Random mutagenesis and disulfide bond formation improved thermostability in microbial transglutaminase.

Microbial transglutaminase (MTG) from Streptomyces mobaraensis is widely used in the food and pharmaceutical industries for cross-linking and post-translational modification of proteins. It is believed that its industrial applications could be further broadened by improving its thermostability. In our previous study, we showed that the introduction of structure-based disulfide bonds improved the thermostability of MTG, and we succeeded in obtaining a thermostable mutant, D3C/G283C, with a T50 (incubation temperature at which 50% of the initial activity remains) 9 °C higher than that of wild-type MTG. In this study, we performed random mutations using D3C/G283C as a template and found several amino acid substitutions that contributed to the improvement of thermostability, and investigated a thermostable mutant (D3C/S101P/G157S/G250R/G283C) with three amino acid mutations in addition to the disulfide bond. The T50 of this mutant was 10 °C higher than that of the wild type, the optimal temperature for enzymatic reaction was increased to 65 °C compared to 50 °C for the wild type, and the catalytic efficiency (kcat/Km) at 37.0 °C was increased from 3.3 × 102 M-1 s-1 for the wild type to 5.9 × 102 M-1 s-1. X-ray crystallography of the D3C/G283C MTG showed no major structural differences against wild-type MTG. Structural differences were found that may contribute to thermostabilization and improve catalytic efficiency. KEY POINTS: • Improved heat resistance is essential to broaden the application of MTG. • The MTG mutant D3C/S101P/G157S/G250R/G283C showed improved thermostability. • X-ray crystallography of the disulfide bridge mutant D3C/G283C MTG was elucidated.

Comparison of selenium-mediated regulation of heat shock protein and inflammation in-vitro and in-ovo for heat resistance enhancement in broiler.

Selenium is a heat-stress-reducing substance that improves heat resistance and is being studied for its effective application in the broiler industry. However, research on feed additives is labor-intensive and time-consuming because of the need for feeding experiments. We aimed to compare the effects of selenium under heat stress in vitro and in ovo, specifically examining the gene expression of heat shock proteins (HSP) and inflammatory markers. Two groups were included in the in-vitro study: in-vitro control (TC; selenium 0 µg/ml) and in-vitro selenium (TS; selenium 5 µg/ml). The satellite cells were cultured at 42°C for 48 h after selenium treatment. The in-ovo study comprised 4 groups: in-ovo control and in-ovo selenium 1-3 (OC, OS1, OS2, and OS3; selenium 2.5, 5, and 10 µg/egg, respectively). Selenium was injected on the 18th day after hatching, and heat treatment was performed at 32-34°C from the 14th to the 21st day after hatching, and the leg muscles of the chicks were collected on the 21st day. The gene expression of heat shock proteins (HSP), caspase3, nuclear factor kappa light-chain enhancer of activated B cells (NF-kB), and IL-8 was analyzed in in-vitro and in-ovo experiments, respectively. In-vitro results showed significant increases in HSP90, HSP60, and HSP40 in TS compared to TC, with a significant decrease in HSP70. In the in-ovo study, HSP70, caspase3, NF-kB and IL-8 were significantly increased in OS1. HSP90, HSP60, HSP40, HSP27 and NF-kB were significantly decreased in in-ovo OS2 compared to in-vitro TS, implying a trend in ratio compared to control. Selenium appeared to enhance heat resistance in-vitro and in-ovo by modulating HSPs and inflammation. However, differences in mRNA expression were observed depending on the concentration of selenium. These findings suggest that selenium modulates heat resistance through different mechanisms in-vitro and in-ovo, likely due to the complexity of whole-organism interactions in-ovo compared to the single-cell-type environment in-vitro. Therefore, to directly apply in-vitro results to in-ovo, a concentration comparison study for each additive is necessary.

Enhanced strength, toughness and heat resistance of poly (lactic acid) with good transparency and biodegradability by uniaxial pre-stretching.

