Expression of cGMP-binding cGMP-specific phosphodiesterase (PDE5) in mouse tissues and cell lines using an antibody against the enzyme amino-terminal domain

We have produced a polyclonal antibody that specifically recognizes cGMP-binding cGMP-specific phosphodiesterase (PDES). The antibody was raised in rabbit using as immunogen a fusion protein, in which glutathione S-transferase was coupled to a 171 amino acid polypeptide of the N-terminal region of bovine PDE5. The antibody is able to immunoprecipitate PDES activity from mouse tissues and neuroblastoma extracts while it has no effect on all other PDE isoforms present in the extracts. PDES activity recovered in the immunoprecipitates retains its sensitivity to specific inhibitors such as zaprinast (IC50 = 0.6 muM) and sildenafil (IC50 = 3.5 nM), Bands of the expected molecular mass were revealed when solubilized immunoprecipitates were analysed in Western blots. The antibody selectively stained cerebellar Purkinje neurones, which are known to express high levels of PDES mRNA. Western blot analysis of mouse tissues revealed the highest expression signal in mouse lung, followed by heart and cerebellum, while a lower signal was evident in brain, kidney and a very low signal was present in the liver. In the hybrid neuroblastoma-glioma NG108-15 cells the antibody revealed a high PDE5 induction after dibutyryl-cAMP treatment. (C) 2001 Elsevier Science B,V, All rights reserved

Elemental Analyzer/Isotope Ratio Mass Spectrometry (EA/IRMS) as a Tool to Characterize Plastic Polymers in a Marine Environment

In the last 60 years, plastic has become a widely used material due to its versatility and wide range of applications. This characteristic, together with its persistence, makes plastic waste a growing environmental problem, particularly in the marine ecosystems. The production of plant-derived biodegradable plastic polymers is assuming increasing importance. Here, we report the results of a first preliminary characterization of carbon stable isotopes (δ13C) of different plastic polymers (petroleum- and plant-derived) and a first experimental study aimed to determine carbon isotopic shift due to polymer degradation in an aquatic environment. The results showed that the δ13C values determined in different packaging for food uses reflect the plant origin for “BIO” materials and the petroleum-derived source for plastic materials. Considering degradation, δ13C values of both bio bags and HDPE bags showed a gradual decrease toward less negative values when kept immersed in seawater, recording a δ13C variation (Δδ13C) of 1.15 and 1.78‰, respectively. With respect to other analytical methods, the characterization of the plastic polymer composition by isotope ratio mass spectrometry is advantageous due to low cost and rapidity of analysis, small amount of sample required, high sensitivity, and the possibility of analyzing colored samples

Fluorescent Asymmetrically Cobalt-Tipped CdSe@CdS Core@Shell Nanorod Heterostructures Exhibiting Room-Temperature Ferromagnetic Behavior

A colloidal two-step seeded-growth approach has been devised to selectively synthesize three-component magnetic/semiconductor hybrid nanocrystals (HNCs) with a matchstick-like profile and tunable geometric parameters. The newly developed heterostructures individually comprise a single metallic Co head connected to either apexes of one rod-shaped section made of a CdSe core eccentrically embedded in a CdS shell. The specific topological arrangement realized arises from the peculiar anisotropic reactivity of the noncentrosymmetric CdSe@CdS core@shell nanorods that have been used as substrates to seed heterogeneous nucleation of Co in a surfactant-free environment from an organometallic precursor. The HNCs retain appreciable fluorescent emission in spite of photoexcited charge transfer from the semiconductor to the metal domain and exhibit unusual ferromagnetic-like behavior at room temperature

High cadence spectropolarimetry of moving magnetic features observed around a pore

Moving magnetic features (MMFs) are small-size magnetic elements that are seen to stream out from sunspots, generally during their decay phase. Several observational results presented in the literature suggest them to be closely related to magnetic filaments that extend from the penumbra of the parent spot. Nevertheless, few observations of MMFs streaming out from spots without penumbra have been reported. The literature still lacks of analyses of the physical properties of these features. We investigate physical properties of monopolar MMFs observed around a small pore that had developed penumbra in the days preceding our observations and compare our results with those reported in the literature for features observed around sunspots. We analyzed NOAA 11005 during its decay phase with data acquired at the Dunn Solar Telescope in the FeI 617.3$nm and the CaII 854.2$ nm spectral lines with IBIS, and in the G-band. The field of view showed monopolar MMFs of both polarities streaming out from the leading negative polarity pore of the observed active region. Combining different analyses of the data, we investigated the temporal evolution of the relevant physical quantities associated with the MMFs as well as the photospheric and chromospheric signatures of these features. We show that the characteristics of the investigated MMFs agree with those reported in the literature for MMFs that stream out from spots with penumbrae. Moreover, observations of at least two of the observed features suggest them to be manifestations of emerging magnetic arches.Comment: Accepted by A&

Circulating tumor cells as early predictors of metastatic spread in breast cancer patients with limited metastatic dissemination.

