bluuebunny/crossref_metadata_2025_split huggingface dataset:
Investigation of telomerase activity in inflammatory and noninflammatory breast cancer.
Authors: Salem Ayman Maher , El-Shinawi Mohamed , Sabet Salwa Farouk 
Published: March 2020

e21055 Background: Inflammatory breast cancer (IBC) is an aggressive type of breast cancer disease that has a high incidence in Egypt than western countries. It is characterized by rapid progression, involvement of dermal lymphatic emboli and extensive lymph node involvement. Basic and translational studies needed to define IBC disease biology and identify specific biomarkers have been limited by the paucity of patient samples. Hence, the current study aimed to introduce the telomerase activity level as a novel diagnostic marker for breast cancer and specifically for IBC to be differentiated from non-IBC. Methods: Breast cancer patients were enrolled from Ain Shams University hospitals in Cairo, divided into two groups: IBC (n=26) and non-IBC (n=27). Tissue samples were collected during modified radical mastectomy. TRAP (Telomerase repeat amplification protocol) assay was used to assess the telomerase activity in inflammatory and non inflammatory breast cancer tissue samples and their representative cell lines, SUM149 and MCF-7 respectively. Immunohistochemistry was used to investigate the expression of hTERT subunit of telomerase in paraffin embedded tissue samples of both types of patients. MTT assay was used to compare the effect of AZT (3’-azido-3’-deoxythymidine) on the growth of cell lines SUM149 and MCF-7. Results: : IBC showed Telomerase activity ranged from 12 to 376 units with a mean value of 75 and a median value of 43, while telomerase activity in non-IBC ranged from 6 to 109 units with a mean value of 35 and a median value of 24. On the other hand, normal tissues showed telomerase activity below 5 (P > 0.001). Using immunohistochemistry, the hTERT expression was higher in IBC than non-IBC and no expression at all in normal tissues. Also, a positive correlation was found between the telomerase activity and the number of metastatic lymph nodes in both IBC (r =0.511) and non-IBC (r =0.622). Besides, IBC cell line showed a faster response (53%) to telomerase inhibition by AZT after 24 hrs than non-IBC cell line (13%) at the same conditions. Conclusions: Telomerase could be a promising marker at the diagnostic and therapeutic levels in breast cancer and specifically in IBC.
Mitosis interference of cancer cells during anaphase by electric field from NovoTTF-100A: An update.
Authors: Lee Sze Xian , Tunkyi Adrian , Wong Eric , Swanson Kenneth D. 
Published: March 2020

e21078 Background: The NovoTTF-100A is a device that emits an alternating electric field (TTField) that kills mitotic cells and has been FDA-approved for the treatment of recurrent glioblastoma. Exposure to the TTField resulted in impaired cytokinesis and asymmetric chromosome segregation. To further elucidate its mechanism of action, we performed experiments using synchronized tumor cells and TTField-treated xenografts. Methods: Biochemical analysis of cell cycle-specific proteins, timelapse movies, and immunoflourescence microsopy were used to examine the effects of TTField on synchronized tumor cells. Xenografts treated with TTField were analyzed for the presence of phospho-Histone H3 and cell cycle-specific epitopes. Results: Cells exposed to TTField as they transited through M-phase exhibited persistence of phospho-Histone H3 while Securin levels decreased, suggesting that these cells transit through metaphase to anaphase but were delayed in mitotic exit. Immunoflourescence of treated cells showed evidence of impaired cytokinesis together with disordered separation of chromosomes from metaphase plates. Lagging chromosomes, dispersion of chromosomes, chromosomes decondensation in the absence of cytokinesis, and asymmetric chromosomal segregation were also observed in treated samples. The number of 4N cells increased following TTField exposure compared to parallel cultures that had entered the G1-phase of the cell cycle indicating a failure of mitosis. Exposed cells showed no detectable p53 induction indicating that cell death is achieved by a p53-independent mechanism. Lastly, immunostaining of treated xenografts for phospho-Histone H3 revealed the presence of significantly higher numbers of immunopositive mitotic figures compared to untreated samples. Conclusions: These data suggest that TTField exposure resulted in defective entry into anaphase and interfered with orderly exit from mitosis, a mechanism of mitotic perturbation that differs from currently used spindle poisons. NovoTTF-100A may therefore exhibit synergism when combined with conventional cancer treatments. Supported in part by a grant from NovoCure, Inc. and A Reason To Ride research fund.
Association of IL17-expressing γδ t cells with acute radiation-induced pneumonitis.
Authors: Liao Winston W. P. , Chao K. S. Clifford , Hei Tom K , Cheng Simon 
Published: March 2020

