Signaling System Research Articles

Analyzing immune cell infiltrates in skeletal muscle of infantile-onset Pompe disease using bioinformatics and machine learning

Pompe disease, a severe lysosomal storage disorder, is marked by heart problems, muscle weakness, and respiratory difficulties. This study aimed to identify novel markers for infantile-onset Pompe disease by analyzing key genes and immune cells infiltrating affected skeletal muscles, building on existing research linking its progression to the immune cell infiltration. The datasets GSE38680 and GSE159062 were downloaded and differential expression genes (DEGs) were identified. Potential markers were screened using support vector machine recursive feature elimination (SVM-RFE) analysis and least absolute shrinkage and selection operator (LASSO) regression models. To analyze DEGs and immune processes, 22 immune cell types in affected tissues were examined using CIBERSORT. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed significant enrichment in the calcium signaling pathway, phosphatidylinositol signaling system, ubiquitin-mediated proteolysis and JAK-STAT signaling pathway. Machine learning identified GPNMB, CALML6 and TRIM7 as key genes. Immune cell infiltration analysis showed that the expression levels of the above three genes were closely related to immune cells in our target population under study, including T cells regulatory (Tregs), monocytes, macrophages M0, resting and activated dendritic cells, naive and memory B cells. Overall, our study results may provide new clues for exploring markers related to Pompe disease by highlighting key genes and their relationship with immune infiltration, offering insights into the disease’s development. However, previous studies have shown limited evidence of significant immune cell infiltration in muscle biopsies from Pompe patients, and this lack of direct evidence necessitates further investigation and experimental validation in laboratory settings to confirm these associations.

Accumulation of Bioactive Lipid Species in LPS-Induced Neuroinflammation Models Analysed with Multi-Modal Mass Spectrometry Imaging

Neuroinflammation is a complex biological process related to a variety of pathologies, often requiring better understanding in order to develop new, targeted therapeutic interventions. Within this context, multimodal Mass Spectrometry Imaging (MSI) has been used to characterise molecular changes in neuroinflammation for biomarker discovery not possible to other techniques. In this study, molecules including bioactive lipids were detected across inflamed regions of the brain in rats treated with lipopolysaccharide (LPS). The detected lipids may be acting as inflammatory mediators of the immune response. We identified that N-acyl-phosphatidylethanolamine (NAPE) species accumulated in the inflamed area. The presence of these lipids could be related to the endocannabinoid (eCB) signalling system, mediating an anti-inflammatory response from microglial cells at the site of injury to balance pro-inflammation and support neuronal protection. In addition, polyunsaturated fatty acids (PUFAs), specifically n-3 and n-6 species, were observed to accumulate in the area where LPS was injected. PUFAs are directly linked to anti-inflammatory mediators resolving inflammation. Finally, acylcarnitine species accumulated around the inflammation region. Accumulation of these molecules could be due to a deficient β-oxidation cycle.

Advanced Applications of Polymer Hydrogels in Electronics and Signal Processing

The current state of affairs in the field of using polymer hydrogels for the creation of innovative systems for signal and image processing, of which computing is a special case, is analyzed. Both of these specific examples of systems capable of forming an alternative to the existing semiconductor-based computing technology, but assuming preservation of the used algorithmic basis, and non-trivial signal converters, the nature of which requires transition to fundamentally different algorithms of data processing, are considered. It is shown that the variability of currently developed information processing systems based on the use of polymers, including polymer hydrogels, leads to the need to search for complementary algorithms. Moreover, the well-known thesis that modern polymer science allows for the realization of functional materials with predetermined properties, at the present stage, receives a new sounding: it is acceptable to raise the question of creating systems built on a quasi-biological basis and realizing predetermined algorithms of information or image processing. Specific examples that meet this thesis are considered, in particular, promising information protection systems for UAV groups, as well as systems based on the coupling of neural networks with holograms that solve various applied problems. These and other case studies demonstrate the importance of interdisciplinary cooperation for solving problems arising from the need for further modernization of signal processing systems.

