Potency and specificity of amiloride and its analogues on branchial sodium fluxes in freshwater trout and goldfish.

There is a consensus that electroneutral Na+/H+ exchangers (NHEs) are important in branchial Na+ uptake in freshwater fish. There is also widespread belief, based on mammalian data, that EIPA [5-(N-ethyl-N-isopropyl)-amiloride]], and HMA [5-(N,N-hexamethylene)-amiloride)] are more potent and specific in blocking Na+ uptake than amiloride. We evaluated this idea by testing the three drugs at 10-7 to 10-4 M, i.e. 0.1 to 100 µM in two model species, rainbow trout (Oncorhynchus mykiss) and goldfish (Carassius auratus), using 22Na+ to measure unidirectional Na+ influx and efflux rates. In both species, the potency order for inhibiting unidirectional Na+ influx was HMA > amiloride > EIPA (IC50 values in the 10-70 µM range), very different from in mammals. At 100 µM, all three drugs inhibited Na+ influx by >90% in both species, except for amiloride in goldfish (65%). However, at 60-100 µM, all three drugs also stimulated unidirectional Na+ efflux rates, indicating non-specific effects. In trout, HMA and EIPA caused significant increases (2.1- to 2.3-fold) in efflux rates, whereas in goldfish, significant efflux elevations were greater (3.1- to 7.2-fold) with all three drugs. We conclude that the inhibitory potency profile established in mammals does not apply to the NHEs in fish gills, that non-specific effects on Na+ efflux rates are a serious concern, and that EIPA and HMA offer no clear benefits in terms of potency or specificity. Considering its much lower cost, we recommend amiloride as the drug of choice for in vivo experiments on freshwater fishes.

Use of antioxidants to retard aging of bitumen: A review.

Oxidative aging of bitumen is an inevitable and irreversible phenomenon. Exposure to detrimental factors such as sunlight, oxygen, and UV radiations accelerates the aging of bitumen and bituminous pavement. The aging process induces hardening and embrittlement in bitumen, leading to premature pavement failure. Therefore, for constructing sustainable long-lasting pavements anti-aging additives are used. Among the available additives, the use of antioxidants has emerged as a promising solution to mitigate the aging of bitumen. The current review aims to summarise the existing literature for a comprehensive understanding of the effectiveness of these additives as aging inhibitors. It provides an overview of the chemical pathway involved during bitumen oxidation and various quantification techniques to measure the effect of aging. This review also highlights the potential use of antioxidants in bitumen and elaborates on the working mechanism of different types of antioxidants to prevent bitumen aging. Further, the effect of modification in bitumen at micro, macro, and mixture levels are discussed. Additionally, cost analysis and future prospects on the use of antioxidants for bitumen are presented.

Transcriptomic responses of Mediterranean sponges upon encounter with symbiont microbial consortia.

BACKGROUND: Sponges (phylum Porifera) constantly interact with microbes. They graze on microbes from the water column by filter-feeding and they harbor symbiotic partners within their bodies. In experimental setups, sponges take up symbionts at lower rates compared with seawater microbes. This suggests that sponges have the capacity to differentiate between microbes and preferentially graze in non-symbiotic microbes, although the underlying mechanisms of discrimination are still poorly understood. Genomic studies showed that, compared to other animal groups, sponges present an extended repertoire of immune receptors, in particular NLRs, SRCRs, and GPCRs, and a handful of experiments showed that sponges regulate the expression of these receptors upon encounter with microbial elicitors. We hypothesize that sponges may rely on differential expression of their diverse repertoire of poriferan immune receptors to sense different microbial consortia while filter-feeding. To test this, we characterized the transcriptomic response of two sponge species, Aplysina aerophoba and Dysidea avara, upon incubation with microbial consortia extracted from A. aerophoba in comparison with incubation with seawater microbes. The sponges were sampled after 1 h, 3 h, and 5 h for RNA-Seq differential gene expression analysis. RESULTS: D. avara incubated with A. aerophoba-symbionts regulated the expression of genes related to immunity, ubiquitination, and signaling. Within the set of differentially-expressed immune genes we identified different families of Nucleotide Oligomerization Domain (NOD)-Like Receptors (NLRs). These results represent the first experimental evidence that different types of NLRs are involved in microbial discrimination in a sponge. In contrast, the transcriptomic response of A. aerophoba to its own symbionts involved comparatively fewer genes and lacked genes encoding for immune receptors. CONCLUSION: Our work suggests that: (i) the transcriptomic response of sponges upon microbial exposure may imply "fine-tuning" of baseline gene expression as a result of their interaction with microbes, (ii) the differential response of sponges to microbial encounters varied between the species, probably due to species-specific characteristics or related to host's traits, and (iii) immune receptors belonging to different families of NLR-like genes played a role in the differential response to microbes, whether symbionts or food bacteria. The regulation of these receptors in sponges provides further evidence of the potential role of NLRs in invertebrate host-microbe interactions. The study of sponge responses to microbes exemplifies how investigating different animal groups broadens our knowledge of the evolution of immune specificity and symbiosis.

