In-depth analysis of isochorismate synthase-derived metabolism in plant immunity: identification of meta-substituted benzoates and salicyloyl-malate.

Isochorismate-derived metabolism enables biosynthesis of the plant defence hormone salicylic acid (SA) and its derivatives. In Arabidopsis thaliana, the stress-induced accumulation of SA depends on ISOCHORISMATE SYNTHASE1 (ICS1), and also requires the presumed isochorismate transporter ENHANCED DISEASE SUSCEPTIBILITY5 (EDS5) and the GH3 enzyme avrPphB SUSCEPTIBLE3 (PBS3). By comparative metabolite and structural analyses, we identified several hitherto unreported ICS1- and EDS5-dependent, biotic stress-inducible Arabidopsis metabolites. These involve meta-substituted SA derivatives (5-formyl-SA, 5-carboxy-SA, 5-carboxymethyl-SA), their benzoic acid (BA) analogues (3-formyl-BA, 3-carboxy-BA, 3-carboxymethyl-BA) and, besides the previously detected salicyloyl-aspartate (SA-Asp), the ester conjugate salicyloyl-malate (SA-Mal). SA functions as a biosynthetic precursor for SA-Mal and SA-Asp, but not for the meta-substituted SA- and BA-derivatives, which accumulate to moderate levels at later stages of bacterial infection. Interestingly, Arabidopsis leaves possess oxidising activity to effectively convert meta-formyl- into meta-carboxy-SA/BAs. In contrast to SA, exogenously applied meta-substituted SA/BA-derivatives and SA-Mal exert moderate impact on plant immunity and defence-related gene expression. While the isochorismate-derived metabolites are negatively regulated by the SA receptor NON-EXPRESSOR OF PR GENES1, SA conjugates (SA-Mal, SA-Asp, SA-glucose conjugates) and meta-substituted SA/BA-derivatives are oppositely affected by PBS3. Notably, our data indicate a PBS3-independent path to isochorismate-derived SA at later stages of bacterial infection, which does not considerably impact immune-related characteristics. Moreover, our results argue against a previously proposed role of EDS5 in the biosynthesis of the immune signal N-hydroxypipecolic acid and associated transport processes. We propose a significantly extended biochemical scheme of plant isochorismate metabolism that involves an alternative generation mode for benzoate- and salicylate-derivatives.

Sex-dependent effects of monomeric α-synuclein on calcium and cell death of lateral hypothalamic mouse neurons are altered by orexin.

Parkinson's Disease (PD) patients experience sleeping disorders in addition to the disease-defining symptomology of movement dysfunctions. The prevalence of PD is sex-based and presence of sleeping disorders in PD also shows sex bias with a stronger phenotype in males. In addition to loss of dopamine-containing neurons in the striatum, arousal-related, orexin-containing neurons in the lateral hypothalamus (LH) are lost in PD, which could contribute to state-related disorders. As orexin has been shown to be involved in sleeping disorders and to have neuroprotective effects, we asked whether orexin could protect sleep-related LH neurons from damage putatively from the protein α-synuclein (α-syn), which is found at high levels in the PD brain and that we have shown is associated with putatively excitotoxic rises in intracellular calcium in brainstem sleep-controlling nuclei, especially in males. Accordingly, we monitored intracellular calcium transients induced by α-syn and whether concurrent exposure to orexin affected those transients in LH cells of the mouse brain slice using calcium imaging. Further, we used an assay of cell death to determine whether LH cell viability was influenced when α-syn and orexin were co-applied when compared to exposure to α-syn alone. We found that excitatory calcium events induced by α-syn were reduced in amplitude and frequency when orexin was co-applied, and when data were evaluated by sex, this effect was found to be greater in females. In addition, α-syn exposure was associated with cell death that was higher in males, and interestingly, reduced cell death was noted when orexin was present, which did not show a sex bias. We interpret our findings to indicate that orexin is protective to α-syn-mediated damage to hypothalamic neurons, and the actions of orexin on α-syn-induced cellular effects differ between sexes, which could underlie sex-based differences in sleeping disorders in PD.

