Accuracy of microRNAs as markers for the detection of neck lymph node metastases in patients with head and neck squamous cell carcinoma

Background: the presence of metastatic disease in cervical lymph nodes of head and neck squamous cell carcinoma (HNSCC) patients is a very important determinant in therapy choice and prognosis, with great impact in overall survival. Frequently, routine lymph node staging cannot detect occult metastases and the post-surgical histologic evaluation of resected lymph nodes is not sensitive in detecting small metastatic deposits. Molecular markers based on tissue-specific microRNA expression are alternative accurate diagnostic markers. Herein, we evaluated the feasibility of using the expression of microRNAs to detect metastatic cells in formalin-fixed paraffin-embedded (FFPE) lymph nodes and in fine-needle aspiration (FNA) biopsies of HNSCC patients.Methods: An initial screening compared the expression of 667 microRNAs in a discovery set comprised by metastatic and non-metastatic lymph nodes from HNSCC patients. the most differentially expressed microRNAs were validated by qRT-PCR in two independent cohorts: i) 48 FFPE lymph node samples, and ii) 113 FNA lymph node biopsies. the accuracy of the markers in identifying metastatic samples was assessed through the analysis of sensitivity, specificity, accuracy, negative predictive value, positive predictive value, and area under the curve values.Results: Seven microRNAs highly expressed in metastatic lymph nodes from the discovery set were validated in FFPE lymph node samples. MiR-203 and miR-205 identified all metastatic samples, regardless of the size of the metastatic deposit. Additionally, these markers also showed high accuracy when FNA samples were examined.Conclusions: the high accuracy of miR-203 and miR-205 warrant these microRNAs as diagnostic markers of neck metastases in HNSCC. These can be evaluated in entire lymph nodes and in FNA biopsies collected at different time-points such as pre-treatment samples, intraoperative sentinel node biopsy, and during patient follow-up. These markers can be useful in a clinical setting in the management of HNSCC patients from initial disease staging and therapy planning to patient surveillance.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Universidade Federal de São Paulo, Dept Biol Sci, Lab Canc Mol Biol, BR-04039032 São Paulo, SP, BrazilBarretos Canc Hosp, Mol Oncol Res Ctr, BR-14784400 Barretos, SP, BrazilBarretos Canc Hosp, Dept Pathol, BR-14784400 Barretos, SP, BrazilBarretos Canc Hosp, Dept Head & Neck Surg, BR-14784400 Barretos, SP, BrazilDuke NUS Grad Med Sch, Canc Stem Cell Biol Program, Singapore 169857, SingaporeUniversidade Federal de São Paulo, Dept Biol Sci, Lab Canc Mol Biol, BR-04039032 São Paulo, SP, BrazilFAPESP: 2012/14837-7Web of Scienc

TIMP3 and CCNA1 hypermethylation in HNSCC is associated with an increased incidence of second primary tumors

Background: Hypermethylation in the promoter regions is associated with the suppression of gene expression and has been considered a potential molecular marker for several tumor types, including head and neck squamous cell carcinomas (HNSCC).Methods: To evaluate the gene hypermethylation profile as a prognostic marker, this retrospective study used a QMSP approach to determine the methylation status of 19 genes in 70 HNSCC patients.Results: the methylation profile analysis of primary HNSCC revealed that genes CCNA1, DAPK, MGMT, TIMP3 and SFRP1 were frequently hypermethylated, with high specificity and sensitivity. TIMP3 and CCNA1 hypermethylation was significantly associated with lower rates of second primary tumor-free survival (p = 0.007 and p = 0.001; log-rank test, respectively).Conclusion: This study, for the first time, presents CCNA1 and TIMP3 hypermethylation as a helpful tool to identify HNSCC subjects at risk of developing second primary carcinomas.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Universidade Federal de São Paulo, Dept Biol Sci, Lab Mol Canc Biol, BR-04039020 São Paulo, BrazilBarretos Canc Hosp, Stat & Epidemiol Ctr, BR-14784400 Barretos, BrazilAC Camargo Hosp, Dept Head & Neck Surg, BR-01509010 São Paulo, BrazilBarretos Canc Hosp, Dept Head & Neck Surg, BR-14784400 Barretos, BrazilDuke NUS Grad Med Sch, Canc & Stem Cell Biol Program, Singapore 169857, SingaporeUniversidade Federal de São Paulo, Dept Biol Sci, Lab Mol Canc Biol, BR-04039020 São Paulo, BrazilWeb of Scienc

Clinical significance of molecular alterations in histologically negative surgical margins of head and neck cancer patients

