Brush Biopsy Research Articles

Abstract 3537: Targeted deep sequencing of driver mutations in airway epithelial cells from smokers

Abstract Background: Inter-individual variation in risk for lung cancer is based on variation in both exposure to environmental factors and genetic predisposition. As such, it is reasonable to hypothesize that prevalence of mutations in the airway epithelium will vary based on the combined effects of these lung cancer risk factors. Genes most commonly mutated in non-small cell lung cancer were recently reported from The Cancer Genome Atlas (TCGA) project. The purpose of this study was to use targeted deep sequencing to assess the prevalence of these mutations in bronchial epithelial cells (BEC) of individuals at high demographic risk for lung cancer. Methods: We enrolled 19 individuals with varying demographic risk for lung cancer, who were undergoing standard-of-care bronchoscopy into an IRB-approved research study in which we collected 1-5 million BEC via bronchoscopic brush biopsy and extracted genomic DNA. 10 subjects were smokers with lung cancer, 6 subjects were smokers without lung cancer, and 3 subjects were non-smokers without lung cancer. For targeted deep sequencing, we prepared synthetic spike-in competitive internal standards for each of 12 gene loci commonly mutated in lung cancer based on TCGA results and combined them into an IS mixture (ISM). DNA aliquots from each patient were quantified and mixed with equivalent genome equivalents of ISM prior to library preparation to control for technical artifacts associated with library preparation and NGS platform-specific error. The DNA input varied depending on sample quantity; there were 5 samples with 50,000 genome copies loaded, 12 samples with 100,000 genome copies loaded, and 2 samples with 1 million genome copies loaded. This approach enabled reliable measurement of 95% confidence limits of variant allele frequency (VAF), controlled for technical sequencing error, and allowed confident call of variants at a VAF of 0.1% and lower. Libraries for specimens from each of the 19 subjects were first sequenced using the Illumina MiSeq platform to ensure equal coverage across targets and samples. Deeper sequencing was then obtained using the Illumina NextSeq platform. Results: Preliminary results showed that smokers harbored mutations up to 1.6% in gene loci associated with non-small cell lung cancer according to TCGA, while none were identified in non-smoking controls. Conclusion: Initial results suggest that airway epithelial cells in smokers acquire somatic mutations that gradually lead to malignant conversion. Deep sequencing replicate experiments are underway to expand these results. Citation Format: Daniel J. Craig, Thomas M. Blomquist, Erin L. Crawford, James C. Willey. Targeted deep sequencing of driver mutations in airway epithelial cells from smokers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3537.

Etiological verification of infectious exacerbation of chronic obstructive lung disease using molecular genetics methods

Objective: To study the possibilities of the method of determining the 16S rRNA gene of bacteria in the determination of the bacterial spectrum of acute exacerbation of chronic obstructive pulmonary disease (COPD) using a protected material sampling Materials and Methods: 68 Hospitalized patients with acute exacerbation of COPD were prospectively evaluated. Molecular genetic determination of the 16S rRNA gene of bacteria and cultural microbiological techniques from sputum and a bronchial tree material obtained using a protected brush biopsy were used. Results: Bacterial agents were detected in 91,2 % of cases of acute exacerbation of COPD. Pseudomonas aeruginosa was detected in the biopsy material 1.8 times more often than with the use of routine methods. Patterns of COPD with frequent exacerbations, infectious exacerbations and Pseudomonas etiology of exacerbations were determined. Conclusion: Using the method of molecular-genetic determination of the 16S rRNA gene allows to verify the bacterial spectrum of acute exacerbation of COPD.

CDK4, CDK6, cyclin D1 and Notch1 immunocytochemical expression of oral brush liquid-based cytology for the diagnosis of oral leukoplakia and oral cancer.

