Molecular Patterns Research Articles

Metabolic energetic adaptation of Atlantic salmon phagocytes to changes in carbon sources and exposure to PAMPs

Phagocytic cells are pivotal for host homeostasis and infection defense, necessitating metabolic adaptations in glycolysis, the tricarboxylic acid (TCA) cycle, and oxidative phosphorylation (OXPHOS). While mammalian phagocytes shift towards glycolysis and glutaminolysis during polarization, research on fish phagocyte metabolic reprogramming is limited. To address this, the Atlantic salmon phagocytic cell line, SHK-1, serves as a valuable model. Using the Seahorse XFe96 Flux Analyzer, this study compares SHK-1 bioenergetics under glucose-restricted (L-15 medium) and glucose-supplemented (PM) conditions, providing insights into metabolic characteristics and responses to Piscirickettsia salmonis bacterium Pathogen-associated molecular patterns (PAMPs). A standardized protocol for the study of real-time changes in the metabolism study of SHK-1 in PM and L-15 media, determining oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) is shown. Exhibiting metabolic adaptations, SHK-1 cells in the PM medium have higher basal and maximal OCR and spare capacity (SRC), while those grown in the L-15 medium favor OXPHOS, showing minimal glycolytic function. Despite metabolic differences, intracellular ATP levels are comparable, highlighting the metabolic plasticity and adaptability of SHK-1 cells to various carbon sources. Exposure to PAMPs from Piscirickettsia salmonis induces a metabolic shift, increasing glycolysis and OXPHOS, influencing ATP, lactate, glutamine, and glutamate levels. These findings highlight the role of mitochondrial bioenergetics and metabolic plasticity in salmon phagocytes, offering novel nutritional strategies for host-pathogen interventions based on energy metabolism.

Branched oncolytic peptides target HSPGs, inhibit metastasis, and trigger the release of molecular determinants of immunogenic cell death in pancreatic cancer.

Immunogenic cell death (ICD) can be exploited to treat non-immunoreactive tumors that do not respond to current standard and innovative therapies. Not all chemotherapeutics trigger ICD, among those that do exert this effect, there are anthracyclines, irinotecan, some platinum derivatives and oncolytic peptides. We studied two new branched oncolytic peptides, BOP7 and BOP9 that proved to elicit the release of damage-associated molecular patterns DAMPS, mediators of ICD, in pancreatic cancer cells. The two BOPs selectively bound and killed tumor cells, particularly PANC-1 and Mia PaCa-2, but not cells of non-tumor origin such as RAW 264.7, CHO-K1 and pgsA-745. The cancer selectivity of the two BOPs may be attributed to their repeated cationic sequences, which enable multivalent binding to heparan sulfate glycosaminoglycans (HSPGs), bearing multiple anionic sulfation patterns on cancer cells. This interaction of BOPs with HSPGs not only fosters an anti-metastatic effect in vitro, as demonstrated by reduced adhesion and migration of PANC-1 cancer cells, but also shows promising tumor-specific cytotoxicity and low hemolytic activity. Remarkably, the cytotoxicity induced by BOPs triggers the release of DAMPs, particularly HMGB1, IFN-β and ATP, by dying cells, persisting longer than the cytotoxicity of conventional chemotherapeutic agents such as irinotecan and daunorubicin. An in vivo assay in nude mice showed an encouraging 20% inhibition of tumor grafting and growth in a pancreatic cancer model by BOP9.

Molecular mechanisms of emerging inflammasome complexes and their activation and signaling in inflammation and pyroptosis.

