Demonstration of microRNA using in situ hybridisation on formalin fixed paraffin wax samples using conventional oligonucleotide probes: a comparison with the use of locked nucleic acid probes

Background: MicroRNAs (miRNA) regulate the translation of mRNA during gene expression and investigations have highlighted their importance in pathophysiology. qRT-PCR is currently the gold standard method for detecting changes in miRNA expression. However, when used on heterogeneous samples, it cannot identify individual cell types harbouring miRNAs. For this, in situ hybridisation (ISH) can be used. ISH methods using locked nucleic acid (LNA) probes have been shown to give reliable results in formalin fixed paraffin embedded (FFPE) samples. In this study their use has been directly compared with conventional oligonucleotide probes (COP) for ISH. Methods: FFPE samples of colorectal adenocarcinoma, squamous carcinoma of lung and cases of invasive breast carcinoma were used to evaluate COP and LNA methods for the demonstration of miR-126 and miR-205. To demonstrate the utility of the COP method demonstration of miR-21 in 19 Gleason stage 7 prostate biopsy FFPE tissues was also undertaken. The demonstration of miR-21 by ISH in high and low expressing prostate cancer cell lines was also compared with qRT-PCR. Results: Similar results were obtained using the COP and LNA ISH methods for the demonstration of miR-126 and miR-205. miR-21 was successfully demonstrated in the prostate cancer samples by COP ISH and expression levels of the miRNA demonstrated in the cell lines corresponded with qRT-PCR. Conclusion: This study has shown that simplification of ISH protocols by the use of COPs provides equivalent results to the use of LNA methods and it can be used to precisely identify cells in which miRNA are expressed

Dispersion of multi-walled carbon nanotubes in [BMIM]PF 6 for electrochemical sensing of acetaminophen

The influence of functionalized multi-walled carbon nanotubes (fMWCNT) in the presence of 1-butyl-3-methylimidazolium hexafluorophosphate ([BMIM]PF6) in different ratios was investigated on the acetaminophen (ACOP) electrochemical determination. The electrochemical behavior of the ACOP exhibited a pair of well-defined redox peaks, suggesting that the reversibility of ACOP was significantly improved in comparison to irreversible oxidation peak on bare GCE. The redox process was controlled by adsorption, involves two electrons and the value of apparent rate constant (ks) was equal to 14.7 s-1 ± 3.6 s-1. The analytical curves were obtained for concentrations of ACOP ranging from 0.3 to 3.0 μmol L-1. The values of the detection limit were calculated from SWV and found to be 6.73 × 10-8 mol L-1. The proposed electrochemical sensor exhibited good stability and reproducibility and was applied for ACOP determination in tablets (Tylenol® and Tylenol®DC) with satisfactory results.The authors gratefully acknowledge funding provided by the following Brazilian agencies: CNPq-INCT (Proc. 573925/2008-9 and 573548/2008-0), CAPES/Funcap (2133/2012/Proc. 23038.007973/2012-90), CNPq-PVE (Proc. 400223/2014-7 and 303596/2014-7), PRONEX/FUNCAP (Proc. PR2-0101-00030.01.00/15) and CNPq (Proc. 302801/2014-6). R.N. Gomes thanks CAPES and C.P. Sousa thanks CAPES-PNPD for their grants. The authors also are grateful to the Central Analítica-UFC/CT-INFRA/MCTI-SISNANO/Pró-Equipamentos CAPES for technical support and Prof. Pierre Basílio Almeida Fechine and MsC. Davino Machado Andrade Neto for Zeta potential analysis.info:eu-repo/semantics/publishedVersio

Polypharmacy Among The Elderly In The City Of São Paulo, Brazil - Sabe Study [polifarmácia Entre Idosos Do Município De São Paulo - Estudo Sabe]