IntroductionTraditional factors currently used for prognostic stratification do not always predict adequately treatment response and disease evolution in advanced breast cancer patients. Therefore, the use of blood-based markers, such as circulating tumor cells (CTCs), represents a promising complementary strategy for disease monitoring. In this retrospective study, we explored the role of CTC counts as predictors of disease evolution in breast cancer patients with limited metastatic dissemination.Methods492 advanced breast cancer patients who had a CTC count assessed by CellSearch prior to starting a new line of systemic therapy were eligible for this analysis. Using the threshold of 5 cells/7.5 mL of blood, pretreatment CTC counts were correlated in the overall population with metastatic site distribution, evaluated at baseline and at the time of treatment failure, using the Fisher¿s Exact test. Time to visceral progression, as well as, time to the development of new metastatic lesions and sites were estimated in patients with non-visceral metastases and with single-site metastatic disease, respectively, by the Kaplan-Meier method. Survival times were compared among groups according to pretreatment CTC count by log-Rank test.ResultsIn the overall population, pretreatment CTCs¿¿¿5 were associated with increased baseline number of metastatic sites, compared with CTCs

Circulating tumor cells (CTC) are associated with defects in adaptive immunity in patients with inflammatory breast cancer

BACKGROUND: Circulating tumor cells (CTCs) play a crucial role in tumor dissemination and are prognostic in primary and metastatic breast cancer. Peripheral blood (PB) immune cells contribute to an unfavorable microenvironment for CTC survival. This study aimed to correlate CTCs with the PB T-cell immunophenotypes and functions of patients with inflammatory breast cancer (IBC). METHODS: This study included 65 IBC patients treated at the MD Anderson Cancer Center. PB was obtained from patients prior to starting a new line of chemotherapy for CTCs enumeration by CellSearch(®), and T cell phenotype and function by flow cytometry; the results were correlated with CTCs and clinical outcome. RESULTS: At least 1 CTC (≥1) or ≥5 CTCs was detected in 61.5% or 32.3% of patients, respectively. CTC count did not correlate with total lymphocytes; however, patients with ≥1 CTC or ≥5 CTCs had lower percentages (%) of CD3+ and CD4+ T cells compared with patients with no CTCs or <5 CTCs, respectively. Patients with ≥1 CTC had a lower percentage of T-cell receptor (TCR)-activated CD8+ T cells synthesizing TNF-α and IFN-γ and a higher percentage of T-regulatory lymphocytes compared to patients without CTCs. In multivariate analysis, tumor grade and % CD3+ T-cells were associated with ≥1 CTC, whereas ≥5 CTC was associated with tumor grade, stage, % CD3+ and % CD4+ T cells, and % TCR-activated CD8 T-cells synthesizing IL-17. CONCLUSIONS: IBC patients with CTCs in PB had abnormalities in adaptive immunity that could potentially impact tumor cell dissemination and initiation of the metastatic cascade

Circulating tumor cells in newly diagnosed inflammatory breast cancer

Circulating tumor cells (CTCs) are an independent prognostic factor for progression-free survival (PFS) and overall survival (OS) in patients with metastatic breast cancer. Inflammatory breast cancer (IBC) is one of the most aggressive forms of breast cancer. The prognostic value of a CTC count in newly diagnosed IBC has not been established. The aim of this study was to assess the prognostic value of a baseline CTC count in patients with newly diagnosed IBC

Safety and clinical outcomes of abiraterone acetate (aa) after docetaxel (doc) in octogenarians with metastatic castration-resistant prostate cancer (mcrpc)

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Circulating tumor cells in breast cancer: A tool whose time has come of age

Circulating tumor cells (CTCs) are isolated tumor cells disseminated from the site of disease in metastatic and/or primary cancers, including breast cancer, that can be identified and measured in the peripheral blood of patients. As recent technical advances have rendered it easier to reproducibly and repeatedly sample this population of cells with a high degree of accuracy, these cells represent an attractive surrogate marker of the site of disease