Inflammasomes are multi-protein complexes that assemble within the cytoplasm of mammalian cells in response to pathogen-associated molecular patterns (PAMPs) or damage-associated molecular patterns (DAMPs), driving the secretion of the pro-inflammatory cytokines IL-1β and IL-18, and pyroptosis. The best-characterized inflammasome complexes are the NLRP3, NAIP-NLRC4, NLRP1, AIM2, and Pyrin canonical caspase-1-containing inflammasomes, and the caspase-11 non-canonical inflammasome. Newer inflammasome sensor proteins have been identified, including NLRP6, NLRP7, NLRP9, NLRP10, NLRP11, NLRP12, CARD8, and MxA. These inflammasome sensors can sense PAMPs from bacteria, viruses and protozoa, or DAMPs in the form of mitochondrial damage, ROS, stress and heme. The mechanisms of action, physiological relevance, consequences in human diseases, and avenues for therapeutic intervention for these novel inflammasomes are beginning to be realized. Here, we discuss these emerging inflammasome complexes and their putative activation mechanisms, molecular and signaling pathways, and physiological roles in health and disease.

Hybrid modes in multilayer/antenna architecture set side-band-selective molecular Raman scattering

Abstract Technologies with tag-less capacity for analyzing molecules, are poised to make significant advancements in the healthcare industry with a substantial potential to improve sensitivity for individual molecules and binding events. Plasmonics has the potential for enhanced system response when emitters are included in the full architecture. Here, a hybrid multilayer including a sequence of metals and dielectrics has been examined. The surface of the multilayer is covered with 30-μm height features working as plasmonic antennae. Their contribution to the emission of the system has been analyzed. The multilayer has been coupled with a prism to excite the polaritons in an experimental optical setup to measure the reflected and transmitted signals. The measurements demonstrate the surface plasmon polariton/antenna mode hybridization. The optomechanics of the plasmonic resonant multilayer has been studied with reference to the refractive index of the surrounding medium as well as to the incident angle of the exciting beam. Then, the experimental shifts have been measured according to the optomechanical spectrum of the naïve plasmon resonant multilayer. The optomechanics of the plasmonic resonant multilayer, which has shown a natural resonance at ω = 96 MHz , has been coupled with a molecular emitter, e.g. a dye. As the concentration of the red dye increases, the intensity of the peaks at ω > 96 MHz raises, suggesting an important sensitivity in the anti-Stokes domain. On the other side, the decay is equal to κ = 1.1 × 10 − 2 MHz showcasing a high quality factor ( Q = 9.6 × 10 3 ). For validation of increased sensitivity in the anti-Stokes region, bovine serum albumin (BSA), a protein active in the Raman region, has been tested and, through an image-based analysis, the molecular pattern recorded. The demonstrated potential of utilizing optical resonance shifts to investigate molecular patterns is highly promising. Therefore, the proposed sensing method represents a significant advancement in the field, offering new opportunities for the sensitive detection of biomolecules.

Selective degradation of organic pollutants in the aquatic environment by microplastic-derived dissolved organic matter through molecular photoresponse sequence transformation

The photochemical behavior of microplastics (MPs)-dissolved organic matter (PDOM) has received considerable attention, the molecular mechanism by which it undergoes photoresponsive transformation to generate reactive oxygen species (ROS) for the removal of organic pollutants remains unclear. This study combines fluorescence, mass spectrometry, and theoretical models to analyse the differences in the molecular mechanisms of PDOM, biochar-DOM (BDOM) and soil-DOM (SDOM) photoresponsive releasing ROS for selective degradation of organic pollutants. During the photoresponse process, molecular compounds in DOMs mainly undergo demethylation, decarboxylation, deoxygenation reactions to selectively degrade organic pollutants. However, differences in DOMs’ molecular structure composition and transformation patterns lead to varying ROS production. BDOMs containing higher proportion of aromatic compounds (26.68 %) exhibit stronger photoreactivity, resulting in better degradation efficiency of pollutants (>90 %). The high selectivity of PDOM for degradation of organic pollutants stems from the interconversion mode between its high and low oxygen compounds. PDOM degrades large molecular pollutants into less biotoxic small molecular intermediates through both radical and non-radical pathways, thereby reducing environmental hazards (1.5–948 times) posed by pollutants. The results show that although MPs is harmful to aquatic environment, it plays a positive role in regulating the structure of organic carbon pool and reducing environmental risks of other pollutants.

Real-world molecular testing patterns in primary advanced or recurrent endometrial cancer (pA/rEC) in the US.

