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AbstractThe article presents the conceptual design of the solid fuel from the forest and sawmill production waste and agricultural waste biomass-fired water boiler. The project was supported by thermal and flow calculations. The designed WRB-6 boiler is a 3-pass unit with a fully screened furnace chamber and is equipment of a local biomass heating plant. It is fired with biomass, transported over no more than 30 km. The furnace has a double function – dual fuel. Both fuels can be burned simultaneously or alternately, depending on seasonal availability. In the front part there is a channel for feeding and burning large bales of cereal straw. The lower closure of the chamber is a mechanically inclined grate for the combustion of wood biomass in the granulation of chips. As part of the work, calculations of the amount of biomass fuels possible to be obtained on the local market with economy of delivery were also performed. As a result of the calculations, the temperatures of water and flue gas at the outlet, boiler efficiency, slagging and impurity indices for biomass were also presented. [ABSTRACT FROM AUTHOR]

Human β-defensin 3, HBD-3, is a 45-residue antimicrobial and immunomodulatory peptide that plays multiple roles in the host defense system. In addition to interacting with cell membranes, HBD-3 is also a ligand for melanocortin receptors, cytokine receptors and voltage-gated potassium channels. Structural and functional studies of HBD-3 have been hampered by inefficient synthetic and recombinant expression methods. Herein, we report an optimized Fmoc solid-phase synthesis of this peptide using an orthogonal disulfide bonds formation strategy. Our results suggest that utilization of an optimized resin, coupling reagents and pseudoproline dipeptide building blocks decrease chain aggregation and largely improve the amount of the target peptide in the final crude material, making the synthesis more efficient. We also present an alternative synthesis of HBD-3 in which a replacement of a native disulfide bridge with a diselenide bond improved the oxidative folding. Our work enables further biological and pharmacological characterization of HBD-3, hence advancing our understanding of its therapeutic potential. [ABSTRACT FROM AUTHOR]

Abstract: The μ-conotoxin KIIIA is a three disulfide-bridged blocker of voltage-gated sodium channels (VGSCs). The Lys7 residue in KIIIA is an attractive target for manipulating the selectivity and efficacy of this peptide. Here, we report the design and chemical synthesis of μ-conopeptoid analogues (peptomers) in which we replaced Lys7 with peptoid monomers of increasing side-chain size: N-methylglycine, N-butylglycine and N-octylglycine. In the first series of analogues, the peptide core contained all three disulfide bridges; whereas in the second series, a disulfide-depleted selenoconopeptide core was used to simplify oxidative folding. The analogues were tested for functional activity in blocking the Nav1.2 subtype of mammalian VGSCs exogenously expressed in Xenopus oocytes. All six analogues were active, with the N-methylglycine analogue, [Sar7]KIIIA, the most potent in blocking the channels while favouring lower efficacy. Our findings demonstrate that the use of N-substituted Gly residues in conotoxins show promise as a tool to optimize their pharmacological properties as potential analgesic drug leads. [Copyright &y& Elsevier]

Three-disulfide-bridged Ecballium elaterium trypsin inhibitor II (EETI-II) is a 28-residue peptide that belongs to the squash family of canonical trypsin inhibitors. Herein, we report synthesis and biological activity of three EETI-II analogs. In each of analog, a pair of cysteine residues forming a native disulfide bridge was individually replaced by a pair of selenocysteine residues. All selenopeptide analogs were chemically synthesized using the Fmoc protocol and subsequently folded in the presence of oxidized and reduced glutathione. The analogs containing a diselenide bridge displayed association constants with trypsin that ranged from 2.6 × 10 to 5.1 × 10 [M]. Our results suggest that the selenopeptide analogs retained low nanomolar inhibitory potencies, and only the diselenide bridge adjacent to the inhibitory binding loop weakened the interactions with trypsin by approximately fivefold. We discuss these findings in the context of a broader use of selenopeptide analogs as proxies to study cysteine-rich peptides. [ABSTRACT FROM AUTHOR]

Abstract: Disulfide-rich peptides represent a megadiverse group of natural products with very promising therapeutic potential. To accelerate their functional characterization, high-throughput chemical synthesis and folding methods are required, including efficient mapping of multiple disulfide bridges. Here, we describe a novel approach for such mapping and apply it to a three-disulfide-bridged conotoxin, μ-SxlllA (from the venom of Conus striolatus), whose discovery is also reported here for the first time. μ-SxlllA was chemically synthesized with three cysteine residues labeled 100% with [sup15]N/[sup13]C, while the remaining three cysteine residues were incorporated using a mixture of 70%/30% unlabeled/labeled Fmoc-protected residues. After oxidative folding, the major product was analyzed by NMR spectroscopy. Sequence-specific resonance assignments for the isotope-enriched Cys residues were determined with 2D versions of standard tripleresonance ([sup1]H, [sup13]C, [sup15]N) NMR experiments and 2D [[sup13]C, [sup1]H] HSQC. Disulfide patterns were directly determined with cross-disulfide NOEs confirming that the oxidation product had the disulfide connectivities characteristic of μ-conotoxins. μ-SxlllA was found to be a potent blocker of the sodium channel subtype Na[subv]1 .4 (IC[sub50] = 7 nM). These results suggest that differential incorporation of isotope-labeled cysteine residues is an efficient strategy to map disulfides and should facilitate the discovery and structure-function studies of many bioactive peptides. [ABSTRACT FROM AUTHOR]

