bluuebunny/crossref_metadata_2025 huggingface dataset:
Sensitivity Evaluation of Spectral Nudging Schemes in Historical Dynamical Downscaling for South Asia
Authors: Ramzan Mehwish , Ham Suryun , Amjad Muhammad , Chang Eun-Chul , Yoshimura Kei
Published: April 2017

Sensitivity experiments testing two scale-selective bias correction (SSBC) methods have been carried out to identify an optimal spectral nudging scheme for historical dynamically downscaled simulations of South Asia, using the coordinated regional climate downscaling experiment (CORDEX) protocol and the regional spectral model (RSM). Two time periods were selected under the category of short-term extreme summer and long-term decadal analysis. The new SSBC version applied nudging to full wind components, with an increased relaxation time in the lower model layers, incorporating a vertical weighted damping coefficient. An evaluation of the extraordinary weather conditions experienced in South Asia in the summer of 2005 confirmed the advantages of the new SSBC when modeling monsoon precipitation. Furthermore, the new SSBC scheme was found to predict precipitation and wind patterns more accurately than the older version in decadal analysis, which applies nudging only to the rotational wind field, with a constant strength at all heights.
Application of Environmental Scanning Electron Microscope-Nanomanipulation System on Spheroplast Yeast Cells Surface Observation
Authors: Rad Maryam Alsadat , Ahmad Mohd Ridzuan , Nakajima Masahiro , Kojima Seiji , Homma Michio , Fukuda Toshio
Published: April 2017

The preparation and observations of spheroplast W303 cells are described with Environmental Scanning Electron Microscope (ESEM). The spheroplasting conversion was successfully confirmed qualitatively, by the evaluation of the morphological change between the normal W303 cells and the spheroplast W303 cells, and quantitatively, by determining the spheroplast conversion percentage based on the OD800absorbance data. From the optical microscope observations as expected, the normal cells had an oval shape whereas spheroplast cells resemble a spherical shape. This was also confirmed under four different mediums, that is, yeast peptone-dextrose (YPD), sterile water, sorbitol-EDTA-sodium citrate buffer (SCE), and sorbitol-Tris-Hcl-CaCl2(CaS). It was also observed that the SCE and CaS mediums had a higher number of spheroplast cells as compared to the YPD and sterile water mediums. The OD800absorbance data also showed that the whole W303 cells were fully converted to the spheroplast cells after about 15 minutes. The observations of the normal and the spheroplast W303 cells were then performed under an environmental scanning electron microscope (ESEM). The normal cells showed a smooth cell surface whereas the spheroplast cells had a bleb-like surface after the loss of its integrity when removing the cell wall.
Robust and Reversible Audio Watermarking by Modifying Statistical Features in Time Domain
Authors: Xiang Shijun , Yang Le , Wang Yi
Published: April 2017

Robust and reversible watermarking is a potential technique in many sensitive applications, such as lossless audio or medical image systems. This paper presents a novel robust reversible audio watermarking method by modifying the statistic features in time domain in the way that the histogram of these statistical values is shifted for data hiding. Firstly, the original audio is divided into nonoverlapped equal-sized frames. In each frame, the use of three samples as a group generates a prediction error and a statistical feature value is calculated as the sum of all the prediction errors in the frame. The watermark bits are embedded into the frames by shifting the histogram of the statistical features. The watermark is reversible and robust to common signal processing operations. Experimental results have shown that the proposed method not only is reversible but also achieves satisfactory robustness to MP3 compression of 64 kbps and additive Gaussian noise of 35 dB.
A Novel Segmentation Approach Combining Region- and Edge-Based Information for Ultrasound Images
Authors: Luo Yaozhong , Liu Longzhong , Huang Qinghua , Li Xuelong
Published: April 2017

Ultrasound imaging has become one of the most popular medical imaging modalities with numerous diagnostic applications. However, ultrasound (US) image segmentation, which is the essential process for further analysis, is a challenging task due to the poor image quality. In this paper, we propose a new segmentation scheme to combine both region- and edge-based information into the robust graph-based (RGB) segmentation method. The only interaction required is to select two diagonal points to determine a region of interest (ROI) on the original image. The ROI image is smoothed by a bilateral filter and then contrast-enhanced by histogram equalization. Then, the enhanced image is filtered by pyramid mean shift to improve homogeneity. With the optimization of particle swarm optimization (PSO) algorithm, the RGB segmentation method is performed to segment the filtered image. The segmentation results of our method have been compared with the corresponding results obtained by three existing approaches, and four metrics have been used to measure the segmentation performance. The experimental results show that the method achieves the best overall performance and gets the lowest ARE (10.77%), the second highest TPVF (85.34%), and the second lowest FPVF (4.48%).
Optimization of Ultrasound Extraction of Cactus Pear(Opuntia ficus indica)Seed Oil Based on Antioxidant Activity and Evaluation of Its Antimicrobial Activity
Authors: Ortega-Ortega María de los Angeles , Cruz-Cansino Nelly del Socorro , Alanís-García Ernesto , Delgado-Olivares Luis , Ariza-Ortega José Alberto , Ramírez-Moreno Esther , Manríquez-Torres José de Jesús
Published: April 2017