395 Background: Targeted therapy can improve outcomes in women with pA/rEC, but real-world biomarker testing rates are unknown. This study estimated the real-world proportion of patients receiving biomarker testing (within ≤6 months of advanced/recurrent diagnosis) and characterized testing according to sociodemographic and clinical characteristics. Methods: We used the US Flatiron Health electronic health record–derived deidentified database. Records from 1/1/2013 to 8/31/2023 for women ≥18 years old with advanced/recurrent EC were searched to identify testing for DNA mismatch repair/microsatellite instability (MMR/MSI), HER2, and estrogen or progesterone receptors (ER/PR); a subsample dataset from 4/1/2013 to 11/30/2022 was analyzed for additional biomarkers. The subsample has information on additional biomarker testing and was used to evaluate the frequency of testing for these biomarkers. Testing rates were evaluated using descriptive statistics; characteristics associated with testing were identified by multivariate logistic regression. Results: Among persons meeting eligibility criteria (n=2982), the mean age was 65 years; the majority were non-Hispanic White (56%), had commercial insurance (79%), and received care in a community setting (73%). The subsample (n=509) had similar characteristics to the full cohort. Between 2013-2022, 73% (n=2165) of patients received testing for any biomarker. Testing rates for individual biomarkers are shown in the Table. From 2013 to 2021, testing increased for MMR/MSI (17% to 81%), ER/PR (45% to 62%), and HER2 (15% to 43%). The likelihood of testing was higher among patients with primary advanced EC (OR, 1.45; 95% CI, 1.03-2.06), endometrioid carcinoma histology (1.39; 1.01-1.93), commercial insurance (1.51; 1.18-1.93), at an academic facility (1.70; 1.37-2.11), and with no prior surgery (2.29; 1.83-2.86) relative to other attributes. Conclusions: Molecular testing has increased over time, indicating a continuous trend toward personalized treatment decisions. Several factors associated with an increased likelihood of biomarker testing were identified. Understanding these factors can inform approaches to increase access to molecular testing and increase testing rates. Biomarker testing rates, 2013-2022. Biomarker, n (%) 2013 2021 Overall MMR/MSI a 35 (15) 238 (80) 1604 (54) HER2 a 31 (14) 121 (41) 612 (21) ER/PR a 99 (43) 177 (59) 1465 (49) p53/ TP53 b 13 (36) 31 (74) 260 (51) TMB b 3 (8) 14 (33) 91 (18) CTNNB1 b 4 (11) 12 (29) 88 (17) ARID1A b 4 (11) 15 (36) 91 (18) CDK4/6 b 5 (14) 14 (33) 110 (22) POLE b 4 (11) 17 (40) 110 (22) ARID1A, AT-rich interactive domain-containing protein 1A; CDK4/6, cyclin-dependent kinases 4 and 6; CTNNB1, catenin beta-1; POLE, polymerase-epsilon; TMB, tumor mutation burden. a 2013, n=228; 2021, n=298; overall, n=2982. b Rates from subsample dataset (4/1/2013 to 11/30/2022); 2013, n=36; 2021, n=42; overall, n=509.

An array of Zymoseptoria tritici effectors suppress plant immune responses.

Zymoseptoria tritici is the most economically significant fungal pathogen of wheat in Europe. However, despite the importance of this pathogen, the molecular interactions between pathogen and host during infection are not well understood. Herein, we describe the use of two libraries of cloned Z. tritici effectors that were screened to identify effector candidates with putative pathogen-associated molecular pattern (PAMP)-triggered immunity (PTI)-suppressing activity. The effectors from each library were transiently expressed in Nicotiana benthamiana, and expressing leaves were treated with bacterial or fungal PAMPs to assess the effectors' ability to suppress reactive oxygen species (ROS) production. From these screens, numerous effectors were identified with PTI-suppressing activity. In addition, some effectors were able to suppress cell death responses induced by other Z. tritici secreted proteins. We used structural prediction tools to predict the putative structures of all of the Z. tritici effectors and used these predictions to examine whether there was enrichment of specific structural signatures among the PTI-suppressing effectors. From among the libraries, multiple members of the killer protein-like 4 (KP4) and killer protein-like 6 (KP6) effector families were identified as PTI suppressors. This observation is intriguing, as these protein families were previously associated with antimicrobial activity rather than virulence or host manipulation. This data provides mechanistic insight into immune suppression by Z. tritici during infection and suggests that, similar to biotrophic pathogens, this fungus relies on a battery of secreted effectors to suppress host immunity during early phases of colonization.

