bluuebunny/crossref_metadata_2025 huggingface dataset:
Retrospective analysis of cases of intraoperative awareness in a large multi-hospital health system reported in the early postoperative period
Authors: Deis Amanda S. , Schnetz Michael P. , Ibinson James W. , Vogt Keith M.
Published: March 2020

Background

Awareness with recall under general anesthesia remains a rare but important issue that warrants further study.

Methods

We present a series of seven cases of awareness that were identified from provider-reported adverse event data from the electronic anesthesia records of 647,000 general anesthetics.

Results

The low number of identified cases suggests an under-reporting bias. Themes that emerge from this small series can serve as important reminders to anesthesia providers to ensure delivery of an adequate anesthetic for each patient. Commonalities between a majority of our identified anesthetic awareness cases include: obesity, use of total intravenous anesthesia, use of neuromuscular blockade, and either a lack of processed electroencephalogram (EEG) monitoring or documented high depth of consciousness index values. An interesting phenomenon was observed in one case, where adequately-dosed anesthesia was delivered without technical issue, processed EEG monitoring was employed, and the index value suggested an adequate depth of consciousness throughout the case.

Conclusions

Provider-reported adverse event data in the immediate post-operative period are likely insensitive for detecting cases of intraoperative awareness. Though causation cannot firmly be established from our data, themes identified in this series of cases of awareness with recall under general anesthesia provide important reminders for anesthesia providers to maintain vigilance in monitoring depth and dose of anesthesia, particularly with total intravenous anesthesia.
Serum α-synuclein and IL-1β are increased and correlated with measures of disease severity in children with epilepsy: potential prognostic biomarkers?
Authors: Choi Jieun , Kim Soo Yeon , Kim Hunmin , Lim Byung Chan , Hwang Hee , Chae Jong Hee , Kim Ki Joong , Oh Sohee , Kim Eun Young , Shin Jeon-Soo
Published: March 2020

Background

The search for noninvasive biomarkers of neuroinflammation and neurodegeneration has focused on various neurological disorders, including epilepsy. We sought to determine whether α-synuclein and cytokines are correlated with the degree of neuroinflammation and/or neurodegeneration in children with epilepsy and with acquired demyelinating disorders of the central nervous system (CNS), as a prototype of autoimmune neuroinflammatory disorders.

Methods

We analyzed serum and exosome levels of α-synuclein and serum proinflammatory and anti-inflammatory cytokines among 115 children with epilepsy and 10 acquired demyelinating disorders of the CNS and compared to 146 controls. Patients were enrolled prospectively and blood was obtained from patients within 48 h after acute afebrile seizure attacks or relapse of neurological symptoms. Acquired demyelinating disorders of the CNS include acute disseminated encephalomyelitis, multiple sclerosis, neuromyelitis optica spectrum disorders, and transverse myelitis. The controls were healthy age-matched children. The serum exosomes were extracted with ExoQuick exosome precipitation solution. Serum α-synuclein levels and serum levels of cytokines including IFN-β, IFN-γ, IL-1β, IL-6, IL-10 and TNF-α were measured using single and multiplex ELISA kits. Data were analyzed and compared with measures of disease severity, such as age at disease onset, duration of disease, and numbers of antiepileptic drug in use.

Results

Serum α-synuclein levels were significantly increased in patients with epilepsy and acquired demyelinating disorders of the CNS compared to controls (both,p < 0.05) and showed correlation with measures of disease severity both in epilepsy (p < 0.05,r = 0.2132) and in acquired demyelinating disorders of the CNS (p < 0.05,r = 0.5892). Exosome α-synuclein showed a significant correlation with serum α-synuclein (p < 0.0001,r = 0.5915). Serum IL-1β levels were correlated only with the numbers of antiepileptic drug used in children with epilepsy (p < 0.001, r = 0.3428), suggesting drug resistant epilepsy.

Conclusions

This is the first study in children demonstrating that serum α-synuclein levels were significantly increased in children with epilepsy and with acquired demyelinating disorders of the CNS and correlated with measures of disease severity. Serum IL-1β levels showed significant correlation only with drug resistance in children with epilepsy. Thus, these data support that serum levels of α-synuclein and IL-1β are potential prognostic biomarkers for disease severity in children with epilepsy.

CNS, central nervous system.
Incidence trend of type 1 diabetes mellitus in Serbia
Authors: Vojislav Ciric , Natasa Rancic , Milica Pesic , Slobodan Antic , Radivoj Kocic , Danijela Radojkovic , Sasa Radenkovic
Published: March 2020

Background

The incidence of type 1 diabetes mellitus (T1DM) increased worldwide.

The objective of the paper was to compare the incidence trend of T1DM in children and adolescents aged 0–19 and in adults under 30 years of age in Serbia from 2006 to 2017. Additional aim was to compare incidence rates of T1DM and type 2 diabetes mellitus (T2DM) among adults aged 20–24 and 25–29 years of age.

Methods

Trends and annual percentage change (APC) of the incidence rate with corresponding 95% confidence intervals (CI) were calculated by Joinpoint Regression Analyses.

Results

We found a significant increase of incidence in children aged 5–9 with the APC of 5.7% (95%CI: 2.3–9.1), and in children aged 10–14 with the APC of 2.1% (95%CI: 0.6–3.6). A significant decrease of incidence was determined in adolescents aged 15–19 with the APC -4.9% (95%CI: − 8.9 to – 0.7) and in adults aged 25–29 with the APC -7.3% (95%CI: − 12.5 to − 1.8).

