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ALK, ROS1, and RET fusions and MET∆ex14 variant associate with response to targeted therapies in non‐small‐cell lung cancer (NSCLC). Technologies for fusion testing in tissue must be adapted to liquid biopsies, which are often the only material available. In this study, circulating‐free RNA (cfRNA) and extracellular vesicle RNA (EV‐RNA) were purified from liquid biopsies. Fusion and MET∆ex14 transcripts were analyzed by nCounter (Nanostring) and digital PCR (dPCR) using the QuantStudio® System (Applied Biosystems). We found that nCounter detected ALK, ROS1, RET, or MET∆ex14 aberrant transcripts in 28/40 cfRNA samples from positive patients and 0/16 of control individuals (70% sensitivity). Regarding dPCR, aberrant transcripts were detected in the cfRNA of 25/40 positive patients. Concordance between the two techniques was 58%. Inferior results were obtained when analyzing EV‐RNA, where nCounter often failed due to a low amount of input RNA. Finally, results of dPCR testing in serial liquid biopsies of five patients correlated with response to targeted therapy. We conclude that nCounter can be used for multiplex detection of fusion and MET∆ex14 transcripts in liquid biopsies, showing a performance comparable with next‐generation sequencing platforms. dPCR could be employed for disease follow‐up in patients with a known alteration. cfRNA should be preferred over EV‐RNA for these analyses.

Abstract Background Drugs targeting the spindle assembly checkpoint (SAC), such as inhibitors of Aurora kinase B (AURKB) and dual specific protein kinase TTK, are in different stages of clinical development. However, cell response to SAC abrogation is poorly understood and there are no markers for patient selection. Methods A panel of 53 tumor cell lines of different origins was used. The effects of drugs were analyzed by MTT and flow cytometry. Copy number status was determined by FISH and Q-PCR; mRNA expression by nCounter and RT-Q-PCR and protein expression by Western blotting. CRISPR-Cas9 technology was used for gene knock-out (KO) and a doxycycline-inducible pTRIPZ vector for ectopic expression. Finally, in vivo experiments were performed by implanting cultured cells or fragments of tumors into immunodeficient mice. Results Tumor cells and patient-derived xenografts (PDXs) sensitive to AURKB and TTK inhibitors consistently showed high expression levels of BH3-interacting domain death agonist (BID), while cell lines and PDXs with low BID were uniformly resistant. Gene silencing rendered BID-overexpressing cells insensitive to SAC abrogation while ectopic BID expression in BID-low cells significantly increased sensitivity. SAC abrogation induced activation of CASP-2, leading to cleavage of CASP-3 and extensive cell death only in presence of high levels of BID. Finally, a prevalence study revealed high BID mRNA in 6% of human solid tumors. Conclusions The fate of tumor cells after SAC abrogation is driven by an AURKB/ CASP-2 signaling mechanism, regulated by BID levels. Our results pave the way to clinically explore SAC-targeting drugs in tumors with high BID expression.

Background: The EGFR pathway is involved in intrinsic and acquired resistance to a wide variety of targeted therapies in cancer. Vaccination represents an alternative to the administration of anti-EGFR monoclonal antibodies, such as cetuximab or panitumumab. Here, we tested if anti-EGF antibodies generated by vaccination (anti-EGF VacAbs) could potentiate the activity of drugs targeting the ERK/MAPK and PI3K/Akt pathways. Methods: Non-small cell lung cancer (NSCLC), colorectal cancer (CRC) and melanoma cell lines harboring KRAS, NRAS, BRAF and PIK3CA mutations were used. Anti-EGF VacAbs were obtained by immunizing rabbits with a fusion protein containing a synthetic, highly mutated variant of human EGF. Cell viability was determined by MTT, total and phosphorylated proteins by Western blotting, cell cycle distribution and cell death by flow cytometry and emergence of resistance by microscopic examination in low density cultures. Results: Anti-EGF VacAbs potentiated the antiproliferative effects of MEK, KRAS G12C, BRAF, PI3K and Akt inhibitors in KRAS, NRAS, BRAF and PIK3CA mutant cells and delayed the appearance of resistant clones in vitro. The effects of anti-EGF VacAbs were comparable or superior to those of panitumumab and cetuximab. The combination of anti-EGF VacAbs with the targeted inhibitors effectively suppressed EGFR downstream pathways and sera from patients immunized with an anti-EGF vaccine also blocked activation of EGFR effectors. Conclusions: Anti-EGF VacAbs enhance the antiproliferative effects of drugs targeting the ERK/MAPK and PIK3CA/Akt pathways. Our data provide a rationale for clinical trials testing anti-EGF vaccination combined with inhibitors selected according to the patient's genetic profile.