The development of locoregional recurrence is the main reason for treatment failure in head and neck squamous cell carcinomas (HNSCC) and the remaining of tumor cells in surgical margins is associated with recurrence. Surgical margins are considered negative based on histologic assessment of the pathological specimen. However, this method lacks sensitivity in identifying cells that already started malignant transformation but have not yet developed a pathologic phenotype. We investigated the usefulness of assessing the expression of PTHLH, EPCAM, MMP9, LGALS1 and MET for the detection of molecular alterations in histologically negative surgical margins and determine the correlation of these tumor-related alterations with clinical and prognostic parameters. Differential gene expression was determined by quantitative RT-PCR analyses in normal mucosa, HNSCC and negative margin samples. Thirty-eight percent of the histologically negative surgical margins examined were margin-positive for overexpression of at least one of the genes evaluated. Moreover, MMP9 and PTHLH overexpression in the surgical margins was associated with the development of second primary tumors (p = 0.002) and lower rates of local control (log rank test p = 0.022; HR = 4.186; p = 0.035), respectively. These findings demonstrate that the overexpression of tumor-related genes in histologically negative surgical margins is a frequent event. the use of qRT-PCR may be an useful tool in detecting actually negative HNSCC surgical margins and the overexpression of specific genes in these margins could be helpful in the identification of patients with a higher risk of developing second primary tumors and local recurrences, thus aiding the surgeon in the delineation of the HNSCC resection extent and helping in the planning of adjuvant therapy. (C) 2011 Elsevier B.V. All rights reserved.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Universidade Federal de São Paulo, Lab Mol Canc Biol, Dept Biol Sci, BR-04039032 São Paulo, BrazilHosp Canc AC Camargo, Fundacao Antonio Prudente, Dept Head & Neck Surg, BR-01509010 São Paulo, BrazilHosp Canc AC Camargo, Fundacao Antonio Prudente, Dept Pathol, BR-01509010 São Paulo, BrazilHosp Canc Barretos, Dept Head & Neck Surg, Fundacao PIO 12, BR-14784400 Barretos, SP, BrazilUniversidade Federal de São Paulo, Lab Mol Canc Biol, Dept Biol Sci, BR-04039032 São Paulo, BrazilFAPESP: 2007/56245-0FAPESP: 2008/58460-9CNPq: 302360/2008-5Web of Scienc

RESULTADOS ECONÔMICOS DA ATIVIDADE LEITEIRA NA REGIÃO DE LAVRAS (MG) NOS ANOS 2004 E 2005: UM ESTUDO MULTICASOS

Estudou-se a rentabilidade da atividade leiteira de 17 propriedades da região de Lavras/MG e identificaram-se os componentes que exerceram maior influência sobre os custos finais da atividade. Os dados foram coletados mensalmente durante o período de janeiro de 2004 a dezembro de 2005. Considerou-se a margem bruta, a margem líquida e o resultado (lucro ou prejuízo) como indicadores de eficiência econômica. Os itens componentes do custo operacional efetivo que exerceram maior influência sobre os custos da atividade leiteira foram, em ordem decrescente, alimentação (55,9%), mão-de-obra (19,1%), despesas diversas (11,3%), energia (7,1%), sanidade (4,9%), ordenha (0,74%), impostos (0,74%) e inseminação artificial (0,23%). A margem líquida e o resultado positivo indicam que os pecuaristas têm condições de produzir a médio e a longo prazo, com conseqüente capitalização. Os resultados do estudo não podem ser extrapolados por não se tratar de uma amostra probabilística.----------------------------------------------It was studied the profitability of the milk production in 17 properties in the region of Lavras (MG), and the components that had the greatest influence on the final costs of the business were identified. The data were monthly collected from January 2004 to December 2005. The gross margin, net margin, and the result (profit or loss) were taken into consideration as indicators of economic efficiency. The component items of the effective operational cost that had the greatest influence upon the costs of dairy business were, in decreasing order, feeding (55.9%), labor (19.1%), different expenses (11.3%), energy (7.1%), health (4.9%), milking (0.74%), taxes (0.74%), and artificial insemination (0,23%). The net margin and the positive result show the farmers have conditions to produce in middle and long terms, with consequent capitalization. Although, the results of the study can not be extrapolated, since they do not represent a random sampling.bovinocultura de leite, análise de rentabilidade, custo de produção, ponto de equilíbrio, cattle, production cost, breakeven point, profitability analysis, Livestock Production/Industries,

EFEITO DO TIPO DE SISTEMA DE CRIAÇÃO NOS RESULTADOS ECONÔMICOS DE SISTEMAS DE PRODUÇÃO DE LEITE NA REGIÃO DE LAVRAS (MG) NOS ANOS 2004 E 2005