Conclusion

The increase of incidence in children aged 0–14 and its decrease after 15 years of age showed that T1DM is predominantly a metabolic disease of children in Serbia. A significant increase in incidence was recorded in two age groups, namely 5–9 and 10–14 years of age. The highest increase was in children aged 5–9 and the highest incidence rate was in children aged 10–14. An insignificant increasing of T2DM incidence was observed in young adults aged 25–29. The increase in incidence rates in children, but not in young adults, suggests that the precipitating factors of children-onset disease may differ from those of adult-onset T1DM.
Improving medical certification of cause of death: effective strategies and approaches based on experiences from the Data for Health Initiative
Authors: Hart John D. , Sorchik Renee , Bo Khin Sandar , Chowdhury Hafizur R. , Gamage Saman , Joshi Rohina , Kwa Viola , Li Hang , Mahesh Buddhika P. K. , Mclaughlin Deirdre , Mikkelsen Lene , Miki Janet , Napulan Roderick , Rampatige Rasika , Reeve Matthew , Sarmiento Carmina , War Nang Su , Richards Nicola , Riley Ian D. , Lopez Alan D.
Published: March 2020

Background

Accurate and timely cause of death (COD) data are essential for informed public health policymaking. Medical certification of COD generally provides the majority of COD data in a population and is an essential component of civil registration and vital statistics (CRVS) systems. Accurate completion of the medical certificate of cause of death (MCCOD) should be a relatively straightforward procedure for physicians, but mistakes are common. Here, we present three training strategies implemented in five countries supported by the Bloomberg Philanthropies Data for Health (D4H) Initiative at the University of Melbourne (UoM) and evaluate the impact on the quality of certification.

Methods

The three training strategies evaluated were (1) training of trainers (TOT) in the Philippines, Myanmar, and Sri Lanka; (2) direct training of physicians by the UoM D4H in Papua New Guinea (PNG); and (3) the implementation of an online and basic training strategy in Peru. The evaluation involved an assessment of MCCODs before and after training using an assessment tool developed by the University of Melbourne.

Results

The TOT strategy led to reductions in incorrectly completed certificates of between 28% in Sri Lanka and 40% in the Philippines. Following direct training of physicians in PNG, the reduction in incorrectly completed certificates was 30%. In Peru, the reduction in incorrect certificates was 30% after implementation and training on an online system only and 43% after training on both the online system and basic medical certification principles.

Conclusions

The results of this study indicate that a variety of training strategies can produce benefits in the quality of certification, but further improvements are possible. The experiences of D4H suggest several aspects of the strategies that should be further developed to improve outcomes, particularly key stakeholder engagement from early in the intervention and local committees to oversee activities and support an improved culture in hospitals to support better diagnostic skills and practices.
Optimization of perioperative approaches for advanced and late stages of gastric cancer: clinical proposal based on literature evidence, personal experience, and ongoing trials and research
Authors: Beeharry Maneesh Kumarsing , Zhang Tian Qi , Liu Wen Tao , Gang Zhu Zheng
Published: March 2020

Background

The high incidence of gastric cancer (GC) and paradoxical high prevalence of advanced stage GC, amounting to around 2/3 at time of diagnosis, have urged doctors and researchers around the world not only to ameliorate the detection rate of GC at early stages but also to optimize the clinical management of GC at advanced stages.

Content

We hereby recommend a more goal-oriented multimodality approach with objectives to increase survival rate and improve survival status. Based on precision and accurate clinical staging at diagnosis, we suggest that advanced stage GC (AGC) patients should be channeled into different treatment plans according to their disease status where they can be subjected to comprehensive measures involving chemo, radio, immunological, or target therapies depending on the pathophysiological behavior of their tumor. Patients assessed as potentially resectable cT4N + M0 can undergo neoadjuvant chemotherapy with intent of tumor downsizing and downgrading followed by surgery with intraoperative hyperthermic intraperitoneal chemotherapy (HIPEC) to decrease the incidence of peritoneal dissemination due to surgical trauma and adjuvant chemotherapy and radiation in cases of bulky nodal metastasis. In cases with distal metastasis, conversion therapy is recommended with the possibility of surgery of curative intent in case of favorable response. The options of alternate treatment options such as trans-catheter arterial chemoembolization (TACE) for limited liver lesions or neoadjuvant intraperitoneal plus systemic chemotherapy (NIPS) for peritoneal carcinomatosis have to be negotiated. With surgery as the cornerstone for cancer treatment, there is acknowledgment of the significance of perioperative comprehensive approaches but there has not been some consensus guiding clinical application. Henceforth, in this review, based on past literature, current guidelines and ongoing clinical trials, we have shared a proposal of the current treatment modalities in practice for the advanced stages of gastric cancer.

Conclusion

Even though surgery is the golden standard of radical cancer treatment, clinical reality shows that without proper perioperative management, patients undergoing radical resections manifest high rates of recurrence and metastasis. Hence, in this review, we have outlined a clinical agenda to optimize the management of advanced stage GC with objective to improve survival outcome and quality of life of patients.
DJ-1 exerts anti-inflammatory effects and regulates NLRX1-TRAF6 via SHP-1 in stroke
Authors: Peng Li , Zhou Yang , Jiang Ning , Wang Tingting , Zhu Jin , Chen Yanlin , Li Linyu , Zhang Jinyan , Yu Shanshan , Zhao Yong
Published: March 2020

Background

Acute inflammation induced by reactive astrocytes after cerebral ischemia/reperfusion (I/R) injury is important for protecting the resultant lesion. Our previous study demonstrated that DJ-1 is abundantly expressed in reactive astrocytes after cerebral I/R injury. Here, we show that DJ-1 negatively regulates the inflammatory response by facilitating the interaction between SHP-1 and TRAF6, thereby inducing the dissociation of NLRX1 from TRAF6.