Endocannabinoid System and Metabolism: The Influences of Sex

The endocannabinoid system (ECS) is a lipid signaling system involved in numerous physiological processes, such as endocrine homeostasis, appetite control, energy balance, and metabolism. The ECS comprises endocannabinoids, their cognate receptors, and the enzymatic machinery that tightly regulates their levels within tissues. This system has been identified in various organs, including the brain and liver, in multiple mammalian and non-mammalian species. However, information regarding the sex-specific regulation of the ECS remains limited, even though increasing evidence suggests that interactions between sex steroid hormones and the ECS may ultimately modulate hepatic metabolism and energy homeostasis. Within this framework, we will review the sexual dimorphism of the ECS in various animal models, providing evidence of the crosstalk between endocannabinoids and sex hormones via different metabolic pathways. Additionally, we will underscore the importance of understanding how endocrine-disrupting chemicals and exogenous cannabinoids influence ECS-dependent metabolic pathways in a sex-specific manner.

OsPIPK-FAB, A Negative Regulator in Rice Immunity Unveiled by OsMBL1 Inhibition

Phosphatidylinositol signaling system plays a crucial role in plant physiology and development, phosphatidylinositol phosphate kinases (PIPKs) are one of the essential enzymes responsible for catalyzing the synthesis of phosphatidylinositol bisphosphate (PIP2) within this signaling pathway. However, its mechanism of signal transduction remains poorly exploited in plants. OsMBL1, a jacalin-related mannose-binding lectin in rice, plays a crucial role in plant defense mechanisms, acting as a key component of the pattern-triggered immunity (PTI) pathway. Here, a rice phosphatidylinositol-phosphate kinase FAB (OsPIPK-FAB), a member of the rice PIPKs family, as an interacting protein of OsMBL1 through yeast-two-hybrid (Y2H) screening assay. And this interaction was confirmed by using co-immunoprecipitation (Co-IP) and pull-down assay techniques. Furthermore, we demonstrated that the deletion of OsPIPK-FAB gene in plant enhanced resistance against rice blast while overexpression of OsPIPK-FAB increases sensitivity to the fungal infection. Additionally, through determination and measurement of the plant inositol 1,4,5-trisphosphate (IP3) contents and the plant phosphatidylinositol 4-phosphate 5-kinase (PIP5K) activity, we revealed that OsMBL1 inhibits the PIP5K kinase activity of OsPIPK-FAB as well as the plant IP3 contents in rice. Conclusively, these findings indicated that OsPIPK-FAB serves as a novel and critical component that is negatively involved in PTI activation and was inhibited by OsMBL1.

Drug developmental strategies based on functional analysis of pain-regulating molecules in astrocytes under chronic pain

Spinal cord astrocytes are activated in chronic pain models, especially under conditions of prolonged pain. Hence, targeting spinal cord astrocytes for the development of useful analgesics has attracted much attention. In the CNS, connexin43 (Cx43), a membrane protein expressed and functioning exclusively in astrocytes, is well known to be involved in intercellular signaling as a component of gap junction, but also interacts with intracellular molecules via its characteristically long C-terminal region, thereby affecting cellular function. Previously, we found that Cx43 expression was markedly reduced in spinal dorsal horn astrocytes from a mouse model of neuropathic pain. In order to investigate the relationship between reduced Cx43 expression in spinal astrocytes and the onset of pain, we showed that reduced Cx43 expression altered the expression of pain-related molecules such as the glutamate transporter GLT-1 and the pro-inflammatory cytokine interleukin-6 (IL-6). In particular, we focused on the regulation of IL-6 expression by reduced Cx43 expression in both in vivo and in vitro analyses, and found that IL-6 expression is increased through the Akt- glycogen synthase kinase-3β (GSK-3β) signaling system driven by reduced Cx43 expression during neuropathic pain, which in turn triggers pain. These findings suggest that astrocyte Cx43 is involved in pain prolongation by regulating gene expression of nociceptive factors through interactions with intracellular signaling molecules, which is different from its previously known function, and thus raises expectations for its potential as a new drug target for chronic pain.

