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A rodent eyeball is a powerful tool for researching the pathomechanisms of many ophthalmic diseases, such as glaucoma, hypertensive retinopathy, and many more. Preclinical experiments enable researchers to examine the efficacy of novel drugs, develop new methods of treatment, or seek new pathomechanisms involved in the disease's onset or progression. A histological examination provides a lot of information necessary to assess the effects of the conducted experiments and can reveal degeneration, tissue remodeling, infiltration, and many other pathologies. In clinical research, there is rarely any chance of obtaining eye tissue suitable for a histological examination, which is why researchers should take advantage of the opportunity offered by the examination of eyeballs from rodents. This manuscript presents a protocol for the histological preparation of rodent eyeballs' sections. The procedure is presented for the eyeballs of mice and rats and has the following steps: (i) harvesting the eyeball, (ii) preserving the eyeball for further analysis, (iii) processing the tissue in paraffin, (iv) preparing slides, (v) staining with hematoxylin and eosin, (vi) assessing the tissue under a light microscope. With the proposed method, the retina can be easily visualized and assessed in detail.

This study reveals insights into the transurethanization reactions leading to the aliphatic-aromatic non-isocyanate poly(carbonate-urethane)s (NIPCUs) and their structure-property relationships. The crucial impact of the alkyl chain length in 4,4'-diphenylmethylene bis(hydroxyalkyl carbamate) (BHAC) on the process of transurethanization reactions was proved. The strong susceptibility of hydroxyethyl- and hydroxybutyl carbamate moieties to the back-biting side reactions was observed due to the formation of thermodynamically stable cyclic products and urea bonds in the BHACs and NIPCUs. When longer alkyl chains (hydroxypentyl-, hydroxyhexyl-, or hydroxydecyl carbamate) were introduced into the BHAC structure, it was not prone to the back-biting side reaction. Both 1 H and 13 C NMR, as well as FT-IR spectroscopies, confirmed the presence of carbonate and urethane (and urea for some of the samples) bonds in the NIPCUs, as well as proved the lack of allophanate and ether groups. The increase in the alkyl chain length (from 5 to 10 carbon atoms) between urethane groups in the NIPCU hard segments resulted in the increase in the elongation at break and crystalline phase content, as well as the decrease in the T g , tensile strength, and hardness. Moreover, the obtained NIPCUs exhibited exceptional mechanical properties (e.g., tensile strength of 40 MPa and elongation at break of 130%).

Development of hydrophobically modified ethoxylated urethane (HEUR) rheology modifiers enabled the widespread application of waterborne paints and coatings, replacing their environmentally burdening solvent-based predecessors. However, the diisocyanates, required for the conventional synthesis of HEURs, pose severe eco-sustainability threats. In this paper, we demonstrate an innovative approach to avoiding toxic components in the preparation of rheology modifiers by obtaining a new class of water-soluble isocyanate-free hydrophobically modified ethoxylated poly(hydroxy-urethane)s (IFHEURs). The first step in the synthetic pathway was the preparation of CO 2 -based five-membered poly(ethylene glycol) bis(cyclic carbonate) and its subsequent aminolysis using 4,7,10-trioxa-1,13-tridecanediamine, yielding poly(hydroxy-urethane) (PHU) prepolymers terminated with cyclic carbonate groups. The PHU prepolymers were further extended in a reactive extrusion (REX) synthesis using biobased hydrophobic diamine PRIAMINE 1075. The REX technique made it possible to overcome the typical limitations of the aminolysis reaction and to reach the desired conversion within a moderate reaction time. IFHEURs have been structurally elucidated using FT-IR and NMR spectroscopy techniques, MALDI-ToF mass spectrometry, and SEC analysis and applied as rheology modifiers. The study of their associative behavior in aqueous solutions confirmed that the architectural flexibility of the obtained IFHEURs, containing terminal and pendant hydrophobic groups, opens a perspective for tuneable thickening performance., Competing Interests: The authors declare no competing financial interest., (© 2022 The Authors. Published by American Chemical Society.)

The presented research focuses on the synthesis and structure-properties relationship of poly(carbonate-urea-urethane) (PCUU) systems including investigations on shape-memory effect capability. Furthermore, we approached the topic from a broader perspective by conducting extensive analysis of the relationship between the synthesized compounds and the results of computer simulations by means of the Monte Carlo method. For the first time, by using a unique simulation tool, the dynamic lattice liquid model (DLL), all steps of multi-step synthesis of these materials were covered by the simulations. Furthermore, broad thermal, mechanical, and thermomechanical characterization of synthesized PCUUs was performed, as well as determining the shape-memory properties. PCUUs exhibited good mechanical properties with a tensile strength above 20 MPa, elongation at break around 800%, and an exhibited shape-memory effect with shape fixity and shape recovery ratios above 94% and 99%, respectively. The dynamic lattice liquid model was employed to show the products and their molar mass distribution, as well as monomer conversion or the dispersity index for individual reaction steps. The results obtained in the following manuscript allow the planning of syntheses for the PCUUs of various structures, including crosslinked and soluble systems, which can provide a broad variety of applications of these materials, as well as a better understanding of the composition-properties relationship.

