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Antibody therapeutics in oncology can have high cytotoxic potential, which demands superior on-tumor selectivity. Antibody selectivity can be boosted through molecular engineering strategies that exploit the unique characteristics of the tumor and the tumor microenvironment. Strategies, such as superselectivity, dual targeting, conditional binding, inverse targeting, and combinations of molecules, can help boost the selectivity of biologics for tumor cells. Intracellular tumor antigens are becoming increasingly accessible to biologics through approaches including T cell receptor mimicry, intracellular delivery, and transcytosis. Antibodies in oncology are being equipped with toxic cargoes and effector functions that can kill cells at very low concentrations. A key challenge is that most targets on cancer cells are also present on at least some healthy cells. Shared targets can result in off-tumor binding and compromise the safety and potential of therapeutic candidates. In this review, we survey strategies that can help direct biologics to cancer sites more selectively. These strategies are becoming increasingly feasible thanks to advances in molecular design and engineering. The objective is to create therapeutics that exploit changes in cancer and leverage the human body infrastructure, enabling therapeutics that discriminate not just self from non-self but diseased from healthy tissue. [ABSTRACT FROM AUTHOR]

Drawing is an ideal technique to understand children's responses to fictional narratives, including digital ones. In this case, a gamified narrative with augmented reality (AR), based on a picturebook, was designed and used in an intervention in 8 Early Childhood Education classrooms (N = 113), aimed at enhancing their literary competence. This paper analyses the visual responses of the participating students after the intervention, reflected in their drawings and complemented with individual interviews. The methodology is mixed: (a) qualitative, analysing the content of the drawings, and taking into account the narrative‐interpretive and gamification dimensions, through an instrument designed ad hoc; (b) quantitative, through the statistical treatment of the collected data. The results show the predominance of informative and emotional responses, which reveal the understanding of the narrative. Most of the children draw the characters that sparked their interest and recreate key scenes from the story. Drawings emphasise the narrative aspects, and to a lesser extent refer to the game mechanics (challenges and rewards). Finally, it is confirmed that the gamified narrative with AR has the capacity to generate emotional responses and promote the immersion of students in the story and the game. [ABSTRACT FROM AUTHOR]

In this paper, we discuss the requirements and technical challenges within the EHEM project, Enhancement of Heritage Experiences: The Middle Ages, an ongoing research program for the acquisition, analysis, documentation, interpretation, digital restoration, and communication of medieval artistic heritage. The project involves multidisciplinary teams comprising art historians and visual computing experts. Despite the vast literature on digital 3D models in support of Cultural Heritage, the field is so rich and diverse that specific projects often imply distinct, unique requirements which often challenge the computational technologies and suggest new research opportunities. As good representatives of such diversity, we describe the three monuments that serve as test cases for the project, all of them with a rich history of architecture and paintings. We discuss the art historians' view of how digital models can support their research, the expertise and technological solutions adopted so far, as well as the technical challenges in multiple areas spanning geometry and appearance acquisition, color analysis and digital restitution, as well as the representation of the profound transformations due to the alterations suffered over the centuries. [ABSTRACT FROM AUTHOR]

Digital color restitution aims to digitally restore the original colors of a painting. Existing image editing applications can be used for this purpose, but they require a select-and-edit workflow and thus they do not scale well to large collections of paintings or different regions of the same painting. To address this issue, we propose an automated workflow that requires only a few representative source colors and associated target colors as input from art historians. The system then creates a control grid to model a deformation of the CIELAB color space. Such deformation can be applied to arbitrary images of the same painting. The proposed approach is suitable for restituting the color of images from a large photographic campaign, as well as for the textures of 3D reconstructions of a monument. We demonstrate the benefits of our method on a collection of mural paintings from a medieval monument. • Automated workflow for digital color restitution. • Deformation of CIELAB color space to test art historian hypotheses. • Applicable in photographic campaigns and 3D reconstruction. [ABSTRACT FROM AUTHOR]

Adult literacy in diglossic contexts is undergoing a paradigm shift as a result of the circumstances that have arisen during the Covid-19 pandemic. The accelerated adaptation of this educational offer to the digital environment has revealed challenges and opportunities that have an impact on the teaching and social normalisation of minoritized languages, on which it is necessary to reflect. To this end, this article presents a descriptive analysis of four online training programmes dedicated, respectively, to the teaching of four Romance languages: Mirandese, Fala of Xalima, Asturian and Friulian. A qualitative methodology has been employed, based on direct observation and evaluation of the most significant aspects. In particular, an individual characterisation of each training package within its sociolinguistic context is provided, followed by a comparative description highlighting common aspects and discrepancies. Through this research, the contributions of the digital medium to the articulation of a standardising discourse of glocal scope are confirmed, but a marked inequality in the essential conditions of the training projects analysed is also revealed. Likewise, the description of these educational emergency measures shows the efforts of the groups involved to promote minority languages with the means at their disposal, seeking to stimulate them to inaugurate new inter-institutional dialogues aimed at the exchange of their good practices. [ABSTRACT FROM AUTHOR]

Metastatic melanoma is a highly immunogenic tumor with very poor survival rates due to immune system escape-mechanisms. Immune checkpoint inhibitors (ICIs) targeting the cytotoxic T-lymphocyte-associated protein 4 (CTLA4) and the programmed death-1 (PD1) receptors, are being used to impede immune evasion. This immunotherapy entails an increment in the overall survival rates. However, melanoma cells respond with evasive molecular mechanisms. ERK cascade inhibitors are also used in metastatic melanoma treatment, with the RAF activity blockade being the main therapeutic approach for such purpose, and in combination with MEK inhibitors improves many parameters of clinical efficacy. Despite their efficacy in inhibiting ERK signaling, the rewiring of the melanoma cell-signaling results in disease relapse, constituting the reinstatement of ERK activation, which is a common cause of some resistance mechanisms. Recent studies revealed that the combination of RAS–ERK pathway inhibitors and ICI therapy present promising advantages for metastatic melanoma treatment. Here, we present a recompilation of the combined therapies clinically evaluated in patients. [ABSTRACT FROM AUTHOR]

High-end Terrestrial Lidar Scanners are often equipped with RGB cameras that are used to colorize the point samples. Some of these scanners produce panoramic HDR images by encompassing the information of multiple pictures with different exposures. Unfortunately, exported RGB color values are not in an absolute color space, and thus point samples with similar reflectivity values might exhibit strong color differences depending on the scan the sample comes from. These color differences produce severe visual artifacts if, as usual, multiple point clouds colorized independently are combined into a single point cloud. In this paper we propose an automatic algorithm to minimize color differences among a collection of registered scans. The basic idea is to find correspondences between pairs of scans, i.e. surface patches that have been captured by both scans. If the patches meet certain requirements, their colors should match in both scans. We build a graph from such pair-wise correspondences, and solve for the gain compensation factors that better uniformize color across scans. The resulting panoramas can be used to colorize the point clouds consistently. We discuss the characterization of good candidate matches, and how to find such correspondences directly on the panorama images instead of in 3D space. We have tested this approach to uniformize color across scans acquired with a Leica RTC360 scanner, with very good results. [Display omitted] • Algorithm to minimize color differences among scans. • Flow maps are used to quickly find correspondences between pairs of scans. • We estimate the color reliability of the superpixels of each panorama. • Gain factor solved to uniformize color across scans considering patch reliability. • The resulting point clouds exhibit a consistent color. [ABSTRACT FROM AUTHOR]