Study on synergistic effect of multiple physical fields on hot mix asphalt during compaction process.

The multiple physical fields of hot mix asphalt (HMA) during the compaction process have a significant impact on the durability of asphalt pavement, and this research aimed to evaluate the synergistic effect of the HMA field compaction of multi-physical field evolution during the compaction process. First, the temperature field, structural layer thickness variation, and structural layer density variation were monitored during field compaction. Second, the evolution properties of compaction thickness were obtained under the synergistic influence of multi-physical fields by temperature field and compaction thickness. Finally, the evolution properties of compaction density were obtained under the synergistic influence of multi-physical fields based on the temperature field and structural layer density. The results showed that the field compaction process could be characterized by three stages under the synergistic impact of multi-physical fields. The cooling of the temperature field presents two-stage characteristics. There were cubic polynomial evolution properties for the temperature field versus time and the density versus temperature field. There was an exponential relationship between the thickness of the compacted layer and the number of mills. The aggregate particles showed different motion characteristics in the horizontal and vertical directions and vertical directions. The vertical displacement was larger than the horizontal displacement under the synergistic influence of multi-physical fields during the three stages of compaction. The migration and reorganization of aggregate particles affected the evolution of the multi-physics fields of the compaction process under the action of different compaction modes.

R-Napropamide Potentially Regulates Cadmium Accumulation in Arabidopsis Shoots through Transport Channel Modulation.

Heavy metal contamination in soils poses a significant environmental threat to human health. This study examines the effects of the chiral herbicide napropamide (NAP) on Arabidopsis thaliana, focusing on growth metrics and cadmium (Cd) accumulation. R-NAP does not adversely affect plant growth compared to the control, whereas S-NAP significantly reduces root length and fresh weight. Notably, R-NAP markedly increases Cd accumulation in the shoots, exceeding levels observed in the control and S-NAP. This increase coincides with reduced photosynthetic efficiency. Noninvasive electrode techniques reveal a higher net Cd absorption flux in the root mature zone under R-NAP than S-NAP, although similar to the control. Transcriptomic analysis highlights significant stereoisomer differences in Cd transporters, predominantly under R-NAP treatment. SEM and molecular docking simulations support that R-NAP primarily upregulates transporters such as HMA4. The results suggest careful management of herbicides like R-NAP in contaminated fields to avoid excessive heavy metal buildup in crops.

The Effect of Water during the Compaction Process on Surface Characteristics of HMA Pavement.