Hepatocyte activation and liver injury following cerebral ischemia promote HMGB1-mediated hepcidin upregulation in hepatocytes and regulation of systemic iron levels

We previously reported that high mobility group box 1 (HMGB1), a danger-associated molecular pattern (DAMP), increases intracellular iron levels in the postischemic brain by upregulating hepcidin, a key regulator of iron homeostasis, triggering ferroptosis. Since hepatocytes are the primary cells that produce hepcidin and control systemic iron levels, we investigated whether cerebral ischemia induces hepcidin upregulation in hepatocytes. Following middle cerebral artery occlusion (MCAO) in a rodent model, significant liver injury was observed. This injury was evidenced by significantly elevated Eckhoff’s scores and increased serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST). Additionally, total iron levels were significantly elevated in the liver, with intracellular iron accumulation detected in hepatocytes. Hepcidin expression in the liver, which is primarily localized in hepatocytes, increased significantly starting at 3 h after MCAO and continued to increase rapidly, reaching a peak at 24 h. Interestingly, HMGB1 levels in the liver were also significantly elevated after MCAO, with the disulfide form of HMGB1 being the major subtype. In vitro experiments using AML12 hepatocytes showed that recombinant disulfide HMGB1 significantly upregulated hepcidin expression in a Toll-like receptor 4 (TLR4)- and RAGE-dependent manner. Furthermore, treatment with a ROS scavenger and a peptide HMGB1 antagonist revealed that both ROS generation and HMGB1 induction contributed to hepatocyte activation and liver damage following MCAO–reperfusion. In conclusion, this study revealed that cerebral ischemia triggers hepatocyte activation and liver injury. HMGB1 potently induces hepcidin not only in the brain but also in the liver, thereby influencing systemic iron homeostasis following ischemic stroke.

Synergizing autophagic cell death and oxaliplatin-induced immunogenic death by a self-delivery micelle for enhanced tumor immunotherapy

Chemotherapy has become an emerging strategy to activate cytotoxic T cell responses by inducing immunogenic cell death (ICD), but the level of antitumor immunity induced by chemotherapeutic agents, such as oxaliplatin (OXA), is limited due to inadequate tumor antigen presentation and T cell activation. Inducing autophagic cell death (ACD) promotes the release of tumor antigen and the recruitment of dendritic cells, therefore strengthening antitumor immune responses. Here we simultaneously activate ICD and ACD with tumor targeting micelle to achieve enhanced antitumor chemo-immunotherapy. A self-delivery micelle is formulated by conjugating OXA prodrug with tocopherol succinate (TOS) as a hydrophobic segment and further encapsulates autophagy activator SMER28 to afford TOPR/SMER28, which specifically targets αvβ3 on tumor cells with c(RGDfK). Upon cellular internalization, OXA is released from the prodrug in response to the high concentration of reduced glutathione (GSH) in tumor cells, triggering ICD and releasing associated molecular patterns (DAMPs) signaling molecules to stimulate immunity. Meanwhile, SMER28 over-activates autophagy to induce autophagic cell death, which further leads to the maturation of dendritic cells and ultimately activates anti-tumor immune response. In the 4T1 tumor-bearing mice, the combination of OXA and SMER28 effectively inhibits tumor growth and activates antitumor immune responses. The tumor targeted micelle releases OXA and SMER28 in an on-demand profile and strengthens tumor chemo-immunotherapy by synergizing ICD and ACD, providing an alternative for antitumor immunotherapy. Statement of significanceChemotherapy induces immunogenic cell death (ICD) to activate anti-tumor immunity. However, the efficacy is limited by low levels of antigen presentation and T cell activation. To strengthen the antitumor immune responses induced by ICD, we first combine autophagic cell death (ACD) with ICD by formulating a glutathione-responsive oxaliplatin prodrug micelle co-encapsulating the autophagy activator SMER28. The activated autophagic level by SMER28 enhances the release of antigen and the recruitment of APCs, and ultimately bolsters T cell-mediated antitumor immune responses. We provide a potential strategy to amplify antitumor immune effects by combining autophagy activation with chemotherapy.