Sustainable poly (lactic acid) (PLA) with excellent strength, toughness, heat resistance, transparency, and biodegradability was achieved by uniaxial pre-stretching at 70 °C. The effect of pre-stretched ratio (PSR) on the microstructure and properties of the PLA was investigated. The undrawn PLA was brittle. However, after pre-stretching, the elongation at break was increased significantly. The maximum value of 161.2 % was obtained at pre-stretching ratio (PSR) of 1.0. With the increase of PSR, the modulus and strength were improved obviously (from 1601 MPa and 60.2 MPa for undrawn PLA to 2932 MPa and 106.3 MPa for the ps-PLA at PSR =3.0). Meanwhile, the heat resistance of PLA was improved obviously with the increase of PSR. For the ps-PLA3.0, there were almost no deformation and shrink at 140 °C. Interestingly, after pre-stretching, the PLA still maintained the good transparency and biodegradability. The brittleness for undrawn PLA was attributed to the network structure of cohesional entanglements. After pre-stretching, the destruction of the network structure and formation of the orientation, mesophase and oriented nanosized crystalline phase lead to the increased the toughness, strength and heat resistance without sacrificing the transparency and biodegradability. This work provides a significant guidance for the fabrication of PLA material with excellent comprehensive performance including strength, toughness, heat resistance, transparency, and biodegradability.

Thermal stability and in vitro digestive behavior of Pickering emulsion stabilized by high-amylose starch nanocrystals.

In this study, high-amylose starch (HAS) was processed using sulfuric acid-ultrasonic cross-linking to produce high-amylose starch nanocrystals (HASNC). These nanocrystals were used to stabilize Pickering emulsions and assess their effectiveness in encapsulating ß-carotene. Normal starch nanocrystals (NSNC) were prepared similarly for comparison. The HASNC retained key HAS properties, such as heat and enzyme resistance, providing several advantages to HASNC-stabilized emulsions. First, after exposure to 100 °C heat and in vitro tests simulating the mouth and stomach, the HASNC-stabilized emulsions demonstrated significantly greater stability and higher ß-carotene retention compared to the NSNC-stabilized emulsions. This enhanced stability is attributed to the lower gelatinization degree and increased resistance to α-amylase hydrolysis of HASNC, which provides stronger steric stabilization of the oil droplets. Second, during in vitro small intestine tests, the greater enzyme resistance of HASNC allowed for the formation of a denser barrier around the oil droplets, effectively preventing lipase and bile salts from contacting the oil droplets. This led to a reduced rate and extent of lipid digestion and facilitated a sustained-release effect. Consequently, HASNC, as a starch-based emulsifier, show great potential as an effective delivery system for the sustained release of bioactive compounds.

Methods for Screening and Isolating Extremely Heat-Resistant Escherichia coli from Meat Sources.

Meat animals harbor diverse E. coli populations in their digestive tracts and can serve as sources of pathogenic E. coli. The consumption of meat and produce contaminated with virulent E. coli from animal sources is associated with human illnesses and outbreaks. Heat treatment is an antimicrobial intervention that is commonly used during meat processing to ensure effective reductions in microbial load. Extreme heat resistance (XHR) has been reported among meat-borne E. coli and is mainly attributed to an ~15-19 kb genetic element known as the transmissible locus of stress tolerance (tLST). XHR E. coli can resist treatments used during meat processing and cooking. Therefore, the detection of heat-resistant E. coli is important for devising effective control measures to prevent meat spoilage and ensure meat safety. Here, we present methods used to (1) screen for tLST genes by multiplex PCR and (2) screen and isolate XHR E. coli from meat sources. The mode of heat exposure affects the outcome of XHR testing. Hence, the protocols were optimized to achieve maximum agreement between the tLST genotype and the XHR phenotype.

Potential improvement in the mechanical performance and thermal resistance of geopolymer with appropriate microplastic incorporation: A sustainable solution for recycling and reusing microplastics.

