RESUMO
BACKGROUND: Over the past few years, next-generation sequencing (NGS) has become reliable and cost-effective, and its use in clinical practice has become a reality. A relevant role for NGS is the prediction of response to anti-EGFR agents in metastatic colorectal cancer (mCRC), where multiple exons from KRAS, NRAS, and BRAF must be sequenced simultaneously. METHODS: We optimized a 14-amplicon NGS panel to assess, in a consecutive cohort of 219 patients affected by mCRC, the presence and clinico-pathological associations of mutations in the KRAS, NRAS, BRAF, and PIK3CA genes from formalin-fixed, paraffin-embedded specimens collected for diagnostics and research at the time of diagnosis. RESULTS: We observed a statistically significant association of RAS mutations with sex, young age, and tumor site. We demonstrated that concomitant mutations in the RAS/RAF pathway are not infrequent in mCRC, and as anticipated by whole-genome studies, RAS and PIK3CA tend to be concurrently mutated. We corroborated the association of BRAF mutations in right mCRC tumors with microsatellite instability. We established tumor side as prognostic parameter independently of mutational status. CONCLUSIONS: To our knowledge, this is the first monocentric, consecutively accrued clinical mCRC cancer cohort tested by NGS in a real-world context for KRAS, NRAS, BRAF, and PIK3CA. Our study has highlighted in clinical practice findings such as the concomitance of mutations in the RAS/RAF pathway, the presence of multiple mutations in single gene, the co-occurrence of RAS and PIK3CA mutations, the prognostic value of tumor side and possible associations of sex with specific mutations.
Assuntos
Neoplasias Colorretais/genética , Neoplasias Colorretais/secundário , Mutação/genética , Proteínas Proto-Oncogênicas B-raf/genética , Transdução de Sinais , Proteínas ras/genética , Idoso , Neoplasias Colorretais/patologia , Feminino , Genes Neoplásicos , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Estimativa de Kaplan-Meier , Masculino , Pessoa de Meia-Idade , Transdução de Sinais/genéticaRESUMO
AIM: Healthcare professionals have an important role to play both as advisers influencing smoking cessation and as role models. Despite this many of them continue to smoke. In this pilot study we have evaluated the reliability and validity of the Global Health Professional Students Survey questionnaire to examine smoking prevalence, in the Piedmont region. METHODS: Reliability analysis was tested and content validity was evaluated using Cronbach's alpha to check internal consistency with the intention to obtain no misunderstanding with the results. The questionnaire composed of 6 sections for a total of 36 items, was administered among nursing students in Piedmont's hospitals and data were collected in the period between January and July 2013. Statistical analysis was performed through SPSS Statistics for Windows version 19 (IBM Corp. Released 2010. IBM SPSS Statistics for Windows, Version 19.0.Armonk, NY: IBM Corp, USA). RESULTS: Questionnaire was administered to 265 nursing students: 77 (29.1%) men and 188 (70.9%) women. Only 37 (14%) students were over 30 year old, 57 (21.5) were in a range of 25-29 and 171 (64.5%) were in a range of 18-24 years old. Students had attended the 3th year of the course. The prevalence of current smokers was 25.3% (67). The higher value of Cronbach's Alpha resulted on the selection of 14 items (0.841), while on the other all sections (34 items in total) resulted in a value (0.786) meaning that the questionnaire has a satisfactory internal validity. CONCLUSIONS: This pilot study demonstrated that the questionnaire has very good reliability properties in the study and this needs to be taken into account for future extensive studies needed due to the high percentage of students in nursing who currently smoke (about a quarter of the total in our study). In addition, this study revealed that the education of students on smoking cessation is highly associated with their positive perception of their role as health model. This is a fact which may indicate the importance of adopting a core-curriculum related to the prevention and treatment of tobacco addiction among nursing students.
Assuntos
Atitude Frente a Saúde , Comportamentos Relacionados com a Saúde , Fumar , Estudantes de Enfermagem , Inquéritos e Questionários/normas , Adolescente , Adulto , Distribuição por Idade , Feminino , Conhecimentos, Atitudes e Prática em Saúde , Humanos , Itália/epidemiologia , Masculino , Projetos Piloto , Prevalência , Reprodutibilidade dos Testes , Fatores de Risco , Fumar/epidemiologia , Estudantes de Enfermagem/estatística & dados numéricosRESUMO
A straightforward, low-cost, and scalable solid-state mechanochemical protocol for the synthesis of silver nanoparticles (AgNP) based on the use of the highly reducing agri-food by-product pecan nutshell (PNS) is reported herein. Under optimized conditions (180 min, 800 rpm, PNS/AgNO3 ratio = 55/45 w/w), a complete reduction in silver ions was achieved, leading to a material containing ca. 36% w/w Ag0 (X-ray diffraction analysis). Dynamic light scattering and microscopic analysis showed a uniform size distribution (15-35 nm average diameter) of the spherical AgNP. The 2,2-Diphenyl-1-picrylhydrazyl (DPPH) assay revealed lower-although still absolutely high (EC50 = 5.8 ± 0.5 mg/mL)-antioxidant properties for PNS for the further incorporation of AgNP, supporting the efficient reduction of Ag+ ions by PNS phenolic compounds. Photocatalytic experiments indicated that AgNP-PNS (0.4 mg/mL) was able to induce the >90% degradation of methylene blue after 120 min visible light irradiation, with good recycling stability. Finally, AgNP-PNS demonstrated high biocompatibility and significantly light-enhanced growth inhibition properties against Pseudomonas aeruginosa and Streptococcus mutans at concentrations as low as 250 µg/mL, also eliciting an antibiofilm effect at 1000 µg/mL. Overall, the adopted approach allowed to reuse a cheap and abundant agri-food by-product and required no toxic or noxious chemicals, making AgNP-PNS a sustainable and easy-to-access multifunctional material.