Methods

We used oxygen-glucose deprivation/reoxygenation (OGD/R) in vitro in primary astrocyte cultures and transient middle cerebral artery occlusion/reperfusion (MCAO/R) in vivo to mimic I/R insult.

Results

The inhibition of DJ-1 expression increased the expression of the inflammatory cytokines TNF-α, IL-1β, and IL-6. DJ-1 knockdown facilitated the interaction between NLRX1 and TRAF6. However, the loss of DJ-1 attenuated the interaction between SHP-1 and TRAF6. In subsequent experiments, a SHP-1 inhibitor altered the interaction between SHP-1 and TRAF6 and facilitated the interaction between NLRX1 and TRAF6 in DJ-1-overexpressing astrocytes.

Conclusion

These findings suggest that DJ-1 exerts an SHP-1-dependent anti-inflammatory effect and induces the dissociation of NLRX1 from TRAF6 during cerebral I/R injury. Thus, DJ-1 may be an efficacious therapeutic target for the treatment of I/R injury.
High-resolution modeling of the selection on local mRNA folding strength in coding sequences across the tree of life
Authors: Peeri Michael , Tuller Tamir
Published: March 2020

Background

mRNA can form local secondary structure within the protein-coding sequence, and the strength of this structure is thought to influence gene expression regulation. Previous studies suggest that secondary structure strength may be maintained under selection, but the details of this phenomenon are not well understood.

Results

We perform a comprehensive study of the selection on local mRNA folding strengths considering variation between species across the tree of life. We show for the first time that local folding strength selection tends to follow a conserved characteristic profile in most phyla, with selection for weak folding at the two ends of the coding region and for strong folding elsewhere in the coding sequence, with an additional peak of selection for strong folding located downstream of the start codon. The strength of this pattern varies between species and organism groups, and we highlight contradicting cases.

To better understand the underlying evolutionary process, we show that selection strengths in the different regions are strongly correlated, and report four factors which have a clear predictive effect on local mRNA folding selection within the coding sequence in different species.

Conclusions

The correlations observed between selection for local secondary structure strength in the different regions and with the four genomic and environmental factors suggest that they are shaped by the same evolutionary process throughout the coding sequence, and might be maintained under direct selection related to optimization of gene expression and specifically translation regulation.
Comparative analysis of structural and functional aspects of phytoene synthase from Meiothermus taiwanensis strain RP
Authors: Mukherjee Trinetra , Mukhopadhyay Subhra Kanti
Published: March 2020

Purpose

The aim of this study is the in silico characterization of the structure and function of the phytoene synthase (PSY) of a red carotenoid producing thermophile Meiothermus taiwanensis strain RP with a comparative approach.

Methods

PSYs from M. taiwanensis strain RP and other groups of thermophilic, mesophilic and psychrophilic bacteria, plants, protozoa, and algae were analyzed by ExPASy ProtParam, NCBI Conserved Domain Search, SOPMA, PSIPRED, Robetta server, ProQ, and QMEAN, with the superposition of 3-D structures in PyMOL.

Results

RP PSY shows the highest (97.5 %) similarity with M. ruber and the lowest with the psychrophile Gelidibacter algens (36.7 %). The amino acid sequence of RP PSY is one of the shortest, with 275 residues. The instability index of RP PSY is much lower compared with plant sequences. Alanine, arginine, glycine, and leucine residues are the highest in Meiothermus sp., and they have a high amount of alpha-helix. Most of the 32 active site residues are conserved in all the sequences. However, some residues are more prone to substitutions in other PSYs except M. ruber. The three-dimensional structures of M. taiwanensis strain RP, Gelidibacter algens, Thermus thermophiles, Meiothermus ruber, and Brassica napus PSYs were homology modeled, validated, and submitted to Protein Model Databank. The superposition of the 3-D structures shows that their active site region structure is identical.

Conclusion

RP PSY is one of the most stable PSYs and knowledge of its individual properties, similarities, and dissimilarities with other PSYs may be useful for genetic engineering and purification of the protein for improved carotenoid production.
Quantum Hermite–Hadamard inequality by means of a Green function
Authors: Adil Khan Muhammad , Mohammad Noor , Nwaeze Eze R. , Chu Yu-Ming
Published: March 2020

The purpose of this work is to present the quantum Hermite–Hadamard inequality through the Green function approach. While doing this, we deduce some novel quantum identities. Using these identities, we establish some new inequalities in this direction. We contemplate the possibility of expanding the method, outlined herein, to recast the proofs of some known inequalities in the literature.
Effects of school climate and teacher self-efficacy on job satisfaction of mostly STEM teachers: a structural multigroup invariance approach
Authors: Zakariya Yusuf F.
Published: March 2020

Background

Identification and retention of effective teachers in STEM education play cardinal roles in teacher recruitment exercises worldwide. Studies on factors that characterize effective teachers have therefore gained popularity in recent times. Teacher self-efficacy, job satisfaction and school climate are among other factors that have attracted global attention. Thus, proper understanding of the relations between these factors is equally important. The purpose of this study is to validate and cross-validate a model of direct/indirect effects of school climate and teacher self-efficacy on job satisfaction.

