Sequential Colocalization of ERa, PR, and AR Hormone Receptors Using Confocal Microscopy Enables New Insights into Normal Breast and Prostate Tissue and Cancers

Multiplex immunohistochemistry (mIHC) use markers staining different cell populations applying widefield optical microscopy. Resolution is low not resolving subcellular co-localization. We sought to colocalize markers at subcellular level with antibodies validated for clinical diagnosis, including the single secondary antibody (combination of anti-rabbit/mouse-antibodies) used for diagnostic IHC with any primary antibody, and confocal microscopy. We explore colocalization in the nucleus (ColNu) of nuclear hormone receptors (ERa, PR, and AR) along with the baseline marker p63 in paired samples of breast and prostate tissues. We established ColNu mIHCF as a reliable technique easily implemented in a hospital setting. In ERa+ breast cancer, we identified different colocalization patterns (nuclear or cytoplasmatic) with PR and AR on the luminal epithelium. A triple-negative breast-cancer case expressed membrane-only ERa. A PR-only case was double positive PR/p63. In normal prostate, we identified an ERa+/p63+/AR-negative distinct population. All prostate cancer cases characteristically expressed ERa on the apical membrane of the AR+ epithelium. We confirmed this using ERa IHC and needle-core biopsies. ColNu mIHCF is feasible and already revealed a new marker for prostate cancer and identified sub-patterns in breast cancer. It could be useful for pathology as well as for functional studies in normal prostate and breast tissues

Modeling of the binodal curve of ionic liquid/salt aqueous systems

Ionic Liquid-based Aqueous Two Phase Systems (ILATPS) are an innovative technique to separate biomolecules that combines the advantages of liquid–liquid extraction and hydrophilic ionic liquids. Most ILATPS are based on ionic liquids and conventional inorganic salts, and the phase envelope, described by the binodal curve, is usually modeled by empirical equations that are used to determine the phase compositions and assess the ionic liquid recyclability. However, these empirical equations may provide a poor extrapolation ability or low accuracy at the extreme regions of the binodal curve or suffer from problems of convergence. Therefore, the aim of this work is the analysis of the binodal curve equations, comparing the models reported in the literature to describe ILATPS and proposing alternative equations to improve accuracy or to reduce the mathematical complexity. For this purpose, a database compiling binodal experimental data of 100 ILATPS has been built, so that the analysis could make it possible to obtain representative conclusions for all these systems. Several models were developed, and different statistical criteria were used to assess the advantages and disadvantages of each one of these models for the binodal curve. The results show that, when accuracy is critical, a proposed model with just an additional parameter reduced more than 25% the residual mean squared error (RMSE) with respect to the commonly used equation, without losing the statistical significance of the parameters. For complex problems where an explicit equation in both the concentration of ionic liquid and of salt is needed, the use of an explicit model developed with 3 adjusted parameters that kept high accuracy (R2 > 0.996 and RMSE < 0.66) is proposed. Finally, the analysis also revealed that a fitting method based on the minimization of relative errors is recommended to increase the accuracy of the binodal curve at high salt concentrations, which is the crucial region for assessing the recyclability of the ionic liquid.This work was developed in the scope of the project CICECO Aveiro Institute of Materials (Ref. FCT UID/CTM/50011/2013), financed by national funds through the FCT/MEC and co-financed by FEDER under the PT2020 Partnership Agreement. The authors also acknowledge FCT for the Post-doctoral grant SFRH/BPD/79263/2011 of S.P.M. Ventura

On the phonon dissipation contribution to nanoscale friction by direct contact

CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQCOORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPESFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DO RIO GRANDE DO SUL - FAPERGSFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESPThe friction phenomenon is a ubiquitous manifestation of nature. Models considering phononic, electronic, magnetic, and electrostatic interactions are invoked to explain the fundamental forces involved in the friction phenomenon. In order to establish the incidence of the phonon prompting at the nanoscale friction by direct contact, we study a diamond spherical dome sliding on carbon thin films containing different amount of deuterium and hydrogen. The friction coefficient decreases by substituting hydrogen by deuterium atoms. This result is consistent with an energy dissipation vibration local mechanism from a disordered distribution of bond terminators.718CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQCOORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPESFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DO RIO GRANDE DO SUL - FAPERGSFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESPCONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQCOORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPESFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DO RIO GRANDE DO SUL - FAPERGSFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP554336/2010-3087/113189032012/10127-5The authors are grateful to UCS, INCT-INES-CNPq (Grant #554336/2010-3), CAPES (Grant #Brafitec 087/11, FAPERGS and SUMA2 Network Project - 7th Framework Program of the European Commission (IRSES Project #318903) for financial support. SRSM is FAPERGS fellow. CMM is CAPES fellow. MEHMC, FLF, FA, and CAF are CNPq fellows. FA was supported by FAPESP project 2012/10127-5

Arsenic remediation by formation of arsenic sulfide minerals in a continuous anaerobic bioreactor

Arsenic (As) is a highly toxic metalloid that has been identified at high concentrations in groundwater in certain locations around the world. Concurrent microbial reduction of arsenate (AsV) and sulfate (SO42-) can result in the formation of poorly soluble arsenic sulfide minerals (ASM). The objective of this research was to study As biomineralization in a minimal iron environment for the bioremediation of As-contaminated groundwater using simultaneous AsV and SO42- reduction. A continuous-flow anaerobic bioreactor was maintained at slightly acidic pH (6.25-6.50) and fed with AsV and SO42-, utilizing ethanol as an electron donor for over 250 d. A second bioreactor running under the same conditions but lacking SO42- was operated as a control to study the fate of As (without S). The reactor fed with SO42- removed an average 91.2% of the total soluble As at volumetric rates up to 2.9 mg As/(L∙h), while less than 5% removal was observed in the control bioreactor. Soluble S removal occurred with an S to As molar ratio of 1.2, suggesting the formation of a mixture of orpiment- (As2S3) and realgar-like (AsS) solid phases. Solid phase characterization using K-edge X-Ray absorption spectroscopy confirmed the formation of a mixture of As2S3 and AsS. These results indicate that a bioremediation process relying on the addition of a simple, low-cost electron donor offers potential to promote the removal of As from groundwater with naturally occurring or added sulfate by precipitation of ASM.12 month embargo; published online: 2 September 2015This item from the UA Faculty Publications collection is made available by the University of Arizona with support from the University of Arizona Libraries. If you have questions, please contact us at [email protected]

