Recognition Of Molecular Patterns Research Articles

Ss4368: Pathogen-Associated Molecular Pattern for Inducing Plant Cell Death and Resistance to Phytophthora capsici.

Plant recognition of pathogen-associated molecular patterns (PAMPs) is pivotal in triggering immune responses, highlighting their potential as inducers of plant immunity. However, the number of PAMPs identified and applied in such contexts remains limited. In this study, we characterize a novel PAMP, designated Ss4368, which is derived from Scleromitrula shiraiana. Ss4368 is specifically distributed among a few fungal genera, including Botrytis, Monilinia, and Botryotinia. The transient expression of Ss4368 elicits cell death in a range of plant species. The signaling peptides, three conserved motifs, and cysteine residues (C46, C88, C112, C130, and C148) within Ss4368 are crucial for inducing robust cell death. Additionally, these signaling peptides are essential for the protein's localization to the apoplast. The cell death induced by Ss4368 and its homologous protein, Bc4368, is independent of the SUPPRESSOR OF BIR1-1 (SOBIR1), BRI1-ASSOCIATED KINASE-1 (BAK1), and salicylic acid (SA) pathways. Furthermore, the immune responses triggered by Ss4368 and Bc4368 significantly enhance the resistance of Nicotiana benthamiana to Phytophthora capsici. Therefore, we propose that Ss4368, as a novel PAMP, holds the potential for developing strategies to enhance plant resistance against P. capsici.

HIF-1α is required to differentiate the neonatal Macrophage protein secretome from adults

The immune response to stress diverges with age, with neonatal macrophages implicated in tissue regeneration versus tissue scarring and maladaptive inflammation in adults. Integral to the macrophage stress response is the recognition of hypoxia and pathogen-associated molecular patterns (PAMPs), which are often coupled. The age-specific, cell-intrinsic nature of this stress response remains vague. To uncover age-defined divergences in macrophage crosstalk potential after exposure to hypoxia and PAMPs, we interrogated the secreted proteomes of neonatal versus adult macrophages via non-biased mass spectrometry. Through this approach, we newly identified age-specific signatures in the secretomes of neonatal versus adult macrophages in response to hypoxia and the prototypical PAMP, lipopolysaccharide (LPS). Neonatal macrophages secreted proteins most consistent with an anti-inflammatory, regenerative phenotype protective against apoptosis and oxidative stress, dependent on hypoxia inducible transcription factor-1α (HIF-1α). In contrast, adult macrophages secreted proteins consistent with a pro-inflammatory, glycolytic phenotypic signature consistent with pathogen killing. Taken together, these data uncover fundamental age and HIF-1α dependent macrophage responses that may be targeted to calibrate the innate immune response during stress and inflammation.

Phenotypic and Functional Outcomes in Macrophages Exposed to an Environmentally Relevant Mixture of Organophosphate Esters in Vitro.

Organophosphate esters (OPEs) are flame retardants and plasticizers used in consumer products. OPEs are found ubiquitously throughout the environment with high concentrations in indoor house dust. Exposure to individual OPEs is associated with immune dysfunction, particularly in macrophages. However, OPEs exist as complex mixtures and the effects of environmentally relevant mixtures on the immune system have not been investigated. The objectives of this study were to evaluate the toxicity of an environmentally relevant mixture of OPEs that models Canadian house dust on macrophages using phenotypic and functional assessments in vitro. High-content live-cell fluorescent imaging for phenotypic biomarkers of toxicity in THP-1 macrophages treated with the OPE mixture was undertaken. We used confocal microscopy and cholesterol analysis to validate and expand on the observed OPE-induced lipid phenotype. Then, we used flow cytometry and live-cell imaging to conduct functional tests and uncover mechanisms of OPE-induced phagocytic suppression. Finally, we validated our THP-1 findings in human primary peripheral blood mononuclear cells (hPBMC) derived macrophages. Exposure to non-cytotoxic dilutions of the OPE mixture resulted in higher oxidative stress and disrupted lysosome and lipid homeostasis in THP-1 and primary macrophages. We further observed that phagocytosis of apoptotic cells in THP-1 and primary macrophages was lower in OPE-exposed cells vs. controls. In THP-1 macrophages, phagocytosis of both Gram-positive and Gram-negative bacteria was also lower in OPE-exposed cells vs. controls. Additionally, the OPE mixture altered the expression of phagocytic receptors linked to the recognition of phosphatidylserine and pathogen-associated molecular patterns. The results of this in vitro study suggested that exposure to an environmentally relevant mixture of OPEs resulted in higher lipid retention in macrophages and poor efferocytic response. These effects could translate to enhanced foam cell generation resulting in higher cardiovascular mortality. Furthermore, bacterial phagocytosis was lower in OPE-exposed macrophages in an in vitro setting, which may indicate the potential for reduced bacterial clearance in models of infections. Taken together, our data provide strong evidence that mixtures of OPEs can influence the biology of macrophages and offer new mechanistic insights into the impact of OPE mixtures on the immune system. https://doi.org/10.1289/EHP13869.