Introduction: As recently evidenced by the ADAURA trial, most patients with stages IB to IIIA of resected EGFR-mutant lung adenocarcinoma benefit from osimertinib as adjuvant therapy. Nevertheless, predictive markers of response and recurrence are still an unmet need for more than 10% of these patients. Some circular RNAs (circRNAs) have been reported to play a role in tumor growth and proliferation. In this project, we studied circRNA expression levels in formalin-fixed, paraffin-embedded lung tumor samples to explore their biomarker potential and develop a machine learning (ML)-based signature that could predict the benefit of adjuvant EGFR tyrosine kinase inhibitors in patients with EGFR-mutant NSCLC. Methods: Patients with surgically resected EGFR mutant-positive, stages I to IIIB NSCLC were recruited from February 2007 to December 2015. Formalin-fixed, paraffin-embedded tumor samples were retrospectively collected from those patients with a follow-up period of more than or equal to 36 months (N = 76). Clinicopathologic features were annotated. Total RNA was purified and quantified prior nCounter processing with our circRNA custom panel. Data analysis and ML were performed taking into consideration circRNA expression levels and recurrence-free survival (RFS). RFS was defined from the day of surgery to the day when recurrence was detected radiologically or the death owing to any cause. Results: Of the 76 patients with EGFR mutation-positive NSCLC included in the study, 34 relapsed within 3 years after resection. The median age of the relapsing cohort was 71.5 (range: 49–89) years. Most patients were nonsmokers (n = 21; 61.8%) and of female sex (n = 21; 61.8%). Most cases (n = 17; 50%) presented an exon 21 mutation, whereas 15 and four patients had an exon 19 and exon 18 mutation, respectively. Differential expression analysis revealed that circRUNX1, along with circFUT8 and circAASDH, was up-regulated in relapsing patients (p < 0.05 and >2 fold-change). A ML-based circRNA signature predictive of recurrence in patients with EGFR mutation-positive NSCLC, comprising circRUNX1, was developed. Our final model including selected 6-circRNA signature with random forest algorithm was able to classify relapsing patients with an accuracy of 83% and an area under the receiver operating characteristic curve of 0.91.RFS was significantly shorter not only for the subgroup of patients with high versus low circRUNX1 expression but also for the group classified as recurrent by the ML circRNA signature when compared with those classified as nonrecurrent. Conclusions: Our findings suggest that circRUNX1 and the presented ML-developed signature could be novel tools to predict the benefit of adjuvant EGFR tyrosine kinase inhibitors with regard to RFS in patients with EGFR-mutant NSCLC. The training and validation phases of our ML signature will be conducted including bigger independent cohorts.

Genetic and drug sensitivity assays on primary cultures are not only of basic but also of translational interest and could eventually aid oncologists in the selection of treatments. However, cancer cells need to be identified and differentiated from the non-tumor cells always present in primary cultures. Also, successive passages can change the proportions of these two subpopulations. In this study, we propose fluorescence in situ hybridization (FISH) analysis on cell smears to determine the presence of tumor cells in primary cultures obtained from patients carrying translocations or copy number gains. FISH proved to be an easy, fast, economic, and reliable method of characterizing cell populations, which could be used repeatedly at different passages to monitor variations and to confirm the maintenance of translocations and copy number gains throughout the culture process.

Lung cancer screening programs, particularly in the UK, have shown a decrease in lung cancer-related deaths among individuals who underwent low-dose computed tomography (CT) screening. Researchers are now focusing on evaluating cell-free DNA through various methods to determine if pre-diagnostic mutations can be detected years before clinical diagnosis. This could help identify individuals at high risk of developing lung cancer. However, while this approach has successfully identified precursors of follicular lymphoma, the presence of occult lung preneoplasia in non-small-cell lung cancer still requires further investigation.The TRACERx consortium is conducting extensive research to comprehensively assess the detection and progression of non-small cell lung cancers (NSCLC). Liquid biopsy is being used in advanced stages of the disease to monitor disease progression, predict treatment response, and identify targetable driver oncogenic mutations and fusion genes. Intense research is also underway to identify numerous diagnostic gene signatures with high accuracy for early-stage lung cancer. However, a more focused clinical approach is needed, with a mechanistic focus on the key pathways of cancer development.Loss of liver kinase B1 (LKB1) function and deactivation due to 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL), a metabolite of tobacco-specific carcinogens, could potentially be traced and contribute to the development of new biomarkers. This testing could complement machine-learning approaches. The discovery of epidermal growth factor receptor (EGFR) and Kirsten rat sarcoma viral oncogene homolog (KRAS) mutations in healthy lung tissues by TRACERx investigators may also lead to the development of novel diagnostic tools. Tumor protein 53 (TP53) loss should also be considered as a marker that could contribute to malignant transformation. Intercepting aggressive non-small-cell lung cancer is a pressing priority.In this review, we discuss our experience and explore other research on exosomes and plasma circular RNA as potential biomarkers. Circular RNAs, formed through non-sequential back-splicing of pre-mRNA transcripts, play a role in epithelial-mesenchymal transition, with many of them regulated by the RNA-binding protein Quaking. Platelet RNA has shown promise in detecting early and late-stage cancer. The extensive exploration of liquid biopsy aims to provide affordable methods for tracing circulating precursors of non-small-cell lung cancer, highlighting the importance of its mission.