Results

The data used for the current study are extracted from a publicly available data of Teaching and Learning International Survey (TALIS) 2018 survey. Structural equation modeling approach was used in the analyses coupled with robust maximum likelihood to ensure accurate estimations in the models. The results of the validated models show a strong direct impact of school climate on job satisfaction, a direct impact of teacher self-efficacy on job satisfaction and a mediating effect of teacher self-efficacy between school climate and job satisfaction. This model exhibits structural invariance in factor loadings, intercepts and regression weights across two independent samples from a population of 3951 lower secondary school teachers in Norway.

Conclusion

The findings of this study do provide empirical evidence for the relations between teacher self-efficacy, job satisfaction and school climate among Norwegian lower secondary school teachers. The cross-validation of these relations was also established using an independent sample to enhance generalization of the findings. Two methodological observations concerning recoding of some items as well as an addition of item cross-loading in the measurement model of the job satisfaction scales are raised and addressed. It is therefore recommended that researchers who will be using TALIS 2018 data should take note of these observations.
Intravenous leiomyomatosis with intracardiac extension managed by one-stage surgical excision: a case presentation
Authors: Bhoil Rohit , Thakur Vijay , Verma Yashwant Singh , Bhoil Sabina , Tomar Ashwani , Pathania Rajnish , Sharma Sanjeev
Published: March 2020

Background

Intravenous leiomyomatosis is characterized by the extension of pathologically benign smooth muscle tumor into venous channels. Its extension into the heart is a very rare occurrence, which may lead to heart failure and occasionally sudden death. Due to its rarity, the condition is poorly understood and often misdiagnosed.

Case presentation

We report a case of intravenous leiomyomatosis with intracardiac extension in a 40 years old woman who underwent hysterectomy for uterine leiomyoma (one year earlier), managed successfully through one-stage surgical approach with main focus on its imaging features.

Conclusion

The diagnosis of intravenous leiomyomatosis should be considered in women who present with cardiac symptoms associated with a right atrial mass and who also have a pelvic mass or have previously undergone hysterectomy because of uterine leiomyoma.
p53 expression in breast cancer is more likely to predict survival in African-American than Hispanic or white women
Authors: Dookeran K. A. , Ferrer K. , Sekosan M. , Gao X. , Wang Y. , De La Torre R. , Lukaszczyk B. , Diaz de Leon L. , Roman G. , Zaren H. A.
Published: March 2020

10001

Background: Abnormal nuclear accumulation of p53 protein may predict poorer survival & aggressive tumor biology in African-American (AA) women with breast cancer. We hypothesized that compared to Hispanics or whites, survival would be worse for AA women with p53 positive tumors. Methods: Tumors from 195 consecutive AA, 46 Hispanic & 52 white women were evaluated for p53 expression with IHC, using antibody Pab1801 on paraffin embedded tissue, at the SHCC MBCCOP, in Chicago. Staining was scored on the UK Receptor Group 0–8 system with score ≥2 read as positive. Survival was computed by the Kaplan-Meier method & correlated with p53 expression. Univariate analyses were done with two-sided log-rank test & multivariate analysis was done with Cox proportional-hazards model. Results: There were no significant differences between racial groups regarding tumor size, grade, node status, stage, estrogen receptor (ER) status, treatment & p53 expression. Hispanics were significantly younger (p=0.014). For AA, Hispanic & white women respectively, p53 expression was 29.2%, 34.8% & 18.8%. High grade (p=0.0001) & negative ER status (p=0.006) were significantly associated with p53 expression in AA women; similar results were seen in whites (p=0.014 & p=0.031 respectively), however although the trend for grade was similar, only negative ER status was significantly associated with p53 expression in Hispanics. AA women with p53 positive tumors had significantly worse survival compared to AA women with p53 negative tumors (Hazard Ratio=1.945; 95% CI=1.040–3.638; p=0.037). However, comparison of survival between races for women with p53 positive tumors (p=0.793) and also for women with p53 negative tumors (p=0.4027) was not different. Multivariate analysis demonstrated that there was no interaction (p=0.523) between p53 expression & stage as predictors of poor survival in AA women. Conclusions: Analyses between racial groups demonstrated that AA, Hispanic & white women with breast tumors with abnormal p53 expression had similar associations with high tumor grade, negative ER status and survival. However within racial groups, abnormal p53 expression was only able to predict significantly worse survival in AA women. This association appeared to be independent of stage.

[Table: see text]
ERCC1 and RRM1 but not EGFR gene expression is predictive of shorter survival in advanced non-small cell lung cancer (NSCLC)
Authors: Selvaggi G. , Ceppi P. , Volante M. , Saviozzi S. , Novello S. , Calogero R. , Papotti M. , Scagliotti G. V.
Published: March 2020