The accumulation of microplastics (MPs) has been a major threat to the natural environment and human health. However, incineration and landfilling may not be appropriate for the management of MPs. This paper evaluated the feasibility of incorporating MPs with diverse dimensions (50 to 500 µm) and contents (2.5 % to 10 %) into geopolymer cured under different temperatures (40 and 80 °C). The compressive (fc) and flexural strength (ff) after curing and thermal exposure (200 to 600 °C) were determined. When cured at 40 °C, fc and ff decreased with percentages of MPs incorporated. By contrast, when cured at 80 °C, the addition of 2.5 % MPs increased fc and ff by up to 33 % (from 52.2 to 69.4 MPa) and 18 % (from 8.2 to 9.7 MPa), depending on MPs' sizes. The XRD and TGA results suggested that the observed increases in mechanical properties can be attributed to the formation of more calcium alumino (silicate) hydrates (C-(A)-S-H gels) induced by the incorporation of a small quantity of MPs (2.5 %). The SEM images also showed better adhesion between MPs and geopolymeric products when cured under 80 °C, potentially inhibiting crack development. After being exposed to evaluated temperatures (200 and 400 °C), fc of the specimens with 2.5 % MPs and cured at 80 °C was higher than that without MPs. The fc dropped dramatically due to the degradation of MPs between 400 and 600 °C. The increase in strength and heat resistance (up to 400 °C) of MPs-incorporated geopolymer cured under 80 °C indicated the potential recycling and reuse of MPs for geopolymer materials.

Fabrication of liquid-free ionic conductive elastomer (ICE) from cellulose-rosin derived poly(esterimide) towards temperature-tolerant and solvent-resistant UV shadowless adhesive and sensor.

Recently, the utilization of the cellulose to fabricate the multifunctional materials with aim to replace the petroleum-based product, is receiving significant attentions. However, the development of cellulose-based multifunctional materials with high mechanical strength and temperature resistance is still a challenge. Herein, the intrinsic feature and property of cellulose and rosin were creatively employed to fabricate a novel cellulose-rosin based poly(esterimide) (PEI) by esterification reaction and imidization reaction, and the obtained cellulose-rosin derived PEI exhibits superior thermal stability. Then the as-prepared cellulose-rosin derived PEI was dissolved in polymerizable deep eutectic solvents (PDES) and in-situ formed the ionic conductive elastomer (ICE) with via UV-induced polymerization. These cellulose-rosin based ICE exhibited excellent mechanical properties, solvent resistance, and temperature tolerance. By adjusting the mass ratio of cellulose-rosin derived PEI and PDES, the as-prepared liquid-free ICE functions as UV shadowless adhesive and wearable sensors. The bonding strength of UV shadowless adhesive could 1.52 MPa, which could be applied to fix the broken glass toy models. Furthermore, wearable sensors based those ICE could monitor the large and subtle movements even under extreme environmental condition, such as being soaked in organic solvent (such as tetrahydrofuran) or at low/high temperature (-25 °C or 80 °C). This work opens a new avenue for the next-generation of multifunctional ICE.

Elastomeric Fire and Heat-Protective Materials Containing Functionally Active Microheterogeneous Systems.

This research aims to explore how functionally active structures affect the physical, mechanical, thermal, and fire-resistant properties of elastomeric compositions using ethylene-propylene-diene rubber as a base. The inclusion of aluminosilicate microspheres, microfibers, and a phosphorus-boron-nitrogen-organic modifier in these structures creates a synergistic effect, enhancing the material's heat-insulating properties by strengthening coke and carbonization processes. This results in a 12-19% increase in heating time for unheated sample surfaces and a 6-17% increase in residual coke compared to existing analogs. Microspheres help counteract the negative impact of microfibers on composition density and thermal conductivity, while the phosphorus-boron-containing modifier allows for controlling the formation of the coke layer.

Design of Co-Cured Multi-Component Thermosets with Enhanced Heat Resistance, Toughness, and Processability via a Machine Learning Approach.

Designing heat-resistant thermosets with excellent comprehensive performance has been a long-standing challenge. Co-curing of various high-performance thermosets is an effective strategy, however, the traditional trial-and-error experiments have long research cycles for discovering new materials. Herein, a two-step machine learning (ML) assisted approach is proposed to design heat-resistant co-cured resins composed of polyimide (PI) and silicon-containing arylacetylene (PSA), that is, poly(silicon-alkyne imide) (PSI). First, two ML prediction models are established to evaluate the processability of PIs and their compatibility with PSA. Then, another two ML models are developed to predict the thermal decomposition temperature and flexural strength of the co-cured PSI resins. The optimal molecular structures and compositions of PSI resins are high-throughput screened. The screened PSI resins are experimentally verified to exhibit enhanced heat resistance, toughness, and processability. The research framework established in this work can be generalized to the rational design of other advanced multi-component polymeric materials.

