Effect of gamma ray dose on granular and molecular structures of gamma ray-irradiated cassava starch and its application in bioplastics.

In this study, the effect of gamma ray irradiation on the granular and molecular structures of cassava starch was examined. Cassava starch was irradiated with various gamma ray doses of 25, 50, 75, and 100 kGy. After irradiation, the starch turned yellow, but its granular morphological characteristics remained intact. However, the inner part and the 'Maltese cross' of the starch granules irradiated with 100 kGy were broken, and its crystallinity decreased considerably. The pH reduction (from 5.6 to 3.7) and carboxyl content increase (up to 0.38 %) confirmed the formation of carboxyl groups on the irradiated starch chains. Gamma ray irradiation caused glycosidic bond cleavages, resulting in shortened amylose chains and debranched amylopectin chains containing terminal carboxyl groups. The irradiated starches with different molecular weights have high potential for use in food and non-food applications, for example, in bioplastics. Thermoplastic-irradiated starch (TPIS) materials, and their blends with poly(lactic acid) (PLA) were prepared via extrusion. Both TPIS and PLA/TPIS blends exhibited considerably increased melt flow index values compared with those from the unirradiated starch at approximate increases of 420-2260% and 2-55%, respectively. The improved melt flow ability and reduced viscosity are advantages for some plastic conversion processes such as injection molding.

Prospects of crude enzymes in replacing pure enzymes for dissolving pulp production.

High-purity cellulose from paper pulp can be obtained after appropriate treatments involving pure xylanases and cellulases/endoglucanases. This study investigated the efficacy of using crude xylanase and cellulase instead of commercial ones to improve process economics. Kraft paper grade pulp produced from veneer waste, hardwood, and non-wood sources was utilized as a more sustainable option. Crude xylanase and cellulase from isolated soil bacteria Bacillus pumilus 3GAH and Bacillus subtilis PJK6 were used for process optimization. The correlation between Fock reactivity, chain scission, and crystallinity after crude-cellulase treatment was established through chemical, FTIR, and XRD analyses. Pentosans in kraft pulp were reduced from an initial 18.7% to 4.9% through sequential treatments with crude xylanase and alkali. Subsequent crude-cellulase treatment, even at 8 U/g o.d. pulp, improved Fock reactivity from 28.2% to 61.2%, fulfilling a major criterion for viscose. Thus, crude enzymes can be effectively used for the efficient and economical upgrading of paper pulp to dissolving pulp.

Optical annealing of peroxo-ferric intermediates in CYP17A1 and product formation.

Human cytochrome P450 CYP17A1 catalyzes the hydroxylation of pregnenolone and progesterone at the C17 position, with subsequent C17-C20 bond scission, to form dehydroepiandrosterone and androstenedione respectively. The first hydroxylation reaction is faster in H2O than in D2O, while the second carbon­carbon bond scission event demonstrates an inverse solvent isotope effect, which is more pronounced for 17-hydroxy pregnenolone. In order to better understand the cause of this difference, we compared the optical absorption spectra of oxygenated CYP17A1 with the four substrates (pregnenolone, progesterone, 17-hydroxy pregnenolone and 17-hydroxy progesterone) in both H2O and D2O. We also studied the temperature-dependent decay of the peroxo-ferric and hydroperoxo-ferric intermediates generated by cryoradiolysis of the corresponding oxygenated heme proteins at 77 K. For both pregnenolone and 17-hydroxypregnenolone, annealing of the peroxo-intermediates was observed at lower temperatures in H2O than in D2O. In contrast, no solvent isotope effect was detected when progesterone or 17-hydroxyprogesterone were used as substrates. These differences are attributed to their different positioning in the P450 active site with respect to the heme bound peroxo (Fe-OO-) moiety, which is in agreement with earlier structural and spectroscopic investigations. Analysis of the samples run in both H2O and in D2O, where 17-hydroxyprogesterone is the substrate, demonstrated significant (∼25%) yield of androstenedione product relative to the oxygenated starting material.

Natural iron-containing minerals catalyze the degradation of polypropylene microplastics: a route to self-remediation learnt from the environment.