Nanoindentation unidirectional sliding and lateral force microscopy: evaluation of experimental techniques to measure friction at the nanoscale

CNPQ - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICOFAPERG - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DO RIO GRANDE DO SULFAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOLateral force microscopy (LFM) is an established technique to assess friction forces at the nanoscale. Nanoindentation followed by unidirectional sliding (NUS) is also used to evaluate friction forces at the micro/nanoscale. However, comparative studies between NUS and LFM evaluating the experimental results at different scales are still missing. In this work, a-C:D/H and a-C:H thin films with different [D]/[C] and [H]/[C] contents were used to analyze the friction forces by NUS and LFM. The results show that the friction behavior assessed by these two techniques in different scales is the same. The correlation between friction forces measured by NUS and LFM depends mainly on a contact area factor that makes invariant the friction force from nanoscale to microscale. Such behavior suggests a similar damping mechanism, probably phonon-coupling phenomena, for the friction force origin.812110CNPQ - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICOFAPERG - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DO RIO GRANDE DO SULFAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOCNPQ - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICOFAPERG - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DO RIO GRANDE DO SULFAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO465423/2014-0sem informação2012/10127-

The young active star SAO 51891 (V383 Lac)

Our aim is investigating surface inhomogeneities of the young late-type star SAO51891, from photosphere to upper chromosphere, analyzing contemporaneous high-resolution spectra and broad-band photometry. The FOCES@CAHA spectral range is used to determine spectral classification and derive vsini and Vrad. The Li abundance is measured to estimate the age. The BVRIJHKs bands are used to construct the SED. The variations of our BV fluxes and Teff are used to infer the presence of photospheric spots and observe their behavior over time. The chromospheric activity is studied applying the spectral subtraction technique to Halpha, CaII H&K, Heps, and CaII IRT lines. We find SAO51891 to be a young K0-1V star with Li abundance close to the Pleiades upper envelope, confirming its youth (~100 Myr), also inferred from its kinematical membership to the Local Association. We detect no IR excess from SED analysis, and rotational modulation of luminosity, Teff, CaII, and Heps total fluxes. A spot model with two active regions, ~240 K cooler than the surrounding photosphere, fits our light/Teff curves, and reproduces the small-amplitude Vrad variations. The anti-correlation of light curves and chromospheric diagnostics indicates plages spatially associated with spots. The large amplitude observed in the Heps-flux curve suggests that this line is very sensitive to the plage presence. Finally, SAO51891 is a young active star, lacking significant amounts of circumstellar dust or any evidence for low mass companions. The spots turn out to be larger and warmer than those in less active MS stars. The Vrad variation produced by spots has an amplitude comparable with those induced by Jupiter-mass planets orbiting close to the star. SAO51891 is a good example of star where the detection of planets may be hampered by the high activity level.Comment: 16 pages, 12 figures, 7 tables, accepted by Astronomy and Astrophysics; abstract here was shortene

Sustainable strategies based on glycine–betaine analogue ionic liquids for the recovery of monoclonal antibodies from cell culture supernatants

Monoclonal antibodies (mAbs) are of crucial interest for therapeutic purposes, particularly in vaccination and immunization, and in the treatment of life-threatening diseases. However, their downstream processing from the complex cell culture media in which they are produced still requires multiple steps, making mAbs extremely high-cost products. Therefore, the development of cost-effective, sustainable and biocompatible purification strategies for mAbs is in high demand to decrease the associated economic, environmental and health burdens. Herein, novel aqueous biphasic systems (ABS) composed of glycine–betaine analogue ionic liquids (AGB-ILs) and K2HPO4/KH2PO4 at pH 7.0, the respective three-phase partitioning (TPP) systems, and hybrid processes combined with ultrafiltration were investigated and compared in terms of performance as alternative strategies for the purification and recovery of anti-human interleukin-8 (anti-IL-8) mAbs, which are specific therapeutics in the treatment of inflammatory diseases, from Chinese Hamster Ovary (CHO) cell culture supernatants. With the studied ABS, mAbs preferentially partition to the IL-rich phase, with recovery yields up to 100% and purification factors up to 1.6. The best systems were optimized in what concerns the IL concentration, allowing to take advantage of IL-based three-phase partitioning approaches where a precipitate enriched in mAbs is obtained at the ABS interface, yielding 41.0% of IgG with a purification factor of 2.7 (purity of 60.9%). Hybrid processes combining the two previous techniques and an ultrafiltration step were finally applied, allowing the recovery of mAbs from the different fractions in an appropriate buffer solution for further biopharmaceutical formulations, while allowing the simultaneous IL removal and reuse. The best results were obtained with the hybrid process combining TPP and ultrafiltration, allowing to obtain mAbs with a purity higher than 60%. The recyclability of the IL was additionally demonstrated, revealing no losses in the purification and recovery performance of these systems for mAbs. The biological activity of anti-IL-8 mAbs is maintained after the several purification and recovery steps, indicating that the novel ABS, three-phase partitioning and hybrid processes comprising AGB-ILs are promising and sustainable strategies in mAbs downstream processing.publishe