This study aimed to investigate the feasibility of using oral liquid-based brush cytology (OLBC) coupled with immunocytochemistry as a minimally invasive approach to stratify the cancer risk in patients with oral leukoplakia. Fifty-five patients diagnosed with either oral leukoplakia (OLK) or oral squamous cell carcinoma (OSCC) were recruited. All patients underwent oral brush biopsy followed by surgical biopsy. 275 liquid-based cytology preparations were made. Pap-stained OLBC slides were assessed using the modified 2014 Bethesda Cytology system. The expression of CDK4, CDK6, cyclin D1, and Notch 1 was immunocytochemically analysed and compared against the histopathological diagnosis. A combined index score of OLBC grading and protein expression was calculated. A significant association was found between the definitive histopathological diagnosis and the cytological interpretation (P=0.0005). The index scores of CDK4, CDK6, and cyclin D1 were significantly associated with the development of disease from non-dysplastic epithelium to OSCC. No significant association was observed between the Notch 1 index score and disease stage. The diagnostic accuracy of OLBC showed the highest values of sensitivity, specificity, positive predictive value, negative predictive value, and accuracy: 84.6%, 70.4%, 73.3%, 82.6%, and 78.8%, respectively, compared with the cumulative protein index, CDK4/6 index, and the combined OLBC grading and CDK4/6 index. This study has also demonstrated the efficacy of the use of OLBC in the detection of OED and OSCC, and showed that the use of CDK4, CDK6, cyclin D1, and Notch 1 immunocytochemistry failed to improve the diagnostic accuracy of OLBC suggesting they are not useful in the early detection of OSCC.

Evidence of genotoxicity and cytotoxicity of X-rays in the oral mucosa epithelium of adults subjected to cone beam computed tomography.

Evidence of genotoxicity and cytotoxicity of X-rays in the oral mucosa epithelium of adults subjected to cone beam computed tomography.

MINIMALLY-INVASIVE ORAL EXFOLIATED CELLS STUDY FOR PREMALIGNANT LESIONS USING RAMAN MICROSPECTROSCOPY

Squamous cell carcinoma of the oral cavity ranks as the 15th most common cancer in the world and the 10th most frequent cancer in males. The present study was undertaken for the development of new methods for early oral cancer detection based on Raman microspectroscopy of exfoliated cells. Exfoliated oral cells were collected by brush biopsy from patients attending Dublin Dental Hospital Dysplasia Clinic (25) and from healthy volunteers (25). Samples of exfoliated cells from normal mucosa and from pre-malignant lesions were collected using an endocervical cytobrush and placed in ThinPrep vials. Slides were prepared using the Thinprep2000 processor with the aim of forming a monolayer of cells for analysis. Raman spectra were acquired from the nucleus and cytoplasm of each cell using an XploRA confocal Raman instrument (HORIBA JobinYvon). As source, a 532 nm laser was focused by a 100X objective onto the sample and the resultant Raman signals were acquired in the 400 to 1800 cm−1 region. Glass spectral contamination was removed using extended multiplicative signal correction Following pre-processing, spectra were subjected to principal component analysis (PCA) and principal component-linear discriminant analysis (PC-LDA). The results show that Raman spectroscopy coupled with PCA could differentiate the nucleus and cytoplasm of the cell, the PC loadings showing that the cytoplasmic regions are dominated by protein bands while thenuclear regions are dominated by DNA bands. Furthermore, patient samples were discriminated from healthy volunteers based on DNA and lipids bands inthe PC loadings. Sensitivities of 91% and 97% and specificites of 98% and 89% were achieved for the cytoplasm and nucleus respectively, using PC-LDA. Thus, the findings of the study support the potential of Raman microspectroscopy for providing molecular level information from oral exfoliated cells and the future potential for screening of minimally invasive brush biopsy samples for oral pre-cancer and cancer.

Indications for Single-Operator Cholangioscopy and Pancreatoscopy: an Expert Review.

Direct endoscopic visualization of the biliary and pancreatic ducts by single-operator cholangioscopy (SOC) and pancreatoscopy (SOP) is an expanding technique with an increasing number of indications. This review provides technical tips and tricks and a literature update on the usefulness of SOC and SOP in clinical practice. SOC has shown a 91.3% and 87.3% diagnostic procedural success in indeterminate strictures and difficult stones respectively, with a 9.4%-16.4% complication rate. Although the interpretation of the images suspected for malignancy can be challenging, new classifications based in mucosal and vascular patterns as well as better devices are in development. A higher sensitivity has been associated to targeted biopsies, compared to brushing and blinded biopsies. Indeed, SOC has recently demonstrated a decrease in the number of procedures and costs when compared with endoscopic retrograde cholangiopancreatography. This technique has also been reported for many other diagnostic and therapeutic indications, such as cholangiocarcinoma staging and foreign body extraction. Similarly, diagnostic and therapeutic SOP in indeterminate strictures or pancreatic stones, and intraductal papillary mucinous neoplasms, has been successfully reported. Research to date supports the use of cholangioscopy as a complementary examination in patients with indeterminate strictures, difficult stones and specific situations. In a near future, there will probably be new established indications, a better understanding of the imaging findings and a place of SOC/SOP as first line in selected scenarios.