Virgin and environmentally aged polypropylene (PP) micropowders (V-PP and E-PP, respectively) were used as reference microplastics (MPs) in comparative photo- and thermo-oxidative ageing experiments performed on their mixtures with a natural ferrous sand (NS) and with a metal-free silica sand (QS). The ferrous NS was found to catalyze the photo-oxidative degradation of V-PP after both UV and simulated solar light irradiation. The catalytic activity in the V-PP/NS mixture was highlighted by the comparatively higher fraction of photo-oxidized PP extracted in dichloromethane, and the higher carbonyl index of the bulk polymer extracted with boiling xylene, when compared with the V-PP/QS mixture. Similarly, NS showed a catalytic effect on the thermal degradation (at T = 60 °C) of E-PP. The results obtained indicate that, under suitable environmental conditions (in this case, an iron-containing sediment or soil matrix, combined with simulated solar irradiation), the degradation of some types of MPs could be much faster than anticipated. Given the widespread presence of iron minerals (including the magnetite and iron-rich serpentine found in NS) in both coastal and mainland soils and sediments, a higher than expected resilience of the environment to the contamination by this class of pollutants is anticipated, and possible routes to remediation of polluted natural environments by eco-compatible iron-based minerals are envisaged.

Comparing Multifunctional Viral and Eukaryotic Proteins for Generating Scission Necks in Membranes.

Deterministic formation of membrane scission necks by protein machinery with multiplexed functions is critical in biology. A microbial example is M2 viroporin, a proton pump from the influenza A virus that is multiplexed with membrane remodeling activity to induce budding and scission in the host membrane during viral maturation. In comparison, the dynamin family constitutes a class of eukaryotic proteins implicated in mitochondrial fission, as well as various budding and endocytosis pathways. In the case of Dnm1, the mitochondrial fission protein in yeast, the membrane remodeling activity is multiplexed with mechanoenzyme activity to create fission necks. It is not clear why these functions are combined in these scission processes, which occur in drastically different compositions and solution conditions. In general, direct experimental access to changing neck sizes induced by individual proteins or peptide fragments is challenging due to the nanoscale dimensions and influence of thermal fluctuations. Here, we use a mechanical model to estimate the size of scission necks by leveraging small-angle X-ray scattering structural data of protein-lipid systems under different conditions. The influence of interfacial tension, lipid composition, and membrane budding morphology on the size of the induced scission necks is systematically investigated using our data and molecular dynamic simulations. We find that the M2 budding protein from the influenza A virus has robust pH-dependent membrane activity that induces nanoscopic necks within the range of spontaneous hemifission for a broad range of lipid compositions. In contrast, the sizes of scission necks generated by mitochondrial fission proteins strongly depend on lipid composition, which suggests a role for mechanical constriction.

Dealkenylative Functionalizations: Conversion of Alkene C(sp3)-C(sp2) Bonds into C(sp3)-X Bonds via Redox-Based Radical Processes.

This review highlights the history and recent advances in dealkenylative functionalization. Through this deconstructive strategy, radical functionalizations occur under mild, robust conditions. The reactions described proceed with high efficiency, good stereoselectivity, tolerate many functional groups, and are completed within a matter of minutes. By cleaving the C(sp3)-C(sp2) bond of terpenes and terpenoid-derived precursors, rapid diversification of natural products is possible.

Cellular remodeling and JAK inhibition promote zygotic gene expression in the Ciona germline.

Transcription control is a major determinant of cell fate decisions in somatic tissues. By contrast, early germline fate specification in numerous vertebrate and invertebrate species relies extensively on RNA-level regulation, exerted on asymmetrically inherited maternal supplies, with little-to-no zygotic transcription. However delayed, a maternal-to-zygotic transition is nevertheless poised to complete the deployment of pre-gametic programs in the germline. Here, we focus on early germline specification in the tunicate Ciona to study zygotic genome activation. We first demonstrate that a peculiar cellular remodeling event excludes localized postplasmic Pem-1 mRNA, which encodes the general inhibitor of transcription. Subsequently, zygotic transcription begins in Pem-1-negative primordial germ cells (PGCs), as revealed by histochemical detection of elongating RNA Polymerase II, and nascent Mef2 transcripts. In addition, we uncover a provisional antagonism between JAK and MEK/BMPRI/GSK3 signaling, which controls the onset of zygotic gene expression, following cellular remodeling of PGCs. We propose a 2-step model for the onset of zygotic transcription in the Ciona germline and discuss the significance of germ plasm dislocation and remodeling in the context of developmental fate specification.