The purpose of the present study was to determine the optimal ultrasound conditions (amplitude level and time) for the extraction of cactus pear seed oil with the highest antioxidant activity using a closed system. Seed oil was analyzed for yield, antioxidant activity by ABTS and DPPH, and antimicrobial activity. Conventional extraction methods were assessed for comparison. Amplitude level significantly affected antioxidant activity in linear terms (p< 0.0001 DPPH andp< 0.001 ABTS, resp.) so, at lower amplitudes, the higher antioxidant activity was achieved. The optimum ultrasound extraction conditions were of 78% amplitude for 10 min and yielded antioxidant activity values of 66.25 mg AAE/100 g and 289 µmol TE/100 g for ABTS and DPPH, respectively. Compared with conventional extraction methods, ultrasound exhibited lower oil yield and antioxidant activity but had the potential to achieve comparable results if multiple ultrasound extractions are performed in the time needed by conventional methods. Seed oils showed similar antimicrobial activity despite the extraction method and were more effective againstEscherichia coli. The results demonstrated that ultrasound can be an alternative extraction method of seed oils from fruits such as cactus pear.
tRF/miR-1280 Suppresses Stem Cell–like Cells and Metastasis in Colorectal Cancer
Authors: Huang Bingqing , Yang Huipeng , Cheng Xixi , Wang Dan , Fu Shuyu , Shen Wencui , Zhang Qi , Zhang Lijuan , Xue Zhenyi , Li Yan , Da Yurong , Yang Qing , Li Zesong , Liu Li , Qiao Liang , Kong Ying , Yao Zhi , Zhao Peng , Li Min , Zhang Rongxin
Published: April 2017

Several studies have shown that tRNAs can be enzymatically cleaved to generate distinct classes of tRNA-derived fragments (tRF). Here, we report that tRF/miR-1280, a 17-bp fragment derived from tRNALeu and pre-miRNA, influences Notch signaling pathways that support the function of cancer stem-like cells (CSC) in colorectal cancer progression. tRF/miR-1280 expression was decreased in human specimens of colorectal cancer. Ectopic expression of tRF/miR-1280 reduced cell proliferation and colony formation, whereas its suppression reversed these effects. Mechanistic investigations implicated the Notch ligand JAG2 as a direct target of tRF/miR-1280 binding through which it reduced tumor formation and metastasis. Notably, tRF/miR-1280–mediated inactivation of Notch signaling suppressed CSC phenotypes, including by direct transcriptional repression of the Gata1/3 and miR-200b genes. These results were consistent with findings of decreased levels of miR-200b and elevated levels of JAG2, Gata1, Gata3, Zeb1, and Suz12 in colorectal cancer tissue specimens. Taken together, our results established that tRF/miR-1280 suppresses colorectal cancer growth and metastasis by repressing Notch signaling pathways that support CSC phenotypes. Furthermore, they provide evidence that functionally active miRNA can be derived from tRNA, offering potential biomarker and therapeutic uses. Cancer Res; 77(12); 3194–206. ©2017 AACR.
Landscape of Combination Immunotherapy and Targeted Therapy to Improve Cancer Management
Authors: Colli Leandro M. , Machiela Mitchell J. , Zhang Han , Myers Timothy A. , Jessop Lea , Delattre Olivier , Yu Kai , Chanock Stephen J.
Published: April 2017

Cancer treatments composed of immune checkpoint inhibitors and oncogene-targeted drugs might improve cancer management, but there has been little investigation of their combined potential as yet. To estimate the fraction of cancer cases that might benefit from such combination therapy, we conducted an exploratory study of cancer genomic datasets to determine the proportion with somatic mutation profiles amenable to either immunotherapy or targeted therapy. We surveyed 13,349 genomic profiles from public databases for cases with specific mutations targeted by current agents or a burden of exome-wide nonsynonymous mutations (NsM) that exceed a proposed threshold for response to checkpoint inhibitors. Overall, 8.9% of cases displayed profiles that could benefit from combination therapy, which corresponded to approximately 11.2% of U.S. annual incident cancer cases. Frequently targetable mutations were in PIK3CA, BRAF, NF1, NRAS, and PTEN. We also noted a high burden of NsM in cases with targetable mutations in SMO, DDR2, FGFR1, PTCH1, FGFR2, and MET. Our results indicate that a significant proportion of solid tumor patients are eligible for immuno-targeted combination therapy, and they suggest prioritizing specific cancers for trials of certain targeted and checkpoint inhibitor drugs. Cancer Res; 77(13); 3666–71. ©2017 AACR.
The National Cancer Institute ALMANAC: A Comprehensive Screening Resource for the Detection of Anticancer Drug Pairs with Enhanced Therapeutic Activity
Authors: Holbeck Susan L. , Camalier Richard , Crowell James A. , Govindharajulu Jeevan Prasaad , Hollingshead Melinda , Anderson Lawrence W. , Polley Eric , Rubinstein Larry , Srivastava Apurva , Wilsker Deborah , Collins Jerry M. , Doroshow James H.
Published: April 2017

To date, over 100 small-molecule oncology drugs have been approved by the FDA. Because of the inherent heterogeneity of tumors, these small molecules are often administered in combination to prevent emergence of resistant cell subpopulations. Therefore, new combination strategies to overcome drug resistance in patients with advanced cancer are needed. In this study, we performed a systematic evaluation of the therapeutic activity of over 5,000 pairs of FDA-approved cancer drugs against a panel of 60 well-characterized human tumor cell lines (NCI-60) to uncover combinations with greater than additive growth-inhibitory activity. Screening results were compiled into a database, termed the NCI-ALMANAC (A Large Matrix of Anti-Neoplastic Agent Combinations), publicly available at https://dtp.cancer.gov/ncialmanac. Subsequent in vivo experiments in mouse xenograft models of human cancer confirmed combinations with greater than single-agent efficacy. Concomitant detection of mechanistic biomarkers for these combinations in vivo supported the initiation of two phase I clinical trials at the NCI to evaluate clofarabine with bortezomib and nilotinib with paclitaxel in patients with advanced cancer. Consequently, the hypothesis-generating NCI-ALMANAC web-based resource has demonstrated value in identifying promising combinations of approved drugs with potent anticancer activity for further mechanistic study and translation to clinical trials. Cancer Res; 77(13); 3564–76. ©2017 AACR.
Conjunctival Ultraviolet Autofluorescence as a Measure of Past Sun Exposure in Children
Authors: Sun Cong , Pezic Angela , Mackey David A. , Carlin John B. , Kemp Andrew , Ellis Justine A. , Cameron Fergus J. , Rodda Christine P. , Dwyer Terence , Coroneo Minas T. , Ponsonby Anne-Louise
Published: April 2017

Background: Conjunctival ultraviolet autofluorescence (CUVAF) area detected from UVAF photographs is a recently developed potential marker for past sun exposure, but its relationship with sun-related factors has not been fully investigated.