In this article we report an easy synthetic route towards hyperbranched polyglycerols (Amm-HBPGs) containing trimethylammonium groups and siloxane or hydroxyl end-groups. Siloxane derivatives of Amm-HBPGs were synthesized in an efficient five-step procedure including an anionic ring opening copolymerization of the phthalimide-epoxy monomer with glycidol, followed by reactions with allyl bromide, hydrosililation with hydrogenheptamethyltrisiloxane, hydrazinolysis of phthalimide groups and quaternization of resulting amine groups with methyl iodide. Hydroxyl derivatives were obtained by quaternization of previously reported aminated HBPG's with methyl iodide. Polymeric products were characterized using various NMR techniques, FTIR, and elemental analysis. Both Amm-HBPGs were shown to be effective in catalysis of addition of CO 2 to oxirane. The hydrophilic catalysts showed higher efficiency but synthesis of ethylene carbonate was accompanied by formation of small amounts of ethylene glycol. The siloxane-containing catalyst was easily separable from reaction mixture showing high potential in the process of converting carbon dioxide into valuable chemical raw materials.

The nine identified human homologues of E. coli AlkB 2-oxoglutarate (2OG) and Fe(II)-dependent dioxygenase, ALKBH1-8 and FTO, display different substrate specificities and diverse biological functions. Here we discovered the combined overexpression of members of the ALKBH family in head and neck squamous cell carcinomas (HNSCC). We found direct correlation of ALKBH3 and FTO expression with primary HNSCC tumor size. We observed unidentified thus far cytoplasmic localization of ALKBH2 and 5 in HNSCC, suggesting abnormal role(s) of ALKBH proteins in cancer. Further, high expression of ALKBHs was observed not only in HNSCC, but also in several cancerous cell lines and silencing ALKBH expression in HeLa cancer cells resulted in dramatically decreased survival. Considering the discovered impact of high expression of ALKBH proteins on HNSCC development, we screened for ALKBH blockers among newly synthetized anthraquinone derivatives and demonstrated their potential to support standard anticancer therapy.

Introduction: M2 macrophages are predominant in the immune infiltrates of resected tumours, but little is known about macrophage phenotype in the local lung cancer environment, which may be evaluated by bronchoalveolar lavage fluid (BALF)., Aim of the Study: To find differences between BALF from lung affected by cancer (clBALF) and hlBALF from the opposite, healthy lung, as a control, from the same patient, regarding their individual macrophage polarization and their correlation with IL-10 and TGF-β., Material and Methods: Eighteen patients with confirmed lung cancer were investigated. Macrophage subtyping was performed by immunofluorescence with antibodies anti-CCR7 and CD163 (M1 and M2, respectively)., Results: We found five populations of macrophages: cells with a single reaction: only for CCR7+ or CD163+, a double reaction (CCR7+CD163+), cells with a stronger CD163 (CCR7 low CD163+), and cells with a stronger CCR7 (CCR7+CD163 low ). The main population in the clBALF was composed of cells with a phenotype similar to M2 (CCR7 low CD163+), while in the hlBALF the predominating phenotype was the one similar to M1 (CCR7+CD163 low ). The median proportion of TGF-β1 concentration was higher in the clBALF and hlBALF supernatant than in the serum., Conclusions: In this study we confirmed the usefulness of the immunofluorescence method with CCR7 and CD163 in the evaluation of BALF macrophage polarization in lung cancer., Competing Interests: The authors declare no conflict of interest., (Copyright © 2019 Termedia.)