Abstract Background Intercellular communication is mediated by extracellular vesicles (EVs), as they enclose selectively packaged biomolecules that can be horizontally transferred from donor to recipient cells. Because all cells constantly generate and recycle EVs, they provide accurate timed snapshots of individual pathophysiological status. Since blood plasma circulates through the whole body, it is often the biofluid of choice for biomarker detection in EVs. Blood collection is easy and minimally invasive, yet reproducible procedures to obtain pure EV samples from circulating biofluids are still lacking. Here, we addressed central aspects of EV immunoaffinity isolation from simple and complex matrices, such as plasma. Methods Cell-generated EV spike-in models were isolated and purified by size-exclusion chromatography, stained with cellular dyes and characterized by nano flow cytometry. Fluorescently-labelled spike-in EVs emerged as reliable, high-throughput and easily measurable readouts, which were employed to optimize our EV immunoprecipitation strategy and evaluate its performance. Plasma-derived EVs were captured and detected using this straightforward protocol, sequentially combining isolation and staining of specific surface markers, such as CD9 or CD41. Multiplexed digital transcript detection data was generated using the Nanostring nCounter platform and evaluated through a dedicated bioinformatics pipeline. Results Beads with covalently-conjugated antibodies on their surface outperformed streptavidin-conjugated beads, coated with biotinylated antibodies, in EV immunoprecipitation. Fluorescent EV spike recovery evidenced that target EV subpopulations can be efficiently retrieved from plasma, and that their enrichment is dependent not only on complex matrix composition, but also on the EV surface phenotype. Finally, mRNA profiling experiments proved that distinct EV subpopulations can be captured by directly targeting different surface markers. Furthermore, EVs isolated with anti-CD61 beads enclosed mRNA expression patterns that might be associated to early-stage lung cancer, in contrast with EVs captured through CD9, CD63 or CD81. The differential clinical value carried within each distinct EV subset highlights the advantages of selective isolation. Conclusions This EV isolation protocol facilitated the extraction of clinically useful information from plasma. Compatible with common downstream analytics, it is a readily implementable research tool, tailored to provide a truly translational solution in routine clinical workflows, fostering the inclusion of EVs in novel liquid biopsy settings.

Although many studies highlight the implication of circular RNAs (circRNAs) in carcinogenesis and tumor progression, their potential as cancer biomarkers has not yet been fully explored in the clinic due to the limitations of current quantification methods. Here, we report the use of the nCounter platform as a valid technology for the analysis of circRNA expression patterns in non‐small cell lung cancer (NSCLC) specimens. Under this context, our custom‐made circRNA panel was able to detect circRNA expression both in NSCLC cells and formalin‐fixed paraffin‐embedded (FFPE) tissues. CircFUT8 was overexpressed in NSCLC, contrasting with circEPB41L2, circBNC2, and circSOX13 downregulation even at the early stages of the disease. Machine learning (ML) approaches from different paradigms allowed discrimination of NSCLC from nontumor controls (NTCs) with an 8‐circRNA signature. An additional 4‐circRNA signature was able to classify early‐stage NSCLC samples from NTC, reaching a maximum area under the ROC curve (AUC) of 0.981. Our results not only present two circRNA signatures with diagnosis potential but also introduce nCounter processing following ML as a feasible protocol for the study and development of circRNA signatures for NSCLC.

Abstract EGFR tyrosine kinase inhibitor (TKI) resistance in non-small cell lung cancer (NSCLC) patients is inevitable. Identification of resistance mechanisms and corresponding targeting strategies can lead to more successful later-line treatment in many patients. Using spectrometry-based proteomics, we identified increased fibroblast growth factor receptor 1 (FGFR1) expression and Akt activation across erlotinib, gefitinib, and osimertinib EGFR-TKI-resistant cell line models. We show that while combined EGFR-TKI and FGFR inhibition showed some efficacy, simultaneous inhibition of FGFR and Akt or PI3K induced superior synergistic growth inhibition of FGFR1-overexpressing EGFR-TKI-resistant NSCLC cells. This effect was confirmed in vivo. Only dual FGFR and Akt inhibition completely blocked the resistance-mediating signaling pathways downstream of Akt. Further, increased FGFR1 expression was associated with significantly lower PFS in EGFR-TKI-treated NSCLC patients, and increased FGFR1 were demonstrated in a few post- vs. pre-EGFR-TKI treatment clinical biopsies. The superior therapeutic benefit of combining FGFR and Akt inhibitors provide the rationale for clinical trials of this strategy.

MET inhibitors have shown activity in non‐small‐cell lung cancer patients (NSCLC) with MET amplification and exon 14 skipping (METΔex14). However, patient stratification is imperfect, and thus, response rates have varied widely. Here, we studied MET alterations in 474 advanced NSCLC patients by nCounter, an RNA‐based technique, together with next‐generation sequencing (NGS), fluorescence in situ hybridization (FISH), immunohistochemistry (IHC), and reverse transcriptase polymerase chain reaction (RT–PCR), exploring correlation with clinical benefit. Of the 474 samples analyzed, 422 (89%) yielded valid results by nCounter, which identified 13 patients (3%) with METΔex14 and 15 patients (3.5%) with very‐high MET mRNA expression. These two subgroups were mutually exclusive, displayed distinct phenotypes and did not generally coexist with other drivers. For METΔex14, 3/8 (37.5%) samples positive by nCounter tested negative by NGS. Regarding patients with very‐high MET mRNA, 92% had MET amplification by FISH and/or NGS. However, FISH failed to identify three patients (30%) with very‐high MET RNA expression, among which one received MET tyrosine kinase inhibitor treatment deriving clinical benefit. Our results indicate that quantitative mRNA‐based techniques can improve the selection of patients for MET‐targeted therapies.