Subcellular localization requirements and specificities for plant immune receptor Toll-interleukin-1 receptor signaling.

Recent work shed light on how plant intracellular immune receptors of the nucleotide-binding leucine-rich repeat (NLR) family are activated upon pathogen effector recognition to trigger immune responses. Activation of Toll-interleukin-1 receptor (TIR) domain-containing NLRs (TNLs) induces receptor oligomerization and close proximity of the TIR domain, which is required for TIR enzymatic activity. TIR-catalyzed small signaling molecules bind to EDS1 family heterodimers and subsequently activate downstream helper NLRs, which function as Ca2+ permeable channel to activate immune responses eventually leading to cell death. Subcellular localization requirements of TNLs and signaling partners are not well understood, although they are required to understand fully the mechanisms underlying NLR early signaling. TNLs show diverse subcellular localization while EDS1 shows nucleocytosolic localization. Here, we studied the impact of TIR and EDS1 mislocalization on the signaling activation of different TNLs. In Nicotiana benthamiana, our results suggest that close proximity of TIR domains isolated from flax L6 and Arabidopsis RPS4 and SNC1 TNLs drives signaling activation from different cell compartments. Nevertheless, both Golgi-membrane anchored L6 and nucleocytosolic RPS4 have the same requirements for EDS1 subcellular localization in Arabidopsis thaliana. By using mislocalized variants of EDS1, we found that autoimmune L6 and RPS4 TIR domain can induce seedling cell death when EDS1 is present in the cytosol. However, when EDS1 is restricted to the nucleus, both induce a stunting phenotype but no cell death. Our data point out the importance of thoroughly investigating the dynamics of TNLs and signaling partners subcellular localization to understand TNL signaling fully.

Host-Specific and Homologous Pairs of Melampsora larici-populina Effectors Unveil Novel Nicotiana benthamiana Stromule Induction Factors.

The poplar rust fungus Melampsora larici-populina is part of one of the most devastating group of fungi (Pucciniales) and causes important economic losses to the poplar industry. Because M. larici-populina is a heteroecious obligate biotroph, its spread depends on its ability to carry out its reproductive cycle through larch and then poplar parasitism. Genomic approaches have identified more than 1,000 candidate secreted effector proteins (CSEPs) from the predicted secretome of M. larici-populina that are potentially implicated in the infection process. In this study, we selected CSEP pairs (and one triplet) among CSEP gene families that share high sequence homology but display specific gene expression profiles among the two distinct hosts. We determined their subcellular localization by confocal microscopy through expression in the heterologous plant system Nicotiana benthamiana. Five out of nine showed partial or complete chloroplastic localization. We also screened for potential protein interactors from larch and poplar by yeast two-hybrid assays. One pair of CSEPs and the triplet shared common interactors, whereas the members of the two other pairs did not have common targets from either host. Finally, stromule induction quantification revealed that two pairs and the triplet of CSEPs induced stromules when transiently expressed in N. benthamiana. The use of N. benthamiana eds1 and nrg1 knockout lines showed that CSEPs can induce stromules through an eds1-independent mechanism. However, CSEP homologs shared the same impact on stromule induction and contributed to discovering a new stromule induction cascade that can be partially and/or fully independent of eds1. [Formula: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Clinical-Genomic Analysis of 1261 Patients with Ehlers-Danlos Syndrome Outlines an Articulo-Autonomic Gene Network (Entome).

Systematic evaluation of 80 history and 40 history findings diagnosed 1261 patients with Ehlers-Danlos syndrome (EDS) by direct or online interaction, and 60 key findings were selected for their relation to clinical mechanisms and/or management. Genomic testing results in 566 of these patients supported EDS relevance by their differences from those in 82 developmental disability patients and by their association with general rather than type-specific EDS findings. The 437 nuclear and 79 mitochondrial DNA changes included 71 impacting joint matrix (49 COL5), 39 bone (30 COL1/2/9/11), 22 vessel (12 COL3/8VWF), 43 vessel-heart (17FBN1/11TGFB/BR), 59 muscle (28 COL6/12), 56 neural (16 SCN9A/10A/11A), and 74 autonomic (13 POLG/25porphyria related). These genes were distributed over all chromosomes but the Y, a network analogized to an 'entome' where DNA change disrupts truncal mechanisms (skin constraint, neuromuscular support, joint vessel flexibility) and produces a mirroring cascade of articular and autonomic symptoms. The implied sequences of genes from nodal proteins to hypermobility to branching tissue laxity or dysautonomia symptoms would be ideal for large language/artificial intelligence analyses.