Antioxidant and Anti-Inflammatory Effects of Curcuma Longa Extracts and Fractions on Skin Inflammation

Addresses the impact of modern society’s rapid industrialization and environmental pollution, leading to increased oxidative stress and immune system disorders. Skin barrier issues have resulted in a rise in inflammatory skin conditions, sparking heightened interest in functional cosmetics to address these concerns and combat aging. The research focused on exploring natural ingredients, particularly specific fractions of <i>Curcuma Longa</i> extracts, known for their antioxidant properties. The results of this study are summarized as follows. In the first stage experiment, DPPH radical scavenging activity and ABTS radical scavenging activity were investigated, and fractions exhibiting higher scavenging activity than extracts were sought. In the 2nd-step experiment, the cell survival rate was examined to determine the concentration that would not affect cytotoxicity. To demonstrate significant anti-inflammatory effects on RAW 264.7 cells, inflammation reactions were induced by LPS and the production of Nitric Oxide (NO) was controlled. As a result, <i>Curcuma Longa</i> extracts and fractions all proved excellent control effects on NO production. In HaCaT cells, skin inflammation was induced with TNF-α/IFN-γ and the control of interleukin (IL)-6 production was measured. As a result, <i>Curcuma Longa</i> extracts and fractions proved to facilitate the immune reaction and control the reaction that would lead to inflammation. In the following 3rd-step experiment, the control of Chemokine’s major factors IL-8, RANTES, and TARC was measured. As a result, it was verified that certain fractions of <i>Curcuma Longa</i> of extract showed excellent Chemokine production control effects and thus that these specific fractions are effective on various inflammatory skin diseases. ICAM-1 expressions were observed to control the initial stage of inflammation in a way of cellular adhesion regulation, and the MAPK path which is important to regulate inflammatory reactions in the signaling system was measured. As a result, the inhibition of phosphorylation, which is important for initial inflammation inhibition and inflammatory reaction control, was verified. Based on the above-stated findings, active ingredients containing certain fractions were analyzed, and color reactions in specific fractions were induced from active ingredient spots of extract. In consideration of future material development and production, the quantitative and qualitative characteristics of active ingredients of certain fractions were examined with the HPLC, and it was verified that certain fractions contained more active ingredients than extracts. In conclusion, certain fractions of natural elements proved the great potentials as a cosmetics ingredient. Therefore, these fractions proved excellent anti-oxidative and anti-inflammatory effects in certain fractions, and the efficiency is quite high with only a small quantity of them. It is expected that these fractions are widely utilized as a cosmetic element, and the future study will examine natural cosmetic elements from these fractions with the aim to contribute to the functional cosmetics industry and anti-aging product market.

Translation profiling of stress-induced small proteins reveals a novel link among signaling systems.

Signaling networks allow adaptation to stressful environments by activating genes that counteract stressors. Small proteins (≤ 50 amino acids long) are a rising class of stress response regulators. Escherichia coli encodes over 150 small proteins, most of which lack phenotypes and their biological roles remain elusive. Using magnesium limitation as a stressor, we identify stress-induced small proteins using ribosome profiling, RNA sequencing, and transcriptional reporter assays. We uncover 17 small proteins with increased translation initiation, several of them transcriptionally upregulated by the PhoQ-PhoP two-component signaling system, crucial for magnesium homeostasis. Next, we describe small protein-specific deletion and overexpression phenotypes, underscoring their physiological significance in low magnesium stress. Most remarkably, we elucidate an unusual connection via a small membrane protein YoaI, between major signaling networks - PhoR-PhoB and EnvZ-OmpR in E. coli, advancing our understanding of small protein regulators in cellular signaling.