Estudou-se a rentabilidade da atividade leiteira na região de Lavras/MG, analisando a influência do tipo de sistema de criação sobre o custo de produção do leite. Foram identificados os componentes com maior influência sobre os custos finais da atividade e calculado o ponto de equilíbrio. Os dados, provenientes de 17 sistemas de produção, representando os sistemas de criação em regime de pastejo, semi-confinado e confinado, foram coletados mensalmente, entre janeiro de 2004 e dezembro de 2005. Considerou-se a margem bruta, a margem líquida e o resultado (lucro ou prejuízo) como indicadores de eficiência econômica. O tipo do sistema de criação influenciou o custo total de produção do leite e, portanto a lucratividade e rentabilidade, sendo os sistemas de produção semi-confinados os que apresentaram os menores custos totais unitários. O tipo do sistema de criação influenciou os “pesos” dos itens componentes do custo operacional efetivo da atividade leiteira, sendo esses diferentes em cada um dos três sistemas estudados. Na análise econômica, por apresentar margem líquida e resultado positivo, nos sistemas de criação confinado e semi confinado, conclui-se que a atividade leiteira tem condições de produzir no longo prazo e os pecuaristas estão se capitalizando. No entanto, no sistema em regime de pastejo os pecuaristas têm condições de produzir no médio prazo, estando se descapitalizando no longo prazo.----------------------------------------------The profitability of the milk activity was studied in the region of Lavras, MG by analyzing the influence of the types of breeding system upon the cost production of milk. It was identified the components with major influence on the final cost of the activity and calculated the breakeven point. Monthly data from 17 production systems representing the grazing regimen, semi-feedlot and feedlot systems were used for the period of January 2004 to December 2005. It was considered the gross margin, the net (profit or loss) as indicators of economical efficiency. The type of breeding system influenced the “weights’’ of the component items of the effective operational cost of dairy business, these being different in each of the three levels studied. In the economical analysis, the feedlot and semi-feedlot presented showing that dairy is profitable in the long-term and that farmers are positive net margin. In the grazing regimen, the conclusion is that milk farmers have conditions to produce in the middle-term. In the long-term; however, milk farmers have been losing money.análise de rentabilidade, bovinocultura de leite, custo de produção, sistema de criação, ponto de equilíbrio, breakeven point, breeding system, dairy cattle, production cost, profitability analysis, Livestock Production/Industries,

The effect of bone marrow-derived stem cells associated with platelet-rich plasma on the osseointegration of immediately placed implants

Stem cells associated with growth factors have been shown to improve bone healing and the osseointegration of dental implants. A Brazilian miniature pig model was used to evaluate the effect of autologous bone marrow-derived mesenchymal stem cells (BM-MS

Diabetes mellitus, metformin and head and neck cancer

Introduction: Diabetes mellitus (DM (Diabetes Mellitus)) is directly associated with some cancers. However, studies on the association between diabetes mellitus and head and neck cancer (HNC (Head and Neck Cancer)) have rendered controversial results. The objective of this study was to evaluate the association between DM and HNC, as well as the impact of metformin use on the risk of HNC. Material and methods: This case-control study was conducted within the framework of the Brazilian Head and Neck Genome Project in 2011-2014. The study included 1021 HNC cases with histologically confirmed squamous cell carcinoma of the head and neck admitted to five large hospitals in Sao Paulo state. A total of 1063 controls were selected in the same hospitals. Odds ratios (OR) and 95% confidence intervals (CI) were estimated using unconditional logistic regression. Results: Diabetic participants had a decreased risk of HNC (OR = 0.68; 95% CI: 0.49-0.95) than nondiabetic participants, and this risk was further decreased among diabetic metformin users (OR = 0.54; 95% CI: 0.29-0.99). Diabetic metformin users that were current smokers (OR = 0.13; 95% CI: 0.04-0.44) or had an alcohol consumption of >40 g/day (OR = 0.31; 95% CI: 0.11-0.88) had lower risk of HNC than equivalent non-diabetic participants. Conclusion: The risk of HNC was decreased among diabetic participants; metformin use may at least partially explain this inverse association. (C) 2016 Elsevier Ltd. All rights reserved.Peer reviewe

Efeito da irrigação de pastagens na rentabilidade de sistemas de produção de leite participantes do programa “Balde Cheio”

