Role of the Polymer Microstructure in Controlling Colloidal and Thermo-Responsive Properties of Nano-Objects Prepared Via RAFT Polymerization in a Non-polar Medium

After having demonstrated their potential in biomedicalapplications,thermo-responsive block copolymers that are able to self-assembleinto nano-objects in response to temperature modifications are becomingmore and more appealing in other sectors, such as the oil and gasand lubricant fields. Reversible addition-fragmentation chaintransfer (RAFT) polymerization-induced self-assembly has been demonstratedas a valuable strategy for producing nano-objects from modular blockcopolymers in non-polar media, required for the mentioned applications.Although the influence of the nature and size of the thermo-responsiveblock of these copolymers on the properties of the nano-objects isextensively studied in the literature, the role of the solvophilicblock is often neglected. In this work, we elucidate the role of themain microstructural parameters, including those of the solvophilicportion, of block copolymers produced by RAFT polymerization in thehydrocarbon blend decane/toluene 50:50 v/v on the thermo-responsivebehavior and colloidal properties of the resulting nano-objects. Twolong-aliphatic chain monomers were employed for the synthesis of fourmacromolecular chain transfer agents (macroCTAs), with increasingsolvophilicity according to the number of units (n) or length of the alkyl side chain (q). Subsequently,the macroCTAs were chain-extended with different repeating units ofdi(ethylene glycol) methyl ether methacrylate (p),leading to copolymers that are able to self-assemble below a criticaltemperature. We show that this cloud point can be tuned by actingon n, p, and q.On the other hand, the colloidal stability, expressed in terms ofarea of the particle covered by each solvophilic segment, is onlya function of n and q, which providesa way for controlling the size distribution of the nano-objects andto decouple it from the cloud point

SOX2 Is a Univocal Marker for Human Oral Mucosa Epithelium Useful in Post-COMET Patient Characterization

Total bilateral Limbal Stem Cells Deficiency is a pathologic condition of the ocular surface due to loss or impairment of corneal stem cell function, altering homeostasis of the corneal epithelium. Cultivated Oral Mucosa Epithelial Transplantation (COMET) is the only autologous treatment for this pathology. During the follow-up, a proper characterization of the transplanted oral mucosa on the ocular surface supports understanding the regenerative process. The previously proposed markers for oral mucosa identification (e.g., keratins 3 and 13) are co-expressed by corneal and conjunctival epithelia. Here, we propose a new specific marker to distinguish human oral mucosa from the epithelia of the ocular surface. We compared the transcriptome of holoclones (stem cells) from the human oral mucosa, limbal and conjunctival cultures by microarray assay. High expression of SOX2 identified the oral mucosa vs. cornea and conjunctiva, while PAX6 was highly expressed in corneal and conjunctival epithelia. The transcripts were validated by qPCR, and immunological methods identified the related proteins. Finally, the proposed markers were used to analyze a 10-year follow-up aniridic patient treated by COMET. These findings will support the follow-up analysis of COMET treated patients and help to shed light on the mechanism of corneal repair and regeneration

Analytic and Dynamic Secretory Profile of Patient-Derived Cytokine-Induced Killer Cells

Adoptive immunotherapy with cytokine induced killer (CIK) cells has shown antitumor activity against several kinds of cancer in preclinical models and clinical trials. CIK cells are a subset of ex vivo expanded T lymphocytes with T-NK phenotype and MHC-unrestricted antitumor activity. The literature provides scant information on cytokines, chemokines and growth factors secreted by CIK cells. Therefore, we investigated the secretory profile of CIK cells generated from tumor patients. The secretome analysis was performed at specific time points (d 1, d 14 and d 21) of CIK cell expansion. Mature CIK cells (d 21) produce a great variety of interleukins and secreted proteins that can be divided into three groups based on their secretion quantity: high (interleukin [IL]-13, regulated on activation normal T cell expressed and secreted [RANTES] chemokine, MIP-1 alpha and 1 beta), medium (IL-1Ra, IL-5, IL-8, IL-10, IL-17, IP-10, INF-gamma, vascular endothelial growth factor [VEGF] and granulocyte-macrophage colony-stimulating factor [GM-CSF]) and low (IL-1 beta, IL-4, IL-6, IL-7, IL-9, IL-12, IL-15, eotaxin, platelet-derived growth factor-bb, basic fibroblast growth factor, G-CSF and monocyte chemoattractant protein [MCP]-1). Moreover, comparing peripheral blood mononuclear cells (PBMCs) (d 1) and mature CIK cells (d 14 and 21) secretomes, we observed that IL-5, IL-10, IL-13, GM-CSF and VEGF were greatly upregulated, while IL-1 beta, IL-6, IL-8, IL-15, IL-17, eotaxin, MCP-1 and RANTES were downregulated. We also performed a gene expression profile analysis of patient-derived CIK cells, showing that mRNA for the different cytokines and secreted proteins was modulated during PBMC-to-CIK differentiation. We highlight previously unknown secretory properties and provide, for the first time, a comprehensive molecular characterization of CIK cells. Our findings provide a rationale to explore the functional implications and possible therapeutic modulation of CIK secretome

Comparison between Cultivated Oral Mucosa and Ocular Surface Epithelia for COMET Patients Follow-Up