Controlled Structural Relaxation of Aramid Nanofibers for Superstretchable Polymer Fibers with High Toughness and Heat Resistance.

Polymer fibers that combine high toughness and heat resistance are hard to achieve, which, however, hold tremendous promise in demanding applications such as aerospace and military. This prohibitive design task exists due to the opposing property dependencies on chain dynamics because traditional heat-resistant materials with rigid molecular structures typically lack the mechanism of energy dissipation. Aramid nanofibers have received great attention as high-performance nanoscale building units due to their intriguing mechanical and thermal properties, but their distinct structural features are yet to be fully captured. We show that aramid nanofibers form nanoscale crimps during the removal of water, which primarily resides at the defect planes of pleated sheets, where the folding can occur. The precise control of such a structural relaxation can be realized by exerting axial loadings on hydrogel fibers, which allows the emergence of aramid fibers with varying angles of crimps. These crimped fibers integrate high toughness with heat resistance, thanks to the extensible nature of nanoscale crimps with rigid molecular structures of poly(p-phenylene terephthalamide), promising as a template for stable stretchable electronics. The tensile strength/modulus (392-944 MPa/11-29 GPa), stretchability (25-163%), and toughness (154-445 MJ/cm3) are achieved according to the degree of crimping. Intriguingly, a toughness of around 430 MJ/m3 can be maintained after calcination below the relaxation temperature (259 °C) for 50 h. Even after calcination at 300 °C for 10 h, a toughness of 310 MJ/m3 is kept, outperforming existing polymer materials. Our multiscale design strategy based on water-bearing aramid nanofibers provides a potent pathway for tackling the challenge for achieving conflicting property combinations.

Microwave De-Icing Efficiency Improvement of Asphalt Mixture with Structural Layer Optimization and Heat-Resistance Design.

The application of microwave de-icing technology in road engineering is constrained by its low energy utilization rate, which can be attributed to low heat production rates and ineffective heat dissipation to the underlying pavement. In this work, asphalt mixtures are designed as an upper layer (heating layer) and a lower layer (thermal-resistance layer). Magnetite slag was selected as a microwave-sensitive source for generating heat, and expanded perlite powder was incorporated into the lower layer as a thermal resistance material. Structural layer optimization and thermal-resistance layer design of the asphalt mixture were carried out by changing the thickness of the upper and lower layers to further improve the heat production rates. The design effectiveness is comprehensively evaluated by factors such as the changing law of the average surface temperature of mixtures, ice-melting time, and cost-effectiveness analyses. The results show that EP possesses better thermal stability, lower microwave energy conversion ability and more excellent heat-resistance potential compared with mineral powder. The heat-resistance layer with EP can prevent heat from being conducted to the lower layer and promote it to concentrate on the specimen surface, which can endow the microwave heating efficiency of specimens to be further improved by up to 26.97% and the de-icing time reduced by 10%, ascribed to the heat-resistance design. Furthermore, the collaborative design of the structural layer optimization and heat-resistance layer can increase energy utilization efficiency and save microwave-absorbing materials while ensuring excellent microwave de-icing efficiency.

Poly(ester imide)s with Low Linear Coefficients of Thermal Expansion and Low Water Uptake (VIII): Structure-Flame Retardancy Relationship.

