Detection and characterization of classical and "uncommon" exon 19 Epidermal Growth Factor Receptor mutations in lung cancer by pyrosequencing

BACKGROUND: The management of advanced stage non-small cell lung cancer is increasingly based on diagnostic and predictive analyses performed mostly on limited amounts of tumor tissue. The evaluation of Epidermal Growth Factor Receptor (EGFR) mutations have emerged as the strongest predictor of response to EGFR-tyrosine kinase inhibitors mainly in patients with adenocarcinoma. Several EGFR mutation detection techniques are available, having both sensitivity and specificity issues, being the Sanger sequencing technique the reference standard, with the limitation of a relatively high amount of mutated cells needed for the analysis. METHODS: A novel nucleotide dispensation order for pyrosequencing was established allowing the identification and characterization of EGFR mutation not definable with commercially and clinically approved kits, and validated in a consecutive series of 321 lung cancer patients (246 biopsies or cytology samples and 75 surgical specimens). RESULTS: 61/321 (19%) mutated cases were detected, 17 (27.9%) in exon 21 and 44 (72.1%) in exon 19, these latter corresponding to 32/44 (72.7%) classical and 12/44 (27.3%) uncommon mutations. Furthermore, a novel, never reported, point mutation, was found, which determined a premature stop codon in the aminoacidic sequence that resulted in a truncated protein in the tyrosine kinase domain, thus impairing the inhibitory effect of specific therapy. CONCLUSIONS: The novel dispensation order allows to detect and characterize both classical and uncommon EGFR mutations. Although several phase III studies in genotypically defined groups of patients are already available, further prospective studies assessing the role of uncommon EGFR mutations are warranted

Poster 6: An Unusual Aspiration Pneumonia Secondary to Diffuse Idiopathic Skeletal Hyperostosis: A Combined Medical and Rehabilitative Approach.

Setting: Outpatient rehabilitation center. Patient: An 84-year-old man presented with an 8-month history of nonsevere dysphagia that was experienced only with solid foods in a patient with diffuse idiopathic skeletal hyperostosis (DISH) or Forestier\u2019s disease of the cervical spine. An episode of pneumonia occurred in the third and sixth months previously and the patient had been admitted to hospital and treated with antibiotics. Case Description: The patient was treated with nonsteroidal anti-inflammatory drugs (10mg aceclofenac QID, maintained for 2wk/mo), a soft diet, and rehabilitation treatment (active exercises to strengthen the swallowing musculature and effortful swallow, exercises for coordinating swallowing and breathing, for strengthening the respiratory, airflow protection maneuvers to produce safer swallowing, improve airway protection, and pharyngeal and laryngeal function, etc). Assessment/Results: At follow-up 6, 12, 18, 24, and 30 months after starting medical and rehabilitation treatment, the patient was still able to maintain an adequate oral food intake with no signs of aspiration or clinical complications. Discussion: This is the first reported case, to our knowledge, which describes a long follow-up in the dysphagia post-DISH. The dysphagia was hypothesized to be caused by mechanical compression and inflammatory changes accompanied by fibrosis in the esophagus wall. The aspiration pneumonia probably had multifactorial etiology: dysphagia, abnormal cough reflex, colonization of the oropharynx and virulent bacteria, etc. We believe that the durable improvement in our patient dysphagia is due to the combined use of medical and rehabilitation treatment. Conclusions: This case illustrates that cervical spinal abnormalities, such as Forestier\u2019s disease, may be potentially treatable causes of aspiration pneumonia

Economia articolare e artrite reumatoide: risultati di uno studio randomizzato controllato