The effect of amyloid beta, membrane, and ER pathways on the fractional behavior of neuronal calcium

Calcium signal transduction is essential for cellular activities such as gene transcription, death, and neuronal plasticity. Dynamical changes in the concentration of calcium have a profound effect on the intracellular activity of neurons. The Caputo fractional reaction-diffusion equation is a useful tool for modeling the intricate biological process involved in calcium concentration regulation. We include the Amyloid Beta, STIM-Orai mechanism, voltage-dependent calcium entry, inositol triphosphate receptor (IPR), endoplasmic reticulum (ER) flux, SERCA pump, and plasma membrane flux in our mathematical model. We use Green's function and Hankel and Laplace integral transforms to solve the membrane flux problem. Our simulations investigate the effects of various factors on the spatiotemporal behavior of calcium levels, with a simulation on the buffers in Alzheimer's disease-affected neurons. We also look at the effects of calcium-binding substances like the S100B protein and BAPTA and EGTA. Our results demonstrate how important the S100B protein Amyloid beta and the STIM-Orai mechanism are, and how important they are to consider when simulating the calcium signaling system. As such, our research indicates that a more realistic and complete model for modeling calcium dynamics may be obtained by using a generalized reaction-diffusion technique.

Directing Nanoparticle Organization in Response to Diverse Chemical Inputs.

Signaling cascades are crucial for transducing stimuli in biological systems, enabling multiple stimuli to regulate a downstream target with precisely controlled timing and amplifying signals through a series of intermediary reactions. Developing a robust signaling system with such capabilities would be pivotal for programming complex behaviors in synthetic DNA-based molecular devices. However, although "software" such as nucleic acid circuits could potentially be harnessed to relay signals to DNA-based nanostructure hardware, such explorations have been limited. Here, we develop a platform for transducing a variety of stimuli via messenger-mediated reactions to regulate the release and reloading of gold nanoparticles (AuNPs) in a 3D DNA framework. In the first step, an in vitro transcription circuit is engineered to sense and amplify chemical stimuli, including arbitrary DNA sequences and proteins, producing RNA. In the second step, the RNA releases the DNA-coated AuNPs from the DNA framework via a strand displacement reaction. AuNP reloading is controlled by a separate step driven by degradation of the RNA. Our platform holds promise for applications requiring dynamic multiagent control over DNA-based devices, offering a versatile tool for advanced molecular device engineering.

Single phage proteins sequester signals from TIR and cGAS-like enzymes.

Prokaryotic anti-phage immune systems use TIR and cGAS-like enzymes to produce 1''-3'-glycocyclic ADP-ribose (1''-3'-gcADPR) and cyclic dinucleotide (CDN) and cyclic trinucleotide (CTN) signalling molecules, respectively, which limit phage replication1-3. However, how phages neutralize thesedistinct and common systems is largely unclear. Here we show that the Thoeris anti-defence proteins Tad14 and Tad25 both achieve anti-cyclic-oligonucleotide-based anti-phage signalling system (anti-CBASS) activity by simultaneously sequestering CBASS cyclic oligonucleotides. Apart from binding to the Thoeris signals 1''-3'-gcADPR and 1''-2'-gcADPR, Tad1 also binds to numerous CBASS CDNsandCTNs with high affinity, inhibiting CBASS systems that use these molecules in vivo and in vitro. The hexameric Tad1 has six binding sites for CDNs or gcADPR, which are independent of the two high-affinity binding sites for CTNs. Tad2 forms a tetramer that also sequesters various CDNs in addition to gcADPR molecules, using distinct binding sites to simultaneously bind to these signals. Thus, Tad1 and Tad2 are both two-pronged inhibitors that, alongside anti-CBASS protein 2 (Acb26-8), establish a paradigm of phage proteins that use distinct binding sites to flexibly sequester a considerable breadth of cyclic nucleotides.

Examining dermatology residency applicant profiles for the 2023-2024 cycle: a cross-sectional analysis.

