Aberrant MUC1 accumulation in salivary glands of Sjögren's syndrome patients is reversed by TUDCA in vitro.

OBJECTIVES: Xerostomia in SS patients has been associated with low quality and quantity of salivary mucins, which are fundamental for the hydration and protection of the oral mucosa. The aim of this study was to evaluate if cytokines induce aberrant mucin expression and whether tauroursodeoxycholic acid (TUDCA) is able to counteract such an anomaly. METHODS: Labial salivary glands from 16 SS patients and 15 control subjects, as well as 3D acini or human submandibular gland cells stimulated with TNF-α or IFN-γ and co-incubated with TUDCA, were analysed. mRNA and protein levels of Mucin 1 (MUC1) and MUC7 were determined by RT-qPCR and western blot, respectively. Co-immunoprecipitation and immunofluorescence assays for mucins and GRP78 [an endoplasmic reticulum (ER)-resident protein] were also performed. mRNA levels of RelA/p65 (nuclear factor-κB subunit), TNF-α, IL-1ß, IL-6, SEL1L and EDEM1 were determined by RT-qPCR, and RelA/p65 localization was evaluated by immunofluorescence. RESULTS: MUC1 is overexpressed and accumulated in the ER of labial salivary gland from SS patients, while MUC7 accumulates throughout the cytoplasm of acinar cells; however, MUC1, but not MUC7, co-precipitated with GRP78. TUDCA diminished the overexpression and aberrant accumulation of MUC1 induced by TNF-α and IFN-γ, as well as the nuclear translocation of RelA/p65, together with the expression of inflammatory and ER stress markers in 3D acini. CONCLUSION: Chronic inflammation alters the secretory process of MUC1, inducing ER stress and affecting the quality of saliva in SS patients. TUDCA showed anti-inflammatory properties decreasing aberrant MUC1 accumulation. Further studies are necessary to evaluate the potential therapeutic effect of TUDCA in restoring glandular homeostasis in SS patients.

Sedation-analgesia with propofol and remifentanil: concentrations required to avoid gag reflex in upper gastrointestinal endoscopy.

BACKGROUND: The purpose of this study was to identify optimal target propofol and remifentanil concentrations to avoid a gag reflex in response to insertion of an upper gastrointestinal endoscope. METHODS: Patients presenting for endoscopy received target-controlled infusions (TCI) of both propofol and remifentanil for sedation-analgesia. Patients were randomized to 4 groups of fixed target effect-site concentrations: remifentanil 1 ng•mL (REMI 1) or 2 ng•mL (REMI 2) and propofol 2 µg•mL (PROP 2) or 3 µg•mL (PROP 3). For each group, the other drug (propofol for the REMI groups and vice versa) was increased or decreased using the "up-down" method based on the presence or absence of a gag response in the previous patient. A modified isotonic regression method was used to estimate the median effective Ce,50 from the up-down method in each group. A concentration-effect (sigmoid Emax) model was built to estimate the corresponding Ce,90 for each group. These data were used to estimate propofol bolus doses and remifentanil infusion rates that would achieve effect-site concentrations between Ce,50 and Ce,90 when a TCI system is not available for use. RESULTS: One hundred twenty-four patients were analyzed. To achieve between a 50% and 90% probability of no gag response, propofol TCIs were between 2.40 and 4.23 µg•mL (that could be achieved with a bolus of 1 mg•kg) when remifentanil TCI was fixed at 1 ng•mL, and target propofol TCIs were between 2.15 and 2.88 µg•mL (that could be achieved with a bolus of 0.75 mg•kg) when remifentanil TCI was fixed at 2 ng•mL. Remifentanil ranges were 1.00 to 4.79 ng•mL and 0.72 to 3.19 ng•mL when propofol was fixed at 2 and 3 µg•mL, respectively. CONCLUSIONS: We identified a set of propofol and remifentanil TCIs that blocked the gag response to endoscope insertion in patients undergoing endoscopy. Propofol bolus doses and remifentanil infusion rates designed to achieve similar effect-site concentrations can be used to prevent gag response when TCI is not available.

Cell membrane CD44v6 levels in squamous cell carcinoma of the lung: association with high cellular proliferation and high concentrations of EGFR and CD44v5.

Membranous CD44v6 levels in tumors and surrounding samples obtained from 94 patients with squamous cell lung carcinomas were studied and compared to clinical stage, cellular proliferation, membranous CD44v5 levels, epidermal growth factor receptor EGFR and cytoplasmatic concentrations of CYFRA 21.1. CD44v6 positive values were observed in 33/38 non-tumor samples and in 76/94 tumor samples, but there were not statistically significant differences between both subgroups. In CD44v6 positive tumor samples, CD44v6 was not associated with clinical stage, histological grade, ploidy and lymph node involvement, but significant association was found with high cellular proliferation. Likewise, CD44v6 positive tumors had significantly higher levels of EGFR and CD44v5. In patients with squamous cell lung carcinomas and clinical stage I, positive CD44v6 cases were associated with the same parameters. Furthermore, positive CD44v5 squamous tumors were associated significantly with histological grade III and lower levels of CYFRA21.1. Our findings support the value of CD44v6 as a possible indicator of poor outcome in patients with squamous lung carcinomas.