Bryonia dioica Jacq. , Evernia prunastri (L.) Ach. , Telephium imperati L. , and Aristolochia longa L. are species widely used in traditional medicine to treat several diseases including cancer. Conjugation of two or more extracts is an approach to improve the effectiveness of their pharmacological activities. To evaluate the synergistic anticancer and anti-angiogenic effects of medicinal plants and edible species combinations. In this work, B. dioica, E. prunastri, Telephium imperati, and Aristolochia longa extracts were conjugated to form four mixtures. The antiproliferative effect of mixtures on several carcinoma cells was examined by MTT assay, and the antiangiogenic activity was estimated through Hen's egg test in vivo. Moreover, in an Ovo model, 35 fertilized Ross eggs were used to test the embryotoxicity of mixtures. At the highest concentration of 200 μg/mL, both mixtures exerted an important cytotoxic effect against human carcinoma cells. The mixture BETE (Bryonia Evernia Telephium Extract) significantly reduced HT-29, PC-3, and A-549 cell viability. Likewise, this mixture strongly suppressed vascularization in vivo at 200 μg/mL. Interestingly, no signs of toxicity on Perdix embryos were recorded within 21 days of treatment. More importantly, the mixture did not have any cytotoxic effect on non cancerous cells. Taken together, our results suggest that the synergy between B. dioica, E. prunastri and T. imperati may be promising for developing new anti-cancer treatments. [Display omitted] • The mixture BETE (Bryonia Evernia Telephium Extract) exerted a significant cytotoxic effect. • The mixture BETE significantly inhibited the growth of cancer cells. • The mixture BETE was non toxic against non-cancerous human cells CCD18-Co. • The mixture exhibited significant anti-angiogenic activity. [ABSTRACT FROM AUTHOR]

Although cancer has been the main reason for death for decades, until now, no safe certified drugs have been discovered to efficiently inhibit its progression. Currently, we aim to employ an in-silico model to predict the action mechanism of phenolic components that we previously demonstrated as inhibitors of colon, prostate and lung cancer. The work was performed by testing pharmacokinetics and pharmacodynamics proprieties of selected molecules. Four compounds: Kaempferol, chrysin, vanillin and p-hydroxybenzoic acid widely obeyed the five rules and the ADME/T criteria with minimum toxicity. Docking prediction recorded an intense affinity between kaempferol-CDK1(−17.72 ± 0.02 kcal/mol), chrysin-CDK1 (−17.61 ± 0.01 kcal/mol) and kaempferol-caspase 8 (−9.30 ± 0.00 kcal/mol), chrysin-caspase 8 (−8.18 ± 0.12 kcal/mol), vanillin-MMP9 (−9.83 ± 0.04 kcal/mol) and p-hydroxybenzoic acid-Ca2 (−8.91 ± 0.01 kcal/mol). Likewise, MMGBSA results demonstrated that these compounds are evidently bound to the targets’ active sites with solid interactions. The passed compounds might target the responsible factors for transcription as inactivators. The molecular mechanism based on kaempferol and vanillin actions was predicted to target CDK1 and MMP9 via cell cycle arrest and apoptosis. All results were confirmed through RMSD analysis suggesting that kaempferol and vanillin are the best ligands to suppress cancer cell proliferation. Thus, these findings allowed us to imagine a potential molecular mechanism of these binaries, phenolic compounds/target proteins to lead further trails on cancer, of which the synergistic activity between phenolic compounds could promote the therapeutic capacity. [ABSTRACT FROM AUTHOR]

Trastuzumab deruxtecan (T-DXd) is an antibody-drug conjugate (ADC) consisting of a humanised, anti-human epidermal growth factor receptor 2 (HER2) monoclonal antibody covalently linked to a topoisomerase I inhibitor cytotoxic payload (DXd). The high drug-to-antibody ratio (8:1) ensures a high DXd concentration is delivered to target tumour cells, following internalisation of T-DXd and subsequent cleavage of its tetrapeptide-based linker. DXd's membrane-permeable nature enables it to cross cell membranes and potentially exert antitumour activity on surrounding tumour cells regardless of HER2 expression. T-DXd's unique mechanism of action is reflected in its efficacy in clinical trials in patients with HER2-positive advanced breast cancer (in heavily pretreated populations and in those previously treated with a taxane and trastuzumab), as well as HER2-low metastatic breast cancer. Thus, ADCs such as T-DXd have the potential to change the treatment paradigm of targeting HER2 in metastatic breast cancer, including eventually within the adjuvant/neoadjuvant setting. [Display omitted] • Antibody-drug conjugates have significantly improved breast cancer (BC) treatment. • Trastuzumab deruxtecan's (T-DXd) structure allows large payload delivery to targets. • T-DXd has demonstrated unprecedented efficacy in patients with HER2+ advanced BC. • T-DXd has significantly improved survival rates in heavily pretreated BC patients. • T-DXd may change the treatment paradigm of targeting HER2 in metastatic BC. [ABSTRACT FROM AUTHOR]

The neuregulins and their ErbB/HER receptors play essential roles in mammalian development and tissue homeostasis. In addition, deregulation of their function has been linked to the pathogenesis of diseases such as cancer or schizophrenia. These circumstances have stimulated research into the biology of this ligand-receptor system. Here we show the identification of programmed cell death protein-4 (PDCD4) as a novel neuregulin-ErbB signaling mediator. Phosphoproteomic analyses identified PDCD4 as protein whose phosphorylation increased in cells treated with neuregulin. Mutagenesis experiments defined serine 67 of PDCD4 as a site whose phosphorylation increased upon activation of neuregulin receptors. Phosphorylation of that site promoted degradation of PDCD4 by the proteasome, which depended on exit of PDCD4 from the nucleus to the cytosol. Mechanistic studies defined mTORC1 and ERK1/2 as routes implicated in neuregulin-induced serine 67 phosphorylation and PDCD4 degradation. Functionally, PDCD4 regulated several important biological functions of neuregulin, such as proliferation, migration, or invasion. [ABSTRACT FROM AUTHOR]