Methods: The study included 339 healthy children ages 5 to 15 years in Melbourne, Australia. Data were collected by questionnaire and examination at school. CUVAF area was measured using a computer program and analyzed as a continuous and dichotomous outcome (any/none).

Results: Fifty-three children (15.6%) had detectable CUVAF, and the youngest age at which a child showed sun damage was 8 years. Compared with silicone skin cast score, there was good inter-grader agreement on CUVAF grading, with Cohen kappa 0.85 [95% confidence interval (CI), 0.65–1.00] for total CUVAF area using both eye photographs. Perfect intra-grader agreement was achieved. Fairer pigmentation, including medium/fair skin color [adjusted odds ratio (AOR), 3.42; 95% CI, 1.02–11.48 vs. dark/olive] and blue/gray eye color (AOR, 4.07; 95% CI, 1.73–9.55 vs. brown) was associated with increased odds of CUVAF. Increasing lifetime sunburn number (e.g., AOR, 2.89; 95% CI, 1.14–7.35 and 4.29; 1.04–17.76 for sunburns 2 to 4 and ≥ 5 times, respectively, vs. no sunburns, trend P = 0.004) and freckling by the end of last summer were associated with increased odds of CUVAF.

Conclusions: CUVAF area can be an a priori objective measure of past sun exposure in pediatric populations for future research.

Impact: To our knowledge, this is the first pediatric study that evaluated associations of sun-related risk factors with CUVAF. Cancer Epidemiol Biomarkers Prev; 26(7); 1146–53. ©2017 AACR.
Comparison of the Natural History of Genital HPV Infection among Men by Country: Brazil, Mexico, and the United States
Authors: Sudenga Staci L. , Torres B. Nelson , Silva Roberto , Villa Luisa L. , Lazcano-Ponce Eduardo , Abrahamsen Martha , Baggio Maria Luiza , Salmeron Jorge , Quiterio Manuel , Giuliano Anna R.
Published: April 2017

Background: Male genital human papillomavirus (HPV) prevalence and incidence has been reported to vary by geographical location. Our objective was to assess the natural history of genital HPV by country among men with a median of 48 months of follow-up.

Methods: Men ages 18–70 years were recruited from United States (n = 1,326), Mexico (n = 1,349), and Brazil (n = 1,410). Genital specimens were collected every 6 months and HPV genotyping identified 37 HPV genotypes. Prevalence of HPV was compared between the three countries using the Fisher exact test. Incidence rates and 95% confidence intervals were calculated. The median time to HPV clearance among men with an incident infection was estimated using the Kaplan–Meier method.

Results: The prevalence and incidence of the genital HPV types known to cause disease in males (HPV 16 and 6) was significantly higher among men from Brazil than men from Mexico. Prevalence and incidence of those genital HPV types in the United States varied between being comparable with those of Mexico or Brazil. Although genital HPV16 duration was significantly longer in Brazil (P = 0.04) compared with Mexico and the United States, HPV6 duration was shortest in Brazil (P = 0.03) compared with Mexico and the United States.

Conclusions: Men in Brazil and Mexico often have similar, if not higher prevalence of HPV compared with men from the United States.

Impact: Currently, there is no routine screening for genital HPV among males and while HPV is common in men, and most naturally clear the infection, a proportion of men do develop HPV-related diseases. Men may benefit from gender-neutral vaccine policies. Cancer Epidemiol Biomarkers Prev; 26(7); 1043–52. ©2017 AACR.
Cellular Expression of PD-L1 in the Peripheral Blood of Lung Cancer Patients is Associated with Worse Survival
Authors: Boffa Daniel J. , Graf Ryon P. , Salazar Michelle C. , Hoag Jessica , Lu David , Krupa Rachel , Louw Jessica , Dugan Lyndsey , Wang Yipeng , Landers Mark , Suraneni Mahipal , Greene Stephanie B. , Magaña Marisa , Makani Samir , Bazhenova Lyudmila , Dittamore Ryan V. , Nieva Jorge
Published: April 2017

Background: Lung cancer treatment has become increasingly dependent upon invasive biopsies to profile tumors for personalized therapy. Recently, tumor expression of programmed death-ligand 1 (PD-L1) has gained interest as a potential predictor of response to immunotherapy. Circulating biomarkers present an opportunity for tumor profiling without the risks of invasive procedures. We characterized PD-L1 expression within populations of nucleated cells in the peripheral blood of lung cancer patients in hopes of expanding the role of liquid biopsy in this setting.

Methods: Peripheral blood samples from a multi-institutional prospective study of patients with clinical diagnosis of lung cancer were subjected to cytomorphometric and immunohistochemical evaluation using single-cell, automated slide-based, digital pathology. PD-L1 expression was determined by immunofluorescence.

