Evidence of a noncoding transcript of the RIPK2 gene overexpressed in head and neck tumor

Receptor-interacting proteins are a family of serine/threonine kinases, which integrate extra and intracellular stress signals caused by different factors, including infections, inflammation and DNA damage. Receptor-interacting serine/threonine-protein kinase 2 (RIP-2) is a member of this family and an important component of the nuclear factor NF-kappa-B signaling pathway. The corresponding human gene RIPK2 generates two transcripts by alternative splicing, the full-length and a short transcript. The short transcript has a truncated 5? sequence, which results in a predicted isoform with a partial kinase domain but able to transduce signals through its caspase recruitment domain. In this study, the expression of RIPK2 was investigated in human tissue samples and, in order to determine if both transcripts are similarly regulated at the transcriptional level, cancer cell lines were submitted to temperature and acid stresses. We observed that both transcripts are expressed in all tissues analyzed, with higher expression of the short one in tumor samples, and they are differentially regulated following temperature stress. Despite transcription, no corresponding protein for the short transcript was detected in tissues and cell lines analyzed. We propose that the shorter transcript is a noncoding RNA and that its presence in the cell may play regulatory roles and affect inflammation and other biological processes related to the kinase activity of RIP-2.Fil: Mancini Villagra, Ulises Maximiliano. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Biotecnología y Biología Molecular. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Instituto de Biotecnología y Biología Molecular; ArgentinaFil: da Cunha, Bianca R.. Universidade de Sao Paulo; BrasilFil: Polachini, Giovana M.. No especifíca;Fil: Tiago, Tiago Henrique. No especifíca;Fil: Carlos H. T. P. da Silva. Universidade de Sao Paulo; BrasilFil: Feitosa, Olavo A.. Universidade de Sao Paulo; BrasilFil: Fukuyama, Erica E.. Arnaldo Vieira de Carvalho Cancer Institute; BrasilFil: López, Rossana V. M.. No especifíca;Fil: Dias Neto, Emmanuel. Universidade de Sao Paulo; BrasilFil: Nunes, Fabio D.. Universidade de Sao Paulo; BrasilFil: Severino, Patricia. Hospital Israelita Albert Einstein; BrasilFil: Tajara, Eloiza Helena Tajara. Universidade de Sao Paulo; Brasi

Evidence of a noncoding transcript of the RIPK2 gene overexpressed in head and neck tumor

Receptor-interacting proteins are a family of serine/threonine kinases, which integrate extra and intracellular stress signals caused by different factors, including infections, inflammation and DNA damage. Receptor-interacting serine/threonine-protein kinase 2 (RIP-2) is a member of this family and an important component of the nuclear factor NF-kappa-B signaling pathway. The corresponding human gene RIPK2 generates two transcripts by alternative splicing, the full-length and a short transcript. The short transcript has a truncated 5’ sequence, which results in a predicted isoform with a partial kinase domain but able to transduce signals through its caspase recruitment domain. In this study, the expression of RIPK2 was investigated in human tissue samples and, in order to determine if both transcripts are similarly regulated at the transcriptional level, cancer cell lines were submitted to temperature and acid stresses. We observed that both transcripts are expressed in all tissues analyzed, with higher expression of the short one in tumor samples, and they are differentially regulated following temperature stress. Despite transcription, no corresponding protein for the short transcript was detected in tissues and cell lines analyzed. We propose that the shorter transcript is a noncoding RNA and that its presence in the cell may play regulatory roles and affect inflammation and other biological processes related to the kinase activity of RIP-2.Instituto de Biotecnologia y Biologia Molecula

Bias corrections of GOSAT SWIR XCO₂ and XCH₄ with TCCON data and their evaluation using aircraft measurement data