Exploitation of the protein degradation machinery as a therapeutic strategy to degrade oncogenic proteins is experiencing revolutionary advances with the development of proteolysis targeting chimeras (PROTACs). PROTACs are heterobifunctional structures consisting of a ligand that binds a protein to be degraded and a ligand for an E3 ubiquitin ligase. The bridging between the protein of interest and the E3 ligase mediated by the PROTAC facilitates ubiquitination of the protein and its proteasomal degradation. In this review we discuss the molecular medicine behind PROTAC mechanism of action, with special emphasis on recent developments and their potential translation to the clinical setting. [ABSTRACT FROM AUTHOR]

Multiple Myeloma (MM) prognosis has recently improved thanks to the incorporation of new therapies to the clinic. Nonetheless, it is still a non-curable malignancy. Targeting cancer cells with agents inducing cell death has been an appealing alternative investigated over the years, as is the case of TRAIL, an agonist of DR4 and DR5 death receptors. This pathway, involved in apoptosis triggering, has demonstrated efficacy on MM cells. In this research, we have investigated the sensitivity of a panel of MM cells to this agent and generated TRAIL-resistant models by continuous culture of sensitive cells with this peptide. Using genomic and biochemical approaches, the mechanisms underlying resistance were investigated. In TRAIL-resistant cells, a strong reduction in cell-surface receptor levels was detected and impaired the apoptotic machinery to respond to the treatment, enabling cells to efficiently form the Death Inducing Signalling Complex. In addition, an upregulation of the inhibitory protein c-FLIP was detected. Even though the manipulation of these proteins was able to modify cellular responses to TRAIL, it was not complete, pointing to other mechanisms involved in TRAIL resistance. [ABSTRACT FROM AUTHOR]

This work is intended to validate the importance of silence in communicating processes, proposing an interdisciplinary theoretical reflection concerning its use as an essential resource under the following oppressive contexts, located in the Italian social fabric: the omertà Mafia's code, the homosexual omertà related to homophobia and the mutism of irregular immigrants because of popular refusal. Three very silent silences, which remain stranded in a society that today venerates Word. [ABSTRACT FROM AUTHOR]

This work is intended to validate the importance of silence in communicating processes, proposing an interdisciplinary theoretical reflection concerning its use as an essential resource under the following oppressive contexts, located in the Italian social fabric: the omertà Mafia's code, the homosexual omertà related to homophobia and the mutism of irregular immigrants because of popular refusal. Three very silent silences, which remain stranded in a society that today venerates Word. [ABSTRACT FROM AUTHOR]

Highlights • Current chemotherapy has strong limitations due to the side effects and resistance. • Medicinal plants provide molecules with anticolorectal cancer activities, both in vitro and in vivo. • Apoptosis, autophagy and cell cycle arrest are induced by the natural products. • Different signaling pathways are activated or inhibited by the herbal products. Abstract Colorectal cancer is one of the most frequent malignancies in the world. Although recent advances in chemotherapy have improved management and survival of colorectal cancer patients, side effects and resistance to chemotherapy have shown the limitations of current chemotherapy and led to the search for alternative treatments. In this context, medicinal plants provide a large number of molecules with proven cytotoxic and apoptogenic activities against several types of cancers including colorectal cancer. These molecules belong to various phytochemical families and trigger different signaling pathways. Here, we review the recent findings regarding the anti-colorectal cancer activities of several plants, both in vitro and in vivo , and the phytochemical molecules possibly responsible for these activities. Besides, their effects on several cancer signaling pathways are discussed. This review highlights the importance of medicinal plants as promising sources of lead anti-colorectal molecules. [ABSTRACT FROM AUTHOR]

In urban design, estimating solar exposure on complex city models is crucial but existing solutions typically focus on simplified building models and are too demanding in terms of memory and computational time. In this paper, we propose an interactive technique that estimates solar exposure on detailed urban scenes. Given a directional exposure map computed over a given time period, we estimate the sky visibility factor that serves to evaluate the final exposure at each visible point. This is done using a screen-space method based on a two-scale approach, which is geometry independent and has low storage costs. Our method performs at interactive rates and is designer-oriented. The proposed technique is relevant in architecture and sustainable building design as it provides tools to estimate the energy performance of buildings as well as weathering effects in urban environments. [ABSTRACT FROM AUTHOR]

Trastuzumab-emtansine (T-DM1) is an antibody-drug conjugate (ADC) that was approved in 2013 to treat HER2+ breast cancer. Despite its efficacy in the clinic, some patients exhibit intrinsic or acquired resistance to such ADC. To characterize mechanisms of resistance to T-DM1, we isolated several HER2+ resistant clones derived from the HCC1954 HER2+ cell line. The isolated clones were different as per their transcriptomic profiles. However, all the T-DM1-resistant clones showed decreased HER2 levels. Yet, the clones were still oncogenically dependent on HER2, as indicated by knock down experiments. The decrease in HER2 expression caused acquired resistance to T-DM1 and to other anti-HER2 therapies. Antibody array analyses showed that the epidermal growth factor receptor (EGFR) was expressed in these T-DM1-resistant HCC1954 clones. Indeed, therapies targeting EGFR, particularly cetuximab-DM1, demonstrated a strong anti-proliferative action on cells with acquired resistance to T-DM1 and HER2 loss. The expression of EGFR in cells resistant to T-DM1 offers the possibility of using therapies directed to this receptor to combat resistance to anti-HER2 drugs and loss of HER2 overexpression. [Display omitted] • HER2 loss in T-DM1-resistant clones causes resistance to other anti-HER2 therapies. • Cetuximab-DM1 has demonstrated a strong antiproliferative action in those models. • Therapies directed at EGFR combat resistance to anti-HER2 drugs. [ABSTRACT FROM AUTHOR]

The use of thalidomide derivatives (IMIDs) has improved multiple myeloma prognosis, through an unknown mechanism of action. Recently one molecular target, the cereblon (CRBN) protein, has been identified. CRBN acts by binding to DDB1-CUL4-ROC1 forming a ubiquitin ligase multiprotein complex. We have generated antibodies to different regions of CRBN protein, and analyzed the biological consequences of augmenting or decreasing CRBN levels. CRBN was expressed in all the myeloma cell lines tested, independently of their sensitivity to IMIDs, and the CRBN-DDB1-CUL4 complex was efficiently formed. At the molecular level, long-term treatment with IMIDs induced a slight decrease in CRBN levels and a reduction in the CRBN-DDB1-CUL4 complex. Interestingly, treatment with other anti-myeloma drugs downregulated cellular contents of CRBN, and in a much faster fashion. These results suggest that CRBN is an important mediator of the cellular response to IMIDs, but also critical in the maintenance of cell viability and/or proliferation. [ABSTRACT FROM AUTHOR]