Results: PD-L1 expression was detected within peripheral circulating cells associated with malignancy (CCAM) in 26 of 112 (23%) non–small cell lung cancer patients. Two distinct populations of nucleated, nonhematolymphoid, PD-L1–expressing cells were identified; cytokeratin positive (CK+, PD-L1+, CD45−) and cytokeratin negative (CK−, PD-L1+, CD45−) cells, both with cytomorphometric features (size, nuclear-to-cytoplasm ratio) consistent with tumor cells. Patients with >1.1 PD-L1(+) cell/mL (n = 14/112) experienced worse overall survival than patients with ≤1.1 PD-L1(+) cell/mL (2-year OS: 31.2% vs. 78.8%, P = 0.00159). In a Cox model adjusting for stage, high PD-L1(+) cell burden remained a significant predictor of mortality (HR = 3.85; 95% confidence interval, 1.64–9.09; P = 0.002).

Conclusions: PD-L1 expression is detectable in two distinct cell populations in the peripheral blood of lung cancer patients and is associated with worse survival.

Impact: These findings could represent a step forward in the development of minimally invasive liquid biopsies for the profiling of tumors. Cancer Epidemiol Biomarkers Prev; 26(7); 1139–45. ©2017 AACR.
Enrichment of PI3K-AKT–mTOR Pathway Activation in Hepatic Metastases from Breast Cancer
Authors: Pierobon Mariaelena , Ramos Corinne , Wong Shukmei , Hodge K. Alex , Aldrich Jessica , Byron Sara , Anthony Stephen P. , Robert Nicholas J. , Northfelt Donald W. , Jahanzeb Mohammad , Vocila Linda , Wulfkuhle Julia , Gambara Guido , Gallagher Rosa I. , Dunetz Bryant , Hoke Nicholas , Dong Ting , Craig David W. , Cristofanilli Massimo , Leyland-Jones Brian , Liotta Lance A. , O'Shaughnessy Joyce A. , Carpten John D. , Petricoin Emanuel F.
Published: April 2017

Purpose: Little is known about the molecular signatures associated with specific metastatic sites in breast cancer. Using comprehensive multi-omic molecular profiling, we assessed whether alterations or activation of the PI3K–AKT–mTOR pathway is associated with specific sites of breast cancer metastasis.

Experimental Design: Next-generation sequencing–based whole-exome sequencing was coupled with reverse-phase protein microarray (RPPA) functional signaling network analysis to explore the PI3K–AKT–mTOR axis in 32 pretreated breast cancer metastases. RPPA-based signaling data were further validated in an independent cohort of 154 metastatic lesions from breast cancer and 101 unmatched primary breast tumors. The proportion of cases with PI3K–AKT–mTOR genomic alterations or signaling network activation were compared between hepatic and nonhepatic lesions.

Results: PIK3CA mutation and activation of AKT (S473) and p70S6K (T389) were detected more frequently among liver metastases than nonhepatic lesions (P < 0.01, P = 0.056, and P = 0.053, respectively). However, PIK3CA mutations alone were insufficient in predicting protein activation (P = 0.32 and P = 0.19 for activated AKT and p70S6K, respectively). RPPA analysis of an independent cohort of 154 tumors confirmed the relationship between pathway activation and hepatic metastasis [AKT (S473), mTOR (S2448), and 4EBP1 (S65); P < 0.01, P = 0.02, and P = 0.01, respectively]. Similar results were also seen between liver metastases and primary breast tumors [AKT (S473) P < 0.01, mTOR (S2448) P < 0.01, 4EBP1 (S65) P = 0.01]. This signature was lost when primary tumors were compared with all metastatic sites combined.

Conclusions: Breast cancer patients with liver metastasis may represent a molecularly homogenized cohort with increased incidence of PIK3CA mutations and activation of the PI3K–AKT–mTOR signaling network. Clin Cancer Res; 23(16); 4919–28. ©2017 AACR.
Sensitivity and Specificity of Cetuximab-IRDye800CW to Identify Regional Metastatic Disease in Head and Neck Cancer
Authors: Rosenthal Eben L. , Moore Lindsay S. , Tipirneni Kiranya , de Boer Esther , Stevens Todd M. , Hartman Yolanda E. , Carroll William R. , Zinn Kurt R. , Warram Jason M.
Published: April 2017

Purpose: Comprehensive cervical lymphadenectomy can be associated with significant morbidity and poor quality of life. This study evaluated the sensitivity and specificity of cetuximab-IRDye800CW to identify metastatic disease in patients with head and neck cancer.

Experimental Design: Consenting patients scheduled for curative resection were enrolled in a clinical trial to evaluate the safety and specificity of cetuximab-IRDye800CW. Patients (n = 12) received escalating doses of the study drug. Where indicated, cervical lymphadenectomy accompanied primary tumor resection, which occurred 3 to 7 days following intravenous infusion of cetuximab-IRDye800CW. All 471 dissected lymph nodes were imaged with a closed-field, near-infrared imaging device during gross processing of the fresh specimens. Intraoperative imaging of exposed neck levels was performed with an open-field fluorescence imaging device. Blinded assessments of the fluorescence data were compared to histopathology to calculate sensitivity, specificity, negative predictive value (NPV), and positive predictive value (PPV).

Results: Of the 35 nodes diagnosed pathologically positive, 34 were correctly identified with fluorescence imaging, yielding a sensitivity of 97.2%. Of the 435 pathologically negative nodes, 401 were correctly assessed using fluorescence imaging, yielding a specificity of 92.7%. The NPV was determined to be 99.7%, and the PPV was 50.7%. When 37 fluorescently false-positive nodes were sectioned deeper (1 mm) into their respective blocks, metastatic cancer was found in 8.1% of the recut nodal specimens, which altered staging in two of those cases.

