Here we describe the establishment of a new canine mammary tumour (CMT) cell line, FR37-CMT that does not show dependence on female hormonal signaling to induce tumour xenografts in NOD-SCID mice. FR37-CMT cell line has a stellate or fusiform shape, displays the ability to reorganize the collagen matrix, expresses vimentin, CD44 and shows the loss of E-cadherin which is considered a fundamental event in epithelial to mesenchymal transition (EMT). The up-regulation of ZEB1, the detection of phosphorylated ERK1/2 and the downregulation of DICER1 and miR-200c are also in accordance with the mesenchymal characteristics of FR37-CMT cell line. FR37-CMT shows a higher resistance to cisplatin (IC50>50 µM) and to doxorubicin (IC50>5.3 µM) compared with other CMT cell lines. These results support the use of FR37-CMT as a new CMT model that may assist the understanding of the molecular mechanisms underlying EMT, CMT drug resistance, fostering the development of novel therapies targeting CMT.

Identification of novel molecules that can selectively inhibit the growth of tumor cells, avoid causing side effects to patients and/or intrinsic or acquired resistance, usually associated with common chemotherapeutic agents, is of utmost importance. Organometallic compounds have gained importance in oncologic chemotherapy, such as organotin(IV) complexes. In this study, we assessed the anti-tumor activity of the cyclic trinuclear organotin(IV) complex with an aromatic oximehydroxamic acid group [nBu2Sn(L)]3(H2L = N,2-dihydroxy-5-[N-hydroxyethanimidoyl]benzamide) – MG85 – and provided further characterization of its biological targets. We have previously shown the high anti-proliferative activity of this complex against human colorectal and hepatocellular carcinoma cell lines and lower cytotoxicity in neonatal non-tumor fibroblasts. MG85 induces tumor cell apoptosis and down-regulation of proteins related to tubulin dynamics (TCTP and COF1). Further characterization included the: (i) evaluation of interference in the cell cycle progression, including the expression of critical genes; (ii) affinity to DNA and the corresponding mode of binding; (iii) genotoxic potential in cells with deficient DNA repair pathways; and (iv) in vivo tumor reduction efficiency using mouse colorectal carcinoma xenografts.

The pH-sensitive affinity pair composed by neutravidin and iminobiotin was used to develop a multilayered Magnetic Resonance Imaging (MRI) nanoprobe responsive to the acidic pH of tumor microenvironment. The multilayer system was assembled on meso-2,3-dimercaptosuccinic acid-coated iron oxide magnetic nanoparticles (MNP), which convey negative MRI contrast enhancement properties to the nanoprobe. The outer stealth PEG-layer is altered in acidic media due to the disruption of interactions between neutravidin–iminobiotin. As a consequence, the positively charged inner layer is exposed and enhances interactions with cells. The nanoprobe uptake by HCT116 cells cultured in vitro under acidic conditions had a 2-fold increase compared to the uptake at physiological pH. The uptake difference is particularly clear in T2-weighted MRI phantoms of cells incubated with the nanoprobes at both pH conditions. This work sets the proof-of-concept of a MNP-based MRI nanoprobe targeting acidic tumor microenvironment through the use of a specific bio-recognition interaction that is pH-sensitive. This tumor targeting strategy is potentially applicable to the generality of tumors since the typical hypoxic conditions and high glycolysis rate in cancer cells create an acidic environment common to the majority of cancer types.

Currently most economical and technological bottlenecks in protein production are placed in the down-stream processes. With the aim of increasing the efficiency and reducing the associated costs, variousaffinity ligands have been developed. Affitins are small, yet robust and easy to produce, proteins derivedfrom the archaeal extremophilic “7 kDa DNA-binding” protein family. By means of combinatorial pro-tein engineering and ribosome display selection techniques, Affitins have shown to bind a diversity oftargets. In this work, two previously developed Affitins (anti-lysozyme and anti-IgG) were immobilizedonto magnetic particles to assess their potential for protein purification by magnetic fishing. The opti-mal lysozyme and human IgG binding conditions yielded 58 mg lysozyme/g support and 165 mg IgG/gsupport, respectively. The recovery of proteins was possible in high yield (≥95{%}) and with high purity,namely ≥95{%} and 81{%}, when recovering lysozyme from Escherichia coli supernatant and IgG from humanplasma, respectively. Static binding studies indicated affinity constants of 5.0 × 104M−1and 9.3 × 105M−1for the anti-lysozyme and anti-IgG magnetic supports. This work demonstrated that Affitins, which canbe virtually evolved for any protein of interest, can be coupled onto magnetic particles creating novelaffinity adsorbents for purification by magnetic fishing.

In Europe, the technology development of high-speed trains is increasingly exposed to societal needs, driven by ICT advancements, external to traditional design. Together with the liberalisation of the rail markets and increase pressures from other transport modes leads to an unprecedented situation where planers, operators and suppliers of high-speed have to take decision in this complex and competitive environment.
In such broadening of elements influencing design and, thus, product development process, from the survey here to be presented, it was not observed technology options assessment or strategic agenda setting from visions shifting in the same way.
For the high-speed train industry this new trend requires going beyond the visions of the past 15 to 20 years’ practices of “sector endogenous” and structurally closed strategic methods approaches to a broader interaction with the widening of societal actors now capable of being active contributors to innovation from digitalization.
This way to understand the European industry readiness for undertaking such supra systemic challenge, this paper presents the results from a survey conducted by the authors to 74 representatives of the high-speed train innovation chain regarding to which extent societal embedding is considered in the drafting of their visions and technology development projects.
This work becomes even more pertinent if considered that the debate is now open in the railway industry (not exclusive to high-speed trains) as they are launching the joint initiative SHIFT2RAIL, revise ERRAC (the European Rail Research Advisory Council) mandate and enter in a new research cycle with the European research framework Horizon 2020.