Trastuzumab-emtansine (T-DM1) is an antibody-drug conjugate (ADC) that was approved in 2013 to treat HER2+ breast cancer. Despite its efficacy in the clinic, some patients exhibit intrinsic or acquired resistance to such ADC. To characterize mechanisms of resistance to T-DM1, we isolated several HER2+ resistant clones derived from the HCC1954 HER2+ cell line. The isolated clones were different as per their transcriptomic profiles. However, all the T-DM1-resistant clones showed decreased HER2 levels. Yet, the clones were still oncogenically dependent on HER2, as indicated by knock down experiments. The decrease in HER2 expression caused acquired resistance to T-DM1 and to other anti-HER2 therapies. Antibody array analyses showed that the epidermal growth factor receptor (EGFR) was expressed in these T-DM1-resistant HCC1954 clones. Indeed, therapies targeting EGFR, particularly cetuximab-DM1, demonstrated a strong anti-proliferative action on cells with acquired resistance to T-DM1 and HER2 loss. The expression of EGFR in cells resistant to T-DM1 offers the possibility of using therapies directed to this receptor to combat resistance to anti-HER2 drugs and loss of HER2 overexpression. [ABSTRACT FROM AUTHOR]

The MAP kinase Erk5 plays important roles in cellular proliferation, and has recently been implicated in the regulation of mitosis. The classic pathway of Erk5 activation involves dual phosphorylation at its TEY microdomain by the upstream regulating kinase MEK5. Here we describe a second pathway that controls Erk5 phosphorylation. This pathway is activated in mitotic cells and involves kinase activities distinct from MEK5. Studies aimed at identifying these kinases suggested that CDK1 activity is required to sustain Erk5 phosphorylation in mitosis, as treatment with RO3306, a CDK1 inhibitor, reversed mitotic phosphorylation of Erk5. Moreover, CDK1 co-precipitated with Erk5 in mitotic cells. The mitotic phosphorylation of Erk5 occurs at multiple sites located at its unique C-terminal region, within an Erk5 subdomain that has formerly been implicated in the control of the subcellular location of Erk5. Furthermore, molecular studies indicated that phosphorylation at these sites may participate in the control of the transit of Erk5 between the cytosol and the nucleus, in addition to regulating its transcriptional activity. Together, our results demonstrate the existence of a second Erk5 phosphorylation pathway, that is activated in mitosis, and that may participate in the regulation of Erk5 functions. [ABSTRACT FROM AUTHOR]

Abstract: The aim of this study was to investigate the survival of a potentially probiotic Lactobacillus plantarum strain in barley, wheat and barley malt extracts. The extracts were produced from three flour/water suspensions, i.e., 5%, 20%, 30% w/w. After inoculation, the cultures were incubated for 24h at 37°C, and were subsequently stored at 4°C for up to seventy days. The lactic acid and reducing sugar concentrations at the beginning of storage were significantly different between the fermented media, ranging from 0.5g/L to 17g/L and from 0.8g/L to 6.5g/L respectively, while the pH ranged between 2.9 and 3.4. It was observed that the cells survived much better in the malt extracts compared to barley and wheat extracts during refrigerated storage. Based on the results from a study using model media and supplemented cereal extracts it was derived that this was most likely due to their higher sugar concentration and the presence of protective unidentified compounds, albeit the fact that the malt extracts contained higher amounts of lactic acid. [Copyright &y& Elsevier]

A molecular hallmark of cancer is the presence of genetic alterations in the tumoral DNA. Understanding how these alterations translate into the malignant phenotype is critical for the adequate treatment of oncologic diseases. Several cancer genome sequencing reports have uncovered the number and identity of proteins and pathways frequently altered in cancer. In this article we discuss how integration of these genomic data with other biological and proteomic studies may help in designing anticancer therapies "a la carte". An important conclusion is that next generation treatment of neoplasias must be based on rational drug combinations that target various pathways and cellular entities that sustain the survival of cancer cells. [ABSTRACT FROM AUTHOR]

Specific pharmacological treatments are currently not available to address problems resulting from fetal ethanol exposure, described as Fetal Alcohol Syndrome or Fetal Alcohol Spectrum Disorders (FASD). The present study evaluated the therapeutic effects of aniracetam against cognitive deficits in a well-characterized and sensitive FASD Sprague–Dawley rat model. Ethanol, administered orally at a moderate dose (4 g/kg/24 h; 38% v/v) during the entire course of pregnancy, caused severe cognitive deficits in offspring. Furthermore, both progeny genders were affected by a spectrum of behavioral abnormalities, such as a delay in the development of the righting reflex, poor novelty seeking behavior, and high anxiety levels in female rats. Cognitive disabilities, monitored in adult rats by a two-way active avoidance task, correlated well with a significant reduction of AMPA (α-amino-3 hydro-5 methyl-isoxazole propionic acid) receptor-mediated miniature excitatory postsynaptic responses (mEPSCs) in the hippocampus. Administration of aniracetam for 10 days (post-natal days (PND) 18–27), at a dose of 50 mg/kg reversed cognitive deficits in both rat genders, indicated by a significant increase in the number of avoidances and the number of ‘good learners’. After the termination of the nootropic treatment, a significant increase in both amplitude and frequency of AMPA receptor-mediated mEPSCs in hippocampal CA-1 pyramidal cells was observed. Significant anxiolytic effects on PND 40 also preceded acquisition improvements in the avoidance task. This study provides evidence for the therapeutic potential of aniracetam in reversing cognitive deficits associated with FASD through positive post-natal modulation of AMPA receptors.Neuropsychopharmacology (2008) 33, 1071–1083; doi:10.1038/sj.npp.1301496; published online 4 July 2007 [ABSTRACT FROM AUTHOR]

Abstract: Nucleocytoplasmic shuttling of multiple signalling proteins is critical in the control of processes such as cell proliferation, differentiation, or apoptosis. One group of proteins whose activity depends on this nucleocytoplasmic traffic includes the mitogen-activated protein kinases. Usually, these kinases reside in the cytoplasm and move to the nucleus upon dual phosphorylation. One of these kinases, Erk5, has been found to reside in the nucleus of breast cancer cells that overexpress the ErbB2 receptor. This raises questions with respect to the mechanisms implicated in Erk5 nuclear location in these cells, as well as the biological consequences of this nuclear residency. In breast cancer cells overexpressing ErbB2, Erk5 dual phosphorylation required ErbB2 tyrosine kinase activity; however, Erk5 nuclear residency did not require ErbB2 activity. Furthermore, translocation of Erk5 from the cytosol to the nucleus occurred in the absence of dual phosphorylation. Nuclear residency of Erk5 in these cells depended on the integrity of a nuclear localization signal present in the unique C-terminus of Erk5. The Erk5 form expressed by these breast cancer cells included the N- and C-terminal cytoplasmic targeting signals, yet Erk5 was nuclear, and remained at this location throughout the interphase without being tightly bound to DNA. Biological studies using a mutant Erk5 that accumulates in the nucleus indicate that nuclear Erk5 favours MEF2-dependent transcriptional activity, and inhibits TRAIL-induced cell death. [Copyright &y& Elsevier]