Sa1731 – A Trans-Nasal, Image-Guided Approach for Brush Biopsy Sampling of the Upper Gi Tract Microbiota

Sa1731 – A Trans-Nasal, Image-Guided Approach for Brush Biopsy Sampling of the Upper Gi Tract Microbiota

Next Steps after Negative Results Obtained by EBUS-TBNA from Patients Suspected Clinically Lung Cancer with Mediastinal Lymphnode Metastasis

背景与目的超声气管镜针吸活检（endobronchial ultrasound guided tranbronchial needle aspiration, EBUS-TBNA）是肺癌诊断和分期的重要手段，但经活检阴性结果的患者后续处理尚无标准流程。本文通过分析来自单中心的临床疑诊肺癌纵隔淋巴结转移但EBUS-TBNA病理结果阴性患者，以探讨此类患者处理方式。方法对北京协和医院2010年9月-2016年12月进行EBUS-TBNA的1, 412例患者资料进行分析，选取临床疑诊肺癌纵隔淋巴结转移但EBUS-TBNA病理诊断阴性的患者51例进行回顾性分析。结果入选51例患者按临床情况和后续处理方式分为以下5组：①经同一次气管镜下其他检查组（9例）：该组患者大多（8例）存在镜下异常表现，通过活检、毛刷、灌洗或经支气管镜肺活检（transbronchial lung biopsy, TBLB）取得明确诊断；②再次EBUS-TBNA组（11例）：该组患者气管粘膜及管腔大致正常，再次行EBUS-TBNA取得诊断；③手术治疗组（6例）：该组患者因EBUS结果除外纵隔淋巴结转移，接受手术治疗。其中5例术后确诊无淋巴结转移癌；④进行其他病理检查组（15例）：该组患者有其他部位转移，针对可能的转移灶进行计算机断层扫描（computed tomography, CT）引导下穿刺、淋巴结活检等确诊。⑤随访组（10例）：该组患者未进行其他有创检查，中位随访时间38个月，其中1例随访中诊断为淋巴瘤。结论对于经EBUS-TBNA未能确定诊断而临床怀疑肺癌的患者，应该根据患者的具体情况，综合多种方式进行诊断。对于暂时无法确诊的患者，仍需要长期随访。

Esophageal IgE, IgG4, and mucosal eosinophilia in individuals with dysphagia.

Eosinophilic esophagitis (EoE) is an inflammatory disease of the esophagus, producing failure to thrive in infants and dysphagia with food impaction in older children and adults. Although most people with EoE manifest atopic/allergic disease, the specific allergens to which immunoglobulin E (IgE) is directed, if any, have not yet been characterized. Mucosal brush biopsy (MBB) and solid tissue biopsy (STB) specimens were prospectively obtained from 25 individuals with dysphagia and suspicion of EoE. Specific IgE (sIgE) against 112 epitopes from airborne and food proteins, antigens known to cause a polyclonal IgE response and IgG4 to food allergens, were measured. There was no difference in total IgE harvested between the 2 biopsy methods (p > 0.05) or between the EoE-positive (N = 12) and EoE-negative (N = 13) groups (p > 0.05). None of the samples in either group contained measurable serum IgE to any of the airborne or food proteins tested, but low levels of IgE specific to Candida and Staphylococcus enterotoxins were detected. Low levels of IgG4 specific to wheat, soy, peanut, and egg were also detected. Both MBB and STB are able to harvest measureable levels of IgE and IgG4 from the esophageal mucosa. Low levels of serum-specific IgE suggest that other inflammatory mechanisms, besides type I, IgE-mediated, allergen-specific hypersensitivity, may act as the primary catalyst for mucosal eosinophilia. Clarifying the role of both IgE-mediated and non-IgE-mediated inflammatory mechanisms will help identify more targeted diagnostic and treatment strategies for individuals who present with dysphagia and esophageal eosinophilia.