A series of ester-linked tetracarboxylic dianhydrides containing multiple para-phenylene units (TA-pPhs) was synthesized to obtain novel modified polyimides, namely poly(ester imide)s (PEsIs). The flame retardancy and film toughness of PEsIs tended to deteriorate with the structural extension of the repeating units (or monomers) via ester groups. To identify the structural factors necessary for achieving the highest flame retardancy rank (UL-94, V-0), we systematically investigated the structure-property relationships of a series of TA-pPh-based PEsIs. Among them, a PEsI derived from para-quaterphenylene-containing TA-pPh (TA-DPQP) and p-phenylenediamine (p-PDA) exhibited the best property combination, featuring an extremely high glass transition temperature (Tg), very low linear coefficient of thermal expansion (CTE), low water uptake (WA), ultralow linear coefficient of humidity (hygroscopic) expansion (CHE), unexpectedly high film toughness, and excellent flame retardancy (V-0 rank). Moreover, we examined the effects of substituents of TA-pPh and discussed the mode of action for the increased film toughness. This study also investigated the structure-property relationship for a series of PEsIs derived from isomeric naphthalene-containing tetracarboxylic dianhydrides. Some of the PEsIs obtained in this study, such as the TA-DPQP/p-PDA system, hold promise as novel high-temperature dielectric substrates for use in flexible printed circuits.

Mitochondrial Fission Is Involved in Heat Resistance in Zebrafish.

Global warming and extreme weather events pose a significant threat to global biodiversity, with rising water temperatures exerting a profound influence on fish conservation and fishery development. In this study, we used zebrafish as a model organism to explore the impact of a heat acclimation period on their survival rates. The results demonstrated that a 2-month heat acclimation period almost completely mitigated heat stress-induced mortality in zebrafish. Subsequent analysis of the surviving zebrafish revealed a predominance of hepatic mitochondria in a fission state. Remarkably, a short-term fasting regimen, which induced hepatic mitochondrial fission, mirrored the outcomes of the protective effect of heat acclimation and augmented animal survival under heat stress. Conversely, treatment with a mitochondrial fission inhibitor within the fasting group attenuated the elevated survival rate. Furthermore, zebrafish embryos subjected to brief heat acclimation also exhibited increased heat resistance, a trait diminished by a chemical intervention inhibiting mitochondrial fission. This suggests a shared mechanism for heat resistance between embryos and adult zebrafish. These findings underscore the potential use of inducing mitochondrial fission to enhance heat resistance in zebrafish, offering promise for fish biodiversity conservation in the face of global warming.

Thermally Stable and Transparent Polyimide Derived from Side-Group-Regulated Spirobifluorene Unit for Substrate Application.

Advancements in flexible electronic technology, especially the progress in foldable displays and under-display cameras (UDC), have created an urgent demand for high-performance colorless polyimide (CPI). However, current CPIs lack sufficient heat resistance for substrate applications. In this work, four kinds of rigid spirobifluorene diamines are designed, and the corresponding polyimides are prepared by their condensation with 5,5'-(perfluoropropane-2,2-diyl) bis(isobenzofuran-1,3-dione) (6FDA) or 9,9-bis(3,4-dicarboxyphenyl) fluorene dianhydride (BPAF). The rigid and conjugated spirobifluorene units endow the polyimides with higher glass transition temperature (Tg) ranging from 356 to 468 °C. Their optical properties are regulated by small side groups and spirobifluorene structure with a periodically twisted molecular conformation. Consequently, a series of CPIs with an average transmittance ranging from 75% to 88% and a yellowness index (YI) as low as 2.48 are obtained. Among these, 27SPFTFA-BPAF presents excellent comprehensive performance, with a Tg of 422 °C, a 5 wt.% loss temperature (Td5) of 562 °C, a YI of 3.53, and a tensile strength (δmax) of 140 MPa, respectively. The mechanism underlying the structure-property relationship is investigated by experimental comparison and theoretical calculation, and the proposed method provides a pathway for designing highly rigid conjugated CPIs with excellent thermal stability and transparency for photoelectric engineering.

Differential impacts of nonsteroidal anti-inflammatory drugs on lifespan and healthspan in aged Caenorhabditis elegans.