Polypharmacy for the elderly and associated factors were assessed. A cross-sectional study was conducted using data from the SABE Study (Health, Well-being and Aging), 2006. 1,115 individuals representing 422,377 elderly aged 65 or more, living in São Paulo City, were interviewed. Polypharmacy was defined as the use of five or more medications. A multivariate regression logistics was used. Polypharmacy prevalence was 36%. Female (OR = 1.7; IC 95%: 1.0; 2.9), age over 75 years (OR = 1.9; CI 95%: 1.3; 2.7), higher income (OR = 1.8; CI 95%: 1.2; 2.8), working (OR = 1.8; CI 95%: 1.1; 2.9), regular self assessed health (OR = 1.6; CI 95%: 1.1; 2.3) or poor (OR = 2.6; CI 95%: 1.4; 4.9), hypertension (OR = 2.0; CI 95%: 1.4; 2.9), diabetes (OR = 4.1; CI 95%: 2.2; 7.5), rheumatic diseases (OR = 2.3; CI 95%: 1.5; 3.6) and cardiac problems (OR = 2.9; CI 95%: 1.9; 4.5) were associated positively with polypharmacy. Using only the public health system (OR = 0.5, 95% CI: 0.3; 0.7) was inversely associated with polypharmacy. Medicines for the cardiovascular system and for the alimentary tract and metabolism were the most used. The knowledge of factors associated with polypharmacy, such as those identified in this study, may be useful to alert health professionals about the importance of identifying and monitoring the elderly groups most vulnerable to polypharmacy.154817827Ramos, L.R., Fatores determinantes do envelhecimento saudável em idosos residentes em centro urbano: Projeto Epidoso, São Paulo (2003) Cad Saúde Pública, 19, pp. 793-798(2002) Active Ageing. A Policy Framework, , http://whqlibdoc.who.int/hq/2002/WHO_NMH_NPH_02.8.pdf, World Health Organization, Geneva: WHO;,. Disponível em, [Acessado em 20 de fevereiro de 2006]Coelho Filho, J.M., Marcopito, L.F., Castelo, A., Medication use patterns among elderly people in urban area in Northeastern Brazil (2004) Rev Saude Publica, 38, pp. 557-564Qato, D.M., Alexander, G.C., Conti, R.M., Johnson, M., Schumm, P., Lindau, S.T., Use of prescription and over-the-counter medications and dietary supplements among older adults in the United States (2008) JAMA, 300, pp. 2867-2878Ribeiro, A.Q., Rozenfeld, S., Klein, C.H., César, C.C., Acurcio Fde, A., Survey on medicine use by elderly retirees in Belo Horizonte, Southeastern Brazil (2008) Rev Saúde Pública, 42, pp. 724-732Flores, L.M., Mengue, S.S., Uso de medicamentos por idosos em região do sul do Brasil (2005) Rev Saúde Pública, 39, pp. 924-929Kaufman, D.W., Kelly, J.P., Rosenberg, L., Anderson, T.E., Michell, A.A., Recent patterns of medication use in the ambulatory adult population of the United States (2002) JAMA, 287, pp. 337-344Safran, D.G., Neuman, P., Schoen, C., Kitchman, M.S., Wilson, I.B., Cooper, B., Prescription drug coverage and seniors: Findings from a 2003 national survey (2005) Health Aff, , (Millwood),Suppl Web Exclusives: W5-152-W5-166Prybys, K.M., Melville, K., Hanna, J., Gee, A., Chyka, P., Polypharmacy in the elderly: Clinical challenges in emergency practice: Part 1 overview, etiology, and drug interactions (2002) Emerg Med Rep, 23, pp. 145-153Rozenfeld, S., Fonseca, M.J.M., Acurcio, F.A., Drug utilization and polypharmacy among the elderly: A survey in Rio de Janeiro City, Brazil (2008) Pan Am J Public Health, 23, pp. 34-43Lebrão, M.L., Duarte, Y.A.O., (2003) SABE-Saúde, Bem Estar e envelhecimento-O projeto