Toll-like receptor 4 (TLR4) is the major pattern recognition receptor triggering the protective effect of a Candida albicans extracellular vesicle-based vaccine prototype in murine systemic candidiasis.

Systemic candidiasis remains a significant public health concern worldwide, with high mortality rates despite available antifungal drugs. Drug-resistant strains add to the urgency for alternative therapies. In this context, vaccination has reemerged as a prominent immune-based strategy. Extracellular vesicles (EVs), nanosized lipid bilayer particles, carry a diverse array of native fungal antigens, including proteins, nucleic acids, lipids, and glycans. Previous studies from our laboratory demonstrated that Candida albicans EVs triggered the innate immune response, activating bone marrow-derived dendritic cells (BMDCs) and potentially acting as a bridge between innate and adaptive immunity. Vaccination with C. albicans EVs induced the production of specific antibodies, modulated cytokine production, and provided protection in immunosuppressed mice infected with lethal C. albicans inoculum. To elucidate the mechanisms underlying EV-induced immune activation, our study investigated pathogen-associated molecular patterns (PAMPs) and pattern recognition receptors (PRRs) involved in EVs-phagocyte engagement. EVs from wild-type and mutant C. albicans strains with truncated mannoproteins were compared for their ability to stimulate BMDCs. Our findings revealed that EV decoration with O- and N-linked mannans and the presence of β-1,3-glucans and chitin oligomers may modulate the activation of specific PRRs, in particular Toll-like receptor 4 (TLR4) and dectin-1. The protective effect of vaccination with wild-type EVs was found to be dependent on TLR4. These results suggest that fungal EVs can be harnessed in vaccine formulations to selectively activate PRRs in phagocytes, offering potential avenues for combating or preventing candidiasis.IMPORTANCESystemic candidiasis is a serious global health concern with high mortality rates and growing drug resistance. Vaccination offers a promising solution. A unique approach involves using tiny lipid-coated particles called extracellular vesicles (EVs), which carry various fungal components. Previous studies found that Candida albicans EVs activate the immune response and may bridge the gap between innate and adaptive immunity. To understand this better, we investigated how these EVs activate immune cells. We demonstrated that specific components on EV surfaces, such as mannans and glucans, interact with receptors on immune cells, including Toll-like receptor 4 (TLR4) and dectin-1. Moreover, vaccinating with these EVs led to strong immune responses and full protection in mice infected with Candida. This work shows how harnessing fungal EVs might lead to effective vaccines against candidiasis.

Impact of chronic muscle use and disuse on innate immune signaling in skeletal muscle