Hemolytic Properties of Fine Particulate Matter (PM2.5) in In Vitro Systems.

Epidemiological studies have suggested that inhalation exposure to particulate matter (PM) air pollution, especially fine particles (i.e., PM2.5 (PM with an aerodynamic diameter of 2.5 microns or less)), is causally associated with cardiovascular health risks. To explore the toxicological mechanisms behind the observed adverse health effects, the hemolytic activity of PM2.5 samples collected during different pollution levels in Beijing was evaluated. The results demonstrated that the hemolysis of PM2.5 ranged from 1.98% to 7.75% and demonstrated a clear dose-response relationship. The exposure toxicity index (TI) is proposed to represent the toxicity potential of PM2.5, which is calculated by the hemolysis percentage of erythrocytes (red blood cells, RBC) multiplied by the mass concentration of PM2.5. In a pollution episode, as the mass concentration increases, TI first increases and then decreases, that is, TI (low pollution levels) < TI (heavy pollution levels) < TI (medium pollution levels). In order to verify the feasibility of the hemolysis method for PM toxicity detection, the hemolytic properties of PM2.5 were compared with the plasmid scission assay (PSA). The hemolysis results had a significant positive correlation with the DNA damage percentages, indicating that the hemolysis assay is feasible for the detection of PM2.5 toxicity, thus providing more corroborating information regarding the risk to human cardiovascular health.

Role of the dynamin-related protein 2 family and SH3P2 in clathrin-mediated endocytosis in Arabidopsis thaliana.

Clathrin-mediated endocytosis (CME) is vital for the regulation of plant growth and development through controlling plasma membrane protein composition and cargo uptake. CME relies on the precise recruitment of regulators for vesicle maturation and release. Homologues of components of mammalian vesicle scission are strong candidates to be part of the scission machinery in plants, but the precise roles of these proteins in this process are not fully understood. Here, we characterised the roles of the plant dynamin-related protein 2 (DRP2) family (hereafter DRP2s) and SH3-domain containing protein 2 (SH3P2), the plant homologue to recruiters of dynamins, such as endophilin and amphiphysin, in CME by combining high-resolution imaging of endocytic events in vivo and characterisation of the purified proteins in vitro. Although DRP2s and SH3P2 arrive similarly late during CME and physically interact, genetic analysis of the sh3p123 triple mutant and complementation assays with non-SH3P2-interacting DRP2 variants suggest that SH3P2 does not directly recruit DRP2s to the site of endocytosis. These observations imply that, despite the presence of many well-conserved endocytic components, plants have acquired a distinct mechanism for CME.

Degradation during Mixing of Silica-Reinforced Natural Rubber Compounds.

The optimal mixing conditions for silica-filled NR compounds dictate the need to proceed at a high temperature, i.e., 150 °C, to achieve a sufficient degree of silanization. On the other hand, natural rubber is prone to degradation due to mechanical shear and thermal effects during mixing, particularly at long exposure times. The present work investigates NR rubber degradation during mixing in relation to prolonged silanization times. The Mooney viscosity and stress relaxation rates, bound rubber content, storage modulus (G'), and delta δ were investigated to indicate the changes in the elastic/viscous responses of NR molecules related to rubber degradation, molecular chain modifications, and premature crosslinking/interaction. In Gum NR (unfilled), an increase in the viscous response with increasing mixing times indicates a major chain scission that causes a decreased molecular weight and risen chain mobility. For silica-filled NR, an initial decrease in the Mooney viscosity with increasing silanization time is attributed to the chain scission first, but thereafter the effect of the degradation is counterbalanced by a sufficient silanization/coupling reaction which leads to leveling off of the viscous response. Finally, the higher viscous response due to degradation leads to the deterioration of the mechanical properties and rolling resistance performance of tire treads made from such silica-filled NR, particularly when the silanization time exceeds 495 s.