e21097 Background: Radiation pneumonitis is a substantial cause of morbidity and mortality with thoracic radiation for lung cancer. Little is known about the crucial mechanisms of the inflammatory response. We seek to determine if a key mediator of organ-specific inflammatory disorders and innate immune response, IL-17+ γδ T cells, is associated with radiation pneumonitis. Methods: C3HBe/FeJ mice (7 mice/group) were sham-irradiated as controls or exposed to a single dose of 15 Gy thoracic X-ray to develop pneumonitis. We have previously shown that TGFβ has an immunosuppressive activity in radiation pneumonitis. To potentiate the radiation pneumonitis, one group of mice was administered anti-TGFβ therapy with inhibitory TGFβ mAb (1D11, i.p.10 mg/kg/wk). Bronchoalveolar lavage fluid was assessed for cytology and inflammatory cytokine level. Lung tissues were examined for cell infiltration and histopathological changes. Cell surface marker and intracellular cytokine staining were performed on lymphocytes from the digested lungs by flow cytometry. Results: At 10 weeks post-irradiation, the lungs of the irradiated mice showed substantially more alveolar wall edema and increased infiltration of inflammatory cells compared with sham controls. Pneumonitis-involved lungs contained more IL-17+ γδ T cells (0.85% ± 0.00%) compared with sham controls (0.33% ± 0.02%), p<0.001. Furthermore increased IL-17+ γδ T cells were associated with potentiated radiation pneumonitis with anti-TGFβ therapy. There was a significant increased alveolar inflammation in irradiated mice injected with anti-TGFβ mAb. Anti-TGFβ irradiated lungs also contained significantly more IL-17+ γδ T cells (1.17% ± 0.13%) compared with irradiated controls (0.72% ± 0.13%), p<0.001. There was no increase of other TGFβ-dependent T cell subtypes such as IFNγ+ αβ T cells (Th1), IL-17+ αβ T cells (Th17), CD25+ Foxp3+ Tregs, nor activated macrophages in the potentiated pneumonitis lungs. Conclusions: Our findings implicate a novel role for IL17-expressing γδ T cells in radiation pneumonitis. This study reveals this innate immune response pathway as a potential target for therapeutic intervention in radiation lung injury
Invasive lobular (ILC) versus invasive ductal (IDC) breast cancer (BC): Clinical-pathologic features and clinical outcomes in monoinstitutional series.
Authors: Ferro Antonella , Eccher Claudio , Caldara Alessia , Triolo Renza , Barbareschi Mattia , Cuorvo Lucia Veronica , DiPasquale MariaChiara , Decarli Nicola L , Mussari Salvatore , Galligioni Enzo 
Published: March 2020

e21102 Background: ILC is less common than IDC and is usually diagnosed at a later stage Purpose:The aim of our study was to investigate different clinico-biological behavior associated to ILC compared to IDC and to evaluate implications on survival outcomes Methods: We analyzed data from 3749 consecutive cases of IBC treated from 1995 to 2008 and classified as ILC, IDC and mixed/other. Relationships with clinical-pathological variables and the impact of ILC/IDC types on Event Free Survival (EFS), Overall Survival (OS) and Post-Progression Survival (PPS) were analyzed. Results: We have identified 445 ILC BC (12%), 3021 IDC (80.5%), 149 mixed (3.9%) and 134 other histotypes (3.6%). Median age was 61 (25-97) years. ILC presented a significantly (p<0.001) larger tumor size (T≥2: 46% vs 34%) and more frequent axilla involvement (43% vs 37%) compared to IDC. It was usually associated with intermediate grade, low ki67, positive ER and negative HER2 status. Mastectomy was more frequently required for ILC (45%) than IDC (37%). Adjuvant hormonal (± previous chemo) therapy was more frequently given (77 vs 64%) to ILC pts due to higher ER expression than IDC. At a median follow up of 77 (0-272) months (ms), there were not significant differences in EFS (81.4 vs 82.1%, p=0.7) and OS (82.8 vs 84.6%; p=0.19) between ILC vs IDC. Local and Distant Relapses were 15 (3.3%) and 50 (11.2%) in ILC vs 179 (5.9%) and 225 (7.4%) in IDC; preferential sites of first distant relapses were similar in both histotypes. Contralateral and second tumors were 9 (2%) and 22 (5%) in ILC vs 84 (2.8%) and 174 (5.7%) in IDC. Median time to first event was 38.3 ms in ILC vs 35.23 in IDC. PPS was 16.5 ms (0-100) in ILC compared to 22.2 (0-187) in IDC. Prognosis in term of OS was consistently worse for ILC pts, compared to IDC, when considering the luminal A (86.9 vs 93.5%; p=0.003), luminal C (70.4 vs 88.5%; p=0.028) and in triple negative subgroups (50 vs 73.1%; p=0.021). No difference in OS was seen for age, size, hormonal and nodal status, except in N>3 ILC pts, in whom a trend of worse prognosis (55.6 vs 60.2%) was observed. Conclusions: ILC pts did not show in our series a better outcome than IDC pts, despite a quite favorable biological pattern.
EBUS-TBNA for identifying biomarkers.
Authors: Chen Kezhong , Yang Fan , Zhao Hui , Zhou Zu li , Jiang Guanchao , Wang Jun 
Published: March 2020

e21103 Background: We assessed the usefulness of endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) to detect EGFR AK-ras AALK mutations from malignant mediastinal nodes of patients with NSCLC. Methods: From February 2010 to December 2011 , 52 specimens diagnosed lung adenocarcinoma by EBUS were used for molecular analysis . Needle puncture site for the EBUS was standard 22G needle, once cytology pathology diagnosed adenocarcinoma, at the same site with the conventional needle puncture method to obtain adequate biopsy specimen for further analysis. The tissue was fixed in 10% formalin and stored in -80°C. When histologic diagnosis confirmed adenocarcinoma, the sample was extracted DNA to detect EGFR mutation by both amplification refractory mutation system( ARMS) and mutation enriched method improved by our department. Partial specimens also extracted DNA or RNA to detect K-ras and ALK mutation. Results: All 52 specimens were successfully extracted DNA, 34 were male and 18 were female, mean age of 62.1 years (34 to 79 years). Somatic mutations of the EGFR gene were detected in 25 of 52 cases (48%), 15 cases of exon 19 mutation, 9 cases of exon 21 mutation, and 1 of T790M mutation in ARMS. The result was of slight difference in mutation enriched method. In the patients who requested other biomarkers analysis, 4 had K-ras mutations and 2 had ALK mutations. 6 patients of 25 cases of patients with EGFR mutations had a radical surgery after neoadjuvant chemotherapy, tumor specimens were tested again with EGFR gene mutation, the same result as the preoperative test results. Conclusions: EBUS-TBNA mediastinal lymph node biopsy in the detection of EGFR AK-ras and ALK mutations in tissue samples was feasible, through the highly sensitive methods, higher detection rate was available, the result can be used as evidence of personalized treatment of lung cancer.
Comprehensive analysis of genetic polymorphisms and irinotecan-induced adverse events in Japanese gastrointestinal cancer patients: A DMET microarray profiling study.
Authors: Kuramochi Hidekazu , Kanno Hitoshi , Uchiyama Tomotaka , Nakajima Go , Saito Kayoko , Hayashi Kazuhiko 
Published: March 2020

e21108 Background: Irinotecan is a key drug in the treatment of colorectal and gastric cancer, that may occasionally cause severe adverse events (AEs), especially neutropenia and diarrhea. Although UDP-glucuronosyltransferase (UGT)1A1 polymorphisms are used as biomarkers for predicting AEs, the effect of UGT1A1 polymorphism in clinical use is limited, suggesting that there is a possibility of the existence of other, unknown biomarkers. Methods: Fourteen gastrointestinal cancer (5 gastric, 9 colorectal) patients who had undergone irinotecan-based chemotherapy were enrolled. DNA extracted from peripheral blood cells was genotyped by the DMET Plus microarray system, and 1,931 gene polymorphisms were investigated. The relationship between AEs and polymorphisms was analyzed statistically. Results: Eleven polymorphisms showed the P value < 0.05 with grade 3,4 neutropenia, but no statistically significant polymorphisms were found after the correction of multiple comparisons. With respect to the association with diarrhea, 12 polymorphisms showed P <0.05, and even after the correction of multiple comparisons, CYP2F1_96G>A(P32P) showed a significant relationship (P<0.00001). With respect to the relationship between UGT1A1*6, *28 polymorphisms and AEs, *6 polymorphism showed significant (P=0.044) association with grade 4 neutropenia. Conclusions: CYP2F1_96G>A showed significant association with diarrhea. The association of UGT1A1*6 polymorphism and neutropenia was confirmed, as in previous reports.
The association of plasma TGFβ1 during radiotherapy and genotypes of TGFβ1 pathway in patients with non-small cell lung cancer.
Authors: Yuan Shuanghu , Yu Jinming , Cai Xuwei , Kong Feng-Ming (Spring) 
Published: March 2020

e21115 Background: Radiation induced thoracic toxicities (RITT) in lung, esophagus and pericardium are dose-limited toxicities in patients with non-small cell lung cancer (NSCLC). Plasma transforming growth factor-beta1 (TGFβ1) and its change during radiotherapy was reported as a biomarker and single nucleotide polymorphisms (SNP) in TGFβ1 pathway was associated with RITT in our previous studies. In order to explore the mechanism of RITT, we tested whether plasma TGFβ1 was associated with genotypes of TGFβ1, tissue plasminogen activator (tPA) and angiotensin converting enzyme (ACE). Methods: Patients with stage I-III NSCLC enrolled in prospective clinical trials were eligible. All patients received radiotherapy with or without concurrent chemotherapy. Platelet-poor plasma was obtained pre-RT and at 4–5 weeks (40–50 Gy) during RT. Plasma TGF-β1 was measured using an enzyme-linked immunosorbent assay. The DNA samples extracted from blood before treatment were analyzed for the following genetic variations: TGFβ1 509C/T, TPA -7351 C/T, and ACE I/D. Results: 76 NSCLC patients received definitive radiotherapy (median dose 66 Gy) were enrolled. For the entire group of patients, the mean pre-RT TGFß1 level was 10.7±2.3 ng/ml, and the mean during-RT TGFß1 level was 6.0±0.7 ng/ml. No significant TGFß1 level differences were found at pre-RT and during-RT in patients with different genotypes of TGFß1 and tPA. Only ACE DD group had marginally higher pre-RT TGFß1 level than II and ID group (DD 23.6ng/ml vs. II 9.0ng/ml vs. ID 7.5ng/ml, p = 0.05), but no difference at during-RT(p=0.346) or during-RT/pre-RT ratio(p = 0.433). However, patients with TGFß1 509CC had higher elevation of plasma TGF ß1 level at fourth week during-RT than T allele carriers (TGFß1 level ratio of during-RT/pre-RT were CC 1.4±0.2 vs. T allele carriers 0.7±0.1, p=0.047). Conclusions: This exploratory study demonstrated that patients with TGFß1 509CC had higher elevation of plasma TGF ß1 level at fourth week during-RT than T allele carriers. This can help explain the correlation of TGF ß1 level elevation and higher risk of radiation induced lung toxicity in patients with NSCLC.
Development of automated quantitative multiparameter immunocytochemical profiling of circulating tumor cells (CTCs) in breast cancer (BC).
Authors: Miller Brandon L. , Deighan Clayton , Chalmers Jeffrey J. , Lustberg Maryam B. 
Published: March 2020

e21124 Background: The currently accepted definition of a CTC is a nucleated cell that is positive by qualitative fluorescence for EpCAM, expresses cytokeratin, and is negative for the pan-hematopoietic marker CD45. Such approach has limitations such as operator bias, difficulty distinguishing true signal from autofluoresence or filter bleed-through, and most importantly, it cannot differentiate expression levels that are low, intermediate or high. This is a problem because CTCs are in fact heterogeneous with respect to phenotypic expression of various markers. Methods: Peripheral blood samples were subjected to our previously described, negative depletion technology. Cytospins of enriched samples were stained with 3 to 5 antibodies conjugated to various Alexa Fluor dyes plus a nuclear dye and then analyzed with both confocal microscopy and a traditional epifluorescence microscope equipped with a Nuance (CRi) multispectral imaging camera and associated software. This imaging system allows not only fluorescent signals to be quantified between specific excitation and emission filters, but also the spectral structure between these boundaries through the use of a liquid crystal that takes images in as little as 10nm increments. Such imaging allows spectra from specific dyes to identify from the contributions of other dyes, filter leakage, and auto fluorescence. Results: Using this technology, and appropriate controls, we are developing continuous variable quantification of the expression of: cytokeratins, vimentin, CD45, Her2, CD44, EpCAM and EGFR in negatively enriched, peripheral blood samples from metastatic breast cancer patients. To date, over 10 metastatic patient samples have been quantified and the study is ongoing. Initial quantification indicates that significant variability in expression levels exists in multiple markers including Her2 neu. Conclusions: Using multispectral technology, we have shown that CTCs are heterogeneous, with different CTCs having varying expression levels of EpCAM, HER2 neu, vimentin and EGFR. Further multiparameter quantitative characterization with up to 6 color analysis on the same cell is in progress.
A retrospective study of immunohistochemical detection of Ki67 and correlation with ER, PR, and HER2-neu status in invasive breast cancer.
Authors: Prakash Deepa Meda , Shivakumar Swarna , Bapsy P P 
Published: March 2020