Mitochondria exhibit high levels of adaptability to various stimuli such as chronic muscle use (i.e. exercise) or disuse, such as immobilization or denervation. These changes in the mitochondrial network ultimately affect the health and functioning of skeletal muscle, which contributes to whole-body metabolism, locomotion, and postural stability. It is now acknowledged that mitochondrial dysfunction can activate inflammatory pathways through the recognition of damage-associated molecular patterns (DAMPs). The objective of this study was to examine how changes in mitochondria brought about by either chronic muscle inactivity or chronic exercise training affect innate immune system activation. We hypothesized that an inverse relationship between mitochondrial content and innate immune signaling would be evident in skeletal muscle. To investigate this, skeletal muscle from the hindlimbs of mice was collected after 7 days of sciatic muscle denervation, or after 4.5 weeks of endurance swim training involving 240 minutes of daily exercise. We analyzed the expression of the NLRP3 inflammasome, innate immune system signaling proteins, mitochondrial markers, and mitochondrial DNA (mtDNA) content using western blotting, cytochrome C oxidase (COX) activity, ELISA and qPCR. Following 7 days of denervation, there was a significant decrease in both COX-IV protein and COX activity, indicating a reduction in mitochondrial content. This was accompanied by an increase (p<0.05) in NLRP3, procaspase-1, gasdermin-D (GSDMD) and its active N-terminal fragment, GSDMD-N, as well as a notable increase in IL-1β. In addition, other possible converging pathways were also upregulated including cGAS-STING, and gasdermin-E. In comparison, following exercise training, there was a significant increase in mitochondrial content, apparent through increases in COX-IV protein, COX activity and mitochondrial RNA content. This was associated with decreases in procaspase-1 and caspase-1 protein expression alongside reduced STING activation. These findings highlight an inverse relationship between mitochondrial content and the activation of several inflammatory pathways that possibly converge upon NLRP3 inflammasome activation and pyroptotic cell death, leading to detriments in skeletal muscle health. This work aims to further the understanding of innate immune signaling pathways within muscle which can potentially highlight therapeutic targets to regulate its activation under divergent metabolic conditions. This work was supported by funds from the Natural Science and Engineering Research Council (NSERC). Priyanka Khemraj is the holder of an Ontario Graduate Scholarship. David A. Hood is the holder of a Canadian Research Chair in Cell Physiology. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Guanosine-specific single-stranded ribonuclease effectors of a phytopathogenic fungus potentiate host immune responses.

Plants activate immunity upon recognition of pathogen-associated molecular patterns. Although phytopathogens have evolved a set of effector proteins to counteract plant immunity, some effectors are perceived by hosts and induce immune responses. Here, we show that two secreted ribonuclease effectors, SRN1 and SRN2, encoded in a phytopathogenic fungus, Colletotrichum orbiculare, induce cell death in a signal peptide- and catalytic residue-dependent manner, when transiently expressed in Nicotiana benthamiana. The pervasive presence of SRN genes across Colletotrichum species suggested the conserved roles. Using a transient gene expression system in cucumber (Cucumis sativus), an original host of C. orbiculare, we show that SRN1 and SRN2 potentiate host pattern-triggered immunity responses. Consistent with this, C. orbiculare SRN1 and SRN2 deletion mutants exhibited increased virulence on the host. In vitro analysis revealed that SRN1 specifically cleaves single-stranded RNAs at guanosine, leaving a 3'-end phosphate. Importantly, the potentiation of C. sativus responses by SRN1 and SRN2, present in the apoplast, depends on ribonuclease catalytic residues. We propose that the pathogen-derived apoplastic guanosine-specific single-stranded endoribonucleases lead to immunity potentiation in plants.

Sexual dimorphism in NLR transcripts and its downstream signaling protein IL-1ꞵ in teleost Channa punctata (Bloch, 1793)

Nucleotide-binding oligomerization domain-like receptors (NOD-like receptors or NLRs) are a family of intracellular pattern recognition receptors (PRRs) that initiates as well as regulate inflammatory responses. NLRs are characterized by a centrally located nucleotide binding domain and a leucine rich repeat domain at the C-terminal responsible for the recognition of intracellular microbe-associated molecular patterns (MAMPs) and danger-associated molecular patterns (DAMPs). In the present study in adult spotted snakehead we have investigated the sex-dependent tissue distribution of NLRs known to be associated with inflammation in teleost namely NOD1, NOD2, NLRC3, NLRC5, and NLRX1. Further, the sexual dimorphism in the expression of NLR transcript as well as the pro-inflammatory protein IL-1β was explored in fish under normal conditions, and in fish exposed to bacterial lipopolysaccharide (LPS). The NLRs show ubiquitous and constitutive expression in all the tissues. Moreover, a prominent disparity between males and females was observed in the basal expression of these genes in various tissues. The sexual dimorphism in NLR expression was also prominent when fish were exposed to LPS. Similarly, IL-1β exhibited sexual dimorphism in both normal as well as LPS-exposed fish.