High-Pressure Limit and Pressure-Dependent Rate Rules for ß-Scission Reaction Class of Hydroperoxyl Alkyl Hydroperoxyl Radicals (•P(OOH)2) in Normal-Alkyl Cyclohexanes Combustion.

Chemical kinetic studies of the ß-scission reaction class of hydroperoxyl alkyl hydroperoxyl radicals (•P(OOH)2) from normal-alkyl cyclohexanes are carried out systematically through high-level ab initio calculations. Geometry optimizations and frequency calculations for all species involved in the reactions are performed at the B3LYP/CBSB7 level of theory. Electronic single-point energy calculations are calculated at the CBS-QB3 level of theory. Rate constants for the reactions of ß-scission, in the temperature range of 500-1500 K and the pressure range of 0.01-100 atm, are calculated using transition state theory (TST) and Rice-Ramsberger-Kassel-Marcus/Master-Equation (RRKM/ME) theory taking asymmetric Eckart tunneling corrections and the one-dimensional hindered rotor approximation into consideration. The rate rules are obtained by averaging the rate constants of the representative reactions of this class. These rate rules can greatly assist in constructing more accurate low-temperature combustion mechanisms for normal-alkyl cyclohexanes.

Analysis of photocatalytic degradation of polyamide microplastics in metal salt solution by high resolution mass spectrometry.

Microplastic pollution has become one of the most concerned focuses in the world. Among many treatment methods, photocatalysis is considered to be one of the most environmentally friendly methods. In this work, the photodegradation behavior of polyamide microplastics is studied by using polyamide 6 PA6) as model microplastics and FeCl3 as catalyst. It is hoped that the PA6 fiber can be effectively degraded by utilizing the strong oxidizing active species that can be produced after FeCl3 is irradiated in water. The results shows that PA6 fiber can be almost completely degraded after 10 days of irradiation in FeCl3 aqueous solution, indicating that it is promising to use this new method to solve the problem of PA6 type microplastics. In addition, the chain scission mechanism and degradation process of PA6 are analyzed in detail by ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS), which provides a new insight for the study of polymer degradation mechanism.

Cholesterol and M2 Rendezvous in Budding and Scission of Influenza A Virus.

The cholesterol of the host cell plasma membrane and viral M2 protein plays a crucial role in multiple stages of infection and replication of the influenza A virus. Cholesterol is required for the formation of heterogeneous membrane microdomains (or rafts) in the budozone of the host cell that serves as assembly sites for the viral components. The raft microstructures act as scaffolds for several proteins. Cholesterol may further contribute to the mechanical forces necessary for membrane scission in the last stage of budding and help to maintain the stability of the virus envelope. The M2 protein has been shown to cause membrane scission in model systems by promoting the formation of curved lipid bilayer structures that, in turn, can lead to membrane vesicles budding off or scission intermediates. Membrane remodeling by M2 is intimately linked with cholesterol as it affects local lipid composition, fluidity, and stability of the membrane. Thus, both cholesterol and M2 protein contribute to the efficient and proper release of newly formed influenza viruses from the virus-infected cells.

Mechanochemistry in Block Copolymers: New Scission Site due to Dynamic Phase Separation.

Mechanochemistry can lead to the degradation of the properties of covalent macromolecules. In recent years, numerous functional materials have been developed based on block copolymers (BCPs), however, like homopolymers, their chains could undergo mechanochemical damage during processing, which could have crucial impact on their performance. To investigate the mechanochemical response of BCPs, multiple polymers comprising different ratios of butyl acrylate and methyl methacrylate were prepared with similar degree of polymerization and stressed in solution via ultrasonication. Interestingly, all BCPs, regardless of the amount of the methacrylate monomer, presented a mechanochemistry rate constant similar to that of the methacrylate homopolymer, while a random copolymer reacted like the acrylate homopolymer. Size-exclusion chromatography showed that, in addition to the typical main peak shift towards higher retention times, a different daughter fragment was produced indicating a secondary selective scission site, situated around the covalent connection between the two blocks. Molecular dynamics modeling using acrylate and methacrylate oligomers were carried out and indicated that dynamic phase separation occurs even in a good solvent. Such non-random conformations can explain the faster polymer mechanochemistry. Moreover, the dynamic model for end-to-end chain overstretching supports bond scission which is not necessarily chain-centered.