DNA cytometry of exfoliated cells in the diagnosis of oral potential malignant disorders

To evaluate the diagnostic efficiency of oral mucosa disease, especially oral squamous cell carcinoma (OSCC) and oral potential malignant disorders (OPMDs) by DNA cytometry compared with histopathological diagnosis, so as to find a convenient, simple and low-invasive method for screening and follow-up. 203 subjects with OSCC, OPMDs and other oral mucosa disease without dysplasia according to the inclusion criteria and exclusion criteria were recruited from Peking University School and Hospital of Stomatology. The mean age was (52.44±13.55) years, 98 males and 105 females. Brush biopsy was taken before scalpel biopsy at the same site. The brush biopsy sample was screened by moticytometer system for DNA cytometry after Feulgen stain, and histopathological examination were taken for the scalpel tissue. Data from DNA cytometry were used to calculate the parameters, such as sensitivity, specificity, positive and negative predictive values, odds ratios, Youden index (YI), positive and negative likelihood ratios, compared with the golden standard, histopathological diagnosis. DNA cytometry and histopathological diagnosis were performed back to back. Totally, 42 OSCC and 4 tumor in situ (TIS), 39 oral leukoplakia (OLK) with dysplasia (17 mild dysplasia, 13 medium dysplasia and 9 severe dysplasia), 29 OLK with hyperplasia, 1 verrucous OLK, 83 oral lichen planus (OLP) and 5 inflammation were included in our research. We grouped the OSCC, TIS and dysplasia as the positive group and others without dysplasia as the negative group, the sensitivity of DNA cytometry was 79.07%, the specificity was 81.20%, and the diagnostic accuracy was 80.30%,We grouped the OSCC and TIS as the tumor group, OLP, OLK with hyperplasia and inflammation as the non-tumor group, The sensitivity of DNA cytometry in diagnosing OSCC and TIS was 95.65%, and the specificity was 81.2%, The diagnostic accuracy was 85.28%. positive predictive values 66.67%, negative predictive values 97.94%, ratio odds 95, positive likelihood ratio 5.09, negative likelihood ratio 0.05, and Youden index 0.77. For the dysplasia, we grouped the different dysplasia together as the dyaplasia group, OLP, OLK with hyperplasia and inflammation as the non-tumor group, the sensitivity of DNA cytometry in diagnosing dyaplasia is 60%, the specificity is 81.2%. The diagnostic accuracy is 75.8%, positive predictive values 52.17%, negative predictive values 85.59%, ratio odds 6.48, positive likelihood ratio 3.19, negative likelihood ratio 0.49, and Youden index 0.41. DNA cytometry is convenient and low-invasive, which can be used as an adjuvant method for screening the early OSCC and OPMDs, monitoring the prognosis of OSCC after surgery. Further large-scale and long period prospective studies are necessary to validate the better value of DNA cytometry.

Comparison of oral brush cytology and tissue biopsy in diagnosing oral lesions

Background: Oral cancer involves cancer of the lips, tongue, floor of mouth, cheeks, soft and hard palate and pharynx. It is the 6th most common cancer in Western world and second most common cancer in some parts of sub-continent . Oral cancers are often diagnosed at a late stage because oral lesions are usually considered benign and clinical examination is not accompanied by specific diagnostic tests. Brush biopsy is a simple and easy test to perform on suspicious lesions before they undergo biopsy.

Main findings and diagnostic yield of bronchoalveolar lavage, bronchial brushing and transbronchial biopsy in HIV-positive patients.