[Usefulness of cytosolic tissue-type plasminogen activator levels in lung adenocarcinomas]. / Utilidad de la determinación del activador del plasminógeno tipo tisular citosólico en el adenocarcinoma de pulmón.

BACKGROUND: We aimed at studying the behavior of cytosol tissue-type plasminogen activator (t-PA) levels in lung adenocarcinomas and their correlation with other clinical and biological parameters. MATERIAL AND METHOD: t-PA cytosol levels were determined using EIA (Boehringer Mannheim. Germany) in 59 samples of lung adenocarcinoma and in 16 samples of normal lung tissue from the same patients. Cathepsin D, CA125, pS2, hyaluronic acid (HA), free beta subunit of chorionic gonadotropin hormone and neuron specific enolase (NSE) cytosol concentrations were determined. We also determined the concentrations of HA, erbB2 oncoprotein, epidermal growth factor receptor (EGFR), CD44s, CD44v5 and CD44v6 in cell surfaces. The following parameters were considered: clinical stage, ploidy, cellular S-phase fraction and histological grade. RESULTS: In adenocarcinomas, t-PA cytosol levels ranged from 0.1 to 14.6 ng/mg prot. (median, 1.4). These levels were lower than those in normal lung tissue (r, 0.1-18.6; median, 2.95 ng/mg prot.) but did not reach statistical significance. On the other hand, t-PA concentrations decreased as the clinical stage increased and were higher in stage I than stage II-III (p = 0.080) and stage III (p = 0.0622). No significant differences of t-PA levels were observed when the histological grade, ploidy and S-phase were considered. Adenocarcinomas with high t-PA values (> 3.7 ng/mg prot., representing the 75th percentile of the whole group), had lower CA125 (p = 0.015) and erbB2 oncoprotein (p = 0.087) concentrations. CONCLUSIONS: These results suggest that cytosol t-PA levels are negatively correlated with tumor size in lung adenocarcinomas. However, the usefulness of t-PA as a prognostic factor needs to be clarified in further studies.

[Cytosolic cathepsin D levels in squamous carcinomas of the lung]. / Significado de la concentración citosólica de catepsina D en el carcinoma escamoso de pulmón.

BACKGROUND AND OBJECTIVES: Cathepsin D is an aspartyl proteinase involved in tumoral invasion. The aim of this work was to study cathepsin D cytosolic levels in squamous carcinomas of the lung and their correlation with several clinical and biological parameters. PATIENTS AND METHOD: The study group included 95 squamous lung carcinomas and 38 normal tissue samples from the same patients. Cathepsin D cytosolic concentrations were determined using an immunoradiometric assay (CIS BioInternational. France). EGFR, erbB2 protein, CD44s, CD44v5 and CD44v6 levels at cell surfaces were determined. The clinical stage, histological grade, ploidy and S-phase cellular fraction (SP) were also considered as variables of the study. RESULTS: Cathepsin D cytosolic levels oscillated between 7.7 and 576 (median: 38.8) pmol/mg protein and were lower (p = 0.001) than those observed in 38 normal lung samples from the same patients. When tumors were classified according to different clinical and biological parameters, we noticed that cathepsin D levels were higher in carcinomas with lower proliferation rates and no nodal involvement, reaching statistical significance in both cases. Moreover, when lung carcinomas were classified according to cathepsin D concentrations, tumors with higher cathepsin D concentrations had lower EGFR levels (p = 0.011) and small global SP values (p = 0.025) and DNA index (p = 0.023). Likewise, they were found to be CD44s positive more frequently (p = 0.001) and SP positive less frequently (p = 0.022). CONCLUSIONS: These results lead us to suggest the following: a) in squamous carcinomas of the lung, cathepsin D cytosolic levels are lower than those observed in normal lung samples from the same patients, and b) in this subtype of lung carcinomas, high cathepsin D levels are associated with tumors without nodal involvement, with low proliferation rates, lower EGFR levels, and a reduced positivity for CD44s, pointing to a possible role of this proteinase as a parameter of good outcome.