Hydrogelation of Regenerated Silk Fibroin via Gamma Irradiation.

Gamma irradiation, which is one of the more conventional sterilization methods, was used to induce the hydrogelation of silk fibroin in this study. The physical and chemical characteristics of the irradiation-induced silk fibroin hydrogels were investigated. Silk fibroin solution with a concentration greater than 1 wt% formed hydrogel when irradiated by gamma rays at a dose of 25 or 50 kGy. The hydrogel induced by 50 kGy of radiation was more thermally stable at 80 °C than those induced by 25 kGy of radiation. When compared to the spontaneously formed hydrogels, the irradiated hydrogels contained a greater fraction of random coils and a lower fraction of ß-sheets. This finding implies that gelation via gamma irradiation occurs via other processes, in addition to crystalline ß-sheet formation, which is a well-established mechanism. Our observation suggests that crosslinking and chain scission via gamma irradiation could occur in parallel with the ß-sheet formation. The irradiation-induced hydrogels were obtained when the solution concentration was adequate to support the radiation crosslinking of the silk fibroin chains. This work has, therefore, demonstrated that gamma irradiation can be employed as an alternative method to produce chemical-free, random coil-rich, and sterilized silk fibroin hydrogels for biomedical applications.

Contribution of free hydroxyl radical to the formation of micro(nano)plastics and release of additives during polyethylene degradation in water.

The omnipresence of secondary microplastics (MPs) in aquatic ecosystems has become an increasingly alarming public health concern. Hydrogen peroxide (H2O2) is an important oxidant in nature and the most stable reactive oxygen species occurred in natural water. In order to explore the contribution of free ˙OH generated from H2O2-driven Fenton-like reactions on the degradation of polyethylene (PE) and generation of micro- and nano-scale plastics in water, a batch experiment was conducted over a period of 620 days in water treated with micromolar H2O2. The incorporation of H2O2 in water induced the formation of flake-like micro(nano)-sized particles due to intensified oxidative degradation of PE films. The presence of ˙OH significantly enhanced the generation of both micro- and nano-scale plastics exhibiting a higher proportion of particles in the range of 200-500 nm compared to the Control. Total organic carbon in the H2O2 treated solution was nearly 174-fold higher than that of the Control indicating a substantial liberation of organic compounds due to the oxidative degradation of native carbon chain of PE and subsequent decomposition of its additives. The highly toxic butylated hydroxytoluene detected from the gas chromatography-mass spectrometry (GC-MS) analysis implied the toxicological behavior of secondary micro(nano)plastics influenced by the oxidation and decomposition processes The findings from this study further expand our understanding of the role of ˙OH in degrading PE micro-scale plastics into nanoparticles as an implication of naturally occurring H2O2 in aquatic environments. In the future, further attention should be drawn to the underlying mechanisms of H2O2-driven in-situ Fenton reaction mediated by natural environmental conditions targeting the alternation of light and darkness on the oxidative degradation of plastics.

Antioxidant and ACE-Inhibitory Activity of Protein Hydrolysates Produced from Atlantic Sea Cucumber (Cucumaria frondosa).