Bacteria renew an OLD protein to cleave host tRNAs and block phage translation

Anti-phage defenses must rapidly sense and respond to diverse viruses. A recent pair of papers in Nature reveal via structural and functional assays how the PARIS defense system, a recently discovered toxin-antitoxin system, senses phage-associated molecular patterns (PhAMPs), thereby activating an endonuclease toxin that cleaves tRNA to block phage replication.

Extracellular adenosine triphosphate: A new gateway for food allergy mechanism research?

Although various studies have been conducted, the detailed mechanisms of food allergy remain a topic of ongoing debate. Recently, researchers have reported that extracellular adenosine triphosphate (eATP), a member of damage-associated molecular patterns secreted by stressed cells, plays a critical role in the progression of asthma and atopic dermatitis. These studies suggest that dysregulated eATP significantly influences various aspects of disease progression, from bodily sensitization to the emergence of clinical manifestations. Given the shared pathogenic mechanisms among asthma, atopic dermatitis, and food allergies, we hypothesize that eATP may also serve as a crucial regulator in the development of food allergies. To elucidate this hypothesis, we first summarize the evidence and limitations of food allergy theories, then discuss the roles of eATP in allergic diseases. We conclude with speculative insights into the potential influence of eATP on food allergy development, aiming to inspire further investigation into the molecular mechanisms of food allergies.

Human iPSC-based disease modeling studies identify a common mechanistic defect and potential therapies for AMD and related macular dystrophies.

Age-related macular degeneration (AMD) and related macular dystrophies (MDs) primarily affect the retinal pigment epithelium (RPE) in the eye. A hallmark of AMD/MDs that drives later-stage pathologies is drusen. Drusen are sub-RPE lipid-protein-rich extracellular deposits, but how drusen forms and accumulates is not known. We utilized human induced pluripotent stem cell (iPSC)-derived RPE from patients with AMD and three distinct MDs to demonstrate that reduced activity of RPE-secreted matrix metalloproteinase 2 (MMP2) contributes to drusen in multiple maculopathies in a genotype-agnostic manner by instigating sterile inflammation and impaired lipid homeostasis via damage-associated molecular pattern molecule (DAMP)-mediated activation of receptor for advanced glycation end-products (RAGE) and increased secretory phospholipase 2-IIA (sPLA2-IIA) levels. Therapeutically, RPE-specific MMP2 supplementation, RAGE-antagonistic peptide, and a small molecule inhibitor of sPLA2-IIA ameliorated drusen accumulation in AMD/MD iPSC-RPE. Ultimately, this study defines a causal role of the MMP2-DAMP-RAGE-sPLA2-IIA axis in AMD/MDs.

P3.06F.06 Spatial Transcriptomic Profiling Reveals Molecular Patterns Associated with Poor Prognosis in ALK-Rearranged NSCLC Patients

P3.06F.06 Spatial Transcriptomic Profiling Reveals Molecular Patterns Associated with Poor Prognosis in ALK-Rearranged NSCLC Patients

Borna disease virus 1 induces ferroptosis, contributing to lethal encephalitis.