Conclusions: Fluorescence imaging of lymph nodes after systemic cetuximab-IRDye800CW administration demonstrated high sensitivity and was capable of identifying additional positive nodes on deep sectioning. Clin Cancer Res; 23(16); 4744–52. ©2017 AACR.
Feasibility of Ultra-High-Throughput Functional Screening of Melanoma Biopsies for Discovery of Novel Cancer Drug Combinations
Authors: Friedman Adam A. , Xia Yun , Trippa Lorenzo , Le Long Phi , Igras Vivien , Frederick Dennie T. , Wargo Jennifer A. , Tanabe Kenneth K. , Lawrence Donald P. , Neuberg Donna S. , Flaherty Keith T. , Fisher David E.
Published: April 2017

Purpose: Successful development of targeted therapy combinations for cancer patients depends on first discovering such combinations in predictive preclinical models. Stable cell lines and mouse xenograft models can have genetic and phenotypic drift and may take too long to generate to be useful as a personalized medicine tool.

Experimental Design: To overcome these limitations, we have used a platform of ultra-high-throughput functional screening of primary biopsies preserving both cancer and stroma cell populations from melanoma patients to nominate such novel combinations from a library of thousands of drug combinations in a patient-specific manner within days of biopsy. In parallel, patient-derived xenograft (PDX) mouse models were created and novel combinations tested for their ability to shrink matched PDXs.

Results: The screening method identifies specific drug combinations in tumor cells with patterns that are distinct from those obtained from stable cell lines. Screening results were highly specific to individual patients. For patients with matched PDX models, we confirmed that individualized novel targeted therapy combinations could inhibit tumor growth. In particular, a combination of multi-kinase and PI3K/Akt inhibitors was effective in some BRAF–wild-type melanomas, and the addition of cediranib to the BRAF inhibitor PLX4720 was effective in a PDX model with BRAF mutation.

Conclusions: This proof-of-concept study demonstrates the feasibility of using primary biopsies directly for combinatorial drug discovery, complementing stable cell lines and xenografts, but with much greater speed and efficiency. This process could potentially be used in a clinical setting to rapidly identify therapeutic strategies for individual patients. Clin Cancer Res; 23(16); 4680–92. ©2017 AACR.
Analytic, Preanalytic, and Clinical Validation of p53 IHC for Detection of TP53 Missense Mutation in Prostate Cancer
Authors: Guedes Liana B. , Almutairi Fawaz , Haffner Michael C. , Rajoria Gaurav , Liu Zach , Klimek Szczepan , Zoino Roberto , Yousefi Kasra , Sharma Rajni , De Marzo Angelo M. , Netto George J. , Isaacs William B. , Ross Ashley E. , Schaeffer Edward M. , Lotan Tamara L.
Published: April 2017

Purpose: TP53 missense mutations may help to identify prostate cancer with lethal potential. Here, we preanalytically, analytically, and clinically validated a robust IHC assay to detect subclonal and focal TP53 missense mutations in prostate cancer.

Experimental Design: The p53 IHC assay was performed in a CLIA-accredited laboratory on the Ventana Benchmark immunostaining system. p53 protein nuclear accumulation was defined as any p53 nuclear labeling in >10% of tumor cells. Fifty-four formalin-fixed paraffin embedded (FFPE) cell lines from the NCI-60 panel and 103 FFPE prostate cancer tissues (88 primary adenocarcinomas, 15 metastases) with known TP53 mutation status were studied. DU145 and VCaP xenografts were subjected to varying fixation conditions to investigate the effects of preanalytic variables. Clinical validation was performed in two partially overlapping radical prostatectomy cohorts.

Results: p53 nuclear accumulation by IHC was 100% sensitive for detection of TP53 missense mutations in the NCI-60 panel (25/25 missense mutations correctly identified). Lack of p53 nuclear accumulation was 86% (25/29) specific for absence of TP53 missense mutation. In FFPE prostate tumors, the positive predictive value of p53 nuclear accumulation for underlying missense mutation was 84% (38/45), whereas the negative predictive value was 97% (56/58). In a cohort of men who experienced biochemical recurrence after RP, the multivariable HR for metastasis among cases with p53 nuclear accumulation compared with those without was 2.55 (95% confidence interval, 1.1–5.91).

Conclusions: IHC is widely available method to assess for the presence of deleterious and heterogeneous TP53 missense mutations in clinical prostate cancer specimens. Clin Cancer Res; 23(16); 4693–703. ©2017 AACR.
The Microbiota: A New Variable Impacting Cancer Treatment Outcomes
Authors: Fessler Jessica L. , Gajewski Thomas F.
Published: April 2017

Preclinical evidence has established that the host commensal microbiota can contribute to therapeutic response in cancer models, a finding supported by early clinical data. This connection between the microbiome and clinical outcome in oncology is cause for new consideration in the administration of antibiotics and microbiota-modulating interventions to improve outcomes. Clin Cancer Res; 23(13); 3229–31. ©2017 AACR.