Differences and similarities in biophysical and biological characteristics between U87 MG glioblastoma and astrocyte cells.

Current cancer studies focus on molecular-targeting diagnostics and interactions with surroundings; however, there are still gaps in characterization based on topological differences and elemental composition. Glioblastoma (GBM cells; GBMCs) is an astrocytic aggressive brain tumor. At the molecular level, GBMCs and astrocytes may differ, and cell elemental/topological analysis is critical for identifying potential new cancer targets. Here, we used U87 MG cells for GBMCS. U87 MG cell lines, which are frequently used in glioblastoma research, are an important tool for studying the various features and underlying mechanisms of this aggressive brain tumor. For the first time, atomic force microscopy (AFM), scanning electron microscopy (SEM) accompanied by energy-dispersive X-ray spectroscopy (EDS), and X-ray photoelectron spectroscopy (XPS) are used to report the topology and chemistry of cancer (U87 MG) and healthy (SVG p12) cells. In addition, F-actin staining and cytoskeleton-based gene expression analyses were performed. The degree of gene expression for genes related to the cytoskeleton was similar; however, the intensity of F-actin, anisotropy values, and invasion-related genes were different. Morphologically, GBMCs were longer and narrower while astrocytes were shorter and more disseminated based on AFM. Furthermore, the roughness values of these cells differed slightly between the two call types. In contrast to the rougher astrocyte surfaces in the lamellipodial area, SEM-EDS analysis showed that elongated GBMCs displayed filopodial protrusions. Our investigation provides considerable further insight into rapid cancer cell characterization in terms of a combinatorial spectroscopic and microscopic approach.

Arabidopsis PHYTOALEXIN DEFICIENT 4 promotes the maturation and nuclear accumulation of immune-related cysteine protease RD19.

Arabidopsis PHYTOALEXIN DEFICIENT 4 (PAD4) has an essential role in pathogen resistance as a heterodimer with ENHANCED DISEASE SUSCEPTIBILITY 1 (EDS1). Here we investigated an additional PAD4 role in which it associates with and promotes the maturation of the immune-related cysteine protease RESPONSIVE TO DEHYDRATION 19 (RD19). We found that RD19 and its paralog RD19c promoted EDS1- and PAD4-mediated effector-triggered immunity to an avirulent Pseudomonas syringae strain, DC3000, expressing the effector AvrRps4 and basal immunity against the fungal pathogen Golovinomyces cichoracearum. Overexpression of RD19, but not RD19 protease-inactive catalytic mutants, in Arabidopsis transgenic lines caused EDS1- and PAD4-dependent autoimmunity and enhanced pathogen resistance. In these lines, RD19 maturation to a pro-form required its catalytic residues, suggesting that RD19 undergoes auto-processing. In transient assays, PAD4 interacted preferentially with the RD19 pro-protease and promoted its nuclear accumulation in leaf cells. Our results lead us to propose a model for PAD4-stimulated defense potentiation. PAD4 promotes maturation and nuclear accumulation of processed RD19, and RD19 then stimulates EDS1-PAD4 dimer activity to confer pathogen resistance. This study highlights potentially important additional PAD4 functions that eventually converge on canonical EDS1-PAD4 dimer signaling in plant immunity.

Leaf abaxial immunity to powdery mildew in Arabidopsis is conferred by multiple defense mechanisms.