ABSTRACTBronchoalveolar lavage, bronchial brushing and transbronchial biopsy are of fundamental importance in the diagnosis of pathologies affecting the lungs of immunosuppressed patients, especially those infected with HIV. This was a descriptive and retrospective study, in which the results of bronchoalveolar lavages, bronchial brushings and transbronchial biopsies of HIV-positive patients attended at the Clinical Hospital of the Federal University of Triangulo Mineiro from 1999 to 2015 were reviewed to determine the most frequent findings in these patients, to evaluate the diagnostic accuracy of these procedures and to correlate bronchoscopy results with clinical and radiological findings. Serological tests for HIV were confirmed and cases with negative or unverified serology were excluded. Medical records were reviewed for correlation with clinical and radiological findings. A total of 1,423 patients with a mean age of 50 years were initially selected; 727 cases had no serology for HIV; 696 had serology for HIV and 64 were positive. Of these, 47 were men, aged 24 to 84 years, and 17 women, aged 31 to 69 years. Biopsies and cytological tests were positive in 20 (31.25%) of the 64 patients and the most frequent diagnosis was pneumocystosis, found in 8 cases (12.5%). Of the 20 bronchofibroscopy-positive patients, only 2 did not show agreement between histopathological and clinical-radiological diagnoses. The analysis of the cytological tests and biopsy specimens obtained by bronchofibroscopy seems to be valuable for the etiological diagnosis of pulmonary infections in HIV- positive patients; however, negative results do not always exclude the diagnosis. In these cases, clinical symptoms and imaging findings may help to guide the best therapy.

Evaluation of Cytomorphologic Changes of Oral Mucosa and Copper Level of Saliva in Occupationally Copper Exposed, Oral Squamous Cell Carcinoma and Normal Population

Introduction: Oral Squamous Cell Carcinoma (SCC) involves half of the oral cavity malignancies. It is very important to find the etiologic factors of oral SCC and its early diagnosis. Copper is one of the most useful metals that have been used by men since ancient time. The level of copper increases when the cancerous changes occur in the body, saliva analysis can be used to discover various illnesses because of its non-invasive methods. In addition to biopsy, an alternative method for examining oral lesions is exfoliative cytology. The aim of this study was to determine the salivary copper level and cytomorphologic changes of oral mucosa among three study groups.Materials and Methods: In this cross-sectional study, 15 individuals with oral SCC, 15 workers exposed to copper and 15 healthy individuals were included. Saliva samples were collected and analyzed by Atomic Absorption Spectrophotometer. The exfoliative smears were prepared by brush biopsy and stained by Papanicolaou and AgNOR staining methods. Data were analyzed by One-way ANOVA and Kruskal-Wallis test. P value <0.05 was considered significant.Result: The results showed a significant difference in mean salivary copper (P=0.008), cytomorphology of oral mucosa and AgNOR among the three groups (P<0.001).Conclusion: Occupational exposure to copper increases the salivary levels of this element and causes changes in mucosal cells. Since this increase in is very high salivary level of copper and nuclear activity are in oral SCC patients, exposure to copper should be considered an important risk factor for oral mucosal changes.

Brush Biopsy Sample Quality: Preliminary Investigation of a Metal Brush Prototype

Objective: To qualitatively investigate whether a prototype brush composed of metal bristles collects oral epithelial cells effectively for cytological evaluation of oral mucosal lesions. Material and Methods: Twenty patients with suspicious oral mucosal lesions were enrolled. Patients were asked to gargle with saline and to deposit the oral rinse into specimen cup. Then, oral mucosal cell samples were collected using a metal oral brush, via sweeping motion. Punch biopsy was performed for histological examination. All samples were evaluated with liquid based cytology (LBC) according to the cellularity, the depth of the epithelial layer, cellular integrity by an oral pathologist. Results: Oral rinse provided samples with 100% cellular integrity and cellularity, mostly from the intermediary layers. With metal brush, both inadequate cellularity and cellular integrity was observed in 25% of the cases. Cellular integrity was adequate in 65%, cellularity was adequate in 45% of the lesions. Samples were dominantly from the intermediary layers, but in one case, metal brush collected cells from the parabasal layer. Conclusion: The narrow spiral pitch and width of metal bristles may have resisted to release the cellular samples collected. With adjustment of the spiral pitch and diameter of metal brush bristles, its’ efficacy could be enhanced.