Dermatology remains highly competitive, with strong USMLE Step 1 scores traditionally crucial for securing residency positions. The 2023-2024 cycle introduced significant changes, including pass/fail USMLE Step 1 score reports and an expanded program signaling system. This study explores dermatology residency applicant profiles within this new context. A survey of 2023-2024 dermatology applicants was conducted via social media to gather demographic and application data. A total of 63 survey responses were collected: 74.6% matched and 25.4% unmatched. The racial distribution was 54% White/Caucasian, 25.4% Asian/Pacific Islander, 9.5% Black/African American, 4.8% Hispanic/Latino, and 6.3% other. The median USMLE Step 2 score was 257 (215-277). Racial differences in USMLE Step 2 scores were significant (P = 0.031), but did not affect match rates (P = 0.116). Letters of recommendation from dermatology program directors were linked to lower match rates (P = 0.036). A positive correlation was found between the number of audition rotations completed and matching at such programs (r²=0.817). Of all matched respondents, 46.8% matched to a program they did not signal; of these, 50.0% matched to a program at which they completed an audition rotation and 40.9% to their home dermatology program affiliation. The mean number of interviews was 8.02, with matched applicants receiving more invitations than unmatched applicants (9.02 vs. 5.06, P = 0.002). The shift to pass/fail USMLE Step 1 scores and expanded program signaling did not notably affect the median USMLE Step 2 score from prior years or match rates among underrepresented minorities. Success in matching continues to depend on a holistic evaluation.

High-Performance Rotating Structure Triboelectric-Electromagnetic Hybrid Nanogenerator for Environmental Wind Energy Harvesting.

As environmental energy harvesting gains increasing importance in self-powered systems and large-scale energy demands, wind energy, as a clean, pollution-free, and renewable source, has garnered widespread attention. However, achieving efficient wind energy collection remains challenging. This study proposes a high-performance rotating structure triboelectric-electromagnetic hybrid nanogenerator designed for environmental wind energy harvesting. By optimizing the magnetic circuit design of the electromagnetic generator, the dispersed radial magnetic field is converted into a unified axial magnetic field, enabling efficient power generation with only a single annular coil, thereby simplifying the generator design and reducing manufacturing and maintenance costs. Additionally, a triboelectric nanogenerator design with soft contact friction between polycarbonate (PC) fur and fluorinated ethylene propylene (FEP) film was implemented, optimizing the spacing between the electrode and friction layers, thus enhancing output performance and device durability. Furthermore, we simulated and experimentally tested the output waveform of the designed hybrid generator structure, with the results showing a high degree of similarity, further validating the rationality of the device design and providing guidance for structural optimization. Subsequently, we achieved efficient energy storage using an energy management circuit (EMC). With the integration of the EMC, the generator successfully powered a Bluetooth temperature and humidity sensor at a wind speed of 10 m/s, achieving wireless transmission, and demonstrating its potential application in traffic signal systems and other natural environmental systems. This research provides an important reference for further exploration of novel wind energy harvesting technologies.

Calcitonin signalling system regulates function and arhythmogenicity of atrial cardiomyocytes

Abstract Background Atrial fibrillation (AF) poses a significant therapeutic challenge due to myocardial remodelling, involving both structural and electrical alterations. Recent investigations have shed light on the role of atrial cardiomyocytes (CMs) in secreting Calcitonin (CT), a factor crucial for maintaining atrial tissue integrity. Dysregulated CT secretion in AF showed to contribute to fibrotic tissue production by atrial fibroblasts, exacerbating arrhythmogenesis [1]. However, the direct impact of CT on CM function and arrhythmogenicity remains unclear, driving the objectives of our study. Methods/Results Serum samples from 20 cardiac surgery patients revealed a noteworthy association between higher pre-operative CT levels and a significant ~2.8-fold reduction in the incidence of post-operative AF (poAF). Using freshly isolated atrial guinea pig CMs, confirmed (by qPCR and immunofluorescence) that CMs express CT-receptor (CTR) to enable CT actions in the cell. Functional studies found that CT administration inhibits spontaneous calcium (Ca2+)-release events induced by pacing and decreases the Ca2+ transients amplitude (at 2 Hz; IonOptix μstep system) in fura2-loaded CMs (n=22 cells) in a concentration-dependent manner. Atrial iPSC-CMs transduced with global RGECO sensor (Genetically encoded Ca2+ indicator) and treated with 15 pM CT showed a reduction in the beat rate when paced at 2Hz, and a significantly increased the time to 50% baseline. Evaluation of the expression and phosphorylation of selected Ca2+ handling proteins, which may potentially account for the observed changes in Ca2+ transients, showed no differences in total or phospho-(ser2808) ryanodine receptor, and SERCA2α in response to CT but an increase in phospho-(Ser16) Phospholamban. Conclusion Our findings highlight the effects of CT on atrial CM function. Maintaining physiological CT levels offer a promising approach for managing AF clinically, potentially through the use of already in clinical use CT-analogues.