Powdery mildew fungi are obligate biotrophic pathogens that only invade plant epidermal cells. There are two epidermal surfaces in every plant leaf: the adaxial (upper) side and the abaxial (lower) side. While both leaf surfaces can be susceptible to adapted powdery mildew fungi in many plant species, there have been observations of leaf abaxial immunity in some plant species including Arabidopsis. The genetic basis of such leaf abaxial immunity remains unknown. In this study, we tested a series of Arabidopsis mutants defective in one or more known defense pathways with the adapted powdery mildew isolate Golovinomyces cichoracearum UCSC1. We found that leaf abaxial immunity was significantly compromised in mutants impaired for both the EDS1/PAD4- and PEN2/PEN3-dependent defenses. Consistently, expression of EDS1-yellow fluorescent protein and PEN2-green fluorescent protein fusions from their respective native promoters in the respective eds1-2 and pen2-1 mutant backgrounds was higher in the abaxial epidermal cells than in the adaxial epidermal cells. Altogether, our results indicate that leaf abaxial immunity against powdery mildew in Arabidopsis is at least partially due to enhanced EDS1/PAD4- and PEN2/PEN3-dependent defenses. Such transcriptionally pre-programmed defense mechanisms may underlie leaf abaxial immunity in other plant species such as hemp and may be exploited for engineering adaxial immunity against powdery mildew fungi in crop plants.

A critical catalogue of SEM-EDS multispectral maps analysis methods and their application to hydrated cementitious materials.

Many methods have been proposed to analyse SEM-EDS hypermaps of hydrated cementitious materials but none can fit all purposes. In this presentation, we review existing methods for phase identification, stoichiometry quantification, and microstructure quantification in cementitious materials and related materials. We first discuss the unique contribution of SEM-EDS with respect to the outstanding scientific challenges towards sustainable construction materials. We then compare the SEM-EDS and image analysis techniques which contribute to answering these challenges. Convergence and divergence in current methods and workflows, and knowledge gaps are highlighted in terms of the specificities of the material (phase assemblage complexity, grain sizes, mixtures, etc.). The discussion is weighted by the required expert knowledge for the sample preparation, microscope operation, and data analysis. We conclude by discussing how microscopy and image analysis integrate into the overall experimental toolkit to investigate and improve cementitious materials.

Assessment of microplastics in coastal ecosystem of Bangladesh.

Microplastics (MPs) pose one of the major environmental threats to marine organisms and ecosystems on a global scale. The present study investigated MPs in surface water, beach sediments, and fish in two coastal areas of Bangladesh namely Cox's Bazar and Kuakata. The MPs were identified and characterized using three different techniques, including the binocular microscope, the ATR-FTIR (Attenuated Total Reflectance-Fourier Transform Infrared Spectroscopy), and SEM-EDS (Scanning Electron Microscopy- Energy Dispersive Spectroscopy). The number of MPs in seawater was 10.1 ± 3.10 and 8.52 ± 3.92 items/100 L and in beach sediment, 13.2 ± 3.68 and 9.48 ± 3.63 items/100 g in Cox's Bazar and Kuakata, respectively. In fish samples, the abundance of MPs was 7.82 ± 1.28 and 6.82 ± 1.87 items/individual species of Cox's Bazar and Kuakata, respectively, where the highest quantities of MP were found in Euthynnus affinisand Sillago sihama and the lowest in Terapon jarbua and Pampus chinensisin Cox's Bazar and Kuakata, respectively. The number of MPs in GITs (Gastrointestinal tracts) was 1.63 ± 0.991 and 1.25 ± 0.546 items/g GIT and in BW (Body Weight) were 0.042 ± 0.014 and 0.037 ± 0.014 items/g BW in Cox's Bazar and Kuakata, respectively. There revealed a positive correlation between MP abundance and GIT weight and body weight in fish species. MPs were predominantly fiber-shaped, white/transparent, and small size. The most common MP polymers were polyethylene and polypropylene. SEM images of MPs demonstrate surface roughness, cracks, mechanical weathering and oxidative weathering, demonstrating their ongoing environmental exposure. The EDS spectrum unearthed that the MPs contained several elements (C, N, O, Na, Al, Fe, and Si). Findings from this study might be useful in coastal plastic particle management and to mitigate the potential risks associated with them.

De-MA.ch: A Web Database for Electron and X-ray Microanalysis to Assist Electron Microscope Lab Managers and Users.