Borna disease virus 1 (BoDV-1) is a neurotropic RNA virus that has been linked to fatal BoDV-1 encephalitis (BVE) in humans. Ferroptosis represents a newly recognized kind of programmed cell death that marked by iron overload and lipid peroxidation. Various viral infections are closely related to ferroptosis. However, the link between BoDV-1 infection and ferroptosis, as well as its role in BVE pathogenesis, remains inadequately understood. Herein, we used primary rat cortical neurons, human microglial HMC3 cells, and Sprague‒Dawley rats as models. BoDV-1 infection induced ferroptosis, as ferroptosis characteristics were detected (iron overload, reactive oxygen species buildup, decreased antioxidant capacity, lipid peroxidation, and mitochondrial damage). Analysis via qRT-PCR and Western blot demonstrated that BoDV-1-induced ferroptosis was mediated through Nrf2/HO-1/SLC7a11/GPX4 antioxidant pathway suppression. Nrf2 downregulation was due to BoDV-1 infection promoting Nrf2 ubiquitination and degradation. Following BoDV-1-induced ferroptosis, the PTGS2/PGE2 signaling pathway was activated, and various intracellular lipid peroxidation products and damage-associated molecular patterns were released, contributing to BVE occurrence and progression. More importantly, inhibiting ferroptosis or the ubiquitin‒proteasome system effectively alleviated BVE. Collectively, these findings demonstrate the interaction between BoDV-1 infection and ferroptosis and reveal BoDV-1-induced ferroptosis as an underlying pathogenic mechanism of BVE.

TLR-2 and TLR-4 mRNA expression in different grades of histopathological lesions of equine endometrium from follicular phase.

Increased synthesis and deposition of collagen(COL) in the extracellular matrix (ECM) of equine endometrium contributes to endometrosis. Toll-like receptors (TLRs) are transmembrane receptors involved in the innate immune response, recognized for their role in antigen recognition and previously associated with equine endometritis. The TLRs not only recognize pathogen-associated molecular patterns but also regulate inflammations, fibrosis and cancer. The aim of this study was to explore the relationship between TLR expression at different stages of Kenney and Doig's (K-D) grading and COL1 expression during the follicular phase of the oestrous cycle. Forty samples of endometrial tissues were collected post-mortem from mares on the follicular phase of the oestrous cycle (10 samples of each K-D category). Relative mRNA transcription of TLR-2, TLR-4 and COL1A2 genes was assessed using qPCR, and COL1 protein expression by Western blot analysis. The COL1A2 transcription increased in category IIB when compared to categories I, IIA and III endometria (p < .01). The relative protein abundance of COL1 showed no significant differences between endometrial categories (p > .05). As for the TLRs mRNA transcription, TLR-2/-4 transcripts increased in IIA when compared to the other K-D endometria categories (p < .05). Our findings suggest that TLRs may be involved in the initiation of the endometrial inflammatory response. Additional studies are needed to explore TLRs' potential role as diagnostic markers for monitoring inflammation progression and fibrosis development, as well as their involvement in the mechanisms underlying fibrotic pathways.

Milk fat globule-epidermal growth factor-VIII–derived oligopeptide 3 (MOP3) attenuates inflammation and improves survival in hepatic ischemia/reperfusion injury