Photodynamic therapy (PDT) has been shown to destroy tumour-associated lymphatic vessels. Therefore, we sought to investigate the functional outcomes of PDT-mediated damage to the lymphatic vessels. We observed that PDT with verteporfin, completely but transiently, blocks the functional lymphatic drainage in the orthotopic mammary tumour models. Sustained inhibition of lymphatic vessels regeneration induced by lenalidomide or the soluble form of vascular endothelial growth factor receptor 3 (sVEGFR3) that neutralises lymphangiogenic vascular endothelial growth factor C (VEGF-C), significantly impaired antitumour efficacy of PDT. Antilymphangiogenic compounds also significantly inhibited the ability of intratumourally inoculated dendritic cells (DCs) to translocate to local lymph nodes and diminished the number of tumour-infiltrating interferon-γ-secreting or tumour antigen-specific CD8 + T cells. Lenalidomide also abrogated antitumour effects of the combination immunotherapy with PDT and anti-programmed death-ligand 1 (PD-L1) antibodies. Altogether, these findings indicate that PDT-mediated damage to the lymphatic vessels negatively affects development of antitumour immunity, and that drugs that impair lymphatic vessel regeneration might not be suitable for the use in combination with PDT., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Endometrial cancer (EC) is the most common gynecological malignancy in Europe and North America. It is classified into two types exhibiting different characteristics and prognosis. Type I is an estrogen-dependent tumor, histologically classified as low grade and low stage, usually with an excellent prognosis. Type II EC is unrelated to estrogen stimulation and is characterized by a poor prognosis. MicroRNAs (miRNAs, miRs) are small non-coding RNA polynucleotides that regulate gene expression post-transcriptionally. Various dysregulations in microRNA expression are often considered to have an impact on the diagnosis, prognosis and overall survival in patients diagnosed with different types of cancers. Recent data suggest that microRNAs play an important role in the pathogenesis of EC. The aim of the study was to evaluate the involvement of matrix metaloprotease 14 (MMP-14) and microRNA-410 in formation of the EC tumor. To this end expression of MMP-14 and microRNA-410 was assessed within the cancer, transient and healthy zones in the histological sections of tumours using immunohistochemical staining and laser capture microdissection (LCM) followed by a quantitative real-time PCR. The results revealed significantly higher expression of MMP-14 in the cancer tissue zone in comparison to the healthy tissue zone, as well as a lower expression of microRNA-410 in the cancer zone compared with the healthy zone. This reverse correlation may suggest a regulatory role of miRNA-410 in modulating levels of MMP-14 in EC. This is the first report on such regulation in human endometrial cancer.

The role of Epstein-Barr virus (EBV) in the biology and clinical characteristics of diffuse large B cell lymphoma (DLBCL) is still poorly defined. A new provisional entity EBV-positive DLBCL of the elderly has been described in Asian population. Its incidence and prognosis remains unknown in middle European patients. Clinical data and tissue samples were collected from 74 Caucasian patients with DLBCL, aged between 23 and 86 years, treated at a single institution. Lymphoma morphology was reassessed, laboratory procedures included in situ hybridization specific for EBV-encoded small RNAs (EBER), immunohistochemical staining for latent membrane protein and serological testing for EBV-specific antibodies. EBER staining revealed 12.2 % of EBV-positive cases, whereas 9.5 % were diagnosed as EBV-positive DLBCL of the elderly. Serologic EBV markers did not correlate with the presence of EBV in tissue samples (P > 0.10). Elderly EBV-positive cases had lower BCL-6 (P = 0.038) and higher CD30 (P = 0.049) expression and were characterized by higher progression risk (median time-to-progression 12.5 months vs not reached; P = 0.029) and a trend towards worse overall survival (median overall survival 24.5 months vs not reached; P = 0.059). EBV-positive DLBCL of the elderly occurs relatively frequently in Polish population and may be associated with inferior prognosis in comparison with DLBCL, not otherwise specified.

Introduction:  Sarcoidosis is a granulomatous disease, which most often affects the lungs. The role of alveolar macrophages (AMs) in granuloma formation in sarcoidosis has been established. Recently, 2 macrophage populations have been described: M1 and M2. In our styudy, we focused on the effect of tobacco smoking on sarcoidosis. The number of AMs in the lungs of smokers is significantly increased; therefore, it is interesting to study the effect of smoking on AM polarization in sarcoidosis. , Objectives:  The aim of the present study was to identify M1 and M2 macrophages in bronchoalveolar lavage (BAL) fluid from patients with sarcoidosis and assess the effect of smoking on these cells. , Patients and Methods:  The study included 36 patients with confirmed sarcoidosis (18 smokers and 18 nonsmokers). Macrophage populations in BAL fluid were assessed by immunocytochemistry using anti-CD40 and anti-CD163 antibodies (for M1 and M2, respectively). The BAL fluid concentration of interleukin 10 (IL-10) was measured using an enzyme-linked immunosorbent assay. , Results:  We identified 3 populations of macrophages stained with anti-CD40 and anti-CD163 antibodies: small strongly positive cells, large weakly positive cells, and negative cells. The median proportions of these macrophages were 61%, 35%, and 2%, respectively, for CD40, and 55.5%, 35%, and 5%, respectively, for CD163; the proportions did not differ significantly between smokers and nonsmokers. Only the proportion of CD163-negative cells was significantly lower in smokers compared with nonsmokers (3.3% vs. 9.5%, P <0.05). The IL-10 concentration in BAL fluid was below the detection limit. , Conclusions:  We did not observe any association between tobacco smoking and macrophage polarization in patients with sarcoidosis. However, our study revealed 2 populations of CD40- and CD163-positive cells, which may indicate that macrophages are involved in granuloma formation and provide direction for future research.