L\u2019economia articolare (EA) \ue8 un insieme di interventi riabilitativi che possono essere utilizzate nei pazienti con artrite reumatoide (AR), con l\u2019obiettivo di diminuire o rallentare i deterioramenti articolari e di superare gli ostacoli che si presentano nelle normali attivit\ue0 della vita quotidiana. Essa comprende l\u2019educazione gestuale, l\u2019addestramento all\u2019uso di tutori, l\u2019adattamento dell\u2019ambiente circostante e la formazione-informazione del malato cronico. In Italia, da quanto ci risulta, questo approccio appare ancora poco diffuso nella pratica riabilitativa; lo scopo di questo studio era, pertanto, quello di verificare la validit\ue0 di un corso di EA in un gruppo di pazienti affetti da AR medio-grave residenti in Veneto. Sono stati reclutati 85 pazienti dell\u2019et\ue0 media di 53.7 anni (DS 11.6, range 34-78 anni), randomizzati in due gruppi: gruppo di trattamento (GT), composto da 36 pazienti dell\u2019et\ue0 media di 54.2 anni (DS 9.8, range 33-68 anni) che partecipavano ad un corso di EA, e gruppo di controllo (GC) composto da 33 pazienti dell\u2019et\ue0 media 46.2 anni (DS 11.9, range 29-64 anni) che seguivano soltanto il trattamento medico. Il corso di EA prevedeva 3 incontri in cui i pazienti con AR erano informati sugli aspetti fisiopatologici ed evolutivi della malattia, addestrati all\u2019esecuzione di esercizi e gestualit\ue0 corrette da praticare nella vita quotidiana insieme a suggerimenti sull\u2019effettuazione di piccole modifiche ambientali e, infine, istruiti sulle normative vigenti a tutela del malato reumatico. La valutazione dei risultati veniva effettuata somministrando, all\u2019inizio e alla fine dello studio, la scala visuo-analogica (VAS) per la valutazione del dolore, l\u2019Health Assessment Questionnaire (HAQ), l\u2019Arthritis Impact Measurement Scale (AIMS) e lo Short Form-36 Health Survey (SF-36) per la valutazione, rispettivamente, dello stato di salute e della disabilit\ue0 nei sui vari aspetti, e l\u2019Health Service Interview (HSI) per la valutazione del grado di conoscenza della propria malattia. Risultava, dopo un tempo medio di 7 mesi, che i pazienti che hanno partecipato al corso di EA presentavano miglioramenti significativamente superiori ai controlli sul sintomo dolore, la disabilit\ue0 motoria e funzionale e lo stato di salute generale; conseguentemente, anche la qualit\ue0 di vita ne risultava migliorata. Infine, dall\u2019analisi dell\u2019HSI risultava che i pazienti presentavano uno scarso grado di conoscenza della patologia reumatica e delle sue conseguenze. In base ai risultati del presente studio, si poteva ritenere che l\u2019EA \ue8 utilmente inseribile tra gli interventi terapeutici nei pazienti con AR

Genome-Wide Analysis of japonica Rice Performance under Limited Water and Permanent Flooding Conditions

A rice GWAS panel of 281 accessions of japonica rice was phenotypically characterized for 26 traits related to phenology, plant and seed morphology, physiology and yield for 2 years in field conditions under permanent flooding (PF) and limited water (LW). A genome-wide analysis uncovered a total of 160 significant marker-trait associations (MTAs), of which 32 were LW-specific, 59 were PF-specific, and 69 were in common between the two water management systems. LW-specific associations were identified for several agronomic traits including days to maturation, days from flowering to maturation, leaf traits, plant height, panicle and seed traits, hundred grain weight, yield and tillering. Significant MTAs were detected across all the 12 rice chromosomes, while clusters of effects influencing different traits under LW or in both watering conditions were, respectively, observed on chromosomes 4, 8, and 12 and on chromosomes 1, 3, 4, 5, and 8. The analysis of genes annotated in the Nipponbare reference sequence and included in the regions associated to traits related to plant morphology, grain yield, and physiological parameters allowed the identification of genes that were demonstrated to affect the respective traits. Among these, three (OsOFP2, Dlf1, OsMADS56) and seven (SUI1, Sd1, OsCOL4, Nal1, OsphyB, GW5, Ehd1) candidate genes were, respectively, identified to co-localize with LW-specific associations and associations in common between the two water treatments. For several LW-specific MTAs, or in common among the two treatments, positional co-localizations with previously identified QTLs for rice adaptation to water shortages were observed, a result that further supports the role of the loci identified in this work in conferring adaptation to LW. The most robust associations identified here could represent suitable targets for genomic selection approaches to improve yield-related traits under LW