During the compaction process of HMA pavement, it is common to spray cold water on the wheel of a road roller to prevent the mixture from sticking to the wheel, which might deteriorate the bonding strength between the asphalt binder and aggregate, and consequently lead to surface polishing of the pavement. This paper aims to demonstrate whether the water used during the compaction process affects the surface performance of HMA pavement. In this study, the black pixel ratio and mass loss ratio were used to evaluate the water effect on the surface performance of asphalt pavement, considering the water consumption, molding temperature and long-term ageing process. The test results indicated that the water used during the compaction process would increase the risk of surface polishing of HMA pavement. This adverse effect became more significant if the HMA samples were prepared using greater water consumption, a greater molding temperature and a long-term ageing process. Moreover, there exists a certain correlation between the black pixel ratio and mass loss ratio, and their relationships were demonstrated by the experimental results in this study. It is recommended that further research concentrates on the influencing mechanism and the treatment strategy for the adverse effect caused by the water used during the compaction process. The use of more types of asphalt binders, aggregate and methodologies is also recommended in further studies.

Genome-wide identification of heavy-metal ATPases genes in Areca catechu: investigating their functionality under heavy metal exposure.

Heavy-metal ATPases (HMAs) play a vital role in plants, helping to transport heavy metal ions across cell membranes.However, insufficient data exists concerning HMAs genes within the Arecaceae family.In this study, 12 AcHMA genes were identified within the genome of Areca catechu, grouped into two main clusters based on their phylogenetic relationships.Genomic distribution analysis reveals that the AcHMA genes were unevenly distributed across six chromosomes. We further analyzed their physicochemical properties, collinearity, and gene structure.Furthermore, RNA-seq data analysis exhibited varied expressions in different tissues of A. catechu and found that AcHMA1, AcHMA2, and AcHMA7 were highly expressed in roots, leaves, pericarp, and male/female flowers. A total of six AcHMA candidate genes were selected based on gene expression patterns, and their expression in the roots and leaves was determined using RT-qPCR under heavy metal stress. Results showed that the expression levels of AcHMA1 and AcHMA3 genes were significantly up-regulated under Cd2 + and Zn2 + stress. Similarly, in response to Cu2+, the AcHMA5 and AcHMA8 revealed the highest expression in roots and leaves, respectively. In conclusion, this study will offer a foundation for exploring the role of the HMAs gene family in dealing with heavy metal stress conditions in A. catechu.

Cadmium and zinc accumulation and tolerance in two Egyptian cultivars (S53 and V120) of Helianthus annuus L. as potential phytoremediator.

One of the most important oil crops in the world, sunflower (Helianthus annuus L.), is recognized to help in soil phytoremediation. Heavy metal (HM) contamination is one of the most abiotic challenges that may affect the growth and productivity of such an important crop plant. We studied the influence of HM-contaminated soils on metal homeostasis and the potential hypertolerance mechanisms in two sunflower Egyptian cultivars (V120 and S53). Both cultivars accumulated significantly higher cadmium concentrations in their roots compared to their shoots during Cd and Zn/Cd treatments. Higher root concentrations of 121 mg g-1 dry weight (DW) and 125 mg g-1 DW were measured in V120 plants compared to relatively lower values of 111 mg g-1 DW and 105 mg g-1 DW in the roots of S53 plants, respectively. Cadmium contamination significantly upregulated the expression of heavy metal ATPases (HaHMA4) in the shoots of V120 plants. On the other hand, their roots displayed a notable expression of HaHMA3. This study indicates that V120 plants accumulated and sequestered Cd in their roots. Therefore, it is advised to cultivate the V120 cultivar in areas contaminated with heavy metals as it is a promising Cd phytoremediator.

The current study confirms and provides new insights into the low Cd and Zn concentration responses of two cultivars of Helianthus annuus as potential HM phytoremediators. HMA3 and HMA4 mediated both root sequestration and reduced root-to-shoot translocation rates. Moreover, high CAT and POX activities may reduce oxidative damage and enhance plant tolerance. The V120 showed higher levels of Cd accumulation in its roots and could be a promising cultivar for the phytoremediation of this heavy metal. This work recalls that Cd tolerance is a trait that may vary among cultivars of the same species and should be taken into consideration in the phytomanagement of heavy metals in contaminated soils.

Utilisation of Waste Sludge from Drinking Water Treatment as a Filler Material in Hot Mix Asphalt.