This research sought to analyze the effect of irrigation of grasslands on the profitability of dairy production by 20 demonstration units (UDs) that participate in the dairy improvement program called “Balde Cheio”. Understudy were the components that exerted the greatest influence on total and effective (cash flow) operating costs and the impact of each one on economic return. The data were processed using PASW 18.0 software and multiple linear regression incorporating the stepwise method to determine the relative effect of each component of production costs on the criteria liquid margin, profitability and investment yield. Pasture irrigation was found to affect the total cost of milk production and, thus, profitability and yield. The stratum of non-irrigating producers had the lowest total cost and effective operating costs. The liquid margins were positive, but the net margins were negative in both strata, thus, the dairy operation is not viable in the long run, due to decapitalization of the producers. The items of effective operating costs with most efforts on the dairy business (in decreasing order) were: in the non-irrigating stratum, feed, sundry expenses and labor, and in the irrigating stratum, feed, labor and energy. In both strata, the most influential components of total cost followed in order: feed, family labor and remuneration of capital invested.Objetivou-se analisar o efeito da irrigaçãode pastagens na rentabilidade de 20 unidades demonstrativas (UD) participantes do programa “Balde Cheio”, no estado do RJ, no período de janeiro a dezembro de 2011. Pretendeu-se, ainda, identificar os componentes que exerceram maior influência sobre os custos total e operacional efetivo e o impacto de cada um deles na receita.Os dados foram analisados utilizando-se o software PASW 18.0. Utilizou-se o modelo de regressão linear múltipla com o método stepwise, para identificar os componentes do custo de produção que mais influenciaram na margem líquida, na lucratividade e na rentabilidade.A irrigação das pastagens influenciou o custo total de produção do leite e, portanto, a lucratividade e rentabilidade. O estrato dos produtores que não irrigam apresentou os menores custos totais e menores custos operacionais efetivos unitários. Por apresentar margem líquida positiva e resultado negativo, nos dois estratos, a atividade leiteira nãoteve condições de produzir no longo prazo, com descapitalização, dos pecuaristas. Os itens componentes do custo operacional efetivo que exerceram maiores representatividades sobre os custos da atividade leiteira, no estrato dos produtores que não irrigam, em ordem decrescente, foram a alimentação, despesas diversas e mão de obra. No estrato daqueles produtores que utilizavam a irrigação, foram a alimentação, mão de obra e energia. Nos dois estratos, os itens componentes do custo total que exerceram maiores representatividades sobre os custos da atividade leiteira, em ordem decrescente, foram a alimentação, mão de obra familiar e remuneração do capital investido

Usefulness of aqueous and vitreous humor analysis in infectious uveitis

OBJECTIVE: To evaluate the role of intraocular fluid analysis as a diagnostic aid for uveitis. METHODS: Twenty-eight samples (27 patients including 3 HIV-infected patients) with active (n=24) or non-active (n=4) uveitis were submitted to aqueous (AH; n=12) or vitreous humor (VH) analysis (n=16). All samples were analyzed by quantitative PCR for herpes simplex virus (HSV), varicella zoster virus (VZV), cytomegalovirus (CMV), Epstein-Barr virus (EBV) and Toxoplasma gondii. RESULTS: The positivity of the PCR in AH was 41.7% (5/12), with 50% (2/4) in immunocompetent and 67% (2/3) in HIV+ patients. The positivity of the PCR in VH was 31.2% (5/16), with 13% (1/8) in immunocompetent and 50% (4/8) in immunosuppressed HIV negative patients. The analysis was a determinant in the diagnostic definition in 58% of HA and 50% of VH. CONCLUSION: Even in posterior uveitis, initial AH analysis may be helpful. A careful formulation of possible clinical diagnosis seems to increase the chance of intraocular sample analysis being meaningful

Effect of technical and management indexes on profitability of dairy production with different types of labour

The effect of some technical and management indexes on economic efficiency was studied in 20 demonstration units (UD), with different types of labour, participants in the «Balde Cheio» program, in Rio de Janeiro State from January to December 2011. The analysis was based on gross margin, liquid margin, net result (profit or loss), rentability and profitability. Data analysis made use of SPSS 18.0 software. Multiple linear regression incorporating the stepwise method was used to identify the technical and management indexes most influential in profitability and rentability. The indexes cows were found to differ among the types of labour and influenced the economic efficiency. The family labour stratum gave the highest percentages of lactating cows/all cows, cows/all animals in the herd, lactating cows/all animals, greatest daily productivity per animal and per hectare-year, but this was not enough to obtain profit. The mixed labour stratum resulted in the highest number of lactating cows/ha and the lowest total labour input, and although it showed a positive net margin, the net result was negative. The contracted labour stratum yielded the best ratios of herd animals to labour and milk production, in addition to being the only one to achieve both positive liquid margin and positive net result