded slides were used to compare PD-L1 status by immunohistochemistry and RT-qPCR. The biopsy cohort confirmed that the MMP-9:TIMP3 ratio was predictive of malignancy and demonstrated that PD-L1 transcript expression was concordant with PD-L1 tumor cell membrane staining in NSCLC (Spearman r=0.636, P<0.0001). This rapid molecular approach can detect malignant cells and using the same single bronchoscopy specimen can generate high-quality unfixed nucleic acid that accurately quantify PD-L1 status and identify multiple oncogenic mutations.

Detection of free intraperitoneal tumour cells in peritoneal lavage fluid from patients with peritoneal metastasis before and after treatment with pressurised intraperitoneal aerosol chemotherapy (PIPAC)

AimsIn this study, we investigated whether free intraperitoneal tumour cells (FITC) were detectable in ascites or peritoneal lavage fluid (PLF) from patients with peritoneal metastasis (PM) before and after treatment...

The HLA ligandome landscape of chronic myeloid leukemia delineates novel T-cell epitopes for immunotherapy

AbstractAntileukemia immunity plays an important role in disease control and maintenance of tyrosine kinase inhibitor (TKI)-free remission in chronic myeloid leukemia (CML). Thus, antigen-specific immunotherapy holds promise for strengthening immune control in CML but requires the identification of CML-associated targets. In this study, we used a mass spectrometry–based approach to identify naturally presented HLA class I– and class II–restricted peptides in primary CML samples. Comparative HLA ligandome profiling using a comprehensive dataset of different hematological benign specimens and samples from CML patients in deep molecular remission delineated a panel of novel frequently presented CML-exclusive peptides. These nonmutated target antigens are of particular relevance because our extensive data-mining approach suggests the absence of naturally presented BCR-ABL– and ABL-BCR–derived HLA-restricted peptides and the lack of frequent tumor-exclusive presentation of known cancer/testis and leukemia-associated antigens. Functional characterization revealed spontaneous T-cell responses against the newly identified CML-associated peptides in CML patient samples and their ability to induce multifunctional and cytotoxic antigen-specific T cells de novo in samples from healthy volunteers and CML patients. Thus, these antigens are prime candidates for T-cell–based immunotherapeutic approaches that may prolong TKI-free survival and even mediate cure of CML patients.

Expression of beta-human chorionic gonadotropin as a prognostic factor in colorectal carcinoma

Background: Colorectal carcinoma (CRC) is a common malignancy in Iraq and carries a high mortality rate. It has been shown that CRCs express high level of beta-human chorionic gonadotropin (β-HCG) as detected by immunohistochemistry. Objectives: The aim of this study was to assess the expression of β-HCG in CRC and to study the association between β-HCG expression and pathological prognostic parameters. Materials and Methods: This study was carried out on 35 colonic specimens consisting of 30 CRC selected from files of Pathologic Laboratory of Al-Kadhymia Teaching Hospital and Teaching Laboratories of the Medical City, Baghdad. Paraffin blocks were cut sections stained with H and E stain and sections for streptavidin-biotin immunohistochemical staining using monoclonal antibody against β-HCG. Immunohistochemical sections were examined for positive or negative staining and positivity assessed as: occasional, focal, and diffuse tumors. Results: Of the 30 CRCs, 60% were male and 40% were female. The most common site of large bowel malignancy was the rectum 13 (43.3%). Most of the tumors were moderately-differentiated adenocarcinoma (83.3%). More than half of the cases were Stage T3, followed by Stage T2, then Stage T4, 3 cases (10%), and no case in Stage T1. Lymph node metastases were found in 19 cases (63.3%). Twelve cases (40%) of CRC show immunoreactivity to β-HCG. β-HCG expression was more frequent in the left side of the large bowel and more common in poorly-differentiated adenocarcinoma (80%). β-HCG was more positive in Stage T4, and there was significant association with lymph node metastasis. Conclusion: Expression of β-HCG in CRC is not an infrequent phenomenon, and no stain was demonstrated in benign and normal colonic specimens, while β-HCG expression associated with poor differentiation, greater local invasion, and regional lymph node metastasis.