Atlantic sea cucumber is a benthic marine echinoderm found in Northwest Atlantic waters and is harvested mainly for its body wall. The body wall, along with internal organs and aquaphyrangeal bulb/flower, is a rich source of proteins, where the latter parts are often considered as processing discards. The objective of this research was to produce protein hydrolysates from sea cucumber tissues (body wall, flower, and internal organs) with bioactive properties associated with antioxidants, DNA and LDL cholesterol oxidation inhibition, and angiotensin-I-converting enzyme (ACE) inhibitory effects. The protein hydrolysates were prepared using food-grade commercial enzymes, namely Alcalase, Corolase, and Flavourzyme, individually and in combination, and found that the combination of enzymes exhibited stronger antioxidant potential than the individual enzymes, as well as their untreated counterparts. Similar trends were also observed for the DNA and LDL cholesterol oxidation inhibition and ACE-inhibitory properties of sea cucumber protein hydrolysates, mainly those that were prepared from the flower. Thus, the findings of this study revealed potential applications of sea cucumber-derived protein hydrolysates in functional foods, nutraceuticals, and dietary supplements, as well as natural therapeutics.

Structure and dynamics of ESCRT-III membrane remodeling proteins by high-speed atomic force microscopy.

Endosomal sorting complex required for transport (ESCRT) proteins assemble on the cytoplasmic leaflet of membranes and remodel them. ESCRT is involved in biological processes where membranes are bent away from the cytosol, constricted, and finally severed, such as in multivesicular body formation (in the endosomal pathway for protein sorting) or abscission during cell division. The ESCRT system is hijacked by enveloped viruses to allow buds of nascent virions to be constricted, severed, and released. ESCRT-III proteins, the most downstream components of the ESCRT system, are monomeric and cytosolic in their autoinhibited conformation. They share a common architecture, a four-helix bundle with a fifth helix that interacts with this bundle to prevent polymerizing. Upon binding to negatively charged membranes, the ESCRT-III components adopt an activated state that allows them to polymerize into filaments and spirals and to interact with the AAA-ATPase Vps4 for polymer remodeling. ESCRT-III has been studied with electron microscopy and fluorescence microscopy; these methods provided invaluable information about ESCRT assembly structures or their dynamics, respectively, but neither approach provides detailed insights into both aspects simultaneously. High-speed atomic force microscopy (HS-AFM) has overcome this shortcoming, providing movies at high spatiotemporal resolution of biomolecular processes, significantly increasing our understanding of ESCRT-III structure and dynamics. Here, we review the contributions of HS-AFM in the analysis of ESCRT-III, focusing on recent developments of nonplanar and deformable HS-AFM supports. We divide the HS-AFM observations into four sequential steps in the ESCRT-III lifecycle: (1) polymerization, (2) morphology, (3) dynamics, and (4) depolymerization.

Lipid oxidation induced protein scission in an oleogel as a model food.

Lipid oxidation induced protein scission was investigated in oleogel using beta-lactoglobulin (whey protein isolate) as gelator. Extracted cleaved peptides were measured using high resolution mass spectrometry (FT-ICR-MS), which was provided by an automatically generated annotation list approach to identify relevant masses and sum formula using the isotopic pattern. The identified oxidized peptides were then further evaluated using partial least squares regression to relevant lipid hydroperoxide formation data, which provide the significance and importance of the peptides toward lipid induced scission. Thereby, the most important peptides are located at the surface of the protein in random coil segments and especially at the ends of the protein sequence. The most important amino acids were cysteine and aliphatic amino acids, which undergo scission mostly by the α-amidation pathway. The findings compare well with studies investigating depletion of amino acids initiated by lipid oxidation in systems containing bovine albumin or gamma-globulin.

Valorization of lignin through reductive catalytic fractionation of fermented corn stover residues.

The fermented corn stover residues are abundant renewable lignin-rich bioresources that show great potential to produce aromatic phenols. However, selective catalytic hydrogenolysis of this residual material still remains challenge to obtain high yields. Herein, a novel strategy to produce monophenolic compounds from the fermented stover over a commercial Pd/C catalyst was proposed. Taking the reaction temperature as the key variable, the highest monomer yield was 28.5 wt% at 220 °C in compaction with that of the pristine corn stover (22.8 wt%). The enhanced monophenol yield was due to the higher contents of lignin and less recalcitrance in the fermented stover. Moreover, the van Krevelen diagram revealed a slight selective CO bond scission of lignin macromolecular during fermentation as well as the dehydration and deoxygenation in hydrogenolysis reaction. Overall, this work opens a new avenue for the valorization of lignin through reductive catalytic fractionation of agricultural wastes.