Hydroxylation of proline residue occurs in specific peptides and proteins derived from plants and animals, but the functional role of this modification has been characterized primarily in collagen. Marine cone snails produce disulfide-rich peptides that have undergone a plethora of posttranslational modifications, including proline hydroxylation. Although Conus snails extensively utilize proline hydroxylation, the consequences of this modification remain largely unexplored. In this work, we investigated the function of 4-hydroxyproline (Hyp) in conotoxins from three distinct gene families: μ-, ω-, and α-conotoxins. Analogues of μ-GIIIA, ω-MVIIC, α-GI, and α-ImI were synthesized with either Pro or Hyp, and their in vitro oxidative folding and biological activity were characterized. For GIIIA, which naturally contains three Hyp residues, the modifications improved the ability to block Nav 1.4 sodium channels but did not affect folding. In contrast, the presence of Hyp in MVIIC had a significant impact on the oxidative folding but not on the biological activity. The folding yields for the MVIIC[Pro7Hyp] analogue were approximately 2-fold higher than for MVIIC under a variety of optimized oxidation conditions. For α-conotoxins ImI and GI, the hydroxylation of the conserved Pro residue improved their folding but impaired their activities against target receptors. Since prolyl-4-hydroxylase and protein disulfide isomerase coexist as a heterotetramer in the ER, we discuss the effects of Hyp on the folding of conotoxins in the context of cis-trans isomerization of Pro and Hyp. Taken together, our data suggest that proline hydroxylation is important for both in vitro oxidative folding and the bioactivity of conotoxins. [ABSTRACT FROM AUTHOR]

This article presents the concept of renewable methanol production in the gas turbine cycle. As part of the work, an analysis was performed, including the impact of changing the parameters in the methanol reactor on the obtained values of power, yield and efficiency of the reactor, and chemical conversion. The aim of this research was to investigate the possibility of integrating the system for the production of renewable methanol and additional production of electricity in the system. The efficiency of the chemical conversion process and the efficiency of the methanol reactor increases with increasing pressure and decreasing temperature. The highest efficiency values, respectively η = 0.4388 and η R = 0.3649, are obtained for parameters in the reactor equal to 160 °C and 14 MPa. The amount of heat exchanged in all exchangers reached the highest value for 14 MPa and 160 °C and amounted to Q ˙   = 2.28 kW. Additionally, it has been calculated that if an additional exchanger is used before the expander (heating the medium to 560 °C), the expander's power will cover the compressor's electricity demand. [ABSTRACT FROM AUTHOR]

• In this work, we study a ferroelectric liquid crystal (FLC) cell as a phase-only modulator. The novelty of the approach is the use of a FLC cell with a large switching rotation angle close to 90° instead of the typical value used in display applications, which is less than 45 ∘. • We show that when the LC layer is a half-wave retarder, this device provides a binary π phase modulation for any input state of polarization and with maximum light conversion efficiency, thus being useful even for unpolarized light. This situation is not possible with the common nematic LC modulators or with standard FLC modulators of smaller rotation angles. • The main physico-chemical properties of the liquid crystalline mixture used to fabricate the FLC modulator are studied. In addition, we perform a reverse engineering procedure aimed at verifying the modulator's physical parameters and the binary π -phase modulation. In this work a ferroelectric liquid crystal (FLC) modulator with a non-standard large switching rotation angle, close to 90 ∘ , is fabricated and characterized. The modulator acts as a switchable wave-plate with an in-plane rotation of the principal axis under the action of a bipolar voltage. In the ideal situation of half-wave retardance, the device is shown to behave as a binary π phase modulator independently of the input state of polarization. We provide physico-chemical properties of the liquid crystalline mixture used to fabricate the FLC modulator with such large switching angle. The characterization method of the device optical properties is presented, which allows the localization of the two LC stable states, the unambiguous determination of the rotation angle, and the evaluation of the spectral retardance function. We demonstrate the polarization-independent π phase shift using an adapted Young's type interferometer for real-time measurements, where we further analyze the operational frequency limits of the device. This FLC operational mode can be exploited to produce binary-phase polarization-independent diffractive optical elements. [ABSTRACT FROM AUTHOR]