In Europe, the technology development of high-speed trains is increasingly exposed to societal needs, driven by ICT advancements, external to traditional design. Together with the liberalisation of the rail markets and increase pressures from other transport modes leads to an unprecedented situation where planers, operators and suppliers of high-speed have to take decision in this complex and competitive environment.
In such broadening of elements influencing design and, thus, product development process, from the survey here to be presented, it was not observed technology options assessment or strategic agenda setting from visions shifting in the same way.
For the high-speed train industry this new trend requires going beyond the visions of the past 15 to 20 years’ practices of “sector endogenous” and structurally closed strategic methods approaches to a broader interaction with the widening of societal actors now capable of being active contributors to innovation from digitalization.
This way to understand the European industry readiness for undertaking such supra systemic challenge, this paper presents the results from a survey conducted by the authors to 74 representatives of the high-speed train innovation chain regarding to which extent societal embedding is considered in the drafting of their visions and technology development projects.
This work becomes even more pertinent if considered that the debate is now open in the railway industry (not exclusive to high-speed trains) as they are launching the joint initiative SHIFT2RAIL, revise ERRAC (the European Rail Research Advisory Council) mandate and enter in a new research cycle with the European research framework Horizon 2020.

Pullulan, a neutral polysaccharide, was chemically modified in order to obtain two charged derivatives: reaction with SO3.DMF complex afforded a sulfate derivative (SP), while reaction with glycidyltrimethylammonium chloride gave a quaternary ammonium salt (AP). The presence of the charged groups was confirmed by FTIR. Assessment of the positions where the reaction took place was based on 1H- and 13C NMR (COSY, HSQC-TOCSY, HSQC-DEPT, and HMBC) experiments. Estimation of the degree of substitution (DS) was made from elemental analysis data, and further confirmed by NMR peak areas in the case of AP. These new derivatives showed the capability to condense with each other, forming nanoparticles with the ability to associate a model protein (BSA) and displaying adequate size for drug delivery applications, therefore making them good candidates for the production of pullulan-based nanocarriers by polyelectrolyte complexation.

In the present work, two drug delivery systems were produced by encapsulating doxorubicin into chitosan and O-HTCC (ammonium-quaternary derivative of chitosan) nanoparticles. The results show that doxorubicin release is independent of the molecular weight and is higher at acidic pH (4.5) than at physiological pH. NPs with an average hydrodynamic diameter bellow 200 nm are able to encapsulate up to 70% and 50% of doxorubicin in the case of chitosan and O-HTCC nanoparticles, respectively. O-HTCC nanoparticles led to a higher amount of doxorubicin released than chitosan nanoparticles, for the same experimental conditions, although the release mechanism was not altered. A burst effect occurs within the first hours of release, reaching a plateau after 24 h. Fitting mathematical models to the experimental data led to a concordant release mechanism between most samples, indicating an anomalous or mixed release, which is in agreement with the swelling behavior of chitosan described in the literature.

In the present work, two drug delivery systems were produced by encapsulating doxorubicin into chitosan and O-HTCC (ammonium-quaternary derivative of chitosan) nanoparticles. The results show that doxorubicin release is independent of the molecular weight and is higher at acidic pH (4.5) than at physiological pH. NPs with an average hydrodynamic diameter bellow 200 nm are able to encapsulate up to 70% and 50% of doxorubicin in the case of chitosan and O-HTCC nanoparticles, respectively. O-HTCC nanoparticles led to a higher amount of doxorubicin released than chitosan nanoparticles, for the same experimental conditions, although the release mechanism was not altered. A burst effect occurs within the first hours of release, reaching a plateau after 24 h. Fitting mathematical models to the experimental data led to a concordant release mechanism between most samples, indicating an anomalous or mixed release, which is in agreement with the swelling behavior of chitosan described in the literature.

Gold nanoparticles functionalized with thiolated oligonucleotides (Au-nanoprobes) have been used in a range of applications for the detection of bioanalytes of interest, from ions to proteins and DNA targets. These detection strategies are based on the unique optical properties of gold nanoparticles, in particular, the intense color that is subject to modulation by modification of the medium dieletric. Au-nanoprobes have been applied for the detection and characterization of specific DNA sequences of interest, namely pathogens and disease biomarkers. Nevertheless, despite its relevance, only a few reports exist on the detection of RNA targets. Among these strategies, the colorimetric detection of DNA has been proven to work for several different targets in controlled samples but demonstration in real clinical bioanalysis has been elusive. Here, we used a colorimetric method based on Au-nanoprobes for the direct detection of the e14a2 BCR-ABL fusion transcript in myeloid leukemia patient samples without the need for retro-transcription. Au-nanoprobes directly assessed total RNA from 38 clinical samples, and results were validated against reverse transcription-nested polymerase chain reaction (RT-nested PCR) and reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The colorimetric Au-nanoprobe assay is a simple yet reliable strategy to scrutinize myeloid leukemia patients at diagnosis and evaluate progression, with obvious advantages in terms of time and cost, particularly in low- to medium-income countries where molecular screening is not routinely feasible.