Lab managers and users of scanning electron microscope or electron probe microanalyzer facilities aiming for qualitative or quantitative X-ray analyses require comprehensive, yet flexible documentation structures for their daily work and available reference material, with a complete X-ray data library, a repository of energy- and wavelength-dispersive spectra, and an instrument scheduling mechanism. An online multilaboratory database system available at https://de-ma.ch is presented with the primary goals of providing information on microanalytical reference materials, analytical setups, characteristic X-ray data, and for managing reservation and training requests. This website is designed for multiuser facilities, where experience ranges from beginners to expert users. Registered users will find these tools useful in developing and maintaining high-quality, reproducible, and efficient analyses, whereas lab managers will keep records of their microanalytical reference material database and analytical protocols. The database also serves an educational purpose by (a) providing information on reference materials, (b) encouraging students to select appropriate X-ray lines to analyze, (c) providing analytical setups for point analysis or mapping, (d) identifying unknown X-ray lines, (e) displaying energy- or wavelength-dispersive spectra, and (f) recalculating mineral formula from quantitative wt% analysis results, based on a number of oxygen atoms or cations.

Atomic-Scale Characterization of Dilute Dopants in Topological Insulators via STEM-EDS Using Registration and Cell Averaging Techniques.

Magnetic dopants in three-dimensional topological insulators (TIs) offer a promising avenue for realizing the quantum anomalous Hall effect (QAHE) without the necessity for an external magnetic field. Understanding the relationship between site occupancy of magnetic dopant elements and their effect on macroscopic property is crucial for controlling the QAHE. By combining atomic-scale energy-dispersive X-ray spectroscopy (EDS) maps obtained by aberration-corrected scanning transmission electron microscopy (AC-STEM) and novel data processing methodologies, including semi-automatic lattice averaging and frame registration, we have determined the substitutional sites of Mn atoms within the 1.2% Mn-doped Sb2Te3 crystal. More importantly, the methodology developed in this study extends beyond Mn-doped Sb2Te3 to other quantum materials, traditional semiconductors, and even electron irradiation sensitive materials.

In vitro studies the influence of Nd: YAG laser on dental enamels.

The present work aimed at assessing chemical, topographical, and morphological changes induced by Nd : YAG laser treatment of dental enamels by means of energy dispersive X-ray spectroscopy (EDS), atomic force microscopy (AFM), and scanning electron microscopy (SEM). Fifteen human enamel specimens were obtained, three of samples were kept untreated as a control while the others twelve samples were equally divided into four groups where each group have a three samples according to treating approach as: G1:(untreated);G2: (treated with Nd:YAG laser, 100 mJ/pulse,10 Hz/1064nm); G3(treated with Nd:YAG laser, 500 mJ/pulse, 10 Hz/1064nm); G4(treated with Nd:YAG laser 1000 mJ/pulse, 10 Hz/1064nm), and finally G5(treated with Nd:YAG laser, 1000 mJ/pulse, 10 Hz/532nm) respectively. Beside many craters and cracks, the AFM results showed fractures with depths of 19.23 nm, 174.7 nm, 216.9 nm, 207.4 nm and 156.5 nm and width of 559.2 nm, 833.4 nm, 1115 nm, 695.0 nm, and 5142 nm for all Groups respectively. The highest surface roughness was found in G5 with 111.4 nm while the lowest surface roughness was found in G1 to be 14.3 nm. The inside surface of the fissures was also rough. The SEM micrographs revealed modifications to the morphology. EDS was used to measure the phosphorous (P), calcium (Ca), oxygen (O), and carbon (C) percentages presented in crater areas and their surroundings, Ca, P, O, and C levels were observed to vary significantly at the crater and its rim, a lower percentage of C wt% were realized corresponding to laser treatment of 1000 mJ/Pulse laser energy. However, it was not feasible to recognize a specific chemical arrangement in the craters. It is also concluded that the higher depth and particular edge of ablated part when teeth were irradiated by laser with 1000 mJ/10Hz/1064nm.

Structure Engineering of Ni/SiO2 Vegetable Oil Hydrogenation Catalyst via CeO2.