Histopathological Study of Non-Cirrhotic Portal Fibrosis (NCPF) With Special Emphasis on Advanced Fibrosis

Background: Non-cirrhotic portal hypertension includes primarily the cases of non-cirrhotic portal fibrosis (NCPF) and extra-hepatic portal vein obstruction, both showing no significant parenchymal disease. NCPF is a disease of uncertain etiology characterized by periportal fibrosis with small and medium sized portal vein branches involvement. Most study done in the past are clinical, hence we decided to analyse its histo-pathological features with clinical correlation. Methods: This is four and half year’s retrospective study of liver biopsies of clinically diagnosed patients of NCPF. Fifteen patients of EHPVO served as controls. Biopsies were stained with Haematoxylin and eosin, reticulin and Masson’s trichrome. Result: Total 978 benign specimens (excluding space-occupying lesions) received, of which 45 diagnosed as NCPF (4.6%). Most patients (68.88%) were between 20-40 years (M:F ratio 1:1.5), commonest symptoms long-standing splenomegaly (95%), anemia (78%) and variceal bleeds (42%) with duration varying from 3 months to 11 years. Commonest histopathological findings were portal tract fibrosis (77.78%), phlebosclerosis (68.89%), portal tract remnants (24.44%), more frequently seen in NCPF than EHPVO, septal fibrosis was seen in five. Follow up available in 91.11% cases showed no mortality or liver failure, but significant morbidity due to repeated episodes of variceal bleeds and hypersplenism. Conclusion: NCPF is still common in India. Distinctive features on histology are portal tract fibrosis, portal tract remnants, with phlebosclerosis which is being statistically significant when comparing with EHPVO. NCPF can show advanced fibrosis, can also present with features of decompensation mimicking cirrhosis, however its course remains stable with good long-term survival.

Cytology of the fallopian tube: A screening model for high-grade serous carcinoma.

Ovarian cancer is a heterogeneous disease having the highest gynecologic fatality in the United States with a 5-year survival rate of 46.5%. Poor overall prognosis is mostly attributed to inadequate screening tools, and the majority of diagnoses occur at late stages of the disease. Due to genetic and biological underpinnings, ovarian high-grade serous carcinomas (HGSC) have etiologic evidence in the distal fallopian tube. Fallopian tube screening modalities are aggressively investigated, but few describe cytological characteristics of benign tubal specimens to help in the comparative detection of HGSC precursor cells. Here, we describe fimbrial cytomorphological and nuclear features of tubal specimens (n = 75) from patients clinically indicated for salpingectomy, bilateral or unilateral salpingo-oophorectomy, and hysterectomies for any diagnosis other than ovarian or peritoneal cancer. Fallopian tube histology was used as the diagnostic reference. A total of 75 samples had benign diagnoses. The benign cytological characteristics of fimbrial tubal specimens included ciliated cells in clustered arrangements with mild nuclear membrane irregularity, mild anisonucleosis, round and/or oval nuclei, hyperchromatic chromatin, and mild nuclear membrane irregularity. In contrast, none of the cytology samples had spindle-shaped nuclei, significantly marked anisonucleosis (n = 1), nor had hypochromasia as a characteristic feature. These cytological characteristics could be a potential area of distinction from HGSC precursor cells. Our study establishes cytomorphological characteristics of nonmalignant tubal cells which help underscore the importance of distinguishing malignant HGSC precursors through fimbrial brush sampling in minimally invasive approach.

Ectopic Thyroid Tissue: Immunohistochemistry and Molecular Analysis.

Ectopic thyroid tissue is rare and controversial. Some experts consider it to always be metastatic thyroid carcinoma, whereas others consider it benign as long as it is restricted to few follicles without cytoarchitectural features of papillary thyroid carcinoma. Immunohistochemistry (IHC) and molecular studies have not yet been performed to further characterize this entity. We retrospectively searched our pathology files for all ectopic thyroid inclusions and reviewed clinicopathologic characteristics and concurrent thyroid pathologic findings. We identified 8 cases from 7 patients. Ectopic thyroid tissue was present in the following locations: neck soft tissue: 3, thymus: 2, neck lymph nodes: 2, perihilar soft tissue: 1. All patients had histologically benign thyroid specimens. BRAFV600E (VE1) IHC, HBME-1 IHC, galectin-3 IHC, BRAFV600E allele-specific polymerase chain reaction (PCR) and NRAS/KRAS pyrosequencing were performed. To assess the sensitivity and specificity of BRAFV600E IHC compared with PCR; we tested 13 cases of primary and metastatic papillary and follicular thyroid carcinomas. All the ectopic cases were HBME-1, galectin-3, BRAFV600E (IHC, PCR), and NRAS/KRAS mutation negative (specificity=100%). Compared with PCR, BRAF IHC had 89% sensitivity and 100% specificity. Lack of common carcinoma-associated mutations supports benign nature of this entity. BRAF, HBME-1, and galectin-3 IHC are accurate and helpful when not enough tissue is available for molecular studies. IHC and molecular studies are more helpful than morphology alone in identifying benign thyroid rests.