We describe a method for removing systematic biases of column-averaged dry air mole fractions of CO2 (XCO2) and CH4 (XCH4) derived from short-wavelength infrared (SWIR) spectra of the Greenhouse gases Observing SATellite (GOSAT). We conduct correlation analyses between the GOSAT biases and simultaneously retrieved auxiliary parameters. We use these correlations to bias correct the GOSAT data, removing these spurious correlations. Data from the Total Carbon Column Observing Network (TCCON) were used as reference values for this regression analysis. To evaluate the effectiveness of this correction method, the tnzuncorrected/corrected GOSAT data were compared to independent XCO2 and XCH4 data derived from aircraft measurements taken for the Comprehensive Observation Network for TRace gases by AIrLiner (CONTRAIL) project, the National Oceanic and Atmospheric Administration (NOAA), the US Department of Energy (DOE), the National Institute for Environmental Studies (NIES), the Japan Meteorological Agency (JMA), the HIAPER Pole-to-Pole observations (HIPPO) program, and the GOSAT validation aircraft observation campaign over Japan. These comparisons demonstrate that the empirically derived bias correction improves the agreement between GOSAT XCO2/XCH4 and the aircraft data. Finally, we present spatial distributions and temporal variations of the derived GOSAT biases

Genomics and proteomics approaches to the study of cancer-stroma interactions

<p>Abstract</p> <p>Background</p> <p>The development and progression of cancer depend on its genetic characteristics as well as on the interactions with its microenvironment. Understanding these interactions may contribute to diagnostic and prognostic evaluations and to the development of new cancer therapies. Aiming to investigate potential mechanisms by which the tumor microenvironment might contribute to a cancer phenotype, we evaluated soluble paracrine factors produced by stromal and neoplastic cells which may influence proliferation and gene and protein expression.</p> <p>Methods</p> <p>The study was carried out on the epithelial cancer cell line (Hep-2) and fibroblasts isolated from a primary oral cancer. We combined a conditioned-medium technique with subtraction hybridization approach, quantitative PCR and proteomics, in order to evaluate gene and protein expression influenced by soluble paracrine factors produced by stromal and neoplastic cells.</p> <p>Results</p> <p>We observed that conditioned medium from fibroblast cultures (FCM) inhibited proliferation and induced apoptosis in Hep-2 cells. In neoplastic cells, 41 genes and 5 proteins exhibited changes in expression levels in response to FCM and, in fibroblasts, 17 genes and 2 proteins showed down-regulation in response to conditioned medium from Hep-2 cells (HCM). Nine genes were selected and the expression results of 6 down-regulated genes (<it>ARID4A</it>, <it>CALR</it>, <it>GNB2L1</it>, <it>RNF10</it>, <it>SQSTM1</it>, <it>USP9X</it>) were validated by real time PCR.</p> <p>Conclusions</p> <p>A significant and common denominator in the results was the potential induction of signaling changes associated with immune or inflammatory response in the absence of a specific protein.</p

Methylation profile of genes CDKN2A (p14 and p16), DAPK1, CDH1, and ADAM23 in head and neck cancer

Hypermethylation in the promoter region has been associated with a loss of gene function that may give a selective advantage to neoplastic cells. In this study, the methylation pattern of genes CDKN2A (alias p14, p14(ARF), p16, p16(INK4a)), DAPK1, CDH1, and ADAM23 was analyzed in 43 samples of head and neck tumors using methylation-specific polymerase chain reaction. In the oropharynx, there was a statistically significant association between hypermethylation of the DAPK1 gene and the occurrence of lymph node metastases, and in the larynx there was statistically significant evidence of an association between hypermethylation of the ADAM23 gene and advanced stages of the tumors. Thus, a correlation was observed between hypermethylation of the promoter region of genes DAPK1 and ADAM23 and the progression of head and neck cancer. (c) 2007 Elsevier B.V. All rights reserved

Análise acústica da voz captada na faringe próximo à fonte glótica através de microfone acoplado ao fibrolaringoscópio Acoustic analysis of voice captured in the pharynx above the glottic source through a microphone on a laryngo-fiberscope