IntroductionHepatic ischemia/reperfusion injury is a severe clinical condition leading to high mortality as the result of excessive inflammation, partially triggered by released damage-associated molecular patterns. Extracellular cold-inducible RNA-binding protein is a new damage-associated molecular pattern. Current clinical management of hepatic ischemia/reperfusion injury is limited to supportive therapy, necessitating the development of novel and effective treatment strategies. Milk fat globule-epidermal growth factor-VIII–derived oligopeptide 3 is a newly invented oligopeptide originating from milk fat globule-epidermal growth factor-VIII. This peptide acts as an opsonic compound that specifically binds to extracellular cold-inducible RNA-binding protein to facilitate its clearance by phagocytes, thereby attenuating inflammation. In this study, we hypothesized that milk fat globule-epidermal growth factor-VIII–derived oligopeptide 3 attenuated hepatic ischemia/reperfusion injury by inhibiting extracellular cold-inducible RNA-binding protein–induced inflammation in Kupffer cells. MethodsWe treated Kupffer cells isolated from male C57BL/6 mice with extracellular cold-inducible RNA-binding protein and various doses of milk fat globule-epidermal growth factor-VIII–derived oligopeptide 3 for 4 hours, then measured cytokines in the culture supernatants. In addition, mice underwent 70% hepatic ischemia for 60 minutes immediately followed by the intravenous administration of either vehicle or milk fat globule-epidermal growth factor-VIII–derived oligopeptide 3. Blood and ischemic liver tissues were collected 24 hours later, and inflammatory markers including cytokines, liver enzymes, chemokines, myeloperoxidase activity, and Z-DNA-binding protein 1 were measured. Hepatic tissue damage and cell death were evaluated histologically. Survival rates were monitored for 10 days posthepatic ischemia/reperfusion. ResultsThe release of interleukin-6 and tumor necrosis factor-α from extracellular cold-inducible RNA-binding protein–challenged Kupffer cells was significantly reduced by milk fat globule-epidermal growth factor-VIII–derived oligopeptide 3 in a dose-dependent manner. In hepatic ischemia/reperfusion mice, milk fat globule-epidermal growth factor-VIII–derived oligopeptide 3 treatment significantly decreased serum levels of extracellular cold-inducible RNA-binding protein, interleukin-6, tumor necrosis factor-α, aspartate aminotransferase, alanine aminotransferase, and lactate dehydrogenase. Milk fat globule-epidermal growth factor-VIII–derived oligopeptide 3 treatment also significantly reduced mRNA levels of interleukin-6, tumor necrosis factor-α, interleukin-1β, Z-DNA-binding protein 1, and chemokine macrophage inflammatory protein-2, as well as myeloperoxidase activity in hepatic tissues. Histologic evaluation demonstrated that treatment with milk fat globule-epidermal growth factor-VIII–derived oligopeptide 3 significantly attenuated tissue damage and cell death in the liver of hepatic ischemia/reperfusion mice. Milk fat globule-epidermal growth factor-VIII–derived oligopeptide 3 treatment significantly improved the survival rate of hepatic ischemia/reperfusion mice. ConclusionMilk fat globule-epidermal growth factor-VIII–derived oligopeptide 3 significantly attenuated inflammation and liver tissue damage and improved survival after hepatic ischemia/reperfusion. Thus, milk fat globule-epidermal growth factor-VIII–derived oligopeptide 3 holds promise as a potential future therapeutic strategy for hepatic ischemia/reperfusion injury.

Role of rs5743708 SNP in the risk of Bacterial Respiratory Infections in Children of Ramadi city/Iraq

Background and Aim: Respiratory Infections are a major global health and financial burden, so among cause the five most common and deadly for respiratory illnesses is acute lower respiratory tract infections (RTIs), these infections claim over 4 million lives annually. Toll-like receptors (TLRs) are key components of the innate immune system that recognize pathogen which associated a molecular patterns) PAMPs) and initiate an immune response. Understanding the role of TLR polymorphisms in susceptibility to respiratory infections in children is important for developing strategies to prevent and treat these infections. So, the current study aims to "Investigate the potential association between TLR2 gene polymorphisms and susceptibility to respiratory infections in children". Methodology: The study included collect of 300 samples with respiratory infection from both sex with range ages(1-12) years, and other 50 healthy control samples. The samples were collected from general and private hospitals in Ramadi city, Iraq. PCR was carried out using primer for gene target and amplification of target gene. Sanger Sequencing conducted with both direction for all samples. Results and Conclusion: The result of sputum culture indicate that 100 of suspected samples were positive culturing while other demonstrate a negative growth. The results analysis of sequencing according to Query and subjected samples indicated that 4 of patient have SNP of rs5743708 G&gt;A, while there is not found in control samples, and the results of genotype frequencies analysis it was found that TLR2 polymorphism was associated with increased risk for bacterial respiratory infections in children according to under study samples, but no statistical significant differences correlation of rs5743708 G&gt;A and infection, with adjusted odds ratio [OD:5.3; 95%Cl: 0.3-101; P: 0.2].