Contained Laparoscopic Power Morcellation for Large Pelvic Masses

Objective: Contained morcellation systems are now considered a mandatory requirement for all myomectomies and large hysterectomies. Although the risk of undiagnosed sarcomas has been largely overestimated (likely only around 1:70), the consequence of upstaging this severe disease through uncontained morcellation can be devastating. Furthermore, the rising incidence of parasitic fibroids after morcellation of benign specimens provides an argument supporting contained extraction techniques for all fibroids. SWEC has previously described the “Sydney Contained In Bag Morcellation” technique. However, it required piercing the bag with an accessory trocar producing the theoretical risk of small volume cell spillage. Whether this equates to a real risk has not yet been established. EMT Medical has produced a fully contained morcellation system designed for laparoscopic power morcellation for large pelvic masses. The objective of the study is to demonstrate contained specimen extraction with laparoscopic power morcellation for large pelvic masses utilising the Espiner Morcellation Containment System.

Incidental Prostate Cancer in Radical Prostatectomy Specimens of Patients Treated for Benign Prostatic Hyperplasia (BPH): Histopathology Analysis

Background: The incidence of prostate cancer (Pca) is the first in malignant tumors among men in the United States, and mortality rate is the second cause of deaths. Incidental prostate cancer is defined as clinically inapparent tumor that is neither palpable nor visible by imaging. Incidental carcinoma is found in 3%-16% of pathology specimens of patients undergoing BPH surgery and it involves less than 5% of resected tissue. Aim: To identify incidentally detected prostate cancer in benign prostatic specimens submitted for histopathology Methods: This is a ten-year retrospective histopathology study of all prostate cancer cases diagnosed between 1991-2000 in the department of Pathology, Ahmadu Bello University Teaching Hospital, Zaria, Nigeria. Haematoxylin &amp; eosin stained histology slides were retrieved and studied. Institution review board (IRB) approval was obtained from the Teaching Hospital to undertake this study. Results: A total of 67 patients had benign prostatic hyperplasia; the patients' age ranged from 30-75 years with a mean age of 52.5 years. The peak age at diagnosis was in the fifth decade. 26 (38.8%) patients had incidental carcinoma. All the tumors were adenocarcinoma and moderately differentiated. Two patients in their fourth decade were found to have incidental prostate cancer. Conclusion: Incidental prostate cancer is common in patients' undergoing radical prostatectomy for BPH, a high index of suspicion, adequate histopathology training and histopathologic evaluation is critical to diagnosis.

Forkhead box O1 as an indicator of prognosis is inactivated in urothelial carcinoma of the upper urinary tract.

The transcription factor forkhead box O1 (FOXO1) can be inactivated via its phosphorylation, resulting in suppression of apoptosis. Using immunohistochemistry, the expression of a phosphorylated form of FOXO1 was assessed in upper urinary tract urothelial carcinoma (UUTUC) specimens. Overall, phospho-FOXO1 (p-FOXO1) was immunoreactive in all 99 UUTUC specimens [12 (12.1%) weak (1+), 46 (46.5%) moderate (2+) and 41 (41.4%) strong (3+)], which was significantly (P=0.018) increased, compared with benign urothelium specimens [77/82 (93.9%): 18 (22.0%) 1+, 41 (50.0%) 2+ and 18 (22.0%) 3+]. Muscle invasion (P=0.031) and lymphovascular invasion (P=0.025) were observed more frequently in p-FOXO1(2+/3+) tumor samples compared with p-FOXO1(1+) tumor samples. No statistically significant associations between p-FOXO1 expression and tumor grade or presence of concurrent carcinoma in situ, hydronephrosis or lymph node metastasis were observed. Furthermore, the levels of p-FOXO1 and estrogen receptor-β expression were significantly (P<0.05) correlated in UUTUC samples [correlation coefficient (CC)=0.244], particularly in tumor samples from male patients (CC=0.330). Additionally, patients with p-FOXO1(3+) tumors had a significantly increased risk of cancer-specific mortality (P=0.043), compared with those with p-FOXO1(1+/2+) tumors. Multivariate analysis further demonstrated a notable, albeit not significant, association between p-FOXO1 expression and cancer-specific survival (hazard ratio=2.204; P=0.053). These findings indicate that FOXO1 is inactivated in UUTUC specimens and p-FOXO1 overexpression may serve as a predictor of poor patient outcomes.