This research investigated the suitability of using sludge from the treatment of drinking water in hot mix asphalt (HMA) as a filler material. The storage and environmental impact of sludge is an enormous problem, especially for countries with large populations. Two different types of sludges, ferric chloride (FC) and aluminium sulphate (AS), were used as a filler material in HMA. The Hamburg Wheel Tracking (HWT) test, which correlates with rutting, and the Indirect Tensile Strength (ITS) test, which indicates the moisture sensitivity of HMA, were carried out at the optimum bitumen content of the mixes to investigate the usability of sludge in HMA. The test results indicate the usability of FC and AS in HMA compared to the reference mixes. However, the AS type of sludge has better rutting resistance than the FC type. Although the results support the usability of both sludges in HMA, it should be noted that the increased cost of the mix containing sludges due to the combustion process and the increased bitumen content during application should be considered.

Enhancing Fatigue Resistance in Asphalt Mixtures with a Novel Additive Derived from Recycled Polymeric Fibers from End-of-Life Tyres (ELTs).

Waste-tire textile fibers (WTTF) represent a challenge for the recycling industry since there are currently very few alternatives for their use. In this study, an evaluation of the effect of a new additive developed in two granular formats from WTTF on the fatigue behavior of asphalt mixtures was performed. For the first format of the WTTF-based additive, its effect was evaluated on hot-mix asphalt (HMA), while for the second format of the additive, the effects were evaluated on stone mastic asphalt (SMA). This second format represents an alternative that allows for the total replacement of the cellulose stabilizing additive used in the reference mix. The evaluation of fatigue damage in the mixes was performed using the four-point bending beam (4PB) test specified in European standard EN 12697-24. The test results show that the asphalt mixtures manufactured with WTTF-based additives exhibited a higher capacity to resist load cycles before failure compared to the reference mixtures. Likewise, once the asphalt mixtures were evaluated in a pavement structure by means of an empirical mechanistic analysis, the pavement structures composed of asphalt mixtures with WTTF-based additives showed significant improvements in their durability for the different load axes evaluated. For an average thickness of 15 cm of asphalt mix of a pavement-type structure, the use of the WTTF additive increases the durability of the structures by up to 129% and 112% compared to the HMA and SMA reference mixtures, respectively. These results show that both formats of the WTTF-based admixture improve the fatigue damage resistance of the HMA and SMA asphalt mixtures.

Increased expression of CD70 in relapsed acute myeloid leukemia after hypomethylating agents.

Acute myeloid leukemia (AML) is the most common acute leukemia in adults. While induction chemotherapy leads to remission in most patients, a significant number will experience relapse. Therefore, there is a need for novel therapies that can improve remission rates in patients with relapsed and refractory AML. CD70 is the natural ligand for CD27 (a member of the TNF superfamily) and appears to be a promising therapeutic target. Consequently, there is considerable interest in developing chimeric antigen receptor (CAR) T-cell therapy products that can specifically target CD70 in various neoplasms, including AML. In this study, we employed routine diagnostic techniques, such as immunohistochemistry and flow cytometry, to investigate the expression of CD70 in bone marrow samples from treatment-naïve and relapsed AML patients after hypomethylating agents (HMA). Also, we evaluated the impact of HMA on CD70 expression and examined CD70 expression in various leukemic cell subsets and normal hematopoietic progenitors.

Predictive value of DNA methylation patterns in AML patients treated with an azacytidine containing induction regimen.