Aging and age-related diseases are intricately associated with oxidative stress and inflammation. Nonsteroidal anti-inflammatory drugs (NSAIDs) have shown their promise in mitigating age-related conditions and potentially extending lifespan in various model organisms. However, the efficacy of NSAIDs in older individuals may be influenced by age-related changes in drug metabolism and tolerance, which could result in age-dependent toxicities. This study aimed to evaluate the potential risks of toxicities associated with commonly used NSAIDs (aspirin, ibuprofen, acetaminophen, and indomethacin) on lifespan, healthspan, and oxidative stress levels in both young and old Caenorhabditis elegans. The results revealed that aspirin and ibuprofen were able to extend lifespan in both young and old worms by suppressing ROS generation and enhancing the expression of antioxidant SOD genes. In contrast, acetaminophen and indomeacin accelerated aging process in old worms, leading to oxidative stress damage and reduced resistance to heat stress through the pmk-1/skn-1 pathway. Notably, the harmful effects of acetaminophen and indomeacin were mitigated when pmk-1 was knocked out in the pmk-1(km25) strain. These results underscore the potential lack of benefit from acetaminophen and indomeacin in elderly individuals due to their increased susceptibility to toxicity. Further research is essential to elucidate the underlying mechanisms driving these age-dependent responses and to evaluate the potential risks associated with NSAID use in the elderly population.

Metal-organic framework adhesives with exceptionally high heat resistance.

We synthesized high-heat-resistant adhesives based on metal - organic frameworks owing to their high decomposition temperature and the absence of a glass transition. Heat-resistance tests were performed on adhesive joints consisting of zeolitic imidazolate framework (ZIF)-67-based adhesives and a copper substrate. The as-synthesized ZIF-67-based adhesive exhibited heat resistances at 600 and 700°C in air and nitrogen atmospheres, respectively, comparable to those of conventional high-heat-resistant polymer-based adhesives. The degradation mechanism of the ZIF-67 adhesives was investigated, and their high heat resistance was attributed to the stable existence of the ZIF-67 qtz phase in the adhesive layer at high temperatures without the formation of voids. Thus, adhesives based on ZIF-67 and other metal - organic frameworks can be applied in high-temperature industrial systems.

By focusing on its high thermal stability and absence of glass transition, the ZIF-67 gel was found to have high potential that is comparable to existing heat-resistant adhesives.

Progress in using cross-linking technologies to increase the thermal stability of colloidal delivery systems.

In recent years, crosslinking technology has been found and widely used in food, textile, pharmaceutical, bioengineering and other fields. Crosslinking is a reaction in which two or more molecules bond to each other to form a stable three-dimensional network structure to improve the strength, heat resistance and other properties of substances. The researchers found that the cross-linking technology has a significant effect on improving the thermal stability of the colloidal delivery system. In this paper, crosslinking techniques that can be used to improve the thermal stability of colloidal delivery systems are reviewed, including enzyme-, ion-, chemical-, and combined cross-linking. Initially, the underlying mechanisms of these crosslinking technologies is reviewed. Then, the impacts of crosslinking on the heat-stability of colloidal delivery systems are discussed. Finally, the application of crosslinked delivery systems in improving the thermal stability of probiotics, polyphenols, pigments, and nutrients in foods and food packaging materials is introduced. The ability of proteins and polysaccharides to form heat-stable colloidal delivery systems can be improved by crosslinking. Nevertheless, more research is required to establish the impact of different crosslinking on the thermal stability of a broader range of different delivery systems, as well as to ensure their safety and efficacy.

Ultrahigh Heat/Fire-Resistant, Mechanically Robust, and Closed-Loop Chemical Recyclable Polycarbonate Enabled by Facile Bond Dissociation Energy Modulation.

Plastics serve as an essential foundation in contemporary society. Nevertheless, meeting the rigorous performance demands in advanced applications and addressing their end-of-life disposal are two critical challenges that persist. Here, an innovative and facile method is introduced for the design and scalable production of polycarbonate, a key engineering plastic, simultaneously achieving high performance and closed-loop chemical recyclability. The bisphenol framework of polycarbonate is strategically adjusted from the low-bond-dissociation-energy bisphenol A to high-bond-dissociation-energy 4,4'-dihydroxydiphenyl, in combination with the incorporation of polysiloxane segments. As expected, the enhanced bond dissociation energy endows the polycarbonate with an extremely high glow-wire flammability index surpassing 1025 °C, a 0.8 mm UL-94 V-0 rating, a high LOI value of 39.2%, and more than 50% reduction of heat and smoke release. Furthermore, the π-π stacking interactions within biphenyl structures resulted in a significant enhancement of mechanical strength by as more as 37.7%, and also played a positive role in achieving a lower dielectric constant. Significantly, the copolymer exhibited outstanding closed-loop chemical recyclability, allowing for facile depolymerization into bisphenol monomers and the repolymerized copolymer retains its high heat and fire resistance. This work provides a novel insight in the design of high-performance and closed-loop chemical recyclable polymeric materials.