Inspired by our finding that metallic Ni particles could be uniformly distributed on a reduced CeO2 surface and stabilized on Ce3+ sites, we suppose a possible improvement in the activity and selectivity of the MgNi/SiO2 vegetable oil hydrogenation catalyst by increasing the surface metal Ni availability via modification by ceria. The proposed approach involved the addition of a CeO2 modifier to the SiO2 carrier and as a catalyst component. Evaluation of the structure, reducibility, and surface and electronic states of the CeO2-doped MgNi/SiO2 catalyst was performed by means of the Powder X-ray diffraction (PXRD), Scanning electron microscopy-energy dispersive spectroscopy (SEM-EDS), and X-ray photoelectron spectroscopy (XPS) combined with High-resolution transmission electron microscopy (HRTEM), Temperature-programmed reduction with hydrogen (H2-TPR), and H2-chemisortion techniques. So far, no studies related to this approach of designing Ni/SiO2 catalysts for the partial hydrogenation of vegetable oil have been reported. The added ceria impact was elucidated by comparing fatty acid compositions obtained by the catalysts at an iodine value of 80. In summary, tuning the hydrogenation performance of Ni-based catalysts can be achieved by structural reconstruction using 1 wt.% CeO2. The introduction mode changed the selectivity towards C18:1-cis and C18:0 fatty acids by applying ceria as a carrier modifier, while hydrogenation activity was improved upon ceria operation as the catalyst dopant.

Exploring the Effect of V2O5 and Nb2O5 Content on the Structural, Thermal, and Electrical Characteristics of Sodium Phosphate Glasses and Glass-Ceramics.

Na-V-P-Nb-based materials have gained substantial recognition as cathode materials in high-rate sodium-ion batteries due to their unique properties and compositions, comprising both alkali and transition metal ions, which allow them to exhibit a mixed ionic-polaronic conduction mechanism. In this study, the impact of introducing two transition metal oxides, V2O5 and Nb2O5, on the thermal, (micro)structural, and electrical properties of the 35Na2O-25V2O5-(40 - x)P2O5 - xNb2O5 system is examined. The starting glass shows the highest values of DC conductivity, σDC, reaching 1.45 × 10-8 Ω-1 cm-1 at 303 K, along with a glass transition temperature, Tg, of 371 °C. The incorporation of Nb2O5 influences both σDC and Tg, resulting in non-linear trends, with the lowest values observed for the glass with x = 20 mol%. Electron paramagnetic resonance measurements and vibrational spectroscopy results suggest that the observed non-monotonic trend in σDC arises from a diminishing contribution of polaronic conductivity due to the decrease in the relative number of V4+ ions and the introduction of Nb2O5, which disrupts the predominantly mixed vanadate-phosphate network within the starting glasses, consequently impeding polaronic transport. The mechanism of electrical transport is investigated using the model-free Summerfield scaling procedure, revealing the presence of mixed ionic-polaronic conductivity in glasses where x < 10 mol%, whereas for x ≥ 10 mol%, the ionic conductivity mechanism becomes prominent. To assess the impact of the V2O5 content on the electrical transport mechanism, a comparative analysis of two analogue series with varying V2O5 content (10 and 25 mol%) is conducted to evaluate the extent of its polaronic contribution.

A kinetic modelling approach to explore mechanism of Cr(VI) detoxification by a novel strain Pseudochrobactrum saccharolyticum NBRI-CRB 13 using response surface methodology.

A novel Pseudochrobactrum saccharolyticum strain NBRI-CRB 13, isolated from tannery sludge, was studied to grow up to 500 mgL-1 of Cr(VI) and showed Cr(VI) detoxification by reducing > 90% of Cr(VI) at different concentrations 25, 50 and 100 mgL-1. Kinetic studies showed that first-order models were fitted (R2 = 0.998) to the time-dependent Cr(VI) reduction with degradation rate constant (k) (1.03-0.429 h-1). Cr(VI) detoxification was primarily related to the extracellular fraction of microbial cells, which showed a maximum extracellular reductase enzyme activity led to 94.6% reduction of Cr(VI). Moreover, the strain showed maximum extracellular polymeric substances (EPS) production at 100 mgL-1 Cr(VI), which is presumably the reason for Cr(VI) removal as EPS serves as the metal binding site for Cr(VI) ions. Further, an optimization study using Box-Behnken design was conducted considering parameters viz., pH, temperature, and initial concentration of Cr(VI). The maximum percent reduction of Cr(VI) was obtained at pH 6.5, temperature 30 °C with 62.5 mgL-1Cr(VI) concentration. Further, the Cr(VI) reduction and adsorption ability of strain P. saccharolyticum NBRI-CRB13 were confirmed by SEM-EDS, FTIR, and XRD analyses. FTIR analysis confirmed the presence of functional groups (-OH, -COOH, -PO4) on bacterial cell walls, which were more likely to interact with positively charged chromium ions. The study elucidated the reduction of Cr(VI) by the novel bacterium within 24 h using the response surface methodology approach and advocated its application in real-time situations.