BACKGROUND: Acute myeloid leukemia (AML) is a heterogeneous disease with a poor prognosis. Dysregulation of the epigenetic machinery is a significant contributor to disease development. Some AML patients benefit from treatment with hypomethylating agents (HMAs), but no predictive biomarkers for therapy response exist. Here, we investigated whether unbiased genome-wide assessment of pre-treatment DNA-methylation profiles in AML bone marrow blasts can help to identify patients who will achieve a remission after an azacytidine-containing induction regimen. RESULTS: A total of n = 155 patients with newly diagnosed AML treated in the AMLSG 12-09 trial were randomly assigned to a screening and a refinement and validation cohort. The cohorts were divided according to azacytidine-containing induction regimens and response status. Methylation status was assessed for 664,227 500-bp-regions using methyl-CpG immunoprecipitation-seq, resulting in 1755 differentially methylated regions (DMRs). Top regions were distilled and included genes such as WNT10A and GATA3. 80% of regions identified as a hit were represented on HumanMethlyation 450k Bead Chips. Quantitative methylation analysis confirmed 90% of these regions (36 of 40 DMRs). A classifier was trained using penalized logistic regression and fivefold cross validation containing 17 CpGs. Validation based on mass spectra generated by MALDI-TOF failed (AUC 0.59). However, discriminative ability was maintained by adding neighboring CpGs. A recomposed classifier with 12 CpGs resulted in an AUC of 0.77. When evaluated in the non-azacytidine containing group, the AUC was 0.76. CONCLUSIONS: Our analysis evaluated the value of a whole genome methyl-CpG screening assay for the identification of informative methylation changes. We also compared the informative content and discriminatory power of regions and single CpGs for predicting response to therapy. The relevance of the identified DMRs is supported by their association with key regulatory processes of oncogenic transformation and support the idea of relevant DMRs being enriched at distinct loci rather than evenly distribution across the genome. Predictive response to therapy could be established but lacked specificity for treatment with azacytidine. Our results suggest that a predictive epigenotype carries its methylation information at a complex, genome-wide level, that is confined to regions, rather than to single CpGs. With increasing application of combinatorial regimens, response prediction may become even more complicated.

The Molecular Docking of MAX Fungal Effectors with Plant HMA Domain-Binding Proteins.

Fungal effector proteins are important in mediating disease infections in agriculturally important crops. These secreted small proteins are known to interact with their respective host receptor binding partners in the host, either inside the cells or in the apoplastic space, depending on the localisation of the effector proteins. Consequently, it is important to understand the interactions between fungal effector proteins and their target host receptor binding partners, particularly since this can be used for the selection of potential plant resistance or susceptibility-related proteins that can be applied to the breeding of new cultivars with disease resistance. In this study, molecular docking simulations were used to characterise protein-protein interactions between effector and plant receptors. Benchmarking was undertaken using available experimental structures of effector-host receptor complexes to optimise simulation parameters, which were then used to predict the structures and mediating interactions of effector proteins with host receptor binding partners that have not yet been characterised experimentally. Rigid docking was applied for both the so-called bound and unbound docking of MAX effectors with plant HMA domain protein partners. All bound complexes used for benchmarking were correctly predicted, with 84% being ranked as the top docking pose using the ZDOCK scoring function. In the case of unbound complexes, a minimum of 95% of known residues were predicted to be part of the interacting interface on the host receptor binding partner, and at least 87% of known residues were predicted to be part of the interacting interface on the effector protein. Hydrophobic interactions were found to dominate the formation of effector-plant protein complexes. An optimised set of docking parameters based on the use of ZDOCK and ZRANK scoring functions were established to enable the prediction of near-native docking poses involving different binding interfaces on plant HMA domain proteins. Whilst this study was limited by the availability of the experimentally determined complexed structures of effectors and host receptor binding partners, we demonstrated the potential of molecular docking simulations to predict the likely interactions between effectors and their respective host receptor binding partners. This computational approach may accelerate the process of the discovery of putative interacting plant partners of effector proteins and contribute to effector-assisted marker discovery, thereby supporting the breeding of disease-resistant crops.

A Genome-Wide Identification and Comparative Analysis of the Heavy-Metal-Associated Gene Family in Cucurbitaceae Species and Their Role in Cucurbita pepo under Arsenic Stress.

The heavy-metal-associated (HMA) proteins are a class of PB1-type ATPases related to the intracellular transport and detoxification of metals. However, due to a lack of information regarding the HMA gene family in the Cucurbitaceae family, a comprehensive genome-wide analysis of the HMA family was performed in ten Cucurbitaceae species: Citrullus amarus, Citrullus colocynthis, Citrullus lanatus, Citrullus mucosospermus, Cucumis melo, Cucumis sativus, Cucurbita maxima, Cucurbita moschata, Cucurbita pepo, and Legenaria siceraria. We identified 103 Cucurbit HMA proteins with various members, ranging from 8 (Legenaria siceraria) to 14 (Cucurbita pepo) across species. The phylogenetic and structural analysis confirmed that the Cucurbitaceae HMA protein family could be further classified into two major clades: Zn/Co/Cd/Pb and Cu/Ag. The GO-annotation-based subcellular localization analysis predicted that all HMA gene family members were localized on membranes. Moreover, the analysis of conserved motifs and gene structure (intron/exon) revealed the functional divergence between clades. The interspecies microsynteny analysis demonstrated that maximum orthologous genes were found between species of the Citrullus genera. Finally, nine candidate HMA genes were selected, and their expression analysis was carried out via qRT-PCR in root, leaf, flower, and fruit tissues of C. pepo under arsenic stress. The expression pattern of the CpeHMA genes showed a distinct pattern of expression in root and shoot tissues, with a remarkable expression of CpeHMA6 and CpeHMA3 genes from the Cu/Ag clade. Overall, this study provides insights into the functional analysis of the HMA gene family in Cucurbitaceae species and lays down the basic knowledge to explore the role and mechanism of the HMA gene family to cope with arsenic stress conditions.

Mitotic perturbation is a key mechanism of action of decitabine in myeloid tumor treatment.

Decitabine (DAC) is clinically used to treat myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML). Our genome-wide CRISPR-dCas9 activation screen using MDS-derived AML cells indicates that mitotic regulation is critical for DAC resistance. DAC strongly induces abnormal mitosis (abscission failure or tripolar mitosis) in human myeloid tumors at clinical concentrations, especially in those with TP53 mutations or antecedent hematological disorders. This DAC-induced mitotic disruption and apoptosis are significantly attenuated in DNMT1-depleted cells. In contrast, overexpression of Dnmt1, but not the catalytically inactive mutant, enhances DAC-induced mitotic defects in myeloid tumors. We also demonstrate that DAC-induced mitotic disruption is enhanced by pharmacological inhibition of the ATR-CLSPN-CHK1 pathway. These data challenge the current assumption that DAC inhibits leukemogenesis through DNMT1 inhibition and subsequent DNA hypomethylation and highlight the potent activity of DAC to disrupt mitosis through aberrant DNMT1-DNA covalent bonds.

ZmHMA3, a Member of the Heavy-Metal-Transporting ATPase Family, Regulates Cd and Zn Tolerance in Maize.

The pollution of heavy metals is extremely serious in China, including zinc (Zn), copper (Cu), lead (Pb), and cadmium (Cd). Heavy-metal-transporting ATPase (HMA) belongs to a subfamily of the P-ATPase family, which absorbs and transports Zn, Cu, Pb, and Cd in plants. Here, we describe a ZmHMA-encoding HMA family protein that positively regulates Cd and Zn tolerance. The real-time fluorescence quantification (RT-PCR) results revealed that ZmHMA3 had a high expression in B73, and the expression of ZmHMA3 was sensitive to Cd in yeast cells, which was related to Cd accumulation in yeast. Additionally, the Arabidopsis thaliana homologous mutants of AtHMA2 showed Cd sensitivity compared with WT. The overexpressing ZmHMA3 plants showed higher tolerance under Cd and Zn stresses than the wild type. The overexpression of ZmHMA3 led to higher Cd and Zn accumulation in tissues based on the subcellular distribution analysis. We propose that ZmHMA3 improves maize tolerance to Cd and Zn stresses by absorbing and transporting Cd and Zn ions. This study elucidates the gene function of the ZmHMA3 response to Cd and Zn stress and provides a reference for improving the characteristics of heavy metals enrichment in existing maize varieties and the plant remediation technology of heavy-metal-contaminated soil.