Utilidad de la determinación del activador del plasminógeno tipo tisular citosólico en el adenocarcinoma de pulmón / Usefulness of cytosolic tissue-type plasminogen activator levels in lung adenocarcinomas

FUNDAMENTO: Estudiar el comportamiento del activador del plasminógeno tipo tisular (t-AP) citosólico en adenocarcinomas pulmonares y sus posibles correlaciones con otros parámetros clinicobiológicos. MATERIAL Y MÉTODO: El grupo de estudio incluyó 59 muestras tisulares de adenocarcinomas pulmonares y 16 de tejido normal de los mismos pacientes. El t-AP fue determinado mediante un enzimoinmunoanálisis (EIA) (Boehringer-Mannheim, Alemania). Se determinaron asimismo las concentraciones citosólicas de CA125, catepsina D, pS2, ácido hialurónico (AH), subunidad beta de la hormona gonadotrófica coriónica, y enolasa específica neuronal, así como las del receptor del factor de crecimiento epidérmico (EGFR), proteína oncogénica erbB2/neu, AH, CD44s, CD44v5 y CD44v6 en las membranas celulares. Otros parámetros considerados fueron los siguientes: estadio clínico, grado histológico, ploidía y fase de síntesis celular (FS). RESULTADOS: En los adenocarcinomas, las concentraciones citosólicas de t-AP oscilaron entre 0,1 y 14,6 ng/mg de proteína (mediana, 1,4) y fueron inferiores, pero sin significación estadística, a las observadas en las muestras de pulmón normal (intervalo, 0,1-18,6; mediana, 2,95 ng/mg de proteína). Asimismo, las concentraciones de t-AP disminuyeron conforme se incrementaba el estadio clínico, siendo superiores en el estadio I frente al II-III (p = 0,080) y frente al III (p = 0,0622). No se constataron diferencias significativas en las concentraciones de la proteasa cuando el grado histológico, ploidía y FS fueron considerados. Los tumores con cifras altas de t-AP (> 3,7 ng/mg de proteína, que representan el percentil 75 de todo el grupo) cursaron con menores concentraciones de CA125 (p = 0,015) y proteína erbB2 (p = 0,087). CONCLUSIONES: Los resultados anteriores apuntan a que el t-AP citosólico se relaciona inversamente con el estadio clínico en el adenocarcinoma de pulmón, sin que podamos precisar su interés como indicador pronóstico. (AU)

Significado de la concentración citosólica de catepsina D en el carcinoma escamoso de pulmón / Cytosolic cathepsin D levels in squamous carcinomas of the lung

FUNDAMENTO Y OBJETIVOS: La catepsina D es una aspartilproteasa involucrada en la invasión tumoral. Dada la reducida bibliografía, hemos querido estudiar las concentraciones citosólicas de catepsina D en carcinomas escamosos de pulmón y correlacionarlas con otros parámetros clinicobiológicos. PACIENTES Y MÉTODO: El grupo de estudio incluyó a 95 pacientes afectados de carcinomas escamosos de pulmón y 38 muestras de tejido normal de los mismos pacientes. La catepsina D se determinó en los citosoles del tejido tumoral y normal mediante un método inmunorradiométrico (IRMA, CIS, Biointernacional, Francia). También se dosificaron, mediante métodos inmunológicos, las concentraciones de receptor del factor de crecimiento epidérmico (EGFR), proteína erbB2, CD44s, CD44v5 y CD44v6 en las membranas celulares. Otros parámetros analizados fueron estadio, grado histológico, ploidía y fase de síntesis celular (FS). RESULTADOS: Las concentraciones de catepsina D en los tumores oscilaron entre 7,7 y 576 (mediana, 38,8) pmol/mg de proteína y fueron inferiores (p = 0,001) a las observadas en los tejidos normales (mediana, 74,6; intervalo, 8,9-133,2). Cuando los tumores fueron clasificados en función de los diferentes parámetros clinicobiológicos, pudimos observar que las concentraciones de la proteasa fueron mayores en los casos poco proliferativos y sin afección ganglionar, rozándose en ambos casos la significación estadística. Cuando clasificamos los tumores en función de la concentración de catepsina D, tomando como dinteles las concentraciones de 71,4 y 28,8 pmol/mg de proteína que representan los percentiles 75 y 25 de todo el grupo, pudimos comprobar que los tumores ricos en catepsina presentaron menores concentraciones de EGFR (p = 0,011) y reducidos valores de FS global (p = 0,025) e índice de ADN (p = 0,023). Asimismo fueron más frecuentemente CD44s positivos y menos FS positivos (p = 0,022). CONCLUSIONES: Las concentraciones citosólicas de catepsina D en los carcinomas escamosos de pulmón son menores que las constatadas en el tejido normal de los mismos pacientes. Las cifras elevadas de la proteasa que se asociaron a tumores sin afección ganglionar, poco proliferativos, con menores concentraciones de EGFR y con reducida positividad para el CD44s, apuntan a que tienen un posible valor como indicador de un mejor pronóstico en este tipo de neoformaciones malignas (AU)