In vitro remineralization of adjacent interproximal enamel carious lesions in primary molars using a bioactive bulk-fill composite.

BACKGROUND: Surface remineralization is recommended for the management of active non-cavitated interproximal carious lesions in primary teeth. According to the American Academy of Pediatric Dentistry, a recently recognized category of materials called bioactive restorative materials can be used for remineralization. This study aimed to evaluate the release of fluoride (F), calcium (Ca) and phosphate (P) ions from Predicta® Bioactive Bulk-fill composite compared with EQUIA Forte® and Filtek™ Z350 and to determine the remineralization effect of these 3 restorative materials on adjacent initial interproximal enamel carious lesions. METHODS: The release of F, Ca and P ions from 3 groups ((n = 10/group) (Group 1- Predicta®, Group 2- EQUIA Forte® and Group 3- Filtek™ Z350)) was determined at 1st, 4th, 7th and 14th days. After creating artificial carious lesions, human enamel samples were randomly assigned into 3 groups (n = 13/group) which were placed in contact with occluso-proximal restorative materials and exposed to a 14-day pH cycling period. Surface microhardness was determined using a Knoop microhardness assay at baseline, after artificial carious lesions formation and after pH cycling. The difference in the percentage of surface microhardness recovery (%SMHR) among groups was compared. Mineral deposition was analyzed with energy-dispersive x-ray spectroscopy (EDS) and the enamel surface morphology was evaluated with scanning electron microscopy (SEM). Kruskal-Wallis's test with Dunn's post hoc test and one-way ANOVA with Tukey's post hoc test were used for data analysis. RESULTS: EQUIA Forte® released the highest cumulative amount of F and P ions, followed by Predicta® and Filtek™ Z350. Predicta® released higher amount of Ca ions than EQUIA Forte® and Filtek™ Z350. Predicta® demonstrated the highest %SMHR, followed by EQUIA Forte® and Filtek™ Z350. There was a significant difference in the %SMHR between Predicta® and Filtek™ Z350 (p < 0.05). However, EQUIA Forte® demonstrated the highest fluoride content, followed by Predicta® and Filtek™ Z350. The SEM images of EQUIA Forte® and Predicta® revealed the greater mineral deposition. CONCLUSION: Predicta® demonstrated a marked increase in surface microhardness and fluoride content of adjacent initial interproximal enamel carious lesions in primary molars compared with Filtek™ Z350. Predicta® is an alternative restorative material to remineralize adjacent initial interproximal enamel carious lesions in primary molars, especially in high-risk caries patients.

Cavity surface residues of PAD4 and SAG101 contribute to EDS1 dimer signaling specificity in plant immunity.