A CD6 homolog of Nile tilapia (Oreochromis niloticus) conserved binding bacteria involved in the regulation of Streptococcus agalactiae induced inflammation

As a lymphocyte-specific surface receptor belonging to the cysteine-rich superfamily of scavenger receptors, CD6 acts as a pattern recognition receptor for microbial components and is involved in the regulation of inflammatory responses. However, the characteristics and functions of CD6 molecules in lower vertebrates represented by teleost fish are unknown. In this study, a CD6 homolog (designated OnCD6) was characterized from Nile tilapia (Oreochromis niloticus), and establishing its role as a PRRs that participates in immune recognition. OnCD6 contains an open reading frame of 1872 bp that encodes a peptide of 623 amino acids, and contains two conserved SR domain. Multiple sequence alignment revealed that OnCD6 shares a relatively high level of identity with those of other species. Transcriptional expression analysis revealed that OnCD6 was constitutively expressed in immunes tissues such as head kidney and thymus. The expression level of OnCD6 in mainly immune tissues were found significantly upregulated after the injection of Streptococcus agalactiae (S. agalactiae). Moreover, OnCD6 protein was located in the head kidney and brain, mainly over the plasma membrane of lymphocytes in these immune tissues. In vitro experiments showed that CD6 extracellular protein bound to and aggregated several Gram-positive and -negative bacterial strains through the recognition of bacterial surface conserved components LPS and LTA etc. In vivo experiments demonstrated that overexpression OnCD6 before S. agalactiae challenge significantly improved tilapia survival, and this was concomitant with reduced bacterial load and pro-inflammatory cytokines (IL-1β and TNF-α). Taken together, our results illustrated the function of CD6 molecular pattern recognition receptors (PRRs) is conserved and plays an important role in antibacterial infection.

Design of Multi-Competitors Winner-Take-All Neural Networks Based on DNA Strand Displacement for Molecular Pattern Recognition

DNA strand displacement technology (DSDT) provides flexible and powerful technical support for DNA molecular computing. DNA-based neural networks with Winner-Take-All (WTA) strategy has a great potential for nonlinear calculation. However, so far it has been limited to achieving the simultaneous competition of two competitors. Optimizing the calculation model and reducing system response time to recognize complex and functional molecular patterns remains a huge challenge. Here a novel neural network with WTA strategy based on DSDT was constructed, which allowed three competitors to participate in the competition at the same time. Firstly, the feasibility of the three-competitor WTA neural network was proved by 9-bit pattern recognition. Then the three-competitors WTA neural network was further extended to larger scale pattern recognition, which successfully recognized 64-bit letters A, B, and C and 100-bit handwritten digits 0, 2, and 4, respectively. Simulations showed that when recognizing the same target patterns with same number bits, compared with two-competitors WTA neural network, the three-competitors WTA network only used down to two-thirds DNA strands, and the system response time was reduced by more than ten times. This paper demonstrated the efficient recognition ability of the three-competitor WTA neural network, which is expected to be used to identify more complex information.

Comparative transcriptome analysis of hepatopancreas reveals the potential mechanism of shrimp resistant to Vibrio parahaemolyticus infection