Excessive autophagy-inducing and highly penetrable biomineralized bacteria for multimodal imaging-guided and mild hyperthermia-enhanced immunogenic cell death

The tumor microenvironment, characterized by hypoxia, supports the efficacy of anaerobic bacteria like attenuated S. typhimurium in cancer therapies. These bacteria target and penetrate deep tumor regions, significantly reducing tumor size but often lead to tumor regrowth due to limited long-term efficacy. To enhance the therapeutic impact, a novel biohybrid system, S@UIL, has been developed by coating S. typhimurium with a zirconium-based nanoscale metal–organic framework (UiO-66-NH2) loaded with indocyanine green (ICG) and luteolin (LUT). This system maintains the bacteria’s tumor-targeting ability while increasing the penetration and therapeutic effectiveness through excessive autophagy and mild hyperthermia. In a subcutaneous colon cancer model, the integration of LUT and ICG promotes autophagic cell death and photothermal sensitization, leading to the release of damage-associated molecular patterns (DAMPs). These DAMPs activate immune responses through dendritic cells and T-cells, enhancing immunogenic cell death (ICD) and potentially reducing immune evasion by tumors. This single-administration approach also integrates multimodal imaging capabilities, providing a promising strategy for improved tumor ICD induction and cancer progression inhibition.

LL37-DNA Complex Drives Vitiligo Progression Through TLR9-MyD88 Signaling Pathways.

Vitiligo is an autoimmune disorder characterized by chronic depigmentation and milk-white patches on the skin. Skin infiltration by autoreactive CD8+ T cells causes melanocyte destruction in vitiligo. Multiple risk factors, particularly immune-related inflammatory factors, are involved in the disappearance of melanocytes. LL37 is a classic damage-associated molecular pattern molecule that is involved in the development of various autoimmune diseases. An enhanced expression of LL37 in vitiligo is known; however, the exact role of LL37 in melanocyte loss has not yet been elucidated. In the present study, we detected increased LL37 expression in vitiligo serum and lesions. Furthermore, we confirmed that cultured keratinocytes released LL37 after treatment with H2O2. Moreover, the LL37-DNA complex enhanced the secretion of CXCL9, CXCL10, and CXCL16 from keratinocytes via the TLR9-MyD88 signaling pathway and facilitated the migration of CD8+ T cells. Altogether, our study demonstrates that LL37 released from keratinocytes binds to DNA and contributes to melanocyte destruction under oxidative stress-induced autoimmunity in vitiligo.

Postoperative Immunosuppression Following Breast-Conserving Surgery vs. Mastectomy: The Role of Surgical Injuries and Intraoperative Sympathetic Activation

Breast cancer is the second most prevalent cancer among all types of cancers. Objectives: To evaluate the role of surgical tissue injury and intraoperative sympathetic activation in postoperative immunosuppression after breast-conservative surgery and mastectomy. Methods: This prospective/observational study investigated 36 breast cancer patients in the Department of Surgery Pir Abdul Qadir Shah Jeelani Institute of Medical Sciences, Gambat from June 2022 to May 2023. Patients who were on schedule to undergo either mastectomy or breast-conserving surgery enrolled. Patients were categorized into two groups; Group I (breast-conserving surgery group n=18) and Group II (mastectomy group n=18). The intraoperative sympathetic activation, plasma Damage-associated molecular patterns, and postoperative immune function were compared in both groups. Descriptive statistics were done using SPSS version 28.0. Results: The overall mean age and body mass index of Group I and Group II were 62.8 ± 8.9 vs. 60.6 ± 10.6 years and 26.9 ± 3.8 vs. 25.8 ± 3.7 kg/m2, respectively. The overall duration of surgery (minutes) was 56 ± 18 and 85 ± 22, respectively. The prominent indication for surgery in Group I and Group II was Invasive carcinoma 17 (94.4%) and 11 (61.1%), respectively. The concentration of plasma alarmins and IL-6 was significantly higher in patients who underwent mastectomy as compared to breast-conserving surgery. Conclusions: It was concluded that differences in Damage-associated molecular patterns release and intraoperