Non-small cell lung cancer testing on reference specimens: an italian multicenter experience

Introduction: Biomarker testing is mandatory for the clinical management of patients with advanced non-small cell lung cancer (NSCLC). Myriads of technical platforms are now available for biomarker analysis with differences in terms of multiplexing capability, analytical sensitivity, and turnaround time (TAT). We evaluated the technical performance of the diagnostic workflows of 24 representative Italian institutions performing molecular tests on a series of artificial reference specimens built to mimic routine diagnostic samples. Methods: Sample sets of eight slides from cell blocks of artificial reference specimens harboring exon 19 EGFR (epidermal growth factor receptor) p.E746_AT50del, exon 2 KRAS (Kirsten rat sarcoma viral oncogene homologue) p.G12C, ROS1 (c-ros oncogene 1)-unknown gene fusion, and MET (MET proto-oncogene, receptor tyrosine kinase) Δ exon 14 skipping were distributed to each participating institution. Two independent cell block specimens were validated by the University of Naples Federico II before shipment. Methodological and molecular data from reference specimens were annotated. Results: Overall, a median DNA concentration of 3.3 ng/μL (range 0.1–10.0 ng/μL) and 13.4 ng/μL (range 2.0–45.8 ng/μL) were obtained with automated and manual technical procedures, respectively. RNA concentrations of 5.7 ng/μL (range 0.2–11.9 ng/μL) and 9.3 ng/μL (range 0.5–18.0 ng/μL) were also detected. KRAS exon 2 p.G12C, EGFR exon 19 p.E736_A750del hotspot mutations, and ROS1 aberrant transcripts were identified in all tested cases, whereas 15 out of 16 (93.7%) centers detected MET exon 14 skipping mutation. Conclusions: Optimized technical workflows are crucial in the decision-making strategy of patients with NSCLC. Artificial reference specimens enable optimization of diagnostic workflows for predictive molecular analysis in routine clinical practice

Comparison of ordinations of some apennine forest communities based on different characters and methods

In the course of numerical ordinations of Apennines woodlands, the problem arose of different performance of ordination methods, using different character sets and methods. To reveal underlying trends and relationships, 47 ordinations are jointly examinated, corresponding to combinations of 6 methods and 8 character types. The Spearman formula has been used to measure the similarity between different ordinations. The ordinations based on families are, on the average, the most similar to the other ones irrespective to the method used. The ordination based on detailed life forms (growth forms) are the least similar to the other ordinations. Ordinations based on the same character set and different methods could be different as well as ordinations based on different character sets and the same method. The suggested procedure can be the basis to select among different ordinations those explaining complementary informations

The planarian wound epidermis gene equinox is required for blastema formation in regeneration

AbstractRegeneration often involves the formation of a blastema, an outgrowth or regenerative bud formed at the plane of injury where missing tissues are produced. The mechanisms that trigger blastema formation are therefore fundamental for regeneration. Here, we identify a gene, which we named equinox, that is expressed within hours of injury in the planarian wound epidermis. equinox encodes a predicted secreted protein that is conserved in many animal phyla. Following equinox inhibition, amputated planarians fail to maintain wound-induced gene expression and to subsequently undergo blastema outgrowth. Associated with these defects is an inability to reestablish lost positional information needed for missing tissue specification. Our findings link the planarian wound epidermis, through equinox, to regeneration of positional information and blastema formation, indicating a broad regulatory role of the wound epidermis in diverse regenerative contexts.</jats:p