Vibrio parahaemolyticus carrying a pathogenic plasmid (VPAHPND) is one of the main causative agents of acute hepatopancreatic necrosis disease (AHPND) in shrimp aquaculture. Knowledge about the mechanism of shrimp resistant to VPAHPND is very helpful for developing efficient strategy for breeding AHPND resistant shrimp. In order to learn the mechanism of shrimp resistant to AHPND, comparative transcriptome was applied to analyze the different expressions of genes in the hepatopancreas of shrimp from different families with different resistance to VPAHPND. Through comparative analysis on the hepatopancreas of shrimp from VPAHPND resistant family and susceptible family, we found that differentially expressed genes (DEGs) were mainly involved in immune and metabolic processes. Most of the immune-related genes among DEGs were highly expressed in the hepatopancreas of shrimp from resistant family, involved in recognition of pathogen-associated molecular patterns, phagocytosis and elimination of pathogens, maintenance of reactive oxygen species homeostasis and other immune processes etc. However, most metabolic-related genes were highly expressed in the hepatopancreas of shrimp from susceptible family, involved in metabolism of lipid, vitamin, cofactors, glucose, carbohydrate and serine. Interestingly, when we analyzed the expression of above DEGs in the shrimp after VPAHPND infection, we found that the most of identified immune-related genes remained at high expression levels in the hepatopancreas of shrimp from the VPAHPND resistant family, and most of the identified metabolic-related genes were still at high expression levels in the hepatopancreas of shrimp from the VPAHPND susceptible family. The data suggested that the differential expression of these immune-related and metabolic-related genes in hepatopancreas might contribute to the resistance variations of shrimp to VPAHPND. These results provided valuable information for understanding the resistant mechanism of shrimp to VPAHPND.

Pathogen-Activated Macrophage Membrane Encapsulated CeO2-TCPP Nanozyme with Targeted and Photo-Enhanced Antibacterial Therapy.

Nanozymes with peroxidase-mimic activity have recently emerged as effective strategies for eliminating infections. However, challenges in enhancing catalytic activities and the ability to target bacteria have hindered the broader application of nanozymes in bacterial infections. Herein, a novel nanozyme based on mesoporous CeO2 nanosphere and meso-tetra(4-carboxyphenyl)porphine (TCPP) encapsulated within pathogen-activated macrophage membranes, demonstrates photodynamic capability coupled with photo-enhanced chemodynamic therapy for selective and efficient antibacterial application against infected wounds. Interestingly, the expression of Toll-like receptors accordingly upregulates when macrophages are co-cultured with specific bacteria, thereby facilitating to recognition of the pathogen-associated molecular patterns originating from bacteria. The CeO2 not only serve as carriers for TCPP, but also exhibit intrinsic peroxidase-like catalytic activity. Consequently, Staphylococcus aureus (S. aureus)-activated macrophage membrane-coated CeO2-TCPP (S-MM@CeO2-TCPP) generated singlet oxygen, and simultaneously promoted photo-enhanced chemodynamic therapy, significantly boosting reactive oxygen species (ROS) to effectively eliminate bacteria. S-MM@CeO2-TCPP specifically targeted S. aureus via Toll-like receptor, thereby directly disrupting bacterial structural integrity to eradicate S. aureus in vitro and relieve bacteria-induced inflammation to accelerate infected wound healing in vivo. By selectively targeting specific bacteria and effectively killing pathogens, such strategy provides a more efficient and reliable alternative for precise elimination of pathogens and inflammation alleviation in microorganism-infected wounds.

BTL2 phospho-switch surveils plant immunity.

BRASSINOSTEROID INSENSITIVE1-ASSOCIATED KINASE1 (BAK1) is a co-receptor involved in the recognition of pattern-associated molecular patterns (PAMPs) via plasma membrane-localized pattern recognition receptors (PRRs). Absence of BAK1/SERK4 leads to the activation of autoimmunity in plants. Yu et al. recently showed that BAK-TO-LIFE 2 (BTL2) is required for the surveillance of BAK1/SERK4 integrity to maintain immune homeostasis.

TLR1-10 Protein Expression in Circulating Human White Blood Cells during Bacterial and COVID-19 Infections