Abstract Background Chronic spontaneous urticaria (CSU) negatively impacts patients' sleep, thereby reducing health‐related quality of life (HRQoL). Half of patients with inadequately controlled CSU report sleep interference often or every night, which can lead to depression, anxiety, social, and work‐related problems. Methods This randomized, double‐blind, placebo‐controlled Phase 2b core study (NCT02477332) included adult patients ≥18 years with moderate to severe CSU inadequately controlled with H1‐antihistamines. The current analysis includes patients randomized to receive ligelizumab 72 or 240 mg, omalizumab 300 mg or placebo every 4 weeks (q4w) for five injections over 20 weeks with treatment‐free follow‐up for 24 weeks. Patients could enter the open‐label extension study (NCT02649218) from Week 32 onwards if their weekly urticaria activity score was ≥12, which included an open‐label treatment (52 weeks of ligelizumab 240 mg q4w) and a 48‐week post‐treatment follow‐up. Weekly Sleep Interference Scores (SIS7, range 0 [no interference]–21 [substantial interference]), Weekly Activity Interference Score (AIS7), Dermatology Life Quality Index (DLQI) scores, and Overall Work Impairment were assessed. Results Mean baseline SIS7 scores were balanced between the treatment arms for ligelizumab 72 mg (n = 84) and 240 mg (n = 85), omalizumab 300 mg (n = 85), and placebo (n = 43). By Week 12, patients experienced large improvements in sleep interference, with least square mean (standard error) changes from baseline (CFB) in SIS7 of −7.84 (0.58), −7.55 (0.61), −6.98 (0.60), and −5.85 (0.81), respectively. By Week 12, CFB in AIS7 were −8.25 (0.57), −8.25 (0.59), −7.30 (0.60), and −5.62 (0.79), DLQI scores were −9.79 (0.77), −9.93 (0.81), −8.35 (0.79), and −6.99 (1.11), and Overall Work Impairment scores were −28.96 (3.73), −30.76 (3.71), −25.74 (3.91), and −20.13 (5.10) for ligelizumab 72 and 240 mg, omalizumab 300 mg and placebo, respectively. Improvements in each patient‐reported outcome were sustained with ligelizumab 240 mg treatment during the extension study. Conclusions Ligelizumab showed effective and sustained responses in managing sleep interference in patients with CSU, and numerically higher responses than with omalizumab and placebo. Treating the symptoms of CSU with ligelizumab improved disease burden, HRQoL, and markedly improved sleep quality.

Non-small cell lung cancer with EGFR mutations are known to develop resistance to EGFR tyrosine kinase inhibitors. Here, the authors show AURKB activation to be associated with resistance in EGFR mutant lung cancer cells, and that AURKB is a therapeutic target in resistant tumours that lack the p.T790M or other acquired mutations.

Background: The analysis of liquid biopsies brings new opportunities in the precision oncology field. Under this context, extracellular vesicle circular RNAs (EV-circRNAs) have gained interest as biomarkers for lung cancer (LC) detection. However, standardized and robust protocols need to be developed to boost their potential in the clinical setting. Although nCounter has been used for the analysis of other liquid biopsy substrates and biomarkers, it has never been employed for EV-circRNA analysis of LC patients. Methods: EVs were isolated from early-stage LC patients (n = 36) and controls (n = 30). Different volumes of plasma, together with different number of pre-amplification cycles, were tested to reach the best nCounter outcome. Differential expression analysis of circRNAs was performed, along with the testing of different machine learning (ML) methods for the development of a prognostic signature for LC. Results: A combination of 500 μL of plasma input with 10 cycles of pre-amplification was selected for the rest of the study. Eight circRNAs were found upregulated in LC. Further ML analysis selected a 10-circRNA signature able to discriminate LC from controls with AUC ROC of 0.86. Conclusions: This study validates the use of the nCounter platform for multiplexed EV-circRNA expression studies in LC patient samples, allowing the development of prognostic signatures.

Introduction: DNA repair capacity, as exemplified by BRCA1 gene expression, is related with outcome to EGFR tyrosine kinase inhibitors in patients with EGFR-mutant NSCLC. Olaparib, a PARP inhibitor, reduces BRCA1 expression. Olaparib was tested in combination with gefitinib versus gefitinib single agent, as a first-line therapy for patients with EGFR-mutant NSCLC in the GOAL study (trial registration: NCT01513174). Here, we report the results of the biomarker-related prespecified secondary objectives of the GOAL study. Methods: We evaluated the impact of BRCA1 mRNA expression in 91 patients with EGFR-mutant NSCLC. Of those 91 patients, 51 were randomized to treatment with gefitinib and 40 were randomized to treatment with gefitinib plus olaparib. We explored in vitro whether BRCA1 mRNA levels are related with outcome to gefitinib plus olaparib. The expression levels of 53BP1, CtIP, and AXL were also explored and correlated with the treatment outcome. Results: Overall, as what happened in the GOAL study, no statistically significant difference was observed in median progression-free survival (PFS) between the two treatment arms, for the 91 patients of the present study (p = 0.2419). For patients with high BRCA1 mRNA expression (BRCA1-high group), median PFS was 12.9 months in the gefitinib plus olaparib arm, compared with 9.2 months in the gefitinib arm (p = 0.0449). In the gefitinib arm, median PFS was 9.1 months for the BRCA1-high group and 10.2 months for the BRCA1-low group (p = 0.0193). We observed a more pronounced synergism of gefitinib plus olaparib in cells with higher BRCA1 compared with those with low BRCA1 mRNA expression. Conclusions: High BRCA1 mRNA expression identified patients with NSCLC who benefited from gefitinib plus olaparib in the GOAL phase 2 clinical trial.