The Prevalence of Human Papillomavirus Infection in Benign Sinonasal Lesions: A Single-Center Retrospective Analysis

It is suggested that just like cigarette smoking and occupational exposure, HPV plays a significant role in the etiology of benign sinonasal lesions. It is evident through literature review that low-risk HPV types 6 and 11 are usually confined to benign lesions, whereas the reverse is true for the oncogenic HPV types 16 and 18. Our aim was to seek the prevalence and the types of HPV coinfection in a benign head and neck surgical specimen. Benign specimens from head and neck sites with available HPV wide spectrum testing and immunohistochemistry for HPV and p16 were retrospectively reviewed (January 2011 to October 2015). Thirty-seven sinonasal surgical specimens were identified, requiring direct examination and enough changes warranting a biopsy. On histology, these revealed focal viral cytopathic changes, inverted papillomas, squamous hyperplasia, squamous papilloma, and verrucous hyperplasia type of morphology. The results for HPV wide spectrum, HPV 6, 11, 16, 18, 33, 35, and 45, were collected. Thirteen of 37 benign lesions showed positive results for HPV wide spectrum. Two of 13 came out to be positive for HPV 16 with positive p16 staining on immunohistochemistry. The remaining (11/13) cases were positive by HPV wide spectrum for HPV 6 and HPV 11 (n = 7 with HPV 6, n = 4 with HPV 11) showing focal p16 staining. Twenty-one of 37 showed focal p16 staining, which was interpreted as negative. Current studies link HPV to at least a proportion of benign sinonasal papillomas. HPV 6 and HPV 11 seem to be present in a large number (30%) of these benign lesions in our cohort. This corresponds with the available literature (30%-35%). Although two of our cases did show infection with HPV 16, there was no dysplasia identified in the lesion.

Modified Bethesda system informing cytopathologic adequacy improves malignancy risk stratification in nodules considered benign or atypia(follicular lesion) of undetermined significance

We modified the nondiagnostic/unsatisfactory category of the Bethesda system for reporting thyroid cytopathology to inform cytopathologic adequacy to better stratify the malignancy risk. Malignancy rates from 1,450 cytopathologic specimens not satisfying adequacy criteria from April 2011 to March 2016 were calculated based on sub-classification of the nondiagnostic/unsatisfactory category and sonographic patterns using matched surgical pathology. Rates were compared with those of 1,446 corresponding adequate specimens from July to December 2013. Upon resection, 63.2% of nondiagnostic, 36.7% of unsatisfactory + benign, 72.5% of unsatisfactory + atypia (follicular lesion) of undetermined significance, 98.1% of unsatisfactory + suspicious for malignancy, and 100.0% of unsatisfactory + malignant cases were confirmed to be malignant on surgical pathology. In nodules with inadequate specimens, those with high suspicion sonographic patterns had a malignancy rate (93.2%) higher than the others (45.5%) (p < 0.001). Nodules with unsatisfactory + benign specimens had a higher malignancy rate (36.7%) than satisfactory benign specimens (14.3%) (p = 0.020). For atypia (follicular lesion) of undetermined significance, the malignancy rate of inadequate specimens (72.5%) was higher than that of adequate specimens (51.3%) (p = 0.027). Sparse cellular samples with a few groups of benign follicular cells should not represent a benign lesion. There might be value in qualifying atypia (follicular lesion) of undetermined significance cases less than optimal.

The FOXC1/FBP1 signaling axis promotes colorectal cancer proliferation by enhancing the Warburg effect.

Aberrant expression of Forkhead box (FOX) transcription factors plays vital roles in carcinogenesis. However, the function of the FOX family member FOXC1 in maintenance of colorectal cancer (CRC) malignancy is unknown. Herein, FOXC1 expression in CRC specimens in The Cancer Genome Atlas (TCGA) cohort was analyzed and validated using immunohistochemistry with a tissue microarray. The effect of FOXC1 expression on proliferation of and glycolysis in CRC cells was assessed by altering its expression in vitro and in vivo. Mechanistic investigation was carried out using cell and molecular biological approaches. Our results showed that FOXC1 expression was higher in CRC specimens than in adjacent benign tissue specimens. Univariate survival analyses of the patients from whom the study specimens were obtained, and validated cohorts indicated that ectopic FOXC1 expression was significantly correlated with shortened survival. Silencing FOXC1 expression in CRC cells inhibited their proliferation and colony formation and decreased their glucose consumption and lactate production. In contrast, FOXC1 overexpression had the opposite effect. Furthermore, increased expression of FOXC1 downregulated that of a key glycolytic enzyme, fructose-1,6-bisphosphatase 1 (FBP1). Mechanistically, FOXC1 bound directly to the promoter regions of the FBP1 gene and negatively regulated its transcriptional activity. Collectively, aberrant FBP1 expression contributed to CRC tumorigenicity, and decreased FBP1 expression coupled with increased FOXC1 expression provided better prognostic information than did FOXC1 expression alone. Therefore, the FOXC1/FBP1 axis induces CRC cell proliferation, reprograms metabolism in CRCs, and constitutes potential prognostic predictors and therapeutic targets for CRC.