36 research outputs found
Acid-base and metal ion binding properties of 2-thiocytidine in aqueous solution
The thionucleoside 2-thiocytidine (C2S) occurs in nature in transfer RNAs; it receives attention in diverse fields like drug research and nanotechnology. By potentiometric pH titrations we measured the acidity constants of H(C2S)+ and the stability constants of the M(C2S)2+ and M(C2S−H)+ complexes (M2+=Zn2+, Cd2+), and we compared these results with those obtained previously for its parent nucleoside, cytidine (Cyd). Replacement of the (C2)=O unit by (C2)=S facilitates the release of the proton from (N3)H+ in H(C2S)+ (pK a = 3.44) somewhat, compared with H(Cyd)+ (pK a = 4.24). This moderate effect of about 0.8 pK units contrasts with the strong acidification of about 4 pK units of the (C4)NH2 group in C2S (pK a = 12.65) compared with Cyd (pK a≈16.7); the reason for this result is that the amino-thione tautomer, which dominates for the neutral C2S molecule, is transformed upon deprotonation into the imino-thioate form with the negative charge largely located on the sulfur. In the M(C2S)2+ complexes the (C2)S group is the primary binding site rather than N3 as is the case in the M(Cyd)2+ complexes, though owing to chelate formation N3 is to some extent still involved in metal ion binding. Similarly, in the Zn(C2S−H)+ and Cd(C2S−H)+ complexes the main metal ion binding site is the (C2)S− unit (formation degree above 99.99% compared with that of N3). However, again a large degree of chelate formation with N3 must be surmised for the M(C2S−H)+ species in accord with previous solid-state studies of related ligands. Upon metal ion binding, the deprotonation of the (C4)NH2 group (pK a = 12.65) is dramatically acidified (pK a≈3), confirming the very high stability of the M(C2S−H)+ complexes. To conclude, the hydrogen-bonding and metal ion complex forming capabilities of C2S differ strongly from those of its parent Cyd; this must have consequences for the properties of those RNAs which contain this thionucleosid
Whey protein isolate as a substrate to design Calendula officinalis flower extract controlled-release materials for potential skin application
Proteomic profile alterations in porcine conceptuses during early stages of development
The dynamic embryo development during the early stages of gestation requires precise molecular changes, including proteomic ones. We aimed to find unique proteins for porcine conceptuses specifically during the peri-implantation period, i.e. on days 15–16 of pregnancy. The proteomic profile of these conceptuses was compared with conceptuses at an earlier stage of the development, i.e. collected during maternal recognition of pregnancy on days 12–13 of pregnancy. The 2DE, gel image analysis, and MALDI TOF mass spectrometry were used 500 protein spots were annotated as common to conceptuses harvested during both studied periods. Proteomic profile of the conceptuses collected during the peri-implantation period contains 24 unique proteins. Identified unique for the peri-implantation period proteins are involved in adhesion processes, cadherin, and actin-binding, and actin filament organization, extracellular matrix organization, and cytoskeleton organization. Systemic analysis of identified proteins confirmed their involvement in cell adhesion and cytoskeletal organization as being two major affected functions. The unique proteins might be recognized as factors conditioning the proper peri-implantation embryo development and gaining competences for implantation. In further studies, BRCA1 might be considered as a candidate for a potential marker of embryonic competences for implantation in pigs.The work was funded by a grant from the National Science Centre, Poland (DEC-2012/05/N/NZ4/02343). We acknowledge support to S.S. from Qatar National Research Fund (QNRF) (NPRP9-453-3-089)
Combined systems approaches reveal highly plastic responses to antimicrobial peptide challenge in Escherichia coli
Obtaining an in-depth understanding of the arms races between peptides comprising the innate immune response and bacterial pathogens is of fundamental interest and will inform the development of new antibacterial therapeutics. We investigated whether a whole organism view of antimicrobial peptide (AMP) challenge on Escherichia coli would provide a suitably sophisticated bacterial perspective on AMP mechanism of action. Selecting structurally and physically related AMPs but with expected differences in bactericidal strategy, we monitored changes in bacterial metabolomes, morphological features and gene expression following AMP challenge at sub-lethal concentrations. For each technique, the vast majority of changes were specific to each AMP, with such a plastic response indicating E. coli is highly capable of discriminating between specific antibiotic challenges. Analysis of the ontological profiles generated from the transcriptomic analyses suggests this approach can accurately predict the antibacterial mode of action, providing a fresh, novel perspective for previous functional and biophysical studies
Collagen/Gelatin/Hydroxyethyl Cellulose Composites Containing Microspheres Based on Collagen and Gelatin: Design and Evaluation