Introduction: EGFR mutation-positive lung adenocarcinoma (LUAD) displays impaired phosphorylation of ERK and Src-homology 2 domain-containing phosphatase 2 (SHP2) in comparison with EGFR wild-type LUADs. We hypothesize that SHP2 expression could be predictive in patients positive with resected EGFR mutation versus patients with EGFR wild-type LUAD. Methods: We examined resected LUAD cases from Japan and Spain. mRNA expression levels of AXL, MET, CDCP1, STAT3, YAP1, and SHP2 were analyzed by quantitative reverse transcriptase polymerase chain reaction. The activity of SHP2 inhibitors plus erlotinib were tested in EGFR-mutant cell lines and analyzed by cell viability assay, Western blot, and immunofluorescence. Results: A total of 50 of 100 EGFR mutation-positive LUADs relapsed, among them, patients with higher SHP2 mRNA expression revealed shorter progression-free survival, in comparison with those having low SHP2 mRNA (hazard ratio: 1.83; 95% confidence interval: 1.05–3.23; p = 0.0329). However, SHP2 was not associated with prognosis in the remaining 167 patients with wild-type EGFR. In EGFR-mutant cell lines, the combination of SHP099 or RMC-4550 (SHP2 inhibitors) with erlotinib revealed synergism via abrogation of phosphorylated AKT (S473) and ERK1/2 (T202/Y204). Although erlotinib translocates phosphorylated SHP2 (Y542) into the nucleus, either RMC-4550 alone, or in combination with erlotinib, relocates SHP2 into the cytoplasm membrane, limiting AKT and ERK1/2 activation. Conclusions: Elevated SHP2 mRNA levels are associated with recurrence in resected EGFR mutation-positive LUADs, but not in EGFR wild-type. EGFR tyrosine kinase inhibitors can enhance SHP2 activation, hindering adjuvant therapy. SHP2 inhibitors could improve the benefit of adjuvant therapy in EGFR mutation-positive LUADs.

Background: A Worldwide Antihistamine-Refractory Chronic Urticaria (CU) patient Evaluation (AWARE) is a non-interventional, multicenter study including patients from Europe, Central and Latin America, Asia-Pacific, and the Middle East. AWARE describes real-world evidence for CU, including clinical characteristics, treatment patterns and the impact on quality of life. Methods: Over the 2-year study, therapy changes, angioedema occurrence, and patient-reported outcomes (PROs) were recorded over 9 visits, including dermatology life quality index (DLQI) and 7-day urticaria activity score (UAS7). Data were stratified into subgroups: chronic spontaneous urticaria (CSU), chronic inducible urticaria (CIndU), or CSU + CIndU. Results: Out of 4838 patients analyzed, 9.9% were receiving no treatment for their CU symptoms at baseline, and 20.4% were receiving first-line non-sedating H1-antihistamine at approved doses. The predominant baseline therapy was up-dosed non-sedating H1-antihistamines (25.5%). By Visit 2, omalizumab was the overall most commonly used therapy (29.6%), increasing to 30.1% by the end of the study. Baseline DLQI scores for patients with CSU, CIndU and CSU + CIndU were 8.3, 7.6 and 9.1, respectively; scores decreased over the study for CSU and CSU + CIndU patients, but fluctuated for CIndU patients. Baseline angioedema occurrence was higher in CSU and CSU + CIndU patients, reported in 45.4% and 45.5% of patients, respectively, compared to 17.0% in CIndU patients. By the final visit, angioedema had decreased to 11.9% and 11.2% for CSU and CSU + CIndU, respectively, and 9.6% for CIndU. Conclusion: CU patients are undertreated at baseline; after entering the AWARE study, more patients received appropriate treatment. However, over two thirds are not escalated to third-line treatments.

En la novela Flughunde, publicada en 1995, Marcel Beyer, un autor de la tercera generación nacido en 1965 y, por lo tanto, sin vivencias directas ni de la época del nazismo ni de la posguerra, articula, a partir de una serie de datos históricos relacionados con la muerte de los hijos de Magda y Joseph Goebbels, una ficción en la que se plantea la cuestión de la fiabilidad de los testigos, fundamentalmente, cuando se trata de relatos de perpetradores. El objetivo del artículo es analizar cómo la novela aborda la representación del perpetrador, así como descubrir las estrategias discursivas del texto para desenmascarar el discurso del perpetrador, que intenta ocultar su responsabilidad en los crímenes cometidos durante el nazismo.

Abstract Background European legislation has banned the preservative methylisothiazolinone (MI) from inclusion in leave-on cosmetics. However, the risk for allergic reactions depends on exposure. The aim of this study was to determine the risk of MI in laundry detergents for household machine washing. Methods Different formulations of laundry detergents with commercial MI levels, up to one thousand ppm were used and three different types of clothes were washed in a normal household machine setting one time and 10 times. The level of MI was measured by HPLC. Results While MI could be retrieved in the positive control of clothes drenched with washing powder but not washed afterwards, MI could not be detected in any specimen of clothes washed under household conditions. The detection limit was 0.5 ppm. Conclusion It is important to discuss the difference of risk and hazard. While MI clearly is a high hazard as a strong contact allergen, the risk depends on exposure. Regarding the risk of exposure levels for the consumer to MI in clothes it can be stated that the use of MI in laundry detergents is safe for the consumer if these products are used according to the instructions in the normal household setting machine wash.

EGFR-mutant non-small cell lung cancer are often resistant to EGFR tyrosine kinase inhibitor treatment. In this study, the authors show that resistant tumors display high Akt activation and that a combined treatment with AKT inhibitors causes synergistic tumour growth inhibition in vitro and in vivo.