Toll-like Receptors (TLR) are major sensors of the innate immune system and are involved in the recognition of Pathogen Associated Molecular Patterns (PAMPs) and Damage Associated Molecular Patterns (DAMPs). When activated, TLR signaling pathways trigger an intracellular response that leads to the production of pro-inflammatory cytokines, chemotactic factors, antimicrobial peptides, and interferons. Overexpression of certain TLRs on neutrophils has been associated with increased mortality in patients with infection-induced shock, although the role of TLR4 overexpression is still debated. While TLRs play a role in recognizing in bacterial structures, they can also detect viral patterns, particularly TLR2, TLR3, and TLR7. The activation of the TLR signaling pathway is an early step recognizing viral genomic patterns of SARS-CoV-2 and its spike protein activates TLR4. In this prospective single-center observational cohort study, we conducted a comprehensive analysis of the protein expression of the ten known human TLR in white blood cells from patients with either bacterial and viral infections using flow cytometry, together with CD180, a 95-105 kDa TLR-like glycoprotein expressed on peripheral blood monocytes and B-lymphocytes. 73 patients were included, divided into 4 groups based on infection severity (sepsis, septic shock, moderate and severe COVID-19), along with 7 control subjects. Septic shock was assessed according to the third international consensus definition. COVID-19 patients with confirmed SARS-CoV-2 infection using reverse transcriptase-polymerase chain reaction (RT-PCR), admitted either to the emergency department (ED) or in the ICU were included. This study protocol was reviewed and approved by the local ethic committee, and written informed consent was obtained from participants of their relatives. For endosomal TLRs (TLR3, TLR7, TLR8, TLR9), plasma cell permeabilization was performed using the Perm2 Solution® (BD Biosciences, USA) prior to incubation of monoclonal antibodies staining. Cell surface expression was studied for the other TLRs. In neutrophils, antibody staining for TLR1-10 and CD180 revealed the expression of TLR4, with a large variation in positivity between groups, TLR8, TLR9 were also expressed, while TLR2, TLR3, and TLR7 were weakly expressed. In monocytes, only TLR2, TLR4, and CD180 were detected Notably, we observed a significant reduction in the expression of TLR4 and its co-receptor CD14 in neutrophils of patients with shock compared to controls (MFI=19 273 vs. 111 691, p<0.001 and MFI 2 028 vs. 3 025, p <0.02 respectively). Furthermore, severe COVID-19 patients exhibited a significant increase in TLR3 and TLR7 levels in neutrophils (p=0.003 and p=0.02 respectively) compared to the control group. Patients with shock also showed a similar increase in TLR7 expression in neutrophils, along with elevated intermediate monocytes compared to controls. TLR expression remained unchanged in lymphocytes. We also observed that CD16 expression in neutrophils was significantly lower in septic shock patients compared to healthy donors (p<0.0001), and to a lesser extent in sepsis patients (p<0.01), consistent with previous studies This study provided further insights into the mechanisms of TLR activation in various severe forms of infection. Additional analysis is required to assess their correlation with patient outcomes and evaluate the potential impact of TLR-related therapy on neutrophils activation.

Microbe-associated molecular pattern recognition receptors have little effect on endophytic Arabidopsis thaliana microbiome assembly in the field.

Plant microbiome structure affects plant health and productivity. A limited subset of environmental microbes successfully establishes within plant tissues, but the forces underlying this selectivity remain poorly characterized. Transmembrane pattern recognition receptors (PRRs), used by plants to detect microbe-associated molecular patterns (MAMPs), are strong candidates for achieving this selectivity because PRRs can potentially interact with many members of the microbiome. Indeed, MAMPs found in many microbial taxa, including beneficials and commensals, can instigate a robust immune response that affects microbial growth. Surprisingly, we found that MAMP-detecting PRRs have little effect on endophytic bacterial and fungal microbiome structure in the field. We compared the microbiomes of four PRR knockout lines of Arabidopsis thaliana to wild-type plants in multiple tissue types over several developmental stages and detected only subtle shifts in fungal, but not bacterial, β-diversity in one of the four PRR mutants. In one developmental stage, lore mutants had slightly altered fungal β-diversity, indicating that LORE may be involved in plant-fungal interactions in addition to its known role in detecting certain bacterial lipids. No other effects of PRRs on α-diversity, microbiome variability, within-individual homogeneity, or microbial load were found. The general lack of effect suggests that individual MAMP-detecting PRRs are not critical in shaping the endophytic plant microbiome. Rather, we suggest that MAMP-detecting PRRs must either act in concert and/or are individually maintained through pleiotropic effects or interactions with coevolved mutualists or pathogens. Although unexpected, these results offer insights into the role of MAMP-detecting PRRs in plant-microbe interactions and help direct future efforts to uncover host genetic elements that control plant microbiome assembly.