Squamous cell carcinoma of the head and neck (SCCHN) is the sixth most common cancer in the world. At present several therapeutic approaches, including surgical removal, chemotherapy and radiotherapy, are used. Yet a significant number of patients relapse, often with metastases. In an attempt to improve treatment of SCCHN new targeted therapies are emerging. Among them special interest has been devoted to agents that act on the epidermal growth factor receptor (EGFR) and other receptor tyrosine kinases, or the signal transduction routes used by these receptors to induce tumour cell proliferation. Such treatments include monoclonal antibodies and small molecule inhibitors of either the intracellular tyrosine kinase activity of these receptors or relevant signalling intermediates. Here we review the biological bases of these new targeted treatments, with special emphasis on the clinical results that point to an implementation of these drugs into the therapeutic armamentarium against SCCHN. [ABSTRACT FROM AUTHOR]

The bile tolerance of three strains of lactobacilli, selected on the basis of their ability to ferment cereal substrates and tolerate simulated gastric transit was examined. Lactobacillus reuteri (NCIMB 1195), L. acidophilus (NCIMB 8821), and L. plantarum (NCIMB 8826) were exposed to 2% bile for 4 h in a phosphate-saline buffer at pH 7. The effects of supplementing the assays with cereal extracts, increasing concentrations of glucose, and free amino nitrogen was also examined. In the absence of any supplements L. reuteri showed the greatest resistance to bile, whilst L. acidophilus displayed the greatest sensitivity. Addition of cereal extracts improved the tolerance of all three strains, barley and wheat extracts imparted similar levels of enhancement in viability, whilst the addition of malt showed a greater positive influence. The extent of improvement was related to the concentration of soluble sugars and FAN present in the cereals, this was illustrated further by the addition of glucose to the assays, as the viability of the organisms was progressively improved with increasing concentrations. The presence of free amino nitrogen also increased bacterial resistance to bile but to a lesser extent both glucose and cereal extracts. [Copyright &y& Elsevier]

c-Kit has been shown to be mutated in several types of tumours, and its activity has been correlated with increased proliferation rates in a subset of multiple myeloma (MM) patients. We have investigated the effect of imatinib mesylate (STI571), an inhibitor of c-Kit, on MM cells. STI571 inhibited the proliferation of MM cells by arresting cell cycle progression. Western blotting of cell cycle proteins showed that STI571 increased the levels of p21 and p16. MM cells expressed abl, but its level of tyrosine phosphorylation was low and unaffected by treatment with STI571. c-Kit was also expressed in certain MM cell lines, and its phosphorylation was stimulated by stem cell factor. However, the failure to detect the receptor protein in other MM cell lines in which cell proliferation was inhibited by STI571 suggests that its effect on these c-Kit-negative MM cell lines might be caused by the action of the drug on yet unknown targets. STI571 inhibited the proliferation of MM cells resistant to dexamethasone or melphalan and had an additive effect when combined with dexamethasone. Efforts to understand the action of STI571 in MM cells may help to identify these potentially useful targets in the treatment of this and other disorders. [ABSTRACT FROM AUTHOR]

In this work, the effect of cereal extracts, used as delivery vehicles for potentially probiotic lactic acid bacteria (LAB), on the acid tolerance of the cells was evaluated under conditions that simulate the gastric tract. More specifically, the effect of malt, barley and wheat extracts on the viability of Lactobacillus plantarum, Lactobacillus acidophilus and Lactobacillus reuteri during exposure for 4 h in a phosphate buffer acidified at pH 2.5 was investigated. In the absence of cereal extracts all strains demonstrated a significant reduction in their cell population, particularly L. plantarum. The viability of L. plantarum was improved by approximately 4 log10 cycles in the presence of malt and 3 log10 cycles in the presence of wheat and barley. The survival of L. acidophilus and L. reuteri was increased by more than 1.5 and 0.7 log10 cycle, respectively, upon addition of cereal extracts. In order to evaluate the contribution of the cereal constituents on cell survival, the individual effect of glucose, maltose and free amino nitrogen (FAN), which were added at concentrations that correlated to the reducing sugar and FAN content of the cereal extracts, was examined. The viability of L. plantarum was progressively improved as the maltose or glucose concentration increased; an increase by approximately 2 log10 cycles was observed in the presence of 8.33 g/l sugar. The survival of L. acidophilus increased by more than 1 log10 cycle, even at very low concentrations of maltose and glucose (e.g., 0.67 g/l), while L. reuteri stability was enhanced in the presence of maltose but no appreciable effect was demonstrated in the presence of glucose. Sugar analysis indicated that glycolysis was inhibited in all cases. Addition of tryptone and yeast extract, used as sources of FAN, enhanced L. acidophilus acid tolerance, but did not affect L. reuteri and L. plantarum. The results presented in this study indicate that malt, wheat and barley extracts exhibit a significant protective effect on the viability of L. plantarum, L. acidophilus and L. reuteri under acidic conditions, which could be mainly attributed to the amount of sugar present in the cereal extracts. [Copyright &y& Elsevier]

Gas and solids feeding is a key operation in pneumatic conveying of particulate materials. This article presents an analysis of the interfacing effects between a nozzle gas supplier, a rotary valve solids feeder with dropout box, and the pipeline of a pneumatic conveying test rig for low-velocity dense-phase flow. Experiments were carried out to examine the flow pattern of slugs in different combinations of gas flow conditions and solids loading ratios. The effect of gas and solids feeding on the formation of slugs is analyzed by using both experimental data and computer-modeled results. Solids accumulation and sliding motion at the bottom of the dropout box and near the entrance of the downstream pipe, which happen prior to the bulk motion in the form of a slug, are found important in determining the size of a slug. Gas retention and pressure buildup characteristics in the feed section are also found crucial in influencing the flow patterns of slugs. [ABSTRACT FROM AUTHOR]

An experimental technique has been developed to measure the flow characteristics of slugs in dense phase pneumatic conveying using pressure measurements. This method is based on the unique characteristics of slug flows in pipes, i.e., an axial pressure fluctuation along the pipeline and a pressure difference in the radial direction at the back of a slug. Standard differential pressure transducers were used in this study and the influence of the finite response time of these transducers was considered. Experiments were conducted over a range of gas-solids flow conditions and experimental data were analyzed to describe the behavior of solids slugs through pipes. The calculated slug velocity and length using axial pressure measurements were confirmed by video recordings, and the synthesis between axial and radial pressure signals showed reasonable agreement in flow pattern analysis. This relatively simple measuring technique has been found effective in detecting solids slugs traveling through horizontal pipes and will distinguish various flow regimes. It provides a useful and easily applied tool for system optimizing and benchmarking in industrial applications. [ABSTRACT FROM AUTHOR]