The detection of ALK receptor tyrosine kinase (ALK), ROS proto-oncogen1, receptor tyrosine kinase (ROS1), ret proto-oncogen (RET), and MET proto-oncogen exon 14 skipping (METΔex14) allows for the selection of specific kinase inhibitor treatment in patients with non-small cell lung cancer (NSCLC). Multiplex technologies are recommended in this setting. We used nCounter, a multiplexed technology based on RNA hybridization, to detect ALK, ROS1, RET, and METΔex14 in RNA purified from cytological specimens (n = 16) and biopsies (n = 132). Twelve of the 16 cytological samples (75.0%) were evaluable by nCounter compared to 120 out of 132 (90.9%) biopsies. The geometrical mean (geomean) of the housekeeping genes of the nCounter panel, but not the total amount of RNA purified, was significantly higher in biopsies vs. cytological samples. Among cytological samples, we detected ALK (n = 3), METΔex14 (n = 1) and very high MET expression (n = 1) positive cases. The patient with METΔex14 had a partial response to tepotinib, one of the patients with ALK fusions was treated with crizotinib with a complete response. Cell blocks and cytological extensions can be successfully used for the detection of fusions and splicing variants using RNA-based methods such as nCounter.

Objective: the objective of the present work is to study the effectiveness of treatment with silk fibroin nanoparticles loaded with resveratrol in experimental periodontitis in a diabetic rat model. Introduction: Periodontitis is an inflammatory pathology highly related to other diseases, such as type II diabetes. Both diseases have a specific inflammatory condition, with Interleukin (IL)-6, IL-1β and Transforming Grow Factor (TGF)-1β being the most relevant proinflammatory factors. Silk fibroin (SF) nanoparticles loaded with resveratrol (Res-SFN) are a new alternative as a treatment. Methods: 40 diabetic Sprague Dawley male rats were used and periodontitis was induced by ligation. The animals were divided into 5 treatment groups, and 1 mL of treatment was administered once a day for 4 weeks. The groups were: I: Carboxymethyl cellulose (CMC) 0.8%, II: CMC 0.8% + SF 1%, III: CMC 0.8% + RES-SFN 3 mg/mL, IV: CMC 0.8% + SF 1% + RES-SFN 3 mg/mL, V: Water. A peripheral blood sample was taken every week to quantify the inflammatory profile by ELISA (IL-6, IL-1β and TGF-1β). After 4 weeks the sacrifice was carried out and biopsies of the gum were taken. Results: Treatment with SF and RES-SFN reduced the amount of chemical inflammation mediators (with the exception of IL-1β in comparisons I-IV and II-IV (p > 0.05)), as well as the anatomopathological variables linked to it, in a significant way (p < 0.05). Conclusion: treatment with RES-SFN has reduced local inflammation in this experimental periodontitis model.

El presente artículo aborda la experiencia del exilio retratada en las novelas Transit de Anna Seghers y Zwischenstationen de Vladimir Vertlib a partir de la noción de lo transitorio. Para ello, me centraré no solo en la caracterización de los espacios del exilio como supuestos espacios de tránsito, sino también en el carácter cambiante de la identidad de los protagonistas, que consiguen acercarse, transitar, a las nuevas culturas de los países de acogida. Analizar las experiencias interculturales retratadas en las novelas nos permitirá nuevas lecturas de Transit, una obra ampliamente estudiada, así como constatar cómo la literatura del exilio clásico alemán anticipa ya algunos aspectos de novelas contemporáneas como Zwischenstationen.

En 2003, un año antes de recibir el Premio Nobel de Literatura, la autora austriaca Elfriede Jelinek publicó La muerte y la doncella I-V. Dramas de princesas, un conjunto de cinco obras concebidas como entreactos de otros espectáculos teatrales. Como es habitual en su escritura, las referencias intertextuales son constantes. En el artículo se analizarán los tres primeros dramas del ciclo y se mostrará la capacidad crítica y desestabilizadora del proceso discursivo de la cita. A diferencia de otras lecturas anteriores, no estudiaré la cita de la trama o el pasaje de un texto determinado, sino la forma en que los textos citados, todos ellos del siglo XIX, han abordado la cuestión del sujeto femenino.

Este artículo presenta el estudio llevado a cabo en cinco centros educativos (cuatro de educación infantil y primaria y uno de educación secundaria) con el objetivo de evaluar, desde la perspectiva del profesorado de educación infantil, primaria y secundaria, las fortalezas y debilidades del proceso educativo para establecer propuestas de mejora educativa. El diseño de investigación utilizado fue cuantitativo no experimental de corte descriptivo incluyendo como variables influyentes en el proceso educativo la planificación y organización de la enseñanza, implicación activa del alumnado e interacción entre los alumnos, entre otras. Para la recogida de la información se utilizó el instrumento “Autoevaluación de Centros para la Atención a la Diversidad desde la Inclusión” (ACADI) en su dimensión C referida al Proceso Educativo, cumplimentado por 97 docentes de centros públicos de la Región de Murcia. Los resultados muestran que las principales fortalezas que encuentra el profesorado en su quehacer educativo son las referidas al respeto a los estilos de aprendizaje, planificación y organización de la enseñanza, implicación activa del alumnado en su propio aprendizaje, evaluación tolerante, organización de la acción tutorial, interacción entre los alumnos, y relaciones centro-contexto social. En cambio la atención a los ritmos de aprendizaje y a las características del alumnado son las que reciben las puntuaciones más bajas en todos los centros. Al comparar los resultados de los centros de infantil y primaria con el de secundaria, los docentes de infantil y primaria discrepan de forma significativa con los profesores de secundaria en la organización de la acción tutorial, mientras que en otros aspectos como diversidad metodológica, apoyo colaborativo, conocimiento de los objetivos y contenidos del curriculum por parte del alumnado, las discrepancias no son tan significativas.