A cone snail venom peptide, μO§-conotoxin GVIIJ from Conus geo-graphus, has a unique posttranslational modification, S-cysteinylated cysteine, which makes possible formation of a covalent tether of peptide to its target Na channels at a distinct ligand-binding site. μO§-conotoxin GVIIJ is a 35-aa peptide, with 7 cysteine residues; six of the cysteines form 3 disulfide cross-links, and one (Cys24) is S-cysteinylated. Due to limited availability of native GVIIJ, we primarily used a synthetic analog whose Cys24 was S-glutathionylated (abbreviated GVIIJSSG). The peptide-channel complex is stabilized by a disulfide tether between Cys24 of the peptide and Cys910 of rat (r) Nav1.2. A mutant channel of rNav1.2 lacking a cysteine near the pore loop of domain II (C910L), was >103-fold less sensitive to GVIIJSSG than was wild-type rNav1.2. In contrast, although rNav1.5 was >104-fold less sensitive to GVIIJSSG than Nav1.2, an rNav1.5 mutant with a cysteine in the homologous location, rNav1.5[L869C], was >103-fold more sensitive than wild-type rNav1-5. The susceptibility of rNav1-2 to GVIIJSSG was significantly altered by treating the channels with thiol-oxidizing or disulfide-reducing agents. Furthermore, coexpression of rNavβ2 or rNavβ4, but not that of rNavβ1 or rNavβ3, protected rNav1.1 to -1.7 (excluding Nav1.5) against block by GVIIJSSG- Thus, GVIIJ-related peptides may serve as probes for both the redox state of extracellular cysteines and for assessing which Navβ- and Navα-subunits are present in native neurons. [ABSTRACT FROM AUTHOR]

Abstract: Five serine proteinase inhibitors (Mirabilis jalapa trypsin inhibitors, MJTI I and II and Spinacia oleracea trypsin inhibitors, SOTI I, II, and III) from the garden four-o’clock (M. jalapa) and spinach (S. oleracea) seeds were isolated. The purification procedures included affinity chromatography on immobilized methylchymotrypsin in the presence of 5M NaCl, ion exchange chromatography and/or preparative electrophoresis, and finally RP–HPLC on a C-18 column. The inhibitors, crosslinked by three disulfide bridges, are built of 35 to 37 amino-acid residues. Their primary structures differ from those of known trypsin inhibitors, but showed significant similarity to the antimicrobial peptides isolated from the seeds of M. jalapa (MJ-AMP1, MJ-AMP2), Mesembryanthemum crystallinum (AMP1), and Phytolacca americana (AMP-2 and PAFP-S) and from the hemolymph of Acrocinus longimanus (Alo-1, 2 and 3). The association equilibrium constants (K a) with bovine β-trypsin for the inhibitors from M. jalapa (MJTI I and II) and S. oleracea (SOTI I–III) were found to be about 107 M−1. Fully active MJTI I and SOTI I were obtained by solid-phase peptide synthesis. The disulfide bridge pattern in both inhibitors (Cys1–Cys4, Cys2–Cys5 and Cys3–Cys6) was established after their digestion with thermolysin and proteinase K followed by the MALDI-TOF analysis. [Copyright &y& Elsevier]

The alarming raise of multi-drug resistance among human microbial pathogens makes the development of novel therapeutics a priority task. In contrast to conventional antibiotics, antimicrobial peptides (AMPs), besides evoking a broad spectrum of activity against microorganisms, could offer additional benefits, such as the ability to neutralize toxins, modulate inflammatory response, eradicate bacterial and fungal biofilms or prevent their development. The latter properties are of special interest, as most antibiotics available on the market have limited ability to diffuse through rigid structures of biofilms. Lipidation of AMPs is considered as an effective approach for enhancement of their antimicrobial potential and in vivo stability; however, it could also have undesired impact on selectivity, solubility or the aggregation state of the modified peptides. In the present work, we describe the results of structural modifications of compounds designed based on cationic antimicrobial peptides DK5 and CAR-PEG-DK5, derivatized at their N-terminal part with fatty acids with different lengths of carbon chain. The proposed modifications substantially improved antimicrobial properties of the final compounds and their effectiveness in inhibition of biofilm development as well as eradication of pre-formed 24 h old biofilms of Candida albicans and Staphylococcus aureus. The most active compounds (C5-DK5, C12-DK5 and C12-CAR-PEG-DK5) were also potent against multi-drug resistant Staphylococcus aureus USA300 strain and clinical isolates of Pseudomonas aeruginosa. Both experimental and in silico methods revealed strong correlation between the length of fatty acid attached to the peptides and their final membranolytic properties, tendency to self-assemble and cytotoxicity. [ABSTRACT FROM AUTHOR]