bluuebunny/crossref_metadata_2025_split huggingface dataset:
Antioxidants and oxidative stress in patients with Li-Fraumeni-like syndrome and TP53 p.R337H mutation carriers.
Authors: Macedo Gabriel de Souza , Giacomazzi Juliana , Netto Cristina , Motta Leonardo , Klamt Fábio , Ashton-Prolla Patricia 
Published: March 2020

e21042 Background: Germline mutations in the TP53 gene are the underlying genetic defect of Li-Fraumeni Syndrome (LFS) and its variant, Li-Fraumeni-like (LFL) Syndrome, autosomal dominant disorders characterized by predisposition to multiple early-onset cancers. More recently, p53 is emerging as an important player in redox metabolism and important enzymes involved in defenses against oxidative stress have shown to be regulated by p53, including glutathione peroxidase 1 (GPX1) and mitochondrial superoxide dismutase 2 (SOD2). The aim of the present study was to investigate the redox profile parameters in blood of p.R337H mutation carriers and non-carriers individuals. Methods: A total of 34 individuals were included in the study and they were divided in two groups: mutation carriers (n=17) and non-carriers (n=17). Antioxidant enzyme activities (SOD, CAT, GPx) and oxidative stress parameters (Protein carbonyl content, Sulfhydryl content and TBARS) were measured in plasma, erythrocytes, leukocytes and serum. Results: Erythrocyte SOD and GPx activities, two first-line players in enzimatic antioxidant defense, differed significantly between mutation carriers and non-carriers, with an increased activity in the latter (p>0,05). Plasma Carbonyl content, an indicative of protein damage related to ROS overgeneration, was also increased in carriers (p=0,015). There was no significant difference between groups in all other parameters evaluated. Conclusions: Our findings suggest that TP53 p.R337H mutation carriers present different antioxidant enzyme activities and oxidative stress parameters when compared to non-carriers.
Effect of GDC-0449, a hedgehog (Hh) pathway inhibitor, on tumor activity and on erlotinib and cisplatin in a xenograft model of non-small cell lung cancer (NSCLC) with activated Hh pathway.
Authors: Ahmad Aamir , Sarkar Fazlul H. , Maitah Main , Ali Shadan , Sethi Seema , Aboukameel Amro , Gadgeel Shirish M. 
Published: March 2020

e21057 Background: We have previously shown that chronic exposure of A549 NSCLC cell line to TGF-β1 can induce epithelial-mesenchymal transition (EMT) and can make these cell lines (A549-M) resistant to erlotinib, an EGFR inhibitor and cisplatin. EMT in A549-M cells was associated with significant activation of signaling through the Hh pathway. We therefore evaluated the activity of GDC-0449, an inhibitor of the Hh pathway, and assessed its activity in combination with erlotinib and cisplatin in A549-M cells both in vitro and in vivo. Methods: We assessed the effects of GDC-0449 (20 nM) on clonogenic growth (colony formation assay), cell motility (wound healing assay) and invasion (matrigel assay) in A549-M cells, and also assessed the activity of GDC-0449 in combination with erlotinib and cisplatin both in vitro (MTT assay) and in an experimental lung metastasis animal model in vivo by injecting A549-M cells through tail vein. Results: A549-M cells showed significantly higher rate of clonogenic growth, cell motility and invasion in vitro and a greater propensity to form metastases in vivo compared to parental A549 cells, and also showed higher resistance to erlotinib and cisplatin. A549-M cells showed higher mRNA levels of Shh (sonic hedgehog, a ligand of the Hh pathway) and ZEB1 and reduced levels of E-cadherin than A549 cells. GDC-0449 significantly reduced growth of A-549-M cell in vitro but had no effect on A-549 cells. GDC-0449 also reduced clonogenic growth, cell motility and invasion of A549-M cells. Pre-treatment with GDC-0449 significantly enhanced the activity of erlotinib (5 and 10 mM) and cisplatin (1mM-10mM) in A549-M cells (p < 0.05) but not in A-549 cells. GDC-0449 reduced the number of lung metastases in the xenograft model and enhanced the activity of erlotinib and cisplatin in this model. Conclusions: This data suggests that the EMT phenotype in NSCLC may be associated with the activation of the Hh signaling pathway. It also suggests that GDC-0449 has anti-tumor activity against NSCLC with activated Hh pathway, and can enhance the effects of erlotinib and cisplatin in such tumors.
Vysis ALK non-small cell lung cancer assay performance on archival tissue and cytology specimens.
Authors: Li Gu , Jacobson Kristine B , Matushek Paul E , Perez Thomas B , Herman Christine , Sachs Karen L , Pestova Ekaterina 
Published: March 2020

e21060 Background: Rearrangements of ALK gene on chromosome 2p23 create a constitutively activated tyrosine kinase implicated in the development of NSCLC and targeted by novel therapeutic agents. The Vysis ALK Break Apart (BAP) FISH Probe assay was developed to detect chromosome 2p23 rearrangements in NSCLC FFPE tissue by fluorescence in situ hybridization. Performance of the ALK assay was assessed on FFPE tissue specimens and FFPE cell lines. The ALK probe was also tested on lung cytology cell suspensions. Methods: The ALK FISH assay success rate on primary NSCLC was tested with 30 FFPE tumor resection specimens, four 5-μm section slides each, judged by qualitative rating of fluorescent signal specificity, intensity, and background. Assay performance was further assessed on 15 FFPE biopsy specimens of 4 metastatic sites including mediastinal and axillary lymp nodes, pleural cavity, brain, and adrenal gland. Each specimen was tested in duplicate and evaluated by 2 reviewers. Assay reproducibility on FFPE cell pellets was tested with 3 lots of ALK Negative and ALK Positive Control Slides (cell lines) on 5 non-consecutive days. Slides were enumerated by 3 reviewers to determine % ALK positive cells, to a total of 45 results per control slide type. Additionally, the ALK BAP probe was tested with a modified assay protocol on ThinPrep lung cytology samples from 8 specimens. Results: The assay success rate on FFPE resection specimens was 99.2 % (90% CI 96.1% - 100%). The core biopsy specimens yielded 95% overall success rate. All Control Slides (FFPE cell lines) tested in the reproducibility study were enumerable within assay ranges, and were correctly classified per specifications of 0-8% and ≥20% ALK positive cells in ALK Negative and ALK Positive Control Slides, respectively. All of the lung cytology specimens prepared by ThinPrep hybridized successfully, however 2 specimens had low cellularity for ALK enumeration. Conclusions: The Vysis ALK BAP FISH assay demonstrated reproducible performance on FFPE cell lines and was shown to yield interpretable results on FFPE lung tissue specimens retrieved by resection or biopsy, from primary tumor or metastatic sites. Feasibility using lung cytology cell suspensions was demonstrated.
Ribonucleotide reductase subunit 2 (RRM2) to predict shorter survival in resected stage I-III non-small cell lung cancer (NSCLC) patients.
Authors: Rijavec Erika , Dal Bello Maria Giovanna , Savarino Graziana , Sini Claudio , Barletta Giulia , Genova Carlo , Truini Mauro , Pronzato Paolo , Merlo Domenico Franco , Pfeffer Ulrich , Grossi Francesco 
Published: March 2020

e21065 Background: Biomarkers can help in identifying patients (pts) with early-stage NSCLC with high risk of relapse and poor prognosis. The aim of this study was to investigate the relationship between the levels of expression of 7 biomarkers, various clinicopathological characteristics and their prognostic significance. Methods: Tumor tissue from 82 radically resected stage I-III NSCLC pts were consecutively collected to investigate the mRNA expression and protein levels of the following biomarkers using quantitative reverse transcriptase real-time PCR (qRT-PCR) and immunohistochemistry (IHC) with a tissue microarray technique: excision repair cross-complementation group 1 (ERCC1), breast cancer 1 (BRCA1), ribonucleotide reductase subunit 1 (RRM1), RRM2, p53R2, thymidylate synthase (TS) and class III beta-tubulin (β-Tub-III). Results: On a univariate analysis, p53R2 expression was significantly higher in adenocarcinoma (ADK) compared to squamous cell carcinoma (SSC) samples (p=0.002) and in stage I compared to stage II-III (p≤0.001). ERCC1 expression was significantly higher in females compared to males (p=0.03), and β-Tub-III expression was significantly higher in ADK than in SSC (p=0.03). Pts with lower RRM2 expression survived longer than pts with higher RRM2 expression (p=0.069). The multivariate analysis confirmed RRM2 as an independent prognostic marker of shorter survival (p= 0.031). The comparison between survival curves with qRT-PCR and IHC showed similar results with a trend towards longer survival among ERCC1 negative pts, BRCA1 negative pts, p53R2 positive pts and among pts with low levels of RRM1 and RRM2, although the difference was not statistically significant with both methods. qRT-PCR and IHC have shown that β-Tub-III and TS had no significant impact on survival. Conclusions: This is the first study that identifies RRM2 expression as a negative prognostic factor in resected stage I-III NSCLC. Moreover, we have demonstrated the differential expression of p53R2 and β-Tub-III in ADK compared to SSC and higher expression of p53R2 in pts with stage I compared to stage II-III NSCLC.
Relationship of Ki67 to tumor size and lymph node metastasis in breast cancer.
Authors: Bonta Ioana , Bonta Dacian , Loch Michelle Marie , Eapen Ann , Blanchard Rita A. 
Published: March 2020

e21076 Background: Ki67, a tumor proliferation marker, has demonstrated usefulness in breast cancer prognosis. Prior work with BrdU labeling for cell proliferation in breast cancer has not settled the question whether cell proliferation labeling is independent of other tumor features like tumor size and presence of axillary metastases, see Rew (1992) vs. Thor et al (1999). Methods: We analyzed retrospectively our database of 379 patients for correlation between the and tumor size, presence of axillary lymph node metastases and the percentage of Ki67 positve cells. We used linear and parabolic regression to correlate tumor size with the Ki67 index and receiver operator characteristics curve to correlate the presence of axillary lymph node metastases with the Ki67 index. Results: A very weak linear relationship was detected between tumor size and Ki67 index. The R-squared coefficient was 0.03, indicating that tumor size explained only 3% of the variability in measured Ki67 indexes. The ROC analysis, looking at the correlation between Ki67 and lymph node metastasis, yielded an area under the curve (Az) of 0.53. This indicates a very weak correlation. No relationship between axillary lymph node metastasis would yield an Az of 0.5 and a perfect correlation would yield an Az of 1. Conclusions: Tumor size and axillary lymph node metastasis explain less than 10% of observed Ki67 index variability. Therefore, in breast cancer, the prognostic value of Ki67 is additive to that of tumor size and lymph node involvement.
Prognostic significance of circulating tumor cell (CTC) levels at various times during first- and second-line chemotherapy for metastatic breast cancer (MBC).
Authors: Arteta-Bulos Rafael , Miller Paul J. E. , Overbury Rebecca S. , Schwartzberg Alison , Walker Mark S. , Schwartzberg Lee Steven 
Published: March 2020

e21077 Background: CTCs have prognostic capability in the treatment of MBC based on a dichotomous level of ≥5/7.5 ml blood (Hi) vs. <5/7.5 ml blood (Lo) at baseline and first follow-up. The value of CTCs beyond baseline is not well established, and the implication of values that are reduced but remain high has not been explored. Methods: Chart review of patients (pts) on 1st or 2nd line treatment of MBC, with CTC determinations between 2/2008 and 9/2011, were retrospectively identified at a large community oncology practice. Pts with ≥2 CTCs or 1 CTC ≥5 were eligible. Demographics, treatment regimens and progression-free survival (PFS) were assessed. The Veridex Cell Search system was utilized for all CTC determinations. Qualifying baseline CTCs were those drawn -14 days to +30 days from start of treatment. Results: 221 eligible pts were identified, 71 with baseline CTCs in 1st line, and 84 in 2nd line. Median age was 59.5 (range 26-90), 40.7% were African-American, 17.6% HER2+, and 63%/49% ER/PR+. The median time to baseline CTC was 5 days in the 1st line and 1 day in the 2nd line. 1st line PFS was 8.0 vs. 5.8 mo among baseline Hi vs. Lo groups (p = 0.085). 2nd line PFS was 3.7 vs. 4.0 mo for Hi and Lo groups, respectively, (ns). Follow-up CTCs were obtained at clinical convenience. A 90 day landmark was used for analysis of PFS by four baseline/follow-up CTC pattern groups. 1st line PFS showed medians ranging from 6.0 mo (Hi/Hi) to 10.2 mo (Lo/Lo). In 1st line pts with Hi baseline CTCs (n=24), median PFS was 6.0 vs. 7.0 mo for Hi vs. Lo at follow-up (p = .64). Grouping pts with Hi baseline CTCs by reductions in absolute CTC count at follow-up (<33% vs. ≥33%) showed median PFS 3.5 mo for <33% reduction, 8.8 mo for ≥33% reduction (p = .0179). Sensitivity analyses with other cutoffs in reduction from 20-80% showed similar results. Conclusions: In the community setting, baseline and subsequent CTCs obtained at various times after baseline yield clinically useful prognostic information. Pts with positive baseline CTC levels experiencing a reduction of ≥33% exhibit significantly longer PFS, indicating that a simple high/low approach to the test can be modified to yield more prognostic value.
Pancreatic polypeptide as a biomarker for pancreatic neuroendocrine tumors.
Authors: Alistar Angela Tatiana , Moshier Erin L. , Kim Michelle , Warner Richard , Holcombe Randall F. 
Published: March 2020

e21109 Background: Pancreatic neuroendocrine tumors (PNETs) represent tumors that secrete numerous proteins that could be used as biomarkers. We present a retrospective analysis of patients with PNETs who have had Pancreatic Polypeptide (PP) testing in an attempt to better define PP producing tumors as an entity and the role of PP as a biomarker. To our knowledge, this is the first single center comprehensive review of Pancreatic Polypeptide producing tumors. Methods: A retrospective study of patients with PNETs seen at Mount Sinai from 1980 to 2011. All patients that have had PP concentrations measured at least once were evaluated. Data relating to diagnosis, pathology, surgery, liver directed therapies, chemotherapy and survival outcome were noted. 71 patients with PNETs fulfilled the inclusion criteria. The 71 patients formed 4 subgroups. The 4 subgroups were evaluated and compared for survival: all 71 PP tested patients, 30 patients with PP producing tumors, 8 patients with PPomas, 22 patients with PP producing tumors that were non PP-omas and 41 patients with PP non-producing tumors Results: We identified a trend towards better survival for PNETs patients with PP producing tumors vs. non- PP producing tumors (p=0.19). There was no correlation between survival and a diagnosis of PPoma in relation to other PP producing tumors (p= 0.59) or non-PP producing tumors (p=0.58). Conclusions: Pancreatic Polypeptide is a biomarker that is worth prospective investigation and a standardized assay. Our analysis investigating Pancreatic Polypeptide as a prognostic and or predictive biomarker reveals a trend towards showing these characteristics. Using a standardized test and investigating this biomarker prospectively could lead to the validation of Pancreatic Polypeptide as a biomarker.
Molecular analysis of pancreaticobiliary fine needle aspirate/duct brushing cytocentrifugation supernatant fluid.
Authors: Finkelstein Sydney D. , Schmidt C. Max , Ellsworth Eric M. , Stearns Valerie T. , Smith Dennis M. 
Published: March 2020

e21121 Background: Reliance on minimally invasive sampling techniques (fine needle aspiration (FNA), duct brushing) continues to increase for pancreaticobiliary (PB) pre-cancer/cancer diagnosis. Fundamental limitations include low cellularity, sampling variability, and indeterminate microscopic diagnosis. To address these issues, we developed a novel approach involving DNA mutational profiling of the soluble (or free) DNA in the cytocentrifugation supernatant fluid (supernatant) currently discarded during cytology preparation. Methods: 77 PB specimens underwent combined microscopic/molecular analysis using free DNA in cytology supernatant fluid. Pancreatic FNA and biliary cytobrushes had either surgical pathology or positive cytology as the gold standard. DNA was extracted from 2 mL of the supernatant and from corresponding microdissected cytology cells. Profiling included DNA quantity and quality, and analyses for KRAS mutations and 16 LOH (loss of heterozygosity) mutations. Results: The cell-free supernatant showed significantly higher levels of more intact amplifiable DNA than typical microdissection specimens. All non-neoplastic specimens (n=38) showed no false positive mutations. 37/39 malignant specimens showed mutations (sensitivity 95%, specificity 100%). Mutational profiles of the microdissected cells and the supernatant demonstrated high concordance. However, mutation detection was consistently superior in the supernatant fluid (mean 6.7 mutations) compared to the microdissected cells (mean 4.2). Mutation clonality was also significantly higher in the supernatant. Conclusions: Despite the lack of cells in many PB samples, the post-centrifugation supernatant fluid provides an abundant, robust, and highly representative source of DNA for molecular analysis. Of note, the greater extent and clonality of detected mutations in the supernatant fluid supports it being enriched with neoplastic DNA when cancer is present. This novel approach complements standard cytology practice by increasing sensitivity and specificity of cancer detection while reducing sampling variation.
Adequacy of biopsy samples for molecular testing in patients with non-small cell lung cancer.
Authors: Hrinczenko Borys , Subramonia-Iyer Subramony , Zhang Deng , Olsen Brian 
Published: March 2020

e21133 Background: Testing for a mutated EGFR gene has transformed the care of NSCLC. Although ASCO recommends EGFR mutation testing for cases of NSCLC, there have been no studies to assess adequacy of biopsy samples for molecular testing in non-trial settings. Methods: A retrospective chart review to identify patients diagnosed with NSCLC at our community hospital during 2010. A pathologist then reviewed the adequacy of archivedbiopsy material for EGFR mutation testing based on ASCO criteria (Eberhard, JCO 2008). Results: In 2010, 64 cases of NSCLC were diagnosed. Demographics:Thirty-six (56%) of the patients were male; median age was 69.5 years, 27 were current smokers, 33 were ex-smokers, median survival was 4.9 months. Histologic subtypes: squamous - 28 (44%), adenocarcinoma - 23 (36%), bronchoalveolar - 3 (5%), adeno with mixed bronchoalveolar - 2 (3%), large cell - 3 (5%), and large cell neuroendocrine - 2 (3%). Twenty-eight (44%) were considered adenocarcinoma. Staging: Stage I, II, III, IV - 18 (28%), 2 (3%), 13 (20%), 23 (36%). Eight (13%) had incomplete staging information. Biopsy material: transbronchial FNA in 23 (36%), CT-guided FNA in 13 (20%), definitive surgery in 18 (28%), other - 10 patients (15%). Imaging review: 29 central lesions (45%) and 35 (55%) peripheral. Specimen review: 45/64 (70%) were adequate for EGFR mutation analysis; Twenty cases (30%) were not. Among adenocarcinoma, 19/28 (68%) cases had adequate samples, but 9 (32%) did not. Conclusions: In this series, nearly one-third of all NSCLC cases, including adenocarcinoma, did not yield sufficient specimen from the first biopsy for EGFR mutation analysis. Newer molecular markers (EGFR, EML4-ALK) with targeted therapy such as erlotinib and crizotinib are available in NSCLC so adequate tissue sampling is imperative. Lack of satisfactory tumor tissue sampling may preclude mutation testing thereby denying these patients treatment with potential benefit and lower toxicity. This has implications for planned diagnostic investigations since an appropriate site must be biopsied so as to obtain an ample sample. Efforts to improve clinician awareness about adequate tumor sampling are needed and should be a top priority for multidisciplinary lung cancer teams.
Breast cancer in a patient with multiple endocrine neoplasia type 1 (MEN 1): A case report and review of the literature.
Authors: Inic Zorka Momcilo , Inic Momcilo , Dzodic Radan , Pupic Gordana , Damjanovic Svetozar 
Published: March 2020

e21136 Background: We treated a patient with breast cancer associated with multiple endocrine neoplasia type 1 (MEN 1). There are only few studies of the development of breast cancer in patients diagnosed with MEN1 syndrome. Methods: We found that patient had a pituitary macroadenoma that secretes prolactin (prolactinomas), increased serum concentration of calcium, 4.09 mmol/l and increased serum concentration of PTH, 212 pg/ml. Neck ultrasound and scintigrafy showed suspicious adenoma in lower left parathyroid gland. A computed tomography scan of the abdomen revealed a tumor mass in the body of the pancreas and adenoma in the left suprarenal gland. We diagnosed multiple endocrine neoplasia type 1 (MEN 1). Results: MEN1 occurs as a result of inactivating mutations of the MEN1 gene (MEN1), located on chromosome11q13. Results indicate that menin is a direct activator of ERalpha function. In a clinical study, in Kagawa, with 65 ER-positive breast cancer samples-all of which had been treated with tamoxifen for 2-5 years as adjuvant therapies-menin-positive tumors had a worse outcome than menin-negative ones. This indicated that menin can function as a transcriptional regulator of ERalpha and is a possible predictive factor for tamoxifen resistance.These results demonstrate that allelic deletions of the 13q12-14 region occur in some pituitary adenomas and 16% of parathyroid adenomas. Conclusions: As recent molecular studies have suggested genetic mutations in multiple endocrine neoplasia type 1, this syndrome may possibly predispose patients to breast cancer.This possibility cannot be evaluated definitively on the basis of a single case report; additional observations and studies are necessary to investigate this hypothesis further.