10030

Background: Pivotal studies indicate a role of excision repair cross-complementation 1 (ERCC1) and ribonucleotide reductase M1 (RRM1) in conferring a differential sensitivity to cytotoxic chemotherapy and epidermal growth factor receptor (EGFR) has been recently deeply investigated in NSCLC. Methods: We retrospectively collected 70 formalin-fixed paraffin-embedded (FFPE) bronchoscopic/fine needle aspiration biopsies of NSCLC to investigate the expression levels of ERCC1, RRM1 and EGFR by Real-Time PCR (Lord R et al. Clin Cancer Research 2002, 8:2286–91). Results were correlated with survival using the Kaplan-Meier method. Results: Sixty-one (87%) specimens were successfully amplified. Median age was 62 years (range 26–75), male/ female ratio 44/17, stage III/IV 20/41; 43 patients received cisplatin-based chemotherapy; overall median survival (MS) was 13.3 months over a median follow-up time of 45 months. ERCC1 expression level ranged from 0.70 to 15.12, RRM1 0.60–17.82. By adopting cut-off values according to median expression levels, we found a strong correlation between ERCC1 and RRM1 mRNA levels (r=0.410; p<0.001). MS in patients with low ERCC1 was significantly longer (16.9 vs 11.3 months, p<0.006) as well as in patients with low RRM1 (13.9 vs 10.9 months, p<0.03). Concomitant high expression levels of ERCC1 and RRM1 (n=26) are predictive of a worse outcome (13.9 vs 10.9 months, p<0.05). Among patients treated with cisplatin-based regimens, low ERCC1 levels were also predictive of a significantly longer MS (23.0 vs 11.6 months, p<0.002). A lower median ERCC1 level (3.2 vs 4.7) and a correlation with a better outcome were also observed in females vs males. No correlation between gene expression levels and histology was reported. No significant correlation between EGFR expression levels (range 0.5–85.8) and survival was found, even when different cut-off values were tested. Conclusions: This retrospective study further validates ERCC1 and RRM1 as good candidates genes to customize chemotherapy. Prospective studies based on the selection of patients according to genes expression levels are a research priority in early and advanced stages of NSCLC.

[Table: see text]
Immature myeloid cell (ImC)- mediated immunosuppression in advanced renal cell cancer (RCC)
Authors: Kuebler H. R. , Dannull J. , Tseng T. Y. , Zhang A. , Su Z. , Dahm P. , Vieweg J.
Published: March 2020

10042

Background: RCC affects the immunsystem by inhibiting the process of differentiation of antigen-resenting cells from their myeloid precursors by secreting tumor-derived factors such as VEGF. ImC induce a profound state of immune suppression that foster tumor tolerance by inhibition of tumor specific T cells. In this study, we asked whether increased ImC frequencies can be found in peripheral blood from RCC patients and if ImC frequencies correlated positively with increased levels of tumor-derived serum markers. Methods: ImC frequencies from healthy volunteers and RCC patients were determined by FACS. ImC were isolated by magnetic bead separation techniques and their immunosuppressive activity was determined in IFN-γ ELISPOT analyses, CTL and T-cell proliferation assays. The production of reactive oxygen species by ImC was analyzed utilizing specific probes and inhibitors. Tumor-derived serum markers were quantified by ELISA, enzyme immune assays and cytometric bead arrays. Microarray analyses were performed to identify novel, highly specific ImC markers. Results: RCC patients demonstrate higher ImC frequencies (0.8 - 3.2% of total PBMC) compared to healthy donors. The increased ImC frequencies are positively correlated with serum levels of VEGF, PGE-2, IL-13 and M-CSF in RCC patients and the concentration of byproducts of oxidative burst, including iso-prostane and malondialdehyde, was significantly enhanced. Isolated ImC exhibited profound immunosuppressive effects on CTL and CD4+ T cell response in an antigen specific fashion. Immunosuppression by ImC was mediated by release of ROS, including peroxide and superoxide anions and by generation of nitric oxide radicals as evidenced in functional assays. Results from microarray analyses reveal EP-1, EphA5, and PEX-5 as novel ImC markers. Conclusions: These data suggest that RCC may induce the development of immunosuppressive ImC population through secretion of cytokines such as VEGF, IL-13, M-CSF and PGE-2. Inhibition of specific cytokine activity or the use of differentiating agents may represent strategies to decrease immunosuppressive ImC population. Results from a phase I clinical trial investigating the effects of all-trans retinoic acid (ATRA) on differentiation of ImC in RCC patients will be presented.

No significant financial relationships to disclose.
Detection of colorectal neoplasia associated K-Ras mutations in human urine
Authors: Brenner D. E. , Su Y. , Normolle D. P. , Syngal S. , Bresalier R. , Marcon N. , Baron J. , Block T.
Published: March 2020

1005

Background: Since colorectal neoplasia-associated genes have been detected in human blood, we hypothesized that small DNA fragments containing genetic mutations associated with colorectal neoplasias are filtered and excreted in the urine. If so, genes associated with colorectal cancer will be detected in the urine. K-ras mutations are commonly associated with colorectal neoplasia and do not occur in the urinary tract. Methods: K-ras mutation detection: 200 microl of urine was extracted with guanidine thiocyanate and purifed using a Wizard DNA isolation kit. Codon 12 K-ras mutation detection methods–1: restriction enriched PCR, 20 cycles, with primers that amplify both wild type and mutated DNA but with an artificial BstNI site at the 5’ end of the amplified product (>2 K-ras copies per assay); 2: Peptide nucleic acid clamping real time PCR (>15 K-ras copies per assay). Human subjects: Training set = 20 patients with known K-ras mutations in colorectal cancer tissue. Test set = blinded urine samples from colorectal adenocarcinoma (N=48), adenoma (N=31), hyperplastic polyp (N=12) and endoscopically normal colon and rectum (N=60). Results: 1. Human urine contains 150–250 base pair DNA fragments derived from the circulation. These fragments can be readily distinguished from high molecular weight DNA from sloughed urinary tract cells. 2. Training set for K-ras detection (tissue confirmed K-ras mutations): Serum 6/20 (30%), Plasma (11/20 (55%), Urine: 18/20 (90%), (p<0.15 for plasma, p<0.001 for serum). 3. Test set (blinded): a. >2 copies of mutated K-ras genes were detected in: 16/48 (33%) adenocarcinomas; 23/31 (74%) adenomas; 5/12 (42%) hyperplastic polyps, and 19/60 (31%) non-neoplasia controls. b. >15 copies of mutated K-ras genes were detected in: 12/48 (25%) adenocarinomas; 15/31 (48%) adenomas; 3/12 (25%) hyperplastic polyps, and 11/60 (18%) non-neoplasia controls. Conclusions: Small DNA fragments in human urine contain K-Ras mutations identical to those found in colorectal cancer DNA. The sensitivity for detection of K-ras mutations in urine appears equivalent or superior to K-ras mutation detection in feces or serum. DNA mutations from systemic epithelial neoplasias may be detected in filtered urinary DNA fragments and may be useful for early detection of neoplasia.

