Role of RNA Motifs in RNA Interaction with Membrane Lipid Rafts: Implications for Therapeutic Applications of Exosomal RNAs

RNA motifs may promote interactions with exosomes (EXO-motifs) and lipid rafts (RAFT-motifs) that are enriched in exosomal membranes. These interactions can promote selective RNA loading into exosomes. We quantified the affinity between RNA aptamers containing various EXO- and RAFT-motifs and membrane lipid rafts in a liposome model of exosomes by determining the dissociation constants. Analysis of the secondary structure of RNA molecules provided data about the possible location of EXO- and RAFT-motifs within the RNA structure. The affinity of RNAs containing RAFT-motifs (UUGU, UCCC, CUCC, CCCU) and some EXO-motifs (CCCU, UCCU) to rafted liposomes is higher in comparison to aptamers without these motifs, suggesting direct RNA-exosome interaction. We have confirmed these results through the determination of the dissociation constant values of exosome-RNA aptamer complexes. RNAs containing EXO-motifs GGAG or UGAG have substantially lower affinity to lipid rafts, suggesting indirect RNA-exosome interaction via RNA binding proteins. Bioinformatics analysis revealed RNA aptamers containing both raft- and miRNA-binding motifs and involvement of raft-binding motifs UCCCU and CUCCC. A strategy is proposed for using functional RNA aptamers (fRNAa) containing both RAFT-motif and a therapeutic motif (e.g., miRNA inhibitor) to selectively introduce RNAs into exosomes for fRNAa delivery to target cells for personalized therapy.     

Physicochemical characteristics,proximate analysis and mineral composition of ostrich meat as influenced by muscle

The influence of muscle on the physicochemical characteristics, proximate analysis, and mineral composition of meat from 10 ostriches (10–12 months old), slaughtered according to commercial abattoir procedures, were evaluated. Muscle had no influence (p > 0.05) on L*-values (32.5), a*-values (11.9), water-holding capacity (11.9%), final pH (pH₂₄) values (6.07), and ash contents (1.12 g/100 g edible meat). However, intramuscular lipid contents varied (p < 0.05) from 0.88 (M. fibularis longus) to 1.44 (M. flexor cruris lateralis) g/100 g edible meat, at a mean value of 1.16 g/100 g edible meat for 10 different muscles. Sodium (34.7 mg/100 g edible meat) and iron (3.14 mg/100 g edible meat) contents, both influenced (p < 0.05) by muscle, possessed substantially lower and higher values, respectively, than values reported for beef and chicken.     

Cyclic chronopotentiometry as a detection tool for flowing solution systems

Cyclic chronopotentiometry provides a very simple detection method, which may be particularly useful in capillary electrophoresis (CE) and microseparation systems. It has been shown that for disk microelectrodes it is possible to define safe reduction and oxidation currents that would never lead to the formation of H2 or O2 gas bubbles, even if they are applied for an indefinitely long time period. During end-column CE detection, currents passing through the working microelectrode can be completely controlled by the external electronic circuit and they are not affected by the separation current. Consequently, problems created by the offset potential in CE can be completely eliminated. The detection can be accomplished through a variety of different mechanisms; however, generation of the electrode response as a result of analyte adsorption seems to be most common. The method is applicable to many analytes, which do not have to be electroactive. The analytical signal is obtained by monitoring the change in the average electrode potential (calculated for either a cathodic or an anodic half-cycle) caused by an analyte interacting with the electrode. The analytical signal is proportional to the analyte concentration, within a concentration range extending over approximately 2 orders of magnitude.     

Two-period vs. multi-period model for supply chain disruption management

A novel two-period modelling approach is developed for supply chain disruption mitigation and recovery and compared with a multi-period approach. For the two-period model, planning horizon is divided into two aggregate periods: before disruption and after disruption. The corresponding mitigation and recovery decisions are: (1) primary supply and demand portfolios and production before a disruption, and (2) recovery supply, transshipment and demand portfolios and production after the disruption. In the multi-period model, a multi-period planning horizon is applied to account for a detailed timing of supplies and production. The primary and recovery portfolios are determined simultaneously and for both approaches the integrated decision-making, stochastic mixed integer programming models are developed. While the simplified two-period setting may overestimate (for best-case capacity constraints) or underestimate (for worst-case capacity constraints) the available production capacity, it can be easily applied in practice for a fast, rough-cut evaluation of disruption mitigation and recovery policy. The findings indicate that for both two- and multi-period setting, the developed multi-portfolio approach leads to computationally efficient mixed integer programming models with an embedded network flow structure resulting in a very strong linear programming relaxation.     

Balancing and scheduling of surface mount technology lines

A mixed-integer programming approach to simultaneous or sequential balancing and scheduling of surface mount technology (SMT) lines for printed wiring board (PWB) assembly is presented. The SMT line consists of several processing stages in series separated by finite intermediate buffers, where each stage has one or more identical parallel machines. In the line, different types of PWBs are assembled using various types of electronic components. The components are assigned to feeder slots of a feeder carrier at each placement station. Different types of components occupy a different number of feeder slots. The total number of slots available at each station was limited. The problem objective was to determine an assignment of components to feeder slots at each placement station and to determine an assembly schedule for a mix of board types to complete the boards in minimum time. Numerical examples and some computational results are presented to illustrate applications of the proposed approach.     