Transactivation of the EGF receptor as a novel desensitization mechanism for G protein-coupled receptors, illustrated by dopamine D2-like and β2 adrenergic receptors

Transactivation of epidermal growth factor receptors (EGFR) provides intricate control over multiple regulatory cellular processes that merge the diversity of G protein-coupled receptors (GPCRs) with the robust signaling capacities of receptor tyrosine kinases. Contrary to the typical assertions, our findings demonstrate that EGFR transactivation contributes to the desensitization of GPCRs. Repeated agonist stimulation of certain GPCRs enhanced EGFR transactivation, triggering a series of cellular events associated with GPCR desensitization. This effect was observed in receptors undergoing desensitization (D3R, K149C-D2R, β2AR) but not in those resistant to desensitization (D2R, C147K-D3R, D4R, β2AR mutants lacking GRK2 or GRK6 phosphorylation sites). The EGFR inhibitor AG1478 prevented both desensitization and the associated cellular events. Similarly, these cellular events were also observed when cells were treated with EGF, but only in GPCRs that undergo desensitization. These findings suggest that EGFR transactivation diversifies pathways involved in ERK activation through the EGFR signaling system and also mediates GPCR desensitization. Alongside the widely accepted steric hindrance model, these findings offer new insights into understanding the mechanisms of GPCR desensitization, which occurs through complex cellular processes.

Inhibition of CB1R in the Hypothalamic Paraventricular Nucleus Ameliorates Hypertension Through Wnt/β-Catenin/RAS Pathway.

The hypothalamic paraventricular nucleus (PVN), as an important integrating center, plays a prominent role in the pathogenesis of hypertension, in maintaining the stability of cardiovascular activity through peripheral sympathetic nervous activity and secretion of various humoral factors. Acknowledging that the mechanistic targets of the endocannabinoid type 1 receptor (CB1R) are the key signaling systems involved in the regulation of hypertension, we sought to clarify whether inhibition of CB1R within the PVN ameliorates hypertension through Wnt/β-catenin/RAS pathway. Spontaneously hypertensive rats (SHRs) and Wistar Kyoto rats were randomly assigned to different groups and treated with bilateral PVN injections of AM251 (CB1R antagonist, 10µg/h) or vehicle (artificial cerebrospinal fluid, aCSF) for four weeks. Bilateral PVN injections of AM251 significantly decreased the heart rate, the body weight and the mean arterial pressure in SHRs. AM251 lowered the expression of CB1R, Wnt3, active-β-catenin, p-IKKβ, RAS components, pro-inflammatory cytokines and elevated the expression level of Glycogen synthase kinase3β and Superoxide Dismutase in the PVN of hypertensive rats. Our findings suggest that inhibition of CB1R in the PVN ameliorates hypertension through Wnt/β-catenin/RAS pathway and broaden our current understanding of the pathological mechanism and clinical treatment of hypertension.

Robustness for biochemical networks: Step-by-step approach

We propose two step-by-step approaches to the analysis of robustness in biochemical networks. Our aim is to measure the ability of the network to exhibit step-by-step limited variations on the concentration of a species of interest at varying of the initial concentration of other species. The first approach we propose is reaction-by-reaction, i.e. we compare the states reached by nominal and perturbed networks after they have performed the same number of reactions. We provide a statistical technique allowing for estimating robustness, we implement it in a tool called spebnr (a Simple Python Environment for statistical estimation of Biochemical Network Robustness) and showcase it on three case studies: the EnvZ/OmpR osmoregulatory signaling system of Escherichia Coli, the mechanism of bacterial chemotaxis of Escherichia Coli, and enzyme activity at saturation. Then, we consider a time-by-time approach, in which networks are compared on the basis of the states they reached at the same time point, regardless of how many reactions occurred. This approach is implemented in Stark, and we apply it to the study the robustness of the EnvZ/OmpR osmoregulatory signaling system and the Lotka-Volterra equations.