RESUMENDesde la experiencia en el uso de la herramienta wiki en diversas asignaturas impartidas en la Facultad de Ciencias Jurídicas de la Universitat Rovira i Virgili de Tarragona se pueden extraer algunas conclusiones que permiten la elaboración de una estrategia para la implementación de actividades de trabajo colaborativo, utilizando dicha herramienta. Con carácter previo al diseño de la actividad es necesario contextualizarla, reflexionando sobre algunas cuestiones como los potenciales usuarios, los objetivos de la actividad, el plan de trabajo, los recursos técnicos y humanos necesarios, el sistema de seguimiento del aprendizaje, las vías de comunicación, el sistema y criterios de evaluación. Una vez contextualizada, se puede proceder al diseño de la actividad y de la documentación y a su presentación on line, adoptando decisiones y elaborando materiales sobre las competencias a desarrollar, el objeto del trabajo, los criterios de evaluación, el cronograma, las reglas de estilo, los criterios de formación de los grupos, la metodología, las vías de comunicación, y el diseño y formalización del espacio de trabajo en el campus virtual. Por último, es necesario alertar sobre diversos problemas que pueden aparecer durante la elaboración del trabajo en grupo mediante una wiki, tanto de carácter tecnológico como sociológicos.ABSTRACTFrom the experience in using the wiki tool in several subjects taught at the Faculty of Law at the Universitat Rovira i Virgili, some conclusions can be drawn that allow the development of a strategy to implement collaborative activities using wiki tool. Prior to the design of the activity it is necessary to contextualize them, reflecting on the following questions: What is the wiki for? When will the wiki be used? Who are the potential users? How long do students need to develop this activity? Which are the characteristics of the work developed in the wiki? What role does the teacher play in the activity?, What kind of communication must exist between the participants? What is to be evaluated? What technical and human resources will be needed? Once the activity is contextualised, it is possible to design the activity and the supporting and information documentation and their presentation on line, developing materials and making decisions on: the skills developed, the object of work, the evaluation criteria, the schedule of activities, the style rules, the working methods and communication, and the design and execution of the working space in the virtual campus. Finally, it is necessary to alert to several problems that may arise during the process of developing the group work in the wiki, both of technological and of sociological sort.

Desde la experiencia en el uso de la herramienta wiki en diversas asignaturas impartidas en la Facultad de Ciencias Jurídicas de la Universitat Rovira i Virgili de Tarragona se pueden extraer algunas conclusiones que permiten la elaboración de una estrategia para la implementación de actividades de trabajo colaborativo, utilizando dicha herramienta. Con carácter previo al diseño de la actividad es necesario contextualizarla, reflexionando sobre algunas cuestiones como los potenciales usuarios, los objetivos de la actividad, el plan de trabajo, los recursos técnicos y humanos necesarios, el sistema de seguimiento del aprendizaje, las vías de comunicación, el sistema y criterios de evaluación. Una vez contextualizada, se puede proceder al diseño de la actividad y de la documentación y a su presentación on line, adoptando decisiones y elaborando materiales sobre las competencias a desarrollar, el objeto del trabajo, los criterios de evaluación, el cronograma, las reglas de estilo, los criterios de formación de los grupos, la metodología, las vías de comunicación, y el diseño y formalización del espacio de trabajo en el campus virtual. Por último, es necesario alertar sobre diversos problemas que pueden aparecer durante la elaboración del trabajo en grupo mediante una wiki, tanto de carácter tecnológico como sociológicos.

Wie die meisten Texte Elfriede Jelineks ist auch das Theaterstück Ulrike Maria Stuart reich an intertextuellen Referenzen. Neben Schillers Trauerspiel Maria Stuart werden unter anderem Texte aus den Schriften der RAF (Rote Armee Fraktion) so wie aus journalistischen Veröffentlichungen über die terroristische Vereinigung zitiert, paraphrasiert und umformuliert. Dieser Artikel geht der Frage nach, wie die intertextuellen Referenzen in Jelineks polyphonem Text eine idealistische Auffassung der Welt und der Kunst, die in den in Ulrike Maria Stuart zitierten Prätexten vertreten wird, hinterfragen und dekonstruieren. Gegenstand der Analyse sind Jelineks Text und Nicolas Stemanns Inszenierung von Ulrike Maria Stuart (UA 28. Oktober 2006, Hamburg), die die idealistische Welt mit jener der Postmoderne in dem aktuellen Kontext verflicht

RESUMEN: Para analizar con una cierta perspectiva histórica el asociacionismo gitano, al que algunos gitanos llaman «el movimiento», es necesario remontarse a los años 60.

Para analizar con una cierta perspectiva histórica el asociacionismo gitano, al que algunos gitanos llaman «el movimiento», es necesario remontarse a los años 60.