e21132 Background: Carcinoma breast is a heterogeneous group of disease. Several prognostic factors are known to predict the biological behavior and clinical outcome of breast cancer, including proliferation index, tumor size, age, histological grade, hormone receptors and Her2Neu status. Of theses proliferative index is the most important parameter in predicting tumor aggressiveness and prognosis. Ki67 is a nuclear protein, tightly linked to the cell cycle and is a marker of cell proliferation. High Ki67 usually correlates with a poor prognosis in breast cancer. Methods: This exploratory study is initiated whether Ki67 will be an additional prognostic factor. Immunohistochemical staining of Ki 67 was performed on formalin fixed paraffin embedded blocks of 50 cases of histologically confirmed breast cancer. Ki 67 expression based on distinct nuclear staining of tumor cells was assessed semiquantitatively as, nil (no immunostaining), low (≤10% immunopositivity), high (>10% immunopositivity) by light microscope. ER, PR, Her 2neu status and clinico-pathological parameters were analyzed and correlated with Ki67 immunostaining using the x2 test. Results: Our study showed high Ki 67 staining in 24 cases(48%), low immunostaining in 18 cases(36%) and was nil in 5 cases (10%) and was inconclusive in 3 cases(6%) (core biopsies). There was an inverse correlation of Ki 67 with ER, PR scores in 32 cases (64%) and positive correlation was seen in 28 cases (56%) with Her2 neu status. A good correlation was also observed with mitotic index in 37cases (74%) and histological grade in 29 cases (58%). Good correlation was seen with large tumor size in 24 cases(48%) and metastatic disease 9 cases (64%). Poor correlation was observed with node positive 22 cases (44%). High Ki 67 expression with good treatment response was seen in 13 cases (26%). Conclusions: Based on our study, Ki 67 score may prove to be an important prognostic factor in breast cancer and thereby, useful in making therapeutic decisions. But this is being explored in a large prospective study.
Analyses of the immunologically relevant gene transcript by qRT-PCR in primary cutaneous melanoma by the young and the elderly.
Authors: Hegde Upendra P. , Dadras Soheil Sam , Kerr Phil , Chakraborty Nitya , grant-Kels Jane , Mukherji Bijay 
Published: March 2020

e21135 Background: To understand the dynamics of anti tumor immune responses (IR) in the setting of the ageing immune system, we examined immunologically relevant gene transcripts in formalin fixed paraffin embedded (FFPE) primary cutaneous melanoma (PCM) samples derived from five young (age <50 years) and five elderly (age >or=70 years) patients by qRT-PCR. Methods: A total of 10 FFPE tumor blocks were collected between 2004-2007 of patients with PCM at the University of Connecticut Health Center. Total RNA was extracted from FFPE tissue sections and random primed cDNA was generated. Pre amplification using pooled Taqman assays and Universal PCR Master Mix (Applied Biosystems, USA) was performed for 14 cycles to ensure an adequate amount of starting material for qPCR. Using 15 ng total RNA, the quantification of human IR genes (IL-2, INFg, TNFa, TGFb1, IL-10, Foxp3, and IDO-1) and transcripts of succinate dehydrogenase (SDHA) (as endogenous control) was accomplished by qRT-PCR amplification of a cDNA using TaqMan Universal PCR Master Mix and TaqMan gene expressions assay probes. All reaction assays including that of SDHA were performed in triplicate on an ABI 7500 Fast system (Applied Biosystems). Cycle threshold (Ct) values for each mRNA were normalized to SDHA (DCT) and represented as RQ=2-DCT. The average RQ was calculated for all ten samples in both cohorts and Student’s T-test was performed to evaluate the statistical significance of the IR gene expression. Results: While no significant difference of expression of Th1 cytokines (IL-2, IFNg, TNFa) or Th2/3 cytokines (IL-10, TGFb) could be detected within the two groups, FOXp3 expression was significantly decreased in PCM in the elderly. Conclusions: Our results suggest ageing may be associated with less regulatory responses in situ in PCM and that more expanded analyses of the nature of the IR in PCM are likely to provide information with implications on prognosis and immune based treatment options for elderly PCM subjects.