Objetivo: O objetivo deste trabalho foi estudar a voz próximo à sua fonte produtora, as pregas vocais, através de um microfone miniaturizado de aparelho auditivo que foi adaptado para ser acoplado à extremidade de um fibrolaringoscópio, permitindo a captação da voz durante a laringoscopia direta. Forma de estudo: Experimental. Material e Método: A voz foi estudada em um grupo de 50 indivíduos, 25 homens e 25 mulheres sem doenças, através de um programa de análise acústica MDVP (Multi-Dimensional Voice Program) do laboratório de voz Computerized Speech Lab, Model 4300B, da Kay Elemetrics. Amostras de vogais sustentadas /a/, /i/ e /u/ foram captadas de três formas diferentes, primeiramente com um microfone comum externo a 15 cm da boca, em segundo lugar com o microfone especial na faringe a 1,5 cm acima das pregas vocais e por último com o microfone especial externamente a 2 cm da boca. Doze parâmetros acústicos relacionados a freqüência fundamental, amplitude e ruído de cada uma das vogais foram comparadas estatisticamente conforme à sua forma de captação. Resultados: Os resultados mostraram diferenças estatisticamente significativas entre a voz captada pelo microfone comum externo e o microfone especial, em relação à freqüência fundamental, aos parâmetros de variação de periodicidade de freqüência, amplitude e ruído. Conclusão: A diferença do som da fonte glótica do som da voz externa pode mostrar as modificações sofridas pela voz no decorrer da passagem pelo trato vocal.<br>Aim: The aim of the present study is to examine the voice to its acoustic source - the vocal folds - with a miniature hearing-aid microphone coupled to the extremity of a laryngo-fiberscope allowing the voice to be captured during direct laryngoscopy. Study design: Experimental. Material and Method: The voice of 50 individuals - 25 males and 25 females bearing no pathologies - was collected by the Multi-Dimensional Voice Program (MDVP) by Kay Elemetrics’ Computerized Speech Lab 4300B Model. Samples of the sustained vowels /a/, /i/ and /u/ were picked up in three distinct ways. Firstly, by a common external microphone placed at 15 cm from the mouth. Secondly, a special microphone was placed on the pharynx 1.5 cm above the vocal folds. Lastly, the same special microphone was placed externally at 2 cm from the mouth. Twelve acoustic parameters regarding fundamental frequency, amplitude and noise of each and every vowel were compared statistically as to the way the voice was picked up. Results: Results show statistically significant differences between the voice picked up by the common external microphone and by the special one as regards to the fundamental frequency, frequency and amplitude variability and noise. Conclusion: The difference between the sound coming from the glottic source and the sound from the external voice shows alterations experienced by the voice during its passage through the vocal tract

Proteomic Approaches Identify Members of Cofilin Pathway Involved in Oral Tumorigenesis

<div><p>The prediction of tumor behavior for patients with oral carcinomas remains a challenge for clinicians. The presence of lymph node metastasis is the most important prognostic factor but it is limited in predicting local relapse or survival. This highlights the need for identifying biomarkers that may effectively contribute to prediction of recurrence and tumor spread. In this study, we used one- and two-dimensional gel electrophoresis, mass spectrometry and immunodetection methods to analyze protein expression in oral squamous cell carcinomas. Using a refinement for classifying oral carcinomas in regard to prognosis, we analyzed small but lymph node metastasis-positive versus large, lymph node metastasis-negative tumors in order to contribute to the molecular characterization of subgroups with risk of dissemination. Specific protein patterns favoring metastasis were observed in the “more-aggressive” group defined by the present study. This group displayed upregulation of proteins involved in migration, adhesion, angiogenesis, cell cycle regulation, anti-apoptosis and epithelial to mesenchymal transition, whereas the “less-aggressive” group was engaged in keratinocyte differentiation, epidermis development, inflammation and immune response. Besides the identification of several proteins not yet described as deregulated in oral carcinomas, the present study demonstrated for the first time the role of cofilin-1 in modulating cell invasion in oral carcinomas.</p> </div

Immunodetection of keratin 4 expression in OSCC samples. Immunohistochemistry analysis:

<p>pattern of keratin 4 immunostaining in (A) superficial layers of epithelium in margin showing intense positivity in stratum corneum (A, insert); (B) absence of keratin 4 immunostaining in nests of well differentiated and (C) poorly differentiated areas of OSCC. Scale bar indicates 100 µm. <b>Western blot</b>: (D) tumor samples (lanes 1, 3, 5, 7) and matched margins (lanes 2, 4, 6, 8) from patients with T1N0, T4N2, T4N1 and T4N1 carcinomas, respectively; (E) Surgical margin (lane 1) and tumor samples (lanes 2, 3, 4, 5) from patients with T4N2, T4N2, T4N2, T1N0 and T2N2, respectively. β-actin was used as an internal control. MW, PageRuler™ Prestained Protein Ladder.</p