SABE no município de São Paulo:Uma abordagem inicial [livro na internet], , http://www.opas.org.br/sistema/arquivos/l_saber.pdf, Brasília: Athalaia Bureau;,. Disponível em, [Acessado em 20 de fevereiro de 2006](2006) Anatomical therapeutic chemical (ATC) classification index with defined daily doses (DDDs), , http://www.whocc.no/atcddd/index, World Health Organization, [homepage na Internet]. Genevac2007. Disponível em, [atualizado em 16 de dezembro de, e acessado em 16 de março de 2007]Rao, J.N.K., Scott, A.J., On chi-squared tests for multiway contingency tables with cell proportions estimated from survey data (1984) Annals of Statistics, 12, pp. 46-60(2009) Stata Statistical Software: Release 11, , StataCorp, College Station. Texas: StataCorp LPRollason, V., Vogt, N., Reduction of polypharmacy in the elderly. A systematic review of the role of the pharmacist (2003) Drugs Aging, 20, pp. 817-832Rochon, P.A., Gurwitz, J.H., Optimising drug treatment for elderly people: The prescribing cascade (1997) BMJ, 315, pp. 1096-1099Gurwitz, J.H., Polypharmacy: A new paradigm for quality drug therapy in the elderly? (2004) Arch Int Med, 164, pp. 1957-1959Loyola Filho, A.I., Uchoa, E., Lima-Costa, M.F., A population-based study on use of medication by the elderly in Greater Metropolitan Belo Horizonte, Minas Gerais, Brazil (2006) Cad Saude Publica, 22, pp. 2657-2667Linjakumpu, T., Hartikainen, S., Klaukka, T., Veijola, J., Kivelä, S.L., Isoaho, R., Use of medications and polypharmacy are increasing among the elderly (2002) J Clin Epidemiol, 55, pp. 809-817Jörgensen, T., Johansson, S., Kennerfalk, A., Wallander, M.A., Svärdsudd, K., Prescription drug use, diagnoses, and healthcare utilization among the elderly (2001) Ann Pharmacother, 35, pp. 1004-1009Loyola Filho, A.I., Uchoa, E., Firmo, J.O., Lima-Costa, M.F., A population-based study on use of medications by elderly Brazilians: The Bambuí Health and Aging Study (BHAS) (2005) Cad Saúde Pública, 21, pp. 545-553Loyola Filho, A.I., Uchoa, E., Firmo, J.O., Lima-Costa, M.F., Influence of income on the association between cognitive impairment and polypharmacy: Bambuí Project (2008) Rev Saúde Pública, 42, pp. 89-99Rosholm, J.U., Christensen, K., Relationship between drug use and self-reported health in elderly Danes (1997) Eur J Clin Pharmacol, 53, pp. 179-183Parente, F., Cucino, C., Gallus, S., Bargiggia, S., Greco, S., Pastore, L., Bianchi Porro, G., Hospital use of acid-suppressive medications and its fall-out on prescribing in general practice: A 1-month survey (2003) Aliment Pharmacol Ther, 17, pp. 1503-1506Alves, L.C., Rodrigues, R.N., Determinantes da autopercepção de saúde entre idosos do Município de São Paulo, Brasil (2005) Rev Panam Salud Publica, 17 (5-6), pp. 333-341Woodward, M.C., Deprescribing: Achieving better health outcomes for older people through reducing medications (2003) J Pharm Pract Res, 33, pp. 323-328Stein, C., Moritz, I., (1999) A life course perspective of maintaining independence in older age, , http://whqlibdoc.who.int/hq/1999/WHO_HSC_AHE_99.2_life.pdf, Geneva: WHO.,. Disponível em, [Acessado em 20 de fevereiro de 2006](2006) Dispõe sobre o salário mínimo a partir de 1o de abril de, , http://www010.dataprev.gov.br/sislex/paginas/42/2006/11321.htm, BRASIL, Lei no 11.321, de 7 de julho de 2006, Disponível em, [Acessado em 23 de outubro de 2011