Aims: The overall growth kinetics of four potentially probiotic strains (Lactobacillus fermentum , Lact. reuteri , Lact. acidophilus and Lact. plantarum ) cultured in malt, barley and wheat media were investigated. The objectives were to identify the main factors influencing the growth and metabolic activity of each strain in association with the cereal substrate. Methods and Results: All fermentations were performed without pH control. A logistic-type equation, which included a growth inhibition term, was used to describe the experimental data. In the malt medium, all strains attained high maximum cell populations (8·10–10·11 log10 cfu ml-1 , depending on the strain), probably due to the availability of maltose, sucrose, glucose, fructose (approx. 15 g l-1 total fermentable sugars) and free amino nitrogen (approx. 80 mg l-1 ). The consumption of sugars during the exponential phase (10–12 h) resulted in the accumulation of lactic acid (1·06–1·99 g l-1 ) and acetic acid (0·29–0·59 g l-1 ), which progressively decreased the pH of the medium. Each strain demonstrated a specific preference for one or more sugars. Since small amounts of sugars were consumed by the end of the exponential phase (17–43%), the decisive growth-limiting factor was probably the pH, which at that time ranged between 3·40 and 3·77 for all of the strains. Analysis of the metabolic products confirmed the heterofermentative or homofermentative nature of the strains used, except in the case of Lact. acidophilus which demonstrated a shift towards the heterofermentative pathway. All strains produced acetic acid during the exponential phase, which could be attributed to the presence of oxygen. Lactobacillus plantarum , Lact. reuteri and Lact. fermentum continued to consume the remaining sugars and accumulate metabolic products in the medium, probably due to... [ABSTRACT FROM AUTHOR]

Summary For many centuries fermented foods have been produced and consumed world-wide as they are of such great importance for human nutrition. Fermentation helps to preserve the food, provides a wide variety of textures and flavours and significantly improves the nutritional properties of the raw materials used. In this work we tested the hypothesis that the fermentation process of the Bulgarian cereal-based traditional beverage boza was similar to other cereal-based fermentations. Boza was prepared both from whole-wheat grains and flour, and the effect of the raw material on product quality was studied. The main microorganisms responsible for boza fermentation, yeasts and lactobacilli, were identified and some physical and biochemical changes were monitored during the first 48 h of fermentation. A significant increase in glucose content was observed, while pH, viscosity, free amino nitrogen content and dry matter decreased during the fermentation. The use of wheat flour resulted in a product with higher viscosity and dry matter content when compared to that produced from whole-wheat grains. The effect of temperature on fermentation rate was also tested. [ABSTRACT FROM AUTHOR]

Ocoxin Oral Solution (OOS) and Viusid (VS) are nutritional supplements that include several natural products which affect different cellular functions, such as proliferation or the redox status. In addition, some of their constituent components have been described to exert an antiviral effect. Considering this, it was hypothesized that treatment with OOS and VS could protect from viral infections. In order to evaluate the impact of OOS and VS on viral infection, lentivirus and retrovirus whose genomes coded for green fluorescent protein were used. In addition, and as a second approach to measure viral infection, a hemagglutinin-tagged form of the mitogen-activated protein kinase ERK5 was also inserted in the retroviral vector. Viral particles produced in 293T cells were used to infect HeLa cells in the presence or absence of OOS or VS. It was observed that VS had a minimal effect on the capacity of either lentivirus or retrovirus to infect HeLa cells. However, OOS significantly reduced the infection of HeLa cells with both of these viruses. The effect was dose-dependent, reaching a maximum at a 1:100 dilution of OOS. These results suggested that, in addition to its well-known antitumoral properties, OOS may also inhibit infection with viruses. This effect is relevant since patients receiving oncological therapies are more susceptible to viral infections, and nutritional supplements such as OOS may help in reducing the severity of these potential pathogenic infections. [ABSTRACT FROM AUTHOR]

Defines the mechanism that activates cleavage of membrane protein ectodomains by using pro-transforming growth factor-alpha (pro-TGF-alpha) cleavage as a model system. Preparation of sample cells used; Guanosine 5'-O-3-thiotriphosphate; Heterotrimeric G protein; PMA-sensitive calcium ion-independent protein kinase C; Adenosine triphosphate (ATP) hydrolysis; Absence of cytosol.

The HER3 protein, that belongs to the ErbB/HER receptor tyrosine kinase (RTK) family, is expressed in several types of tumors. That fact, together with the role of HER3 in promoting cell proliferation, implicate that targeting HER3 may have therapeutic relevance. Furthermore, expression and activation of HER3 has been linked to resistance to drugs that target other HER receptors such as agents that act on EGFR or HER2. In addition, HER3 has been associated to resistance to some chemotherapeutic drugs. Because of those circumstances, efforts to develop and test agents targeting HER3 have been carried out. Two types of agents targeting HER3 have been developed. The most abundant are antibodies or engineered antibody derivatives that specifically recognize the extracellular region of HER3. In addition, the use of aptamers specifically interacting with HER3, vaccines or HER3-targeting siRNAs have also been developed. Here we discuss the state of the art of the preclinical and clinical development of drugs aimed at targeting HER3 with therapeutic purposes. [ABSTRACT FROM AUTHOR]

Simple Summary: Breast cancer is a heterogeneous disease that is driven by genetic, epigenetic and phenotypic modifications and is also affected by the microenvironment and the metabolism. In this article we review genetic and non-genetic causes of tumor heterogeneity focusing on the impact that heterogeneity has on resistance to therapy. We will provide examples of personalized medicines and their translation to the clinic. Targeted cancer therapies against oncogenic drivers are actively being developed and tested in clinical trials. Targeting an oncogenic driver may only prove effective if the mutation is present in most tumoral cells. Therefore, highly heterogeneous tumors may be refractory to these therapies. This makes tumor heterogeneity a major challenge in cancer therapy. Although heterogeneity has traditionally been attributed to genetic diversity within cancer cell populations, it is now widely recognized that human cancers are heterogeneous in almost all distinguishable phenotypic characteristics. Understanding the genetic variability and also the non-genetic influences of tumor heterogeneity will provide novel insights into how to reverse therapeutic resistance and improve cancer therapy. [ABSTRACT FROM AUTHOR]

Many of the biological processes of the cell, from its structure to signal transduction, involve protein–protein interactions. On this basis, our aim was to identify cellular proteins that interact with ERK5, a serine/threonine protein kinase with a key role in tumor genesis and progression and a promising therapeutic target in many tumor types. Using affinity chromatography, immunoprecipitation, and mass spectrometry techniques, we unveiled an interaction between ERK5 and the mitochondrial glutaminase GLS in pancreatic tumor cells. Subsequent co-immunoprecipitation and immunofluorescence studies supported this interaction in breast and lung tumor cells as well. Genetic approaches using RNA interference techniques and CRISPR/Cas9 technology demonstrated that the loss of ERK5 function led to increased protein levels of GLS isoforms (KGA/GAC) and a concomitant increase in their activity in tumor cells. It is well known that the tumor cell reprograms its intermediary metabolism to meet its increased metabolic needs. In this sense, mitochondrial GLS is involved in the first step of glutamine catabolism, one of the main energy sources in the context of cancer. Our data suggest that ERK5 contributes to the regulation of tumor cell energy metabolism via glutaminolysis. [ABSTRACT FROM AUTHOR]