No significant financial relationships to disclose.
Lung cancer resection does not change characteristic serum proteomic profiles
Authors: Ward D. G. , Alzetani A. , Starczynski J. , Kaur G. , Trotter S. , James N. , Wakelam M. J. , Wei W. , Billingham L. J. , Johnson P. J. , Martin A.
Published: March 2020

10051

Background: Surface-enhanced laser desorption/ionisation time of flight mass spectrometry (SELDI-TOF-MS) has been proposed as a method for determining characteristic serum proteomic profiles that permit discrimination between cancer and non-cancer patients. The aim of this study was to test in a group of lung cancer patients the hypothesis, that these profiles ‘return to normal’ following tumour resection, as would be expected if they behave as conventional serological tumour makers. Methods: Serum samples were collected from 39 patients with non-small cell lung cancer, 1 day pre-surgery, 1 day post-surgery and, for 17 of these, approximately three months later; samples were also obtained from 38 non-cancer control subjects. All samples were subjected to SELDI-TOF-MS analysis on CM10 chips. Students t-tests were used to identify differentially expressed peaks in the profiles. Results: The SELDI profiles of the lung cancer sera compared to non-lung-cancer controls showed 19 significantly differentially expressed peaks (p<0.01), 14 of which were used to develop a neural network capable of discriminating between the two groups. Analysis of proteomic features showed that despite the absence of cancer in patients 3-months post-surgery, no statistically significant differences could be detected between the pre-treatment and 3-month post-operative samples from the same patients, including the 19 features that were found to discriminate between lung and non-lung cancer. One of the proteins significantly decreased in the serum of lung cancer patients was identified as transthyretin. Conclusions: SELDI can detect differentially expressed protein peaks characteristic of lung cancer compared to non-lung cancer patients but following surgical resection of the cancer, neither the pattern nor the individual peaks revert to the levels observed in non-lung cancer patients. The hypothesis that such differences would revert to normal after tumour removal was not supported. Work funded by Cancer Research UK

No significant financial relationships to disclose.
Detection of clinically significant mutations in the epidermal growth factor receptor missed by direct sequencing using a highly sensitive DNA endonuclease
Authors: Borras A. M. , Kuang Y. , Rogers A. M. , Holmes A. J. , Gallegos Ruiz M. , Joshi V. A. , Distel R. J. , Giaccone G. , Taron M. , Janne P. A.
Published: March 2020

10054

Background: Mutations in the epidermal growth factor receptor (EGFR) are associated with sensitivity and resistance to EGFR inhibitors gefitinib and erlotinib in patients with non-small cell lung carcinoma (NSCLC). Direct sequencing is currently used for mutation detection but sensitivity is limited and requires dissection to obtain a relatively pure population of tumor cells. We examined a DNA endonuclease, SURVEYOR, which cleaves mismatched heteroduplexed DNA, as a more sensitive method for EGFR mutation screening and compared it to direct sequencing. Methods: EGFR exons 18–21 from tumor DNA were amplified using PCR, digested with SURVEYOR, and the products analyzed by HPLC. Specimens that produced digestion products were re-analyzed by size separation or by denaturing HPLC followed by fractionation and sequencing. Tumor specimens from 191 NSCLC patients were analyzed: 61 frozen tumors specimens; 91 dissected formalin fixed paraffin embedded (FFPE) and 39 un-dissected FFPE tumor specimens from patients treated with gefitinib or erlotinib in whom clinical outcome was available. 173 specimens were independently analyzed by direct sequencing. Results: We detected 48 EGFR mutations by sequencing and 61 using SURVEYOR. All EGFR mutations identified by sequencing, including those using un-dissected tumor specimens, were detected by SURVEYOR and none were missed (sensitivity: 100%, negative predictive value: 100%). 13 mutations were detected by SURVEYOR not detected by sequencing. This included 5 mutations (4 exon 19 deletions; 1 L858R) in 7 (71%) patients who clinically had a PR to gefitinib or erlotinib but who were wild type by sequencing. In 4 patients, 2 with clinical acquired resistance to gefitinib, a T790M mutation was found which was undetected by sequencing. Conclusions: SURVEYOR analysis is a more sensitive method for EGFR mutation detection than direct sequencing. It can be used to detect EGFR mutations from un-dissected tumor specimens and can detect clinically significant activating or resistance associated EGFR mutations not detected by direct sequencing.

[Table: see text]
Molecular network analysis of signaling pathways in metastatic breast cancer in patients treated with erlotinib
Authors: Speer R. , Wallwiener D. , Yang S. X. , Swain S. M. , Wulfkuhle J. D. , Liotta L. A. , Petricoin E. F.
Published: March 2020

10064

Erlotinib, an epidermal growth factor receptor tyrosine kinase inhibitor, is under active clinical investigation. It may have profound effects on the cellular molecular network by affecting downstream signaling and inhibiting tumor cell proliferation. Knowledge about the specificity of drug effects on the signaling pathways might enable therapy tailored specifically for each patient’s defect. We examined the activation of cellular signaling pathways involved in cell survival and proliferation in response to erlotinib treatment using a biopsy study set that comprised patient-matched breast cancer metastases from 8 women obtained prior to treatment with erlotinib and after 1 month of therapy. We procured pure cancer cell lysates from very small amounts of frozen tissue by Laser Capture Microdissection (about 60,000 cells). Matching lysates were printed on nitrocellulose and incubated with antibodies against the endpoints of interest. Using the Reverse Phase Protein Microarray technology we were able to generate a portrait of 18 phosphorylated kinase substrate endpoints as well as total expression levels of ERK, EGFR, Her2/neu, and IGF1R that revealed an upregulation of pro-survival pathway signaling in the post-erlotinib treatment set. Especially the measurement of the pro-survival, apoptosis-inhibiting signal transduction pAkt (S473) protein alone showed a statistically significant stronger staining in post-erlotinib treatment samples (p=0.019) in each of the individual patient-matched specimens as well as the entire post-treatment group, showing concordance with the clinical data of the trial, i.e. progressive disease observed for all participating patients. The results indicate the feasibility of this approach to monitor signaling events and molecular pathways in clinical specimens taken during targeted therapy trials.

No significant financial relationships to disclose.
An AS04-containing human papillomavirus (HPV) 16/18 vaccine for prevention of cervical cancer is immunogenic and well-tolerated in women 15–55 years old
Authors: Schwarz T. F.
Published: March 2020

1008

Background: Genital HPV infections can be acquired shortly after sexual debut, and the risk remains throughout a sexually active woman’s lifetime. In women 15–25 years of age, the AS04-containing HPV vaccine was highly immunogenic and conferred 100% protection against HPV-16/18 persistent infection and associated cervical lesions up to 27 months. In the long-term follow-up of this study, sustained vaccine efficacy has been observed up to 48 months. The present phase III study (580299/014) assessed immune responses to the AS04-containing HPV-16/18 vaccine in women 26–55 years old compared with women 15–25 years old. Methods: Healthy women in Germany and Poland between 15 and 55 years of age received 3 doses of HPV-16/18 AS04-containing vaccine at months 0, 1, and 6. The groups were age stratified: 15–25 (n=229), 26–45 (n=226), and 46–55 (n=211). Anti-HPV-16/18 antibody titers were assessed at months 0, 2, 7, and 12 by ELISA (EU/mL). Seropositivity rates and geometric mean antibody titers (GMTs) were calculated for all groups. Safety was assessed after each dose in all participants. Results: All initially seronegative women became seropositive for both HPV 16 and 18 at Month 2. At Month 7, HPV-16 GMTs (95% CI) were 7908.4 (6874.0–9098.5) in 15–25 year olds, 4029.2 (3402.7–4771.0) in 26–45 year olds, and 2566.8 (2181.2–3020.6) in 46–55 year olds. For HPV-18, GMTs were 3499.3 (3098.7–3951.6) in 15–25 year olds, 1837.3 (1602.1–2107.0) in 26–45 year olds, and 1313.0 (1145.6–1504.9) in 46–55 year olds. Overall the vaccine was well-tolerated, and the incidence of local symptoms (within 30 days) tended to be lower in the 46–55 year-old group (69.2% versus 81.6% [26–45] and 85.7% [15–25]). Conclusions: An AS04-containing HPV-16/18 vaccine was immunogenic and generally safe in 15–55 year-old females. As observed with other vaccines, GMTs decreased with age, however, the Month 7 postvaccination antibody levels in the oldest age group (46–55) were still 3–4 times higher than those observed during a separate long-term follow-up study where sustained efficacy has been observed up to 48 months.

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Shc test as a strong prognostic indicator of disease outcome in early stage gastric cancer
Authors: Wanebo H. J. , Frackelton A. R. , Hafer L. J. , Bagdasaryan R. , Lis R. , Sabo E. , Resnick M. B.
Published: March 2020

10090

Background: Treatment planning for gastric cancer is primarily based on clinical staging of disease. Markers predicting likelihood of disease outcome would help guide treatment decisions, especially for early stage disease. The Shc proteins, implicated in many aggressive cancers, and measured in tumor specimens by the immunohistochemical (IHC) Shc Test, have shown strong ability to predict disease outcome in breast cancer. We report here that the Shc Test is a strong prognostic indicator of disease outcome in early stage gastric cancer. Methods: Histopathology was examined in one hundred and seventeen (117) primary gastric cancer patient samples from Rhode Island Hospital in tissue microarray format (21 disease recurrences; 63 disease-specific deaths; average follow-up of 2.7 yrs). IHC staining of the Shc proteins was independently scored on a 0–5 scale by two pathologists, blinded to patient information. Results: Stage I or II gastric cancers (n=62) could be clearly separated at a cutpoint of 1.1 on a 0–5 scale, into good prognosis (16% 4yr relapse risk; demonstrating high PY-Shc) and poor prognosis (46% 4yr relapse risk; showing low PY-Shc) (log-rank, P=0.003). p66 Shc showed similar prognostic abilities. High PY-Shc in patients with early stage disease showed a significant protective effect on overall survival (P=0.003) by univariate log rank analysis. As a continuous variable, PY-Shc had a strong predictive ability (HR = 0.09, P=0.055) that approached significance. By univariate Cox proportional hazards, patients with high PY-Shc had a 5-fold reduction in disease specific death (DSD) compared to patients with low PY-Shc (P=0.002). By multivariate Cox proportional hazards, adjusted for grade, stage, chemotherapy and radiation therapy, only PY-Shc (HR = 0.22, P=0.015) and Intestinal tumor type (HR = 0.38, P=0.046) remained as significant predictors of survival. Conclusions: The Shc Test shows a strong prognostic ability to stratify early stage gastric cancer patients by risk, making it a valuable tool in selecting therapy for these patients.

[Table: see text]
PAI-1 (Plasminogen activator inhibitor-1) expression levels in colorectal cancers and corresponding normal tissues
Authors: Sakakibara T. , Hibi K. , Koike M. , Fujiwara M. , Kodera Y. , Nakao A.
Published: March 2020

10094

Background: We performed a quantitative RT-PCR for Plasminogen Activator Inhibitor-1 (PAI-1) gene in 55 surgical specimens of colorectal cancer (CRC) and corresponding normal mucosa. To test the hypothesis that PAI-1 may serve as a candidate marker for the malignant potential of CRC, we evaluated the possible relationship between PAI-1 gene expression levels and clinicopathological findings in CRC. Next, to investigate the mechanism of PAI-1 overexpression in CRC, we compared the PAI-1 expression levels in these cancers with those in the corresponding normal tissues. Results: A significant increase in PAI-1 expression scores was observed in node-positive CRCs (2.19Å}0.43) compared to negative ones (0.35Å}0.42) (P=0.0037) as well as in CRCs with distant metastasis (3.50Å}1.18) compared to negative ones (0.99Å}0.30). A significant increase in PAI-1 expression scores was observed in lymph node metastasis-positive CRCs (2.19Å}0.43) compared to negative ones (0.35Å}0.42) (P=0.0037) as well as in distant metastasis-positive CRCs (3.50Å}1.18) compared to negative ones (0.99Å}0.30). The PAI-1 expression score markedly increased with the tumor stage (P=0.0063; ANOVA test). Moreover, multivariate analysis revealed the PAI-1 expression score to be a strong and independent prognostic factor for CRC (P=0.0432). There was a significant correlation between CRC and corresponding normal PAI-1 expressions with a Spearman’s rank correlation coefficient of 0.81 (P < 0.0001). Conclusion: These results suggested that PAI-1 might serve as a new parameter for the prediction of prognoses in CRC. Taken together, PAI-1 overexpression in CRCs might originate from higher PAI-1 expression in the corresponding normal tissues and result in a malignant phenotype of these cancers.

No significant financial relationships to disclose.
Expression and function of the four and a half LIM-only protein 2 (FHL2) in breast cancer
Authors: Martin B. T. , Kleiber K. , Kaufmann M. , Strebhardt K.
Published: March 2020

10109

Background: FHL2 belongs to the expanding family of proteins that possess highly conserved double Zn-finger motifs, CX2CX16–23HX2CX2CX2CX16–21CX2 (C/H/D), and are involved in protein-protein interactions and transcriptional regulation. FHL2 was primarily described as “down regulated in rhabdomyosarcama cells LIM domain protein” (DRAL). Further studies also indicated a connection of FHL2 with the tumor biology. Its expression has been shown to be increased in tumors of epithelial and melanoma cell origin. A recent publication describes the interaction of FHL2 with the breast cancer susceptibility gene 1 (BRCA1) but to date little is known about the function of FHL2 in breast cancer. Methods: Tumor samples of 61 breast cancer patients and 8 normal breast tissues were lysed and subjected to Western blot analysis using a monoclonal FHL2-specific antibody; Tissue specimens of normal breast and breast cancer were histologically examined with the monoclonal antibody used for the Western blot analysis; Luciferase assays were carried out in MCF-7 breast cancer cells, using AP1-dependent reporter plasmids. Anchorage-independent growth was assayed by colony formation in soft agar. Results: FHL2 is overexpressed in almost all human breast cancer tissues tested but not in normal breast tissues and only low levels of FHL2 expression were present in four premalignant ductal carcinoma in situ (DCIS). Co-immunoprecipitation assays and GST-pull down experiments revealed that FHL2 interacts with the proto-oncogene c-jun in breast cancer cells. Reporter gene assays demonstrated that FHL2 represses c-jun-mediated activation of an idealized AP-1-regulated reporter gene in a dose-dependent manner. This repression was also seen on the naturally occurring AP-1-dependent IL2 promoter. The repression domain was identified within the N-terminal half of the FHL2 molecule. Finally, we show that stable expression of FHL2 leads to anchorage-independent growth of breast cancer cells in soft agar. Conclusions: Taken together, our data suggest that by modulating the activity of transcription factors such as c-jun, the expression of FHL2 in breast cancer might contribute to the malignant potential of breast tumors.

No significant financial relationships to disclose.