Re-identification and characterization of grass carp Ctenopharyngodon idella TLR20

Toll-like receptors (TLRs) play a crucial role in the recognition of microbial-associated molecular patterns in the innate immune system. Fish TLRs have undergone significant gene expansion to adapt to complex aquatic environments. Among them, TLR20 from the TLR11 family actively responds to viral and bacterial invasions. Previous studies have reported two TLR20s in grass carp (Ctenopharyngodon idella), and in this study, we revised this conclusion. Based on the latest grass carp genome, we identified a new TLR20 member. These three TLR20s are arranged in tandem on chromosome 9, indicating that they are generated by gene duplication events. They were renamed CiTLR20.1 to CiTLR20.3 based on their chromosomal positions. The CiTLR20s in C. idella exhibit higher similarities with those in Danio rerio, Cyprinus carpio, and Megalobrama amblycephala, and lower similarities with those in other distantly related fish species. Selective pressure analysis revealed low conservation and negative evolution of TLR20s during evolution. The 3D structures of the three TLR20s showed significant differences, reflecting functional variations and different downstream adaptor molecule recruitment. Transcriptome data revealed tissue distribution differences of TLR20s, with TLR20.1 showing relatively low expression levels in all the tissues, while TLR20.2 and TLR20.3 showed higher expression in the head kidney, spleen, and gill. Additionally, TLR20.2 and TLR20.3 actively responded to GCRV-II infection, with higher upregulation of TLR20.2 in response to Aeromonas hydrophila challenge. In conclusion, this study corrected the number of grass carp TLR20 members and analyzed TLR20 from an evolutionary and structural perspective, exploring its role in antiviral and antibacterial defense. This study provides reference for future research on fish TLR20.

Sensing and signaling in periodontal inflammation via toll like receptors: A review

Periodontal disease is a chronic inflammatory condition that affects the supporting structures of teeth, including the gingiva, periodontal ligament, and alveolar bone. Toll-like receptors (TLRs) play a crucial role in the recognition of pathogen-associated molecular patterns (PAMPs) from bacteria that cause periodontal disease. TLR activation triggers an inflammatory response that aims to eliminate the bacteria and repair damaged tissue. However, if the inflammation persists, it can lead to further tissue destruction and bone loss. This review discusses the mechanisms of TLRs in periodontal disease progression, including the recognition of bacteria, activation of immune cells, tissue destruction, and inhibition of immune response. Strategies for down-regulating TLRs, such as small-molecule inhibitors, antibodies, natural products, micro RNA, and nutritional interventions, are also discussed. Targeting TLRs is a promising therapeutic approach for reducing chronic inflammation associated with periodontal disease. Further research is needed to develop safe and effective strategies for down-regulating TLRs in humans.

PCAP-1a, an exopolysaccharide from Pectobacterium actinidiae, exerts the dual role of immunogenicity and virulence in plants

Plant defense mechanisms begin with the recognition of microbe-associated molecular patterns or pathogen-associated molecular patterns (MAMPs/PAMPs). Several carbohydrates, such as chitin, were reported to induce plant defenses, acting as elicitors. Regrettably, the structures of polysaccharide elicitors have rarely been characterized, and their recognition receptors in plants remain unknown. In the present study, PCAP-1a, an exopolysaccharide (PCAP-1a) purified from Pectobacterium actinidiae, was characterized and found to induce rapid cell death of dicotyledons, acting as a polysaccharide elicitor to induce plant immunity. A series of pattern-triggered immunity (PTI) responses were triggered, including reactive oxygen species production, phosphorylation of mitogen-activated protein kinases and gene transcriptional reprogramming. Moreover, we confirmed that CERK1 is probably one of the immune coreceptors for plants to recognize PCAP-1a. Notably, PCAP-1a also promotes the infection caused by P. actinidiae. In conclusion, our study supports the potential of PCAP-1a as a toxin that plays a dual role of virulence and immune induction in pathogen-plant interactions.