Involvement of ROS and calcium ions in developing heat resistance and inducing antioxidant system of wheat seedlings under melatonin's effects.

The stress-protective effect of melatonin (N-acetyl-5-methoxytryptamine) on plant cells is mediated by key signaling mediators, in particular calcium ions and reactive oxygen species (ROS). However, the links between changes in calcium and redox homeostasis and the formation of adaptive responses of cultivated cereals (including wheat) to the action of high temperatures have not yet been studied. In the present study, we investigated the possible involvement of ROS and calcium ions as signaling mediators in developing heat resistance in wheat (Triticum aestivum L.) seedlings and activating their antioxidant system. Treatment of 3-day-old etiolated seedlings with melatonin solutions at concentrations 0.01-10 µM increased their survival after exposure to 45 °C for 10 min. The most significant stress-protective effect was exerted by melatonin treatment at 1 µM concentration. Under the influence of melatonin, a transient enhancement of superoxide anion radical (O2•-) generation and an increase in hydrogen peroxide content were observed in roots, with a maximum at 1 h. Four hours after treatment with melatonin, the activity of catalase and guaiacol peroxidase increased in roots, while the activity of superoxide dismutase did not change significantly. After exposure to 45 °C, the activity of catalase and guaiacol peroxidase was higher in the roots of melatonin-treated wheat seedlings, and the indices of ROS generation, content of the lipid peroxidation product malonic dialdehyde, and cell membrane damage were lower than in control seedlings. Melatonin-induced changes in root ROS generation and antioxidant enzyme activities were eliminated by pretreatment with the hydrogen peroxide scavenger dimethylthiourea (DMTU), NADPH oxidase inhibitor imidazole, and calcium antagonists (the extracellular calcium chelator EGTA and phospholipase C inhibitor neomycin). Treatment with DMTU, imidazole, EGTA, and neomycin also abolished the melatonin-induced increase in survival of wheat seedlings after heat stress. The role of calcium ions and ROS, generated with the participation of NADPH oxidase, as signaling mediators in the melatonin-induced antioxidant system and heat stress resistance of wheat seedlings have been demonstrated.

Resistance of an intertidal oyster(Saccostrea mordax)to marine heatwaves and the implication for reef building.

Marine heatwaves (MHWs) have a significant impact on intertidal bivalves and the ecosystems they sustain, causing the destruction of organisms' original habitats. Saccostrea mordax mainly inhabits the intertidal zone around the equator, exhibiting potential tolerance to high temperatures and maybe a species suitable for habitat restoration. However, an understanding about the tolerance mechanism of S. mordax to high temperatures is unclear. It is also unknown the extent to which S. mordax can tolerate repeated heatwaves of increasing intensity and frequency. Here, we simulated the effects of two scenarios of MHWs and measured the physiological and biochemical responses and gene expression spectrum of S. mordax. The predicted responses varied greatly across heatwaves, and no heatwave had a significant impact on the survival of S. mordax. Specifically, there were no statistically significant changes apparent in the standard metabolic rate and the activities of enzymes of the oyster during repeated heatwaves. S. mordax exposed to high-intensity heatwaves enhanced their standard metabolic rate to fuel essential physiological maintenance and increasing activity of SOD and expression of HSP70/90. These strategies are presumably at the expense of functions related to immunity and growth, as best exemplified by significant depressions in activities of enzymes (NaK, CaMg, T-ATP, and AKP) and expression levels of genes (Rab, eEF-2, HMGR, Rac1, SGK, Rab8, etc.). The performance status of S. mordax tends to improve by implementing a suite of less energy-costly compensatory mechanisms at various levels of biological organization when re-exposed to heatwaves. The adaptive abilities shown by S. mordax indicate that they can play a crucial role in the restoration of oyster reefs in tropical seas.