See related article by Galloway-Peña et al., p. 3263
Concordance of Genomic Alterations by Next-Generation Sequencing in Tumor Tissue versus Circulating Tumor DNA in Breast Cancer
Authors: Chae Young Kwang , Davis Andrew A. , Jain Sarika , Santa-Maria Cesar , Flaum Lisa , Beaubier Nike , Platanias Leonidas C. , Gradishar William , Giles Francis J. , Cristofanilli Massimo
Published: April 2017

While identifying genomic alterations in tumor tissue is the current gold-standard technique for molecular profiling, circulating tumor DNA (ctDNA) represents a noninvasive method of assessing genomic alterations using peripheral blood. The concordance of genomic alterations between two commercially available ctDNA and tissue biopsies was compared in 45 patients with breast cancer using paired next-generation sequencing tissue and ctDNA biopsies. Across all genes, concordance between the two platforms was 91.0% to 94.2%. When only considering genomic alterations in either assay (e.g., excluding wild type/wild type genes), concordance was 10.8% to 15.1% with full plus partial concordance of 13.8% to 19.3%. Concordant mutations were associated with significantly higher variant allele frequency. Over half of mutations detected in either technique were not detected using the other biopsy technique. Including variants of unknown significance, the average number of alterations per patient was significantly higher for tissue (4.56) compared with ctDNA (2.16). When eliminating alterations not detectable in the ctDNA assay, mean number of alterations for tissue and ctDNA was similar (2.67 for tissue, 2.16 for ctDNA). Across five representative genes (TP53, PIK3CA, ERBB2, BRCA1, and BRCA2), sensitivity and specificity were 35.7% and 95.0%, respectively. Concordance when genomic alterations was detected in either tissue or ctDNA was low with each technique detecting a significant amount of nonoverlapping mutations. Potential explanations for the lack of concordance include tumor heterogeneity, different sequencing techniques, spatial and temporal factors, and potential germline DNA contamination. The study indicates that both tissue and blood-based NGS may be necessary to describe the complex biology of breast cancer. Mol Cancer Ther; 16(7); 1412–20. ©2017 AACR.
Ref-1/APE1 as a Transcriptional Regulator and Novel Therapeutic Target in Pediatric T-cell Leukemia
Authors: Ding Jixin , Fishel Melissa L. , Reed April M. , McAdams Erin , Czader Magdalena B. , Cardoso Angelo A. , Kelley Mark R.
Published: April 2017

The increasing characterization of childhood acute lymphoblastic leukemia (ALL) has led to the identification of multiple molecular targets but has yet to translate into more effective targeted therapies, particularly for high-risk, relapsed T-cell ALL. Searching for master regulators controlling multiple signaling pathways in T-ALL, we investigated the multifunctional protein redox factor-1 (Ref-1/APE1), which acts as a signaling “node” by exerting redox regulatory control of transcription factors important in leukemia. Leukemia patients' transcriptome databases showed increased expression in T-ALL of Ref-1 and other genes of the Ref-1/SET interactome. Validation studies demonstrated that Ref-1 is expressed in high-risk leukemia T cells, including in patient biopsies. Ref-1 redox function is active in leukemia T cells, regulating the Ref-1 target NF-κB, and inhibited by the redox-selective Ref-1 inhibitor E3330. Ref-1 expression is not regulated by Notch signaling, but is upregulated by glucocorticoid treatment. E3330 disrupted Ref-1 redox activity in functional studies and resulted in marked inhibition of leukemia cell viability, including T-ALL lines representing different genotypes and risk groups. Potent leukemia cell inhibition was seen in primary cells from ALL patients, relapsed and glucocorticoid-resistant T-ALL cells, and cells from a murine model of Notch-induced leukemia. Ref-1 redox inhibition triggered leukemia cell apoptosis and downregulation of survival genes regulated by Ref-1 targets. For the first time, this work identifies Ref-1 as a novel molecular effector in T-ALL and demonstrates that Ref-1 redox inhibition results in potent inhibition of leukemia T cells, including relapsed T-ALL. These data also support E3330 as a specific Ref-1 small-molecule inhibitor for leukemia. Mol Cancer Ther; 16(7); 1401–11. ©2017 AACR.
Cotargeting mTORC and EGFR Signaling as a Therapeutic Strategy in HNSCC
Authors: Swick Adam D. , Prabakaran Prashanth J. , Miller Margot C. , Javaid Amal M. , Fisher Michael M. , Sampene Emmanuel , Ong Irene M. , Hu Rong , Iida Mari , Nickel Kwangok P. , Bruce Justine Y. , Wheeler Deric L. , Kimple Randall J.
Published: April 2017

Head and neck squamous cell carcinomas (HNSCC) are frequently altered along the PI3K/AKT/mTORC signaling axis. Despite excellent preclinical data, the use of compounds targeting this pathway as monotherapy has been underwhelming in initial clinical trials, and identification of predictive biomarkers remains challenging. To investigate mTORC-specific inhibition, we tested catalytic mTORC (AZD8055) and PI3K/mTORC (NVP-BEZ-235) inhibitors ± cetuximab in a panel of HNSCC cell lines and patient-derived xenografts (PDX). Cell lines were assayed for response to all agents and siRNA knockdown of targets by multiple approaches. All cell lines showed similar response to both drug and siRNA inhibition of both PI3K and mTORC pathways, with anti-EGFR combination producing modest additive effect. Five PDX models that presented PIK3CA mutation or intrinsic cetuximab resistance were treated with a combination of cetuximab and AZD8055. In vivo single-agent mTORC inhibition inhibited growth of one PIK3CA-mutant cancer, but had little effect on any PIK3CAWT or a second PIK3CA-mutant model. In all models, the combination therapy showed greater growth delay than monotherapy. The uniform ability of PI3K and mTORC inhibition to suppress the growth of HNSCC cells highlights the pathway's role in driving proliferation. Although single-agent therapy was largely ineffective in vivo, improved response of combination treatment in an array of PDXs suggests the potential for adding a catalytic mTORC inhibitor to cetuximab therapy. Overall, these results add to a growing body of evidence, suggesting that approaches that attempt to match biomarkers to the optimal therapy in HNSCC remain complex and challenging. Mol Cancer Ther; 16(7); 1257–68. ©2017 AACR.
Atypical Clinical and Serological Manifestation of Pemphigus Vegetans: A Case Report and Review of the Literature
Authors: Mergler Rebecca , Kerstan Andreas , Schmidt Enno , Goebeler Matthias , Benoit Sandrine
Published: April 2017