The objective of this study was to develop three-dimensional collagen/gelatin/hydroxyethyl cellulose composites in combination with gelatin or collagen-gelatin loaded microspheres. Microspheres were prepared by an emulsification/crosslinking method. A 1-Ethyl-3-(3-dimethyl-aminopropyl)-carbodiimide (EDC) and N-hydroxysuccinimide (NHS) mixture were used as a crosslinking agent for the obtained materials. The structure of the materials was studied using scanning electron microscopy (SEM) and infrared spectroscopy. Moreover, a Calendula officinalis (pot marigold) flower extract release profile of the microsphere-loaded matrices was assessed in vitro. Additionally, porosity, density, stability, swelling and mechanical properties were tested. On the basis of SEM images, the microspheres exhibited a spherical shape and were irregularly dispersed in the polymer matrix. However, it was found that the addition of microparticles to obtained materials did not significantly change their microstructure. We observed a slight decrease in the swelling properties of matrices and an increase in values of Young’s modulus. Significantly, the addition of microspheres to the polymer matrices led to improved loading capacity of materials and release performance of Calendula officinalis flower extract. This makes the collagen/gelatin/hydroxyethyl cellulose composites containing microspheres a promising and suitable vehicle for biomedical, dermatological, or cosmetic applications
What Influences the Servitization Process the Most? A Perspective of Polish Machinery Manufacturers
The servitization of manufacturing is an increasingly popular way to win market competition and maintain this advantage as well as to ensure more sustainable development of the manufacturing sector. To make this strategy effective, many aspects should be considered. This study mainly aims to identify the most significant factors influencing the process of servitization in the manufacturing sector. An additional research problem that emerged in the research process was aimed at determining the factors perceived by entrepreneurs as more important compared to others, and, correspondingly, the reasons behind such differences of opinion. This article is supported by the results of a literature review, the outcomes of a questionnaire survey (conducted with 150 Polish machinery manufacturers), and the findings of in-depth interviews with experts. In the light of the results of the quantitative research, the most important aspects that determine the servitization process are customer requirements and preferences; economic and financial aspects; added value for the customer and the company; and also the relations, stakeholders and partners of the service processes. More specific determinants, such as long-lasting and stronger relationships, organizational strategy, legal regulations regarding the provision of long-term services, and the economic benefits of service recipients, were indicated by experts as crucial in product–service integration. Also, employees’ competencies were emphasized as vital for extending the service activity of manufacturing companies. The contribution of this study is twofold. First, it identifies the most important aspects for the analysis of a manufacturer starting out on the servitization path. Second, the study reveals differences in the perception of the importance of certain factors that appear to be related to the experience related to the provision of services by machine manufacturers. Hence, some practical guidelines may be drawn for managers of manufacturing companies who plan to extend their service offer
Lyophilized Emulsions in the Form of 3D Porous Matrices as a Novel Material for Topical Application
Researchers are constantly searching for innovations that can be applied to the cosmetic industry. Production of porous materials stored in a lyophilized form and swollen directly before use may be beneficial considering their facilitated packaging, transport and storage. In this study, we propose porous materials based on sodium alginate, gelatin, glycerol and lipids (cottonseed oil and beeswax) obtained by freeze-drying and cross-linking. Material composition with the most promising properties was modified by the addition of PLA microparticles with Calendula officinalis flower extract. The structure and properties of obtained porous materials were analyzed. ATR-FTIR, mechanical properties, residual moisture content, porosity and density were assessed, as well as swelling properties and degradation after their cross-linking. The loading capacity and in vitro release of Calendula officinalis flower extract were performed for samples with incorporated PLA microparticles containing plant extract. The modification of the composition and fabrication method of materials significantly influenced their physicochemical properties. The selected plant extract was successfully incorporated into polymeric microparticles that were subsequently added into developed materials. Prepared materials may be considered during designing new cosmetic formulations
Genes Involved in DNA Repair and Mitophagy Protect Embryoid Bodies from the Toxic Effect of Methylmercury Chloride under Physioxia Conditions
The formation of embryoid bodies (EBs) from human pluripotent stem cells resembles the early stages of human embryo development, mimicking the organization of three germ layers. In our study, EBs were tested for their vulnerability to chronic exposure to low doses of MeHgCl (1 nM) under atmospheric (21%O2) and physioxia (5%O2) conditions. Significant differences were observed in the relative expression of genes associated with DNA repair and mitophagy between the tested oxygen conditions in nontreated EBs. When compared to physioxia conditions, the significant differences recorded in EBs cultured at 21% O2 included: (1) lower expression of genes associated with DNA repair (ATM, OGG1, PARP1, POLG1) and mitophagy (PARK2); (2) higher level of mtDNA copy number; and (3) higher expression of the neuroectodermal gene (NES). Chronic exposure to a low dose of MeHgCl (1 nM) disrupted the development of EBs under both oxygen conditions. However, only EBs exposed to MeHgCl at 21% O2 revealed downregulation of mtDNA copy number, increased oxidative DNA damage and DNA fragmentation, as well as disturbances in SOX17 (endoderm) and TBXT (mesoderm) genes expression. Our data revealed that physioxia conditions protected EBs genome integrity and their further differentiation