A group comparison experiment of two high school classes with pre and post instruction testing has been carried out to study the suitability and advantages of using the active learning of optics and photonics (ALOP) curricula in high schools of developing countries. Two parallel, mixed gender, 12th grade classes of a high school run by the local university were chosen. One course was randomly selected to follow the experimental instruction, based on teacher and student activities contained in the ALOP Manual. The other course followed the traditional, teacher-centered, instruction previously practiced. Conceptual knowledge of the characteristics of image formation by plane mirrors and single convergent and divergent lenses was measured by applying, in both courses, the multiple-choice test, light and optics conceptual evaluation (LOCE). Measurement before instruction showed that initial knowledge was almost null, and therefore equivalent, in both courses. After instruction testing showed that the conceptual knowledge of students following the ALOP curricula more than doubled that achieved by students in the control course, a situation maintained throughout the six conceptual dimensions tested by the 34 questions of the LOCE test used in this experiment. Using a 60% performance level on the LOCE test as the threshold of satisfactory performance, most (about 90%) of the experimental group achieved this level—independent of initial knowledge, while no student following traditional instruction reached this level of understanding. Some considerations and recommendations for prospective users are also included. [ABSTRACT FROM AUTHOR]

Bryonia dioica Jacq. is a climbing perennial herb with tuberous roots which is widely used in traditional medicine in Algeria for the treatment of cancer. The present study aimed to evaluate the apoptogenic activity and phytochemical composition of the aqueous extract of B. dioica roots growing in Algeria. The cytotoxic effect of B. dioica aqueous decoction against breast cancer MDA-MB-231 cells was evaluated by an MTT assay. Apoptosis induction was assessed by an Annexin V-fluorescien iosthiocyanate assay. Propidium iodide staining of cell DNA was used to assess the effects on the cell cycle. In addition to UV–Visible (UV-vis) analysis, the major compounds of the extracts were determined using liquid chromatography-mass spectrometric analyses. Our results showed that the B. dioica aqueous extract induced cell death in a time-dependent manner. The highest inhibitory effect was produced at concentrations of 50 µg/ml or higher after 72 h of treatment (91.15±0.71%). Furthermore, the extract induced apoptosis of MDA-MB-231 cells. At 250 µg/ml, 64.61% of the treated MDA-MB-231 cells were apoptotic. This was accompanied by cell cycle arrest at G2/M phase. The percentage of cells in G2/M increased from 15.7% (untreated cells) to 59.13% (50 µg/ml) and 58.51% (250 µg/ml). The UV-vis absorption spectra of B. dioica aqueous extract showed two absorption bands characteristic of the flavonol skeleton; 350–385 nm (Band I) and 250–280 nm (Band II). Myricetin (2,5,7,3,4,5-pentahydroxylflavonol) was found to be the major compound in the B. dioica aqueous extract. These findings suggest that B. dioica could be considered a promising source for developing novel therapeutics against breast cancer. [ABSTRACT FROM AUTHOR]

Background: Advanced colorectal cancer (CRC) is difficult to treat. For that reason, the development of novel therapeutics is necessary. Here we describe a potentially actionable plasma membrane target, the amino acid transporter protein subunit CD98hc. Methods: Western blot and immunohistochemical analyses of CD98hc protein expression were carried out on paired normal and tumoral tissues from patients with CRC. Immunofluorescence and western studies were used to characterize the action of a DM1-based CD98hc-directed antibody–drug conjugate (ADC). MTT and Annexin V studies were performed to evaluate the effect of the anti-CD98hc-ADC on cell proliferation and apoptosis. CRISPR/Cas9 and shRNA were used to explore the specificity of the ADC. In vitro analyses of the antitumoral activity of the anti-CD98hc-ADC on 3D patient-derived normal as well as tumoral organoids were also carried out. Xenografted CRC cells and a PDX were used to analyze the antitumoral properties of the anti-CD98hc-ADC. Results: Genomic as well proteomic analyses of paired normal and tumoral samples showed that CD98hc expression was significantly higher in tumoral tissues as compared to levels of CD98hc present in the normal colonic tissue. In human CRC cell lines, an ADC that recognized the CD98hc ectodomain, reached the lysosomes and exerted potent antitumoral activity. The specificity of the CD98hc-directed ADC was demonstrated using CRC cells in which CD98hc was decreased by shRNA or deleted using CRISPR/Cas9. Studies in patient-derived organoids verified the antitumoral action of the anti-CD98hc-ADC, which largely spared normal tissue-derived colon organoids. In vivo studies using xenografted CRC cells or patient-derived xenografts confirmed the antitumoral activity of the anti-CD98hc-ADC. Conclusions: The studies herewith reported indicate that CD98hc may represent a novel ADC target that, upon well-designed clinical trials, could be used to increase the therapeutic armamentarium against CRC. [ABSTRACT FROM AUTHOR]

Background: Advanced colorectal cancer (CRC) is difficult to treat. For that reason, the development of novel therapeutics is necessary. Here we describe a potentially actionable plasma membrane target, the amino acid transporter protein subunit CD98hc. Methods: Western blot and immunohistochemical analyses of CD98hc protein expression were carried out on paired normal and tumoral tissues from patients with CRC. Immunofluorescence and western studies were used to characterize the action of a DM1-based CD98hc-directed antibody–drug conjugate (ADC). MTT and Annexin V studies were performed to evaluate the effect of the anti-CD98hc-ADC on cell proliferation and apoptosis. CRISPR/Cas9 and shRNA were used to explore the specificity of the ADC. In vitro analyses of the antitumoral activity of the anti-CD98hc-ADC on 3D patient-derived normal as well as tumoral organoids were also carried out. Xenografted CRC cells and a PDX were used to analyze the antitumoral properties of the anti-CD98hc-ADC. Results: Genomic as well proteomic analyses of paired normal and tumoral samples showed that CD98hc expression was significantly higher in tumoral tissues as compared to levels of CD98hc present in the normal colonic tissue. In human CRC cell lines, an ADC that recognized the CD98hc ectodomain, reached the lysosomes and exerted potent antitumoral activity. The specificity of the CD98hc-directed ADC was demonstrated using CRC cells in which CD98hc was decreased by shRNA or deleted using CRISPR/Cas9. Studies in patient-derived organoids verified the antitumoral action of the anti-CD98hc-ADC, which largely spared normal tissue-derived colon organoids. In vivo studies using xenografted CRC cells or patient-derived xenografts confirmed the antitumoral activity of the anti-CD98hc-ADC. Conclusions: The studies herewith reported indicate that CD98hc may represent a novel ADC target that, upon well-designed clinical trials, could be used to increase the therapeutic armamentarium against CRC. [ABSTRACT FROM AUTHOR]