Optimal versus heuristic scheduling of surface mount technology lines

This paper presents and compares an exact and a heuristic approach for scheduling of printed wiring board assembly in surface mount technology (SMT) lines. A typical SMT line consists of several assembly stations in series and/or in parallel, separated by finite intermediate buffers. The objective of the scheduling problem is to determine the detailed sequencing and timing of all assembly tasks for each individual board, so as to maximize the line's productivity, which is defined in terms of makespan for a mix of board types. The limited intermediate buffers between stations result in a scheduling problem with machine blocking, where a completed board may remain on a machine and block it until a downstream machine becomes available. In addition, limited machine availability due to scheduled downtimes is considered. The exact approach is based on a mixed integer programming formulation that can be used for optimization of assembly schedules by using commercially available software for integer programming, whereas the heuristic approach is designed as a combination of tabu search and a set of dispatching rules. Numerical examples modelled after real-world SMT lines and some computational results are provided to illustrate and compare the two approaches.     

Influence of counter material on friction and wear performance of PTFE–metal binary coatings

The study presented in this paper concerns the influence of the counter materials on wear and friction performance of polytetrafluoroethylene (PTFE) reservoirs arranged in distinct patterns on coated surfaces. Al-bronze and Mo coatings were deposited on a mild steel substrate using an atmospheric plasma spray process. Three patterns of PTFE reservoirs were used. Pins, which served as counter surface, were made of three different materials. Wear tests were carried out in a pin-on-disc test rig at room temperature and under dry contact conditions. The tests were carried out at a constant pressure of 10 MPa. An average linear speed of the test disc was 0.036 m/s. The importance of appropriately matched hardness of two surfaces in sliding contact is emphasised. Due to inadequate hardness of the counter material, performance of Mo coating was adversely affected and expected beneficial action of PTFE reservoirs severely hindered. Al-bronze coating proved to be performing far better than Mo coating. Serious deterioration of Mo coating occurred faster than that for Al-bronze coating.     

Identification of tillering node proteins differentially accumulated in barley recombinant inbred lines with different juvenile growth habits

Barley (Hordeum vulgare L.) is an important cereal crop grown for both the feed and malting industries. The allelic dwarfing gene sdw1/denso has been used throughout the world to develop commercial barley varieties. Proteomic analysis offers a new approach to identify a broad spectrum of genes that are expressed in the living system. Two-dimensional electrophoresis and mass spectrometry were applied to investigate changes in protein abundance associated with different juvenile growth habit as effect of the denso locus in barley homozygous lines derived from a Maresi × Pomo cross combination. A total of 31 protein spots were revealed that demonstrate quantitative differences in protein abundance between the analyzed plants with different juvenile growth habit, and these protein spots were selected to be identified by mass spectrometry. Identification was successful for 27 spots, and functional annotations of proteins revealed that most of them are involved in metabolism and disease/defense-related processes. Functions of the identified proteins and their probable influence on the growth habit in barley are discussed.     

The relationship between grain hardness, dough mixing parameters and bread-making quality in winter wheat

The influence of grain hardness, determined by using molecular markers and physical methods (near-infrared (NIR) technique and particle size index-PSI) on dough characteristics, which in turn were determined with the use of a farinograph and reomixer, as well as bread-making properties were studied. The material covered 24 winter wheat genotypes differing in grain hardness. The field experiment was conducted at standard and increased levels of nitrogen fertilization. Results of molecular analyses were in agreement with those obtained by the use of physical methods for soft-grained lines. Some lines classified as hard (by physical methods) appeared to have the wild-type Pina and Pinb alleles, similar to soft lines. Differences in dough and bread-making properties between lines classified as hard and soft on the basis of molecular data appeared to be of less significance than the differences between lines classified as hard and soft on the basis of physical analyses of grain texture. Values of relative grain hardness at the increased nitrogen fertilization level were significantly higher. At both fertilization levels the NIR parameter determining grain hardness was significantly positively correlated with the wet gluten and sedimentation values, with most of the rheological parameters and bread yield. Values of this parameter correlated with quality characteristics in a higher degree than values of particle size index.     

Effects of nitrogen and phosphorus fertilisers on yield and on inorganic and organic composition of italian ryegrass grown on phosphorus-deficient soil

Italian ryegrass (Lolium multiflorum) S22, grown in a greenhouse in phosphorus-deficient soil in pots was given 40 and 160 mg N/kg of soil (as ammonium nitrate) and 5, 25 and 100 mg P/kg of soil (as calcium tetrahydrogen diorthophosphate). Giving more nitrogen increased yields only when adequate phosphorus was applied. A significant response to added phosphorus was obtained when ryegrass dry matter contained about 0.15% P or less. Nitrogen increased the amount of sugars but decreased that of fructosan. Phosphorus increased the production of both sugars and fructosan. Protein N (expressed as a percentage of total N) increased with increasing amounts of phosphorus given. The largest values for protein N were obtained when grass contained about 0.3 % P in the dry matter. In phosphorus-deficient grass nitrate-N accumulated and the predominant compounds within the free amino acid fraction were asparagine and glutamine. Adding phosphorus decreased the concentrations of both nitrate-N and the amides. The amino acid composition of the grass protein was not affected by the nitrogen and phosphorus treatments.