Review on application of fractional Fourier transform in LinearFrequencyModulation signal and communication system

Traditional Fourier transform often apply to analyze and process stationary signals, however, it is weak for time-varying non-stationary signals, and fractional Fourier transform (FRFT) can better solve such problems. The FRFT can be comprehended as the expressive methods on the fractional Fourier domain constituted by the spinning coordinate axis of the signal anticlockwise about the origin at arbitrarily Angle in the time-frequency plane. In this paper, the improved fractional Fourier transform is combined with other calculation methods to achieve high precision estimation of chirp signal parameters. And the communication system built on weighted fractional Fourier transform and discrete fractional Fourier transform is studied and simulated respectively, which verifies the feasibility and improves the anti-jamming and anti-interception ability of the communication system.

Evaluation of apoptosis markers in patients with atopic dermatitis

The purpose of the study was to analyze indicators of apoptosis in patients with atopic dermatitis (AD). Patients (N = 88) aged from 12 months to 3 years with varying degrees of disease severity were involved into trial. SCORAD indices were: 68.14±2.63 – in patients with severe AD; 32.03±1.43 – with moderate; and 12.12±1.43 – with mild AD, respectively. Determination of apoptosis markers (Caspase 8, Caspase 9, sCD153, sFas-L, Annexin 5) in blood serum was performed using an enzyme-linked immunosorbent assay (ELISA) using commercial kits from Bender MedSystems® (Austria). A study of apoptosis markers in the blood serum of children with AD revealed a significant decrease (p 0.05) in the concentration of Annexin 5 compared to standard values (by 20 times). A statistically significant decrease (p 0.05) in the concentration levels of Caspase 8 and Caspase 9 in the serum of AD patients was revealed. Thus, the concentrations of Caspase 8 were reduced on by 3 times, and Caspase 9 by 8 times compared to the norm. Also In addition, when determining sFas-L indicators, a statistically significant (p 0.05) decrease in concentration was noted compared to standard indicators. The content of sCD153 in the blood serum in AD patients was significantly higher (p 0.05) than the normative values by 5 times (not detected normally). The most pronounced disturbances in apoptosis indicators were observed in patients with severe atopic dermatitis, (p 0.05), highlighting the importance of studying apoptosis in the context of AD and the possible link between disruptions in this process and the severity of clinical manifestations in patients. The study demonstrates the peculiarities of apoptosis processes in children with AD. On the one hand, these patients have significantly increased activation of signaling systems, on the other hand, the elimination of altered immunocompetent cells does not occur properly, which contributes to the progression and chronicity of immune inflammation in the skin.

Structural basis for regulation of a CBASS-CRISPR-Cas defense island by a transmembrane anti-σ factor and its ECF σ partner.

How CRISPR-Cas and cyclic oligonucleotide-based antiphage signaling systems (CBASS) are coordinately deployed against invaders remains unclear. We show that a locus containing two CBASS and one type III-B CRISPR-Cas system, regulated by the transmembrane anti-σ DdvA and its cognate extracytoplasmic function (ECF) σ DdvS, can defend Myxococcus xanthus against a phage. Cryo-electron microscopy reveals DdvA-DdvS pairs assemble as arrow-shaped transmembrane dimers. Each DdvA periplasmic domain adopts a separase/craspase-type tetratricopeptide repeat (TPR)-caspase HetF-associated with TPR (TPR-CHAT) architecture with an incomplete His-Cys active site, lacking three α-helices conserved among CHAT domains. Each active site faces the dimer interface, raising the possibility that signal-induced caspase-like DdvA autoproteolysis in trans precedes RseP-mediated intramembrane proteolysis and DdvS release. Nuclear magnetic resonance reveals a DdvA cytoplasmic CHCC-type zinc-bound three-helix bundle that binds to DdvS σ2 and σ4 domains, undergoing σ4-induced helix extension to trap DdvS. Altogether, we provide structural-mechanistic insights into membrane anti-σ-ECF σ regulation of an antiviral CBASS-CRISPR-Cas defense island.