Expression of high levels of immune cells including neutrophils has been associated with detrimental outcome in several solid tumors and new strategies to decrease their presence and activity are currently under clinical development. Here, we review some of the relevant literature of the role of neutrophils in different stages of the oncogenic process including tumor initiation, growth, proliferation or metastatic spreading and also focus on how neutrophil counts or the neutrophil-to-lymphocyte ratio may be used as a prognostic and predictive biomarker. Strategies to avoid the deleterious effects of neutrophils in cancer and to reduce their activity are discussed. Examples for such strategies include inhibition of CXCR1 and CXCR2 to decrease migration of neutrophils to tumoral areas or the inhibition of granulocyte colony stimulating factor to decrease the amount of neutrophils which has shown efficacy in preclinical models. [ABSTRACT FROM AUTHOR]

Background: The dismal prognosis of advanced ovarian cancer calls for the development of novel therapies to improve disease outcome. In this regard, we set out to discover new molecular entities and to assess the preclinical effectiveness of their targeting. Methods: Cell lines, mice and human ovarian cancer samples were used. Proteome profiling of human phosphokinases, in silico genomic analyses, genetic (shRNA and CRISPR/Cas9) and pharmacological strategies as well as an ex vivo human preclinical model were performed. Results: We identified WNK1 as a highly phosphorylated protein in ovarian cancer and found that its activation or high expression had a negative impact on patients' survival. Genomic analyses showed amplification of WNK1 in human ovarian tumours. Mechanistically, we demonstrate that WNK1 exerted its action through the MEK5–ERK5 signalling module in ovarian cancer. Loss of function, genetic or pharmacological experiments, demonstrated anti‐proliferative and anti‐tumoural effects of the targeting of the WNK1–MEK5–ERK5 route. Additional studies showed that this pathway modulated the anti‐tumoural properties of the MEK1/2 inhibitor trametinib. Thus, treatment with trametinib activated the WNK1–MEK5–ERK5 route, raising the possibility that this effect may limit the therapeutic benefit of ERK1/2 targeting in ovarian cancer. Moreover, in different experimental settings, including an ex vivo patient‐derived model consisting of ovarian cancer cells cultured with autologous patient sera, we show that inhibition of WNK1 or MEK5 increased the anti‐proliferative and anti‐tumour efficacy of trametinib. Conclusions: The present study uncovers the participation of WNK1–MEK5–ERK5 axis in ovarian cancer pathophysiology, opening the possibility of acting on this pathway with therapeutic purposes. Another important finding of the present study was the activation of that signalling axis by trametinib, bypassing the anti‐tumoural efficacy of this drug. That fact should be considered in the context of the use of trametinib in ovarian cancer. [ABSTRACT FROM AUTHOR]

Background: Triple negative breast cancer (TNBC) represents around 15% of all breast tumors. Therapeutic strategies in TNBC are mainly based on classical chemotherapeutic agents, with limited success in the metastatic setting. Here we have explored the action of a novel mithramycin analog named EC-8042 in TNBC. Methods: we used a panel of cell lines representative of triple negative tumors including BT549, HBL100, HCC-1037, HS578T, MDA-MB213, HCC-1187, HCC-3153 and HCC-70. Proliferation studies with EC-8042 were measured by MTT assays. Propidium iodide (PI) staining followed by FACS was used for cell cycle analyses and AnnexinV-fluorescein for analysis of apoptosis. Proteins involved in cell cycle control and apoptosis were evaluated by western blot. Mitochondrial membrane potential was assessed by measuring the fluorescence of tetramethylrhodamineethylester (TMRE) dye. Results: The IC50 values obtained were in the nM range, indicating that the drug had a potent effect on these cell lines. Mechanistically, the drug caused accumulation of cells in the S phase of the cell cycle at doses of 100 and 500 nM in MDA-MB231. Biochemically, EC-8042 caused an increase in cyclin E, cyclin A, cyclin D3, cyclin B, and p27, and decreased cyclin D1 and Rb phosphorylation at serine 780. These data are consistent with an action of EC-8042 on the cell cycle. Moreover, the increase in wee1 and pY15-CDK1 are in line with an effect of the drug blocking cell cycle progression through the S phase. Annexin V staining indicated that treatment with 500 nM of EC8042 provoked apoptosis, which was also revealed by PARP processing. The treatment reduced the amounts of MCL1, BCLXL, and XIAP, but did not affect BCL2 levels. Conclusion: our results indicate that EC-8042 has a strong antitumoral action on TNBC cells, which is caused by increased apoptosis and a decrease in cell cycle progression, and opens the possibility for further development of this drug in TNBC. [ABSTRACT FROM AUTHOR]

Simple Summary: Sarcoma is a heterogeneous group of tumors poorly studied with few therapeutic opportunities. Interestingly, the role of MAPKs still remains unclear in sarcomatous pathology. Here, we describe for the first time the critical role of ERK5 in the biology of soft tissue sarcoma by using in vitro and in vivo approaches in a murine experimental model of chemical sarcomagenesis. Indeed, our observations were extrapolated to a short series of human leiomyosarcoma and rhabdomyosarcomas. Furthermore, transcriptome analysis allows us to demonstrate the critical role of KLF2 in the biological effects of ERK5. Therefore, the data presented here open new windows in the diagnosis and therapy of soft tissue sarcomas. Sarcomas are a heterogeneous group of tumors in which the role of ERK5 is poorly studied. To clarify the role of this MAPK in sarcomatous pathology, we used a murine 3-methyl-cholanthrene (3MC)-induced sarcoma model. Our data show that 3MC induces pleomorphic sarcomas with muscle differentiation, showing an increased expression of ERK5. Indeed, this upregulation was also observed in human sarcomas of muscular origin, such as leiomyosarcoma or rhabdomyosarcoma. Moreover, in cell lines derived from these 3MC-induced tumors, abrogation of Mapk7 expression by using specific shRNAs decreased in vitro growth and colony-forming capacity and led to a marked loss of tumor growth in vivo. In fact, transcriptomic profiling in ERK5 abrogated cell lines by RNAseq showed a deregulated gene expression pattern for key biological processes such as angiogenesis, migration, motility, etc., correlating with a better prognostic in human pathology. Finally, among the various differentially expressed genes, Klf2 is a key mediator of the biological effects of ERK5 as indicated by its specific interference, demonstrating that the ERK5–KLF2 axis is an important determinant of sarcoma biology that should be further studied in human pathology. [ABSTRACT FROM AUTHOR]