Pemphigus vegetans (PVeg) is a rare variant of pemphigus vulgaris characterized by pustules and/or papillomatous vegetations, preferentially affecting intertriginous and periorificial areas. Exceptional manifestations may be misdiagnosed resulting in delayed diagnosis and treatment. Diagnosis is confirmed by immunofluorescence and detection of anti-desmoglein (Dsg) 3 and/or anti-Dsg1 antibodies. We herein report an unusual manifestation of PVeg. At the time of first presentation, lesions were restricted to the right ring finger’s tip. Although mucous membranes were initially not affected, high levels of anti-Dsg3 antibodies were detected while anti-Dsg1 and anti-desmocollin (Dsc) 1, 2, and 3 antibodies were absent. To compare our immunological findings with previous reports, all accessible Anglophone literature published since December 1988 was evaluated. We identified 52 patients suffering from PVeg, 7 of these showed anti-Dsg3 antibodies without any mucous membrane involvement. Notably, the detection of anti-Dsg1 and anti-Dsg3 antibodies does not necessarily correlate with the involvement of skin and/or mucous membranes. This might be due to more specific and complex antibody constellations in nonclassical or atypical pemphigus.
Changing Epidemiological Patterns in Traumatic Brain Injury: A Longitudinal Hospital-Based Study in Belgium
Authors: Peeters Wouter , Majdan Marek , Brazinova Alexandra , Nieboer Daan , Maas Andrew I.R.
Published: April 2017

Background: Various reports have suggested that epidemiological patterns of Traumatic Brain Injury (TBI) are changing in high-income countries, but the evidence to support this is often indirect and only a few longitudinal studies exist. We aimed to explore epidemiological patterns of TBI in Belgium over a 10-year period. Methods: A retrospective analysis of Minimum Hospital Data provided by Statistics Belgium was performed for the period 2003-2012. ICD-9 classification was used to identify TBI and to differentiate subtypes. The annual incidence of hospital admissions and in-hospital mortality rates were calculated and further differentiated for age, gender and cause of injury. Results: The age-adjusted incidence of hospital admissions decreased by 3.6% per year. An increase in the number of elderly patients with TBI and a decrease in the younger age groups were found. Falls now represent the main cause of TBI. A mortality rate of 6.5 per 100,000 population per year was found and did not change significantly over time. Conclusions: This longitudinal study confirms that epidemiological patterns in TBI are changing: overall incidence is steadily decreasing, but in elderly patients, the incidence is increasing. Falls are the leading cause, occurring most frequently in elderly patients. These changes are relevant for prevention.
Safety and Efficacy of a Fully Covered Self-Expandable Metallic Stent in Benign Airway Stenosis
Authors: Fortin Marc , Lacasse Yves , Elharrar Xavier , Tazi-Mezalek Rachid , Laroumagne Sophie , Guinde Julien , Astoul Philippe , Dutau Hervé
Published: April 2017

Background: The use of self-expandable metallic stents (SEMS) in benign airway disease was the object of a boxed warning from the United States Food and Drug Administration in 2005 due to the risk of stent-related complications and difficulties associated with their removal. Third-generation fully covered SEMS have been commercialized since this warning and theoretically should not present the same difficulties associated with removal as they cannot become embedded in the airway mucosa. Objectives: We aimed to examine the safety and efficacy of a specific third-generation SEMS, the Silmet stent. Methods: We reviewed the records of all patients treated for benign airway stenosis with third-generation Silmet SEMS from January 2011 to December 2015 at the North Hospital of Marseilles, France. Results: Forty SEMS were inserted in 30 patients over this period. Twenty (50.0%) stents were removed because of stent-related complications after a median of 77.0 ± 96.6 days (migration 32.5%, granulation tissue formation 7.5%, subjective intolerance 5.0%, mucus plugging 2.5%, laryngeal edema 2.5%). There were no cases of stent-related mortality. All complications were managed successfully endoscopically. Thirty-six stents (90.0%) were removed successfully after a median of 122.0 ± 113.2 days without any complications. The clinical success rate of stent treatment was 40.7%. Conclusion: Third-generation SEMS are a safe treatment option for complex benign airway stenosis, but complications requiring stent removal are frequent. Further studies are needed to compare the performance of third-generation SEMS and silicone stents in benign airway stenosis.
Pruritic Urticarial Papules and Plaques of Pregnancy Occurring Postpartum Treated with Intramuscular Injection of Autologous Whole Blood
Authors: Kim En Hyung
Published: April 2017

Pruritic urticarial papules and plaques of pregnancy (PUPPP) is one of the most common diseases associated with pregnancy. In most cases, the skin lesions develop in the third trimester of primigravidas. There are no systemic alterations seen in PUPPP; however, most patients report severe pruritus. A 34-year-old woman presented 1 week postpartum with typical clinical features of PUPPP. The patient showed good response to intramuscular injection of autologous whole blood with no adverse effects to the patient or her baby. Presentation of PUPPP in the postpartum period is rare. Conservative management with topical corticosteroids and oral antihistamines is commonly used to relieve pruritus. In severe cases, skin lesions and symptoms are controlled with a brief course of systemic corticosteroids. Investigation of new treatment options has been limited by patient concerns about the negative effects of medication on the fetus or breastfeeding. Intramuscular injection of autologous whole blood could be an alternative treatment option for PUPPP, especially for women who worry about the use of medications during pregnancy or breastfeeding.
The Long Non-Coding RNA XIST Controls Non-Small Cell Lung Cancer Proliferation and Invasion by Modulating miR-186-5p
Authors: Wang Haoyou , Shen Qiming , Zhang Xin , Yang Chunlu , Cui Su , Sun Yanbin , Wang Liming , Fan Xiaoxi , Xu Shun
Published: April 2017