Organosilicon Compounds in Hot-Melt Adhesive Technologies.

Hot-melt adhesives (HMAs) are thermoplastic materials that can bond various substrates by solidifying rapidly upon cooling from the molten state, and their modification with organosilicon compounds can result in crosslinking behavior, characteristic of gels. Organosilicon compounds are hybrid molecules that have both inorganic and organic components and can enhance the properties and performance of HMAs. The gel aspect of HMA with and without organosilicon modifiers can be considered in organosilicon-modified systems, the modifiers are often either sol-gel condensation products or their mechanism of action on the adherent surface can be considered of sol-gel type. The purpose of this manuscript is to present the current state of the art on the formulation, characterization, and application of HMAs and optimize their performance with organosilicon compounds for application in various industries such as automotive, construction, and photovoltaics. This review covers articles published within the period of 2018-2022. The article is divided into sections, in which information about hot-melt adhesives is described at the beginning. The following part of the presented review focuses on the composition of hot-melt adhesives, which takes into account the use of organosilicon compounds. The last part of this review outlines the future trends in hot-melt adhesives.

Browsers or Grazers? New Insights into Feral Burro Diet Using a Non-Invasive Sampling and Plant DNA Metabarcoding Approach.

Ungulates play a large role in shaping ecosystems and communities by influencing plant composition, structure, and productivity. We investigated the summer diets of feral burros in two ecosystems in which they are found in the United States: a subtropical desert in Arizona and a temperate juniper shrubland in Utah. Between 24 June and 16 July of 2019, we gathered 50 burro fecal samples from each location and used plant DNA metabarcoding to determine the burros' diets. We found that during our sampling period the burros in the Sonoran Desert consumed a higher proportion of woody browse and had a narrower dietary niche breadth and lower degree of diet diversity compared to the burros in the juniper shrubland ecosystem, where the burros consumed higher proportions of graminoids and forbs and had a higher diet diversity index and broader dietary niche breadth. The burros in the Sonoran Desert relied primarily on Prosopis spp. (mesquite) and Poaceae grasses, whereas the burros in the juniper shrubland relied on a wider variety of forb and grass species, likely due to the greater variability in the forage species temporally and spatially available in that temperate ecosystem. We found that feral burros are highly adaptable with respect to diet and appear to be employing a mixed feeding strategy, similar to their ancestor, the African wild ass, to meet their nutritional needs in whichever ecosystem they are found.

Structural mechanism of heavy metal-associated integrated domain engineering of paired nucleotide-binding and leucine-rich repeat proteins in rice.

Plant nucleotide-binding and leucine-rich repeat (NLR) proteins are immune sensors that detect pathogen effectors and initiate a strong immune response. In many cases, single NLR proteins are sufficient for both effector recognition and signaling activation. These proteins possess a conserved architecture, including a C-terminal leucine-rich repeat (LRR) domain, a central nucleotide-binding (NB) domain, and a variable N-terminal domain. Nevertheless, many paired NLRs linked in a head-to-head configuration have now been identified. The ones carrying integrated domains (IDs) can recognize pathogen effector proteins by various modes; these are known as sensor NLR (sNLR) proteins. Structural and biochemical studies have provided insights into the molecular basis of heavy metal-associated IDs (HMA IDs) from paired NLRs in rice and revealed the co-evolution between pathogens and hosts by combining naturally occurring favorable interactions across diverse interfaces. Focusing on structural and molecular models, here we highlight advances in structure-guided engineering to expand and enhance the response profile of paired NLR-HMA IDs in rice to variants of the rice blast pathogen MAX-effectors (Magnaporthe oryzae AVRs and ToxB-like). These results demonstrate that the HMA IDs-based design of rice materials with broad and enhanced resistance profiles possesses great application potential but also face considerable challenges.