Total bilateral Limbal Stem Cell Deficiency is a pathologic condition of the ocular surface due to the loss of corneal stem cells. Cultivated oral mucosa epithelial transplantation (COMET) is the only autologous successful treatment for this pathology in clinical application, although abnormal peripheric corneal vascularization often occurs. Properly characterizing the regenerated ocular surface is needed for a reliable follow-up. So far, the univocal identification of transplanted oral mucosa has been challenging. Previously proposed markers were shown to be co-expressed by different ocular surface epithelia in a homeostatic or perturbated environment. In this study, we compared the transcriptome profile of human oral mucosa, limbal and conjunctival cultured holoclones, identifying Paired Like Homeodomain 2 (PITX2) as a new marker that univocally distinguishes the transplanted oral tissue from the other epithelia. We validated PITX2 at RNA and protein levels to investigate 10-year follow-up corneal samples derived from a COMET-treated aniridic patient. Moreover, we found novel angiogenesis-related factors that were differentially expressed in the three epithelia and instrumental in explaining the neovascularization in COMET-treated patients. These results will support the follow-up analysis of patients transplanted with oral mucosa and provide new tools to understand the regeneration mechanism of transplanted corneas

Magnolia officinalis L. bark extract and respiratory diseases: From traditional Chinese medicine to western medicine via network target

The understanding of the use of Magnolia officinalis L. (Magnoliaceae) as a possible dietary supplement for supporting the treatment of airway pathologies might be of clinical interest. Two commercially available bark extracts (M. officinalis extract [MOE]) were characterized by quantitation in honokiol and magnolol content by means of high-performance liquid chromatography with UV detection. MOE effects, as well as those of the reference compounds per se, on some targets connected to airway pathologies (antibacterial- and lung and trachea relaxing- activities) were investigated. Results showed that MOE possessed interesting antibacterial activity against Staphylococcus aureus, Pseudomonas aeruginosa, and Streptococcus pneumoniae. This was accompanied by a spasmolytic and antispasmodic activity, possibly owing to its ability to concurrently modulate different targets such as H-1-, beta(2)- and muscarinic receptors and l-type calcium channels involved in bronchodilation. All these effects were directly related to the MOE content in honokiol and magnolol. In conclusion, the properties of MOE highlighted here strongly encourage its application as dietary supplement in the treatment of airway diseases

The Anti-Cholinesterase Potential of Fifteen Different Species of Narcissus L. (Amaryllidaceae) Collected in Spain

Narcissus L. is a renowned plant genus with a notable center of diversity and is primarily located in the Mediterranean region. These plants are widely recognized for their ornamental value, owing to the beauty of their flowers; nonetheless, they also hold pharmacological importance. In Europe, pharmaceutical companies usually use the bulbs of Narcissus pseudonarcissus cv. Carlton to extract galanthamine, which is one of the few medications approved by the FDA for the palliative treatment of mild-to-moderate symptoms of Alzheimer’s disease. The purpose of this study was to evaluate the potential of these plants in Alzheimer’s disease. The alkaloid extract from the leaves of different species of Narcissus was obtained by an acid-base extraction work-up -procedure. The biological potential of the samples was carried out by evaluating their ability to inhibit the enzymes acetyl- and butyrylcholinesterase (AChE and BuChE, respectively). The species N. jacetanus exhibited the best inhibition values against AChE, with IC50 values of 0.75 ± 0.03 µg·mL−1, while N. jonquilla was the most active against BuChE, with IC50 values of 11.72 ± 1.15 µg·mL−1.Programa Iberoamericano de Ciencia y Tecnologia para el Desarrollo (CYTED, 223RT0140)

As representações do movimento de Stonewall nos Estados Unidos (1969): “Stonewall - A Luta Pelo Direito de Amar” (1995) e “Stonewall: Onde o Orgulho Começou” (2015)

O início da história da luta por direitos da comunidade LGBT, por vezes, é atribuído às manifestações contra a invasão do bar Stonewall Inn, no episódio que ficou conhecido como “rebelião de Stonewall”. O presente artigo objetiva pensar como se deu a construção da memória sobre o movimento de Stonewall ocorrido na cidade Nova Iorque, em 1969. Além disso, buscamos analisar de que maneira a experiência LGBT estadunidense é representada no cinema. Dessa forma, abordamos o contexto dos Estados Unidos da época, bem como o que foram os protestos de Stonewall e seu impacto para a comunidade LGBT. A questão da memória foi pensada a partir de fontes audiovisuais, com dois longa-metragens, “Stonewall - A Luta pelo Direito de Amar” (1995) e “Stonewall: Onde o Orgulho Começou” (2015), que tiveram diferentes recepções pelo público. Abordamos como esses dois filmes, apesar de tratarem do mesmo ocorrido, possuem narrativas e leituras diferentes sobre a rebelião de Stonewall

Inhibition of ERK1/2 signaling prevents bone marrow fibrosis by reducing osteopontin plasma levels in a myelofibrosis mouse model

Clonal myeloproliferation and development of bone marrow (BM) fibrosis are the major pathogenetic events in myelofibrosis (MF). The identification of novel antifibrotic strategies is of utmost importance since the effectiveness of current therapies in reverting BM fibrosis is debated. We previously demonstrated that osteopontin (OPN) has a profibrotic role in MF by promoting mesenchymal stromal cells proliferation and collagen production. Moreover, increased plasma OPN correlated with higher BM fibrosis grade and inferior overall survival in MF patients. To understand whether OPN is a druggable target in MF, we assessed putative inhibitors of OPN expression in vitro and identified ERK1/2 as a major regulator of OPN production. Increased OPN plasma levels were associated with BM fibrosis development in the Romiplostim-induced MF mouse model. Moreover, ERK1/2 inhibition led to a remarkable reduction of OPN production and BM fibrosis in Romiplostim-treated mice. Strikingly, the antifibrotic effect of ERK1/2 inhibition can be mainly ascribed to the reduced OPN production since it could be recapitulated through the administration of anti-OPN neutralizing antibody. Our results demonstrate that OPN is a novel druggable target in MF and pave the way to antifibrotic therapies based on the inhibition of ERK1/2-driven OPN production or the neutralization of OPN activity