Arabidopsis pathogen effector-triggered immunity (ETI) is controlled by a family of three lipase-like proteins (EDS1, PAD4, and SAG101) and two subfamilies of HET-S/LOB-B (HeLo)-domain "helper" nucleotide-binding/leucine-rich repeats (ADR1s and NRG1s). EDS1-PAD4 dimers cooperate with ADR1s, and EDS1-SAG101 dimers with NRG1s, in two separate defense-promoting modules. EDS1-PAD4-ADR1 and EDS1-SAG101-NRG1 complexes were detected in immune-activated leaf extracts but the molecular determinants for specific complex formation and function remain unknown. EDS1 signaling is mediated by a C-terminal EP domain (EPD) surface surrounding a cavity formed by the heterodimer. Here we investigated whether the EPDs of PAD4 and SAG101 contribute to EDS1 dimer functions. Using a structure-guided approach, we undertook a comprehensive mutational analysis of Arabidopsis PAD4. We identify two conserved residues (Arg314 and Lys380) lining the PAD4 EPD cavity that are essential for EDS1-PAD4-mediated pathogen resistance, but are dispensable for the PAD4-mediated restriction of green peach aphid infestation. Positionally equivalent Met304 and Arg373 at the SAG101 EPD cavity are required for EDS1-SAG101 promotion of ETI-related cell death. In a PAD4 and SAG101 interactome analysis of ETI-activated tissues, PAD4R314A and SAG101M304R EPD variants maintain interaction with EDS1 but lose association, respectively, with helper nucleotide-binding/leucine-rich repeats ADR1-L1 and NRG1.1, and other immune-related proteins. Our data reveal a fundamental contribution of similar but non-identical PAD4 and SAG101 EPD surfaces to specific EDS1 dimer protein interactions and pathogen immunity.

New Biochemical Principles for NLR Immunity in Plants.

While working for the United States Department of Agriculture on the North Dakota Agricultural College campus in Fargo, North Dakota, in the 1940s and 1950s, Harold H. Flor formulated the genetic principles for coevolving plant host-pathogen interactions that govern disease resistance or susceptibility. His 'gene-for-gene' legacy runs deep in modern plant pathology and continues to inform molecular models of plant immune recognition and signaling. In this review, we discuss recent biochemical insights to plant immunity conferred by nucleotide-binding domain/leucine-rich-repeat (NLR) receptors, which are major gene-for-gene resistance determinants in nature and cultivated crops. Structural and biochemical analyses of pathogen-activated NLR oligomers (resistosomes) reveal how different NLR subtypes converge in various ways on calcium (Ca2+) signaling to promote pathogen immunity and host cell death. Especially striking is the identification of nucleotide-based signals generated enzymatically by plant toll-interleukin 1 receptor (TIR) domain NLRs. These small molecules are part of an emerging family of TIR-produced cyclic and noncyclic nucleotide signals that steer immune and cell-death responses in bacteria, mammals, and plants. A combined genetic, molecular, and biochemical understanding of plant NLR activation and signaling provides exciting new opportunities for combatting diseases in crops. [Formula: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

A Clinical Qualification Protocol Highlights Overlapping Genomic Influences and Neuro-Autonomic Mechanisms in Ehlers-Danlos and Long COVID-19 Syndromes.

A substantial fraction of the 15% with double-jointedness or hypermobility have the traditionally ascertained joint-skeletal, cutaneous, and cardiovascular symptoms of connective tissue dysplasia and its particular manifestation as Ehlers-Danlos syndrome (EDS). The holistic ascertainment of 120 findings in 1261 EDS patients added neuro-autonomic symptoms like headaches, muscle weakness, brain fog, chronic fatigue, dyspnea, and bowel irregularity to those of arthralgia and skin laxity, 15 of these symptoms shared with those of post-infectious SARS-CoV-2 (long COVID-19). Underlying articulo-autonomic mechanisms guided a clinical qualification protocol that qualified DNA variants in 317 genes as having diagnostic utility for EDS, six of them identical (F2-LIFR-NLRP3-STAT1-T1CAM1-TNFRSF13B) and eighteen similar to those modifying COVID-19 severity/EDS, including ADAMTS13/ADAMTS2-C3/C1R-IKBKG/IKBKAP-PIK3C3/PIK3R1-POLD4/POLG-TMPRSS2/TMPRSS6-WNT3/WNT10A. Also, contributing to EDS and COVID-19 severity were forty and three genes, respectively, impacting mitochondrial functions as well as parts of an overlapping gene network, or entome, that are hypothesized to mediate the cognitive-behavioral, neuro-autonomic, and immune-inflammatory alterations of connective tissue in these conditions. The further characterization of long COVID-19 natural history and genetic predisposition will be necessary before these parallels to EDS can be carefully delineated and translated into therapies.