Inflammatory ER stress responses dictate the immunopathogenic progression of systemic candidiasis.

Recognition of pathogen-associated molecular patterns can trigger the inositol-requiring enzyme 1 α (IRE1α) arm of the endoplasmic reticulum (ER) stress response in innate immune cells. This process maintains ER homeostasis and also coordinates diverse immunomodulatory programs during bacterial and viral infections. However, the role of innate IRE1α signaling in response to fungal pathogens remains elusive. Here, we report that systemic infection with the human opportunistic fungal pathogen Candida albicans induced proinflammatory IRE1α hyperactivation in myeloid cells that led to fatal kidney immunopathology. Mechanistically, simultaneous activation of the TLR/IL-1R adaptor protein MyD88 and the C-type lectin receptor dectin-1 by C. albicans induced NADPH oxidase-driven generation of ROS, which caused ER stress and IRE1α-dependent overexpression of key inflammatory mediators such as IL-1β, IL-6, chemokine (C-C motif) ligand 5 (CCL5), prostaglandin E2 (PGE2), and TNF-α. Selective ablation of IRE1α in leukocytes, or treatment with an IRE1α pharmacological inhibitor, mitigated kidney inflammation and prolonged the survival of mice with systemic C. albicans infection. Therefore, controlling IRE1α hyperactivation may be useful for impeding the immunopathogenic progression of disseminated candidiasis.

Association between SNP rs4986790 and COVID-19 infection severity among Baghdad patients

Introduction and Aim: COVID-19, an infectious disease caused by the SARS-CoV-2 coronavirus, is distinguished by the manifestation of severe acute respiratory syndrome. Toll-like receptors (TLRs) are signaling molecules that play crucial roles in the innate immune system through their recognition of pathogen-associated molecular patterns in diverse microorganisms, including coronaviruses. The primary aim of this research was to investigate the plausible association between the TLR4 gene Asp299Gly polymorphism and the degree of infection severity among individuals who contracted COVID-19 in Baghdad, Iraq. Materials and Methods: This cross-sectional prospective study was carried out in Baghdad to investigate the Asp299Gly polymorphism within the TLR4 gene in a cohort of 90 patients diagnosed with Covid-19. Out of the total number of patients, 45 individuals exhibited symptoms indicative of a moderate infection, while the remaining 45 patients presented with a severe illness. The Asp299Gly polymorphism was analyzed using polymerase chain reaction (PCR) technique and restriction fragment length polymorphism (RFLP). Results: Genotyping for the Asp299Gly polymorphism showed only one among the 90 (1.1%) Covid-19 patients tested to be positive. No significant association was seen between covid-19 severity and Asp299Gly (P = 0·31) polymorphism. Conclusion: Our study found no association between SNP rs4986790 of TLR4 gene and COVID-19 severity among Covid-19 patients in Baghdad city.

Automatic Assignment of Molecular Ion Species to Elemental Formulas in Gas Chromatography/Methane Chemical Ionization Accurate Mass Spectrometry.

Gas chromatography-mass spectrometry (GC-MS) usually employs hard electron ionization, leading to extensive fragmentations that are suitable to identify compounds based on library matches. However, such spectra are less useful to structurally characterize unknown compounds that are absent from libraries, due to the lack of readily recognizable molecular ion species. We tested methane chemical ionization on 369 trimethylsilylated (TMS) derivatized metabolites using a quadrupole time-of-flight detector (QTOF). We developed an algorithm to automatically detect molecular ion species and tested SIRIUS software on how accurate the determination of molecular formulas was. The automatic workflow correctly recognized 289 (84%) of all 345 detected derivatized standards. Specifically, strong [M - CH3]+ fragments were observed in 290 of 345 derivatized chemicals, which enabled the automatic recognition of molecular adduct patterns. Using Sirius software, correct elemental formulas were retrieved in 87% of cases within the top three hits. When investigating the cases for which the automatic pattern analysis failed, we found that several metabolites showed a previously unknown [M + TMS]+ adduct formed by rearrangement. Methane chemical ionization with GC-QTOF mass spectrometry is a suitable avenue to identify molecular formulas for abundant unknown peaks.