Combination of Cytopathology and DNA Ploidy Increases the Performance of Oral Epithelial Dysplasia Prediction in Oral Potentially Malignant Disorders

Background: Grading of oral epithelial dysplasia (OED) by a pathologist is currently the key guide used for treatment planning of oral potentially malignant disorders (OPMDs). Conventional oral examination (COE) clinically detects OPMDs but may not predict their risk status to transform to cancer. Therefore, there is a need for a reliable test to predict OED in OPMDs. Aim: This study was conducted to evaluate COE, liquid based cytology (Cytopath) and DNA image cytometry (Ploidy) in predicting OED in OPMDs. Methods: A total of 179 patients from Malaysia, India and Indonesia underwent COE followed by brush biopsies and scalpel biopsies. Brush-biopsy samples were analyzed for cytopathology and DNA ploidy at Dental Faculty, University of Malaya. Histopathological findings of presence/absence of OED were used as the reference standard. Calculations for sensitivity (Sn), specificity (Sp), positive predictive value (PPV), negative predictive value (NPV) and accuracy (A) were done for individual tools and in combinations. The Youden index (Sn+Sp-1) was used as a measure of overall performance. The relevant medical ethics committees of the different research locations approved the study. Results: For COE, the sensitivity (Sn) was high (100%) and the specificity (Sp) was low (5.9%), while both Cytopath and Ploidy showed a low sensitivity (Sn) (28.6% and 22.2%) and high specificity (Sp) (94.3% and 82.3%). All 3 tools individually have high negative predictive value (NPV) for predicting presence of OED (COE-100%, Cytopath-66.7%, Ploidy-78.5%). When combining outcomes from all 3 tools, the best performance indicated by Youden index (42.1) is which defines a positive case when both COE and Cytopath show abnormal. In general, using results from at least 2 tools had better Youden indices than using these tools individually. Conclusion: COE as a screening tool by virtue of its high Sn would be a suitable first level diagnostic test, while the Cytopath and the Ploidy individually with high Sp may be used as a second level test to predict presence of OED. Combining the COE with cytopathology would be the best combination for a high performance of the tools. Cytopathology (when performed by a trained cytologist) would allow for most of the false positives from the first level test to be correctly identified as true negative at the second level. Longitudinal data are needed to assess which of these may correctly identify the malignant potential of OPMDs. Acknowledgment: Grant: High Impact Research - Ministry of Higher Education (HIR-MOHE UM000025/C3)

Sarcomatoid Carcinoma of the Pancreas: An Unusual Cause of Abdominal Pain

Introduction: Sarcomatoid carcinoma of the pancreas (SCP) is a rare and highly malignant variant of pancreas carcinoma that usually has a very poor prognosis. Epithelial-mesenchymal transition (EMT), a transition between different phenotypes, is regarded as an essential process in tumor progression. When EMT occurs, epithelial tumor cells lose their restricted phenotypes, concomitant with the loss of epithelial characteristics and the acquisition of motile behavior. These changes, which contribute to the transition from an epithelial morphology towards a more mesenchymal fibroblastic phenotype, have been reported to occur in some special tumors, such as cholangiocarcinomas; this change is also defined as sarcomatous change. Case Report: A 37-year-old-female with a history of hypertension presented to the hospital with the chief complaint of epigastric abdominal pain. Her abdominal pain progressively worsened and she came back to the hospital 3 weeks later. She also complained of pale, foul-smelling, floating stools that began 3 days prior to presentation with a decrease in her appetite. Routine laboratory showed: hemoglobin, 12.8 g/dL; WBC, 5,100 cells/uL; platelet, 244,000/mm3; with elevation in AST, ALT, alkaline phosphatase; total bilirubin, 4.0 mg/dL; direct bilirubin, 3.1 mg/dL; and LDH 306 u/L and negative viral markers. Tumor marker assays showed elevated CA19-9. Results: CT abdominal scan showed a hypoechoic mass in the uncinate process of the pancreas (Figure 1) with intrahepatic biliary ductal dilatation and dilation of the proximal common bile duct (CBD). The patient underwent an esophagogastroduodenoscopy with brush biopsies of the CBD with stent placement, as well as, an endoscopic ultrasound fine needle aspiration of the pancreatic mass. The patient underwent a pancreaticoduodenectomy to help diagnosis and remove the tumor. The histopathological diagnosis was as follows: ductal adenocarcinoma with sarcomatoid features with the invasion of the tumor in the extra-pancreatic common bile duct, stage IIB (Figure 2A & 2 B).1372_A.tif Figure 1: CT abdomen with contrast showing the hypoechoic mass in the head of the pancreas with intrahepatic biliary ductal dilatation.1372_B.tif Figure 2: Pathology specimen showing adenosquamous carcinomaDiscussion: Adenosquamous carcinoma of the pancreas is associated with high rates of recurrence and metastasis and carries a poor prognosis, much worse than even that of pancreatic adenocarcinoma, which has a reported median overall survival of no more than 5 months. SCP is a rarely appreciated subset of pancreatic malignancy that does not necessarily portend to a uniformly dismal prognosis.1372_C.tif Figure 3: Sarcomatous changes.