Intercultural mediation essentially deals with perceptions. Perceived reality is frequently very different and far removed from actual reality. From our human and cultural position we must endeavor to understand the «other», starting from the principle that our lives are guided by certain norms and principles that are impregnated with cultural values and traditions. Any attempt to find logical reasons for this perception, in other words, the causes and consequences of the behavior and attitudes of those who do not share our culture, will be in vain and take us nowhere. Only by changing our approach can we achieve the fluency of understanding and comprehension that we all strive for. In sum, we must face up to the difficulties of communication by questioning why reality is perceived in this way. This approach activates our perceptive capacity, facilitates attitudes of comprehension, and stimulates the dialogue and communication on which interculturality is grounded. La mediación intercultural esencialmente es una cuestión de percepciones. Desde una posición humana y cultural debemos comprender al «otro» partiendo del principio que nuestras vidas están orientadas por normas y principios que están impregnados de valores y tradiciones culturales. Aplicar «nuestra lógica» buscando las causas y las consecuencias del comportamiento y de las actitudes de aquellos que no comparten nuestra cultura es un intento vano que no nos lleva a ningún lado. Sólo cambiando nuestro enfoque podemos lograr el entendimiento fluido y la comprensión que deseamos. En suma, debemos enfrentarnos a las dificultades de comunicación cuestionándonos por qué la realidad es percibida de esta manera. Esta aproximación aumenta nuestra capacidad de percepción, facilita actitudes de comprensión estimulando el diálogo y la comunicación.

A menudo las fronteras académicas que nos definen y nos separan como investigadores, también delimitan nuestros trabajos hasta el punto de desconectarlos de manera acrítica de las realidades culturales que, por motivos múltiples y no siempre explícitos, nos proponemos estudiar, analizar, transformar, etc. Sabemos que las universidades, como el resto de instituciones sociales, han participado históricamente en el mantenimiento de las fronteras que construimos interesadamente los seres humanos al relacionarnos como tales, tanto a escala individual como colectiva: económicas, políticas, académicas, familiares, personales, etc…

Vitiligo is an acquired skin disease that significantly impacts the quality of life of patients. Medical treatment of vitiligo includes the use of melanocyte transplant, but the results are variable. We have treated 4 patients with either focal or generalized stable vitiligo using a graft of autologous melanocytes' culture on a denuded amniotic membrane (AM). A culture biopsy was obtained in every patient and grown in melanocytes' media for 10–14 days after which cells were transferred to a denuded AM and transplanted into the achromic lesions. Patients were followed for up to 6 months using clinical assessment of achromic lesions. Treated areas ranged between 4 cm2 and 210.6 cm2. Response to treatment was excellent in all patients with 90–95% repigmentation success rate. Our results demonstrate that transplantation of autologous melanocytes cultured on AM is a new, simple, and effective treatment for stable vitiligo.

Abstract: Interventions aimed at changing dietary patterns should be designed based on the main motives underlying the food choices of specific target populations. The aim of the present study was to identify motives underlying food choice and barriers to healthy eating among consumers in two socioeconomic levels in Uruguay. Eleven focus groups were carried out with a total of 76 participants. Six of the groups involved low income participants and the others were conducted with middle income participants. Discussions were held around frequently consumed products, motives underlying food choices and barriers to healthy eating. Results confirmed the strong influence of income level on motives underlying food choice and barriers to the adoption of healthy eating. Low income participants described their choices as mainly driven by economic factors and satiety, whereas convenience was the main determinant of food selection for middle income participants. Implications for the design of public policies targeted at each group are discussed.

Abstract: Marketing of unhealthy products has been identified as one of the main characteristics of the food environment that negatively affects children’s eating patterns. Restrictions on advertising of unhealthy foods to children have already been imposed in different countries. However, marketing strategies are not limited to broadcast and digital advertising, but also include package design. In this context, the current study aimed to describe the food products targeted at children and sold in supermarkets in Montevideo, Uruguay, in terms of package design and nutrient profile. Two supermarkets in Montevideo were selected for data collection. In each supermarket, all products targeted at children were identified. Products were analyzed in terms of package design and nutritional profile, considering the Pan American Health Organization Nutrient Profile Model. A total of 180 unique products were identified, which included a wide range of product categories. The great majority of the products corresponded to ultra-processed products with excessive amounts of sodium, free sugars, total fat, saturated fat, and/or trans fat, which are not recommended for frequent consumption. Several marketing strategies were identified in the design of packages to attract children’s attention and drive their preferences. The most common strategies were the inclusion of cartoon characters, bright colors, childish lettering, and a wide range of claims related to health and nutrition, as well as the products’ sensory and hedonic characteristics. The study’s findings provide additional evidence on the need to regulate packaging of products targeted at children.

Background: Monitoring of adverse events induced by cosmetics performed by health authorities, known as cosmetovigilance, has been relied on the collection of case notifications. Objectives: We aimed to show how a contact dermatitis registry can contribute to the cosmetovigilance of emerging allergens. We used the example of phenylethyl resorcinol, an infrequent allergen with only 6 previous cases reported in Europe and Japan since 2013. Methods: A systematic search in the Spanish Registry of Contact Dermatitis and Cutaneous Allergy (REIDAC) database was performed to identify patients with positive patch test to phenylethyl resorcinol or cosmetics that contains it between June 2018 and January 2023. We collected the main clinical features of these patients and compared them with those of patients recorded in the registry with similar epidemiological features. Results: Thirteen patients with positive patch test to phenylethyl resorcinol were identified. All the patients were women with a mean age of 42 years (range 32–59) and their lesions were mainly in the face. Conclusion: Assessing the importance of infrequent allergens based solely on a case series is difficult. Multicentre registries facilitate the collection of cases and provide appropriate background information for new allergens. [ABSTRACT FROM AUTHOR]