Background/Aims: Long non-coding RNAs (lncRNAs) are key players in the development and progression of human cancers. The lncRNA XIST (X-inactive specific transcript) has been shown to be upregulated in human non-small cell lung cancer (NSCLC); however, its role and molecular mechanisms in NSCLC cell progression remain unclear. Methods: qRT-PCR was conducted to assess the expression of XIST and miR-186. Cell proliferation was detected using MTT assay. Cell invasion and migration were evaluated using transwell assay. Cell cycle distribution and apoptosis rates were analyzed by flow cytometry. Luciferase reporter assay was used to identify the direct regulation of XIST and miR-186. A RNA immunoprecipitation was used to analyze whether XIST was associated with the RNA-induced silencing complex (RISC). Results: We confirmed that XIST was upregulated in NSCLC cell lines and tissues. Functionally, XIST knockdown inhibited cancer cell proliferation and invasion, and induced apoptosis in vitro, and suppressed subcutaneous tumor growth in vivo. Mechanistic investigations revealed a reciprocal repressive interaction between XIST and miR-186-5p. Furthermore, we showed that miR-186-5p has a binding site for XIST. Our data also indicated that XIST and miR-186-5p are likely in the same RNA induced silencing complex. Conclusion: Together, our data revealed that XIST knockdown confers suppressive function in NSCLC and XIST may be a novel therapeutic marker in this disease.
Quantitative Evaluation of Brain Stem Atrophy Using Magnetic Resonance Imaging in Adult Patients with Alexander Disease
Authors: Yoshida Tomokatsu , Yasuda Rei , Mizuta Ikuko , Nakagawa Masanori , Mizuno Toshiki ,   JAPAN Alexander Disease Study Group
Published: April 2017

Brain MRI in adult patients with Alexander disease (AxD) mainly shows atrophy in the medulla oblongata. However, currently there is no quantitative standard for assessing this atrophy. In this study, we quantitatively evaluated the brain stem of AxD patients with glial fibrillary acidic protein (GFAP) mutation using conventional MRI to evaluate its usefulness as an aid to diagnosing AxD in daily clinical practice. Nineteen AxD patients with GFAP mutation were compared with 14 patients negative for GFAP mutation in whom AxD was suspected due to “atrophy of the medulla oblongata.” In the GFAP mutation-positive group, the sagittal diameter of the medulla oblongata, the ratio of the diameter of the medulla oblongata to that of the midbrain (MO/MB), and the ratio of the sagittal diameter of the medulla oblongata to that of the pons (MO/Po) were significantly smaller compared to those of the GFAP mutation-negative group (p < 0.01). The sensitivity and specificity of each parameter were 87.5 and 92.3%, 91.7 and 81.3%, and 88.2 and 100% with a sagittal diameter of the medulla oblongata <9.0 mm, MO/MB <0.60, and sagittal MO/Po <0.46, respectively. These parameters can provide very useful information to differentially diagnose AxD from other disorders associated with brain stem atrophy in adult patients.
Endogenous developmental endothelial locus-1 limits ischaemia- related angiogenesis by blocking inflammation
Authors: Klotzsche - von Ameln Anne , Cremer Sebastian , Hoffmann Jedrzej , Schuster Peggy , Khedr Sherif , Korovina Irina , Troullinaki Maria , Neuwirth Ales , Sprott David , Chatzigeorgiou Antonios , Economopoulou Matina , Orlandi Alessia , Hain Andreas , Zeiher Andreas M. , Deussen Andreas , Hajishengallis George , Dimmeler Stefanie , Chavakis Triantafyllos , Chavakis Emmanouil
Published: April 2017

We have recently identified endothelial cell-secreted developmental endothelial locus-1 (Del-1) as an endogenous inhibitor of β2-integrin–dependent leukocyte infiltration. Del-1 was previously also implicated in angiogenesis. Here, we addressed the role of endogenously produced Del-1 in ischaemia-related angiogenesis. Intriguingly, Del-1–deficient mice displayed increased neovascularisation in two independent ischaemic models (retinopathy of prematurity and hind-limb ischaemia), as compared to Del-1–proficient mice. On the contrary, angiogenic sprouting in vitro or ex vivo (aortic ring assay) and physiological developmental retina angiogenesis were not affected by Del-1 deficiency. Mechanistically, the enhanced ischaemic neovascularisation in Del-1-deficiency was linked to higher infiltration of the ischaemic tissue by CD45+ haematopoietic and immune cells. Moreover, Del-1-deficiency promoted β2-integrin–dependent adhesion of haematopoietic cells to endothelial cells in vitro, and the homing of hematopoietic progenitor cells and of immune cell populations to ischaemic muscles in vivo. Consistently, the increased hind limb ischaemia-related angiogenesis in Del-1 deficiency was completely reversed in mice lacking both Del-1 and the β2-integrin LFA-1. Additionally, enhanced retinopathy-associated neovascularisation in Del-1-deficient mice was reversed by LFA-1 blockade. Our data reveal a hitherto unrecognised function of endogenous Del-1 as a local inhibitor of ischaemia-induced angiogenesis by restraining LFA-1–dependent homing of pro-angiogenic haematopoietic cells to ischaemic tissues. Our findings are relevant for the optimisation of therapeutic approaches in the context of ischaemic diseases.

Supplementary Material to this article is available online at www.thrombosis-online.com.