Synchronous Presentation of Three Distinct Pancreaticobiliary Tumors: Pancreatic Ductal Adenocarcinoma, Cholangiocarcinoma, and a Pancreatic Neuroendocrine Tumor

Introduction: Pancreatic ductal adenocarcinoma (PDAC) accounts for 3% of all cancers in the United States though it comprises approximately 85% of pancreatic cancers. Less common pancreaticobiliary tumors include cholangiocarcinoma and pancreatic neuroendocrine tumors, which have an incidence of approximately 8,000 and 1,000 cases annually in the United States, respectively. Case Presentation: We present a case of a 71 year-old female with a previous history of a cholecystectomy initially presenting with a suspicious mass in the body of the pancreas. CT imaging with pancreas protocol demonstrated a pancreatic neck mass that on EUS biopsy showed pancreatic adenocarcinoma. The CT also showed a second, ill-defined lesion near the common bile duct (CBD) that on ERCP was proximal to the cystic duct remnant. Brush biopsies demonstrated high-grade dysplasia. The patient was treated surgically with a total pancreatectomy including proximal resection of the CBD and portal lymphadenectomy. Her post-operative course was uneventful. Pathology demonstrated a poorly-differentiated PDAC of the neck (T3N1) and a moderately-differentiated cholangiocarcinoma arising from an intraductal papillary neoplasm of bile duct (T2N0). Incidentally, the removed pancreas also demonstrated a sub-centimeter, well-differentiated neuroendocrine tumor. Discussion: Synchronous primary tumors of the pancreas occur in up to 20% of cases, typically involving pancreatic ductal adenocarcinoma and intrapapillary mucinous neoplasms. Synchronous pancreaticobiliary cancer is much less common. After a literature review, we identified fewer than ten reported cases of synchronous pancreatic cancer and gallbladder/bile duct cancer and no prior case reports from patients in the United States. This case provides a unique synchronous presentation of three distinct neoplasms within the pancreaticobiliary region. Although this presentation is extremely rare, it should be considered when assessing multiple pancreaticobiliary lesions.1452.tif Figure 1: CT imaging of the abdomen with contrast using a three-phase pancreas protocol approach. Red circle demonstrates a 1.9 x 1.3 cm mass in the common biliary duct at the level of the upper pancreatic head. Yellow circle demonstrates a 2.9 x 1.9 cm mass in the body of the pancreas.

Cytological and cytometric analysis of oral mucosa in patients with Alzheimer's and Parkinson's disease.

BackgroundAlzheimer’s disease (AD) and Parkinson’s disease (PD) are the two most common neurodegenerative diseases. Recent studies have sought to identify precursor symptoms of AD and PD that occur before the onset of the disease. We evaluated changes in the oral mucosa of patients with AD and PD using a stereological method.Patients and methodsThe study included 29 patients with AD, 30 patients with idiopathic PD, and 30 healthy volunteers. Brush biopsies were obtained from all participants, and the nucleator method was used to estimate the volume of cells obtained from the buccal mucosa.ResultsCytomorphometric analysis revealed that the nuclear volume was 484.39±117.10 µm3 in the AD group, 509.71±132.26 µm3 in PD patients, and 509.30±100.21 µm3 in the control group. The cytoplasmic volume was 115,456.60±30,664.98 µm3 in the AD group, 103,097.93±25,034.65 µm3 in PD patients, and 109,528.45±28,381.43 µm3 in the control group. The nuclear and cytoplasmic volumes were not significantly different among groups (P>0.05).ConclusionThe cytomorphometric analysis revealed no significant differences in the cytoplasmic and nuclear volumes of buccal cells obtained from patients with AD and PD and healthy volunteers.

Abstract 2222: Using rare variants to characterize lung cancer risk

Abstract The human genome is exposed to a variety of exogenous and endogenous assaults capable of inducing DNA damage and contributing to diseases like cancer. Multiple DNA repair mechanisms are tasked with identifying and repairing these aberrant nucleotides, but damage is occasionally missed giving rise to somatic mutations. As we age, the relative abundance of somatic mutations increases, which may increase our risk for developing cancer. In addition, inter-individual variation in somatic mutation prevalence suggests an inter-individual variation in (a) inherited DNA repair capacity and/or (b) exposure to environmental mutagens. Therefore, somatic mutation prevalence may serve as an end-point biomarker for cancer risk, as it provides critical information regarding both. While Next Generation Sequencing (NGS) has provided unparalleled access to the human genome and revolutionized the field of cancer genomics, its implementation into the clinical setting has been limited by technical errors introduced during library preparation and on the sequencing platform. This makes identification of rare variants below 10-4 mutations per nucleotide through NGS a challenge, and the use of somatic mutations to characterize cancer risk unfeasible. To address this issue, unique molecular indexes (UMI) were assigned to each genomic copy, enabling reliable identification of rare somatic mutations versus technical errors. We hypothesize that measurement of somatic mutation prevalence in normal bronchial epithelial cells (NBEC) will serve as an end-point biomarker for lung cancer risk by identifying both (a) inherited sub-optimal DNA repair/protection and (b) exposure to inhaled environmental mutagens including components of cigarette smoke, radon, and/or occupational hazards. In our initial pilot study, we used genomic DNA derived from an A549 cell line to optimize an ERCC5 gene-specific, dual-indexed PCR method for UMI assignment, followed by sequencing on the Illumina MiSeq platform. In parallel analyses, UMI correction using 25,000 genomic copies provided over 10-fold greater sensitivity in identifying somatic mutations compared to no UMI correction. Based on this pilot, we obtained three clinical NBEC specimens via cytology brush biopsy of grossly normal (non-cancerous) airway from (a) heavy smoker with lung cancer, (b) heavy smoker without lung cancer, and (c) non-smoking control. UMIs were assigned to 50,000 genomic copies at 16 loci, increasing our theoretical limit of detection to approximately 1 in 4x107 nucleotides. Results thus far support further application of this approach in studies to assess NBEC somatic mutation prevalence as a biomarker for cancer risk. Citation Format: Daniel J. Craig, Mazzin Elsamaloty, Thomas M. Blomquist, Erin L. Crawford, James C. Willey. Using rare variants to characterize lung cancer risk [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2222.

Chronic E-Cigarette Exposure Alters the Human Bronchial Epithelial Proteome.

E-cigarettes vaporize propylene glycol/vegetable glycerin (PG/VG), nicotine, and flavorings. However, the long-term health effects of exposing lungs to vaped e-liquids are unknown. To determine the effects of chronic vaping on pulmonary epithelia. We performed research bronchoscopies on healthy nonsmokers, cigarette smokers, and e-cigarette users (vapers) and obtained bronchial brush biopsies and lavage samples from these subjects for proteomic investigation. We further employed in vitro and murine exposure models to support our human findings. Visual inspection by bronchoscopy revealed that vaper airways appeared friable and erythematous. Epithelial cells from biopsy samples revealed approximately 300 proteins that were differentially expressed in smoker and vaper airways, with only 78 proteins being commonly altered in both groups and 113 uniquely altered in vapers. For example, CYP1B1 (cytochrome P450 family 1 subfamily B member 1), MUC5AC (mucin 5 AC), and MUC4 levels were increased in vapers. Aerosolized PG/VG alone significantly increased MUC5AC protein in human airway epithelial cultures and in murine nasal epithelia in vivo. We also found that e-liquids rapidly entered cells and that PG/VG reduced membrane fluidity and impaired protein diffusion. We conclude that chronic vaping exerts marked biological effects on the lung and that these effects may in part be mediated by the PG/VG base. These changes are likely not harmless and may have clinical implications for the development of chronic lung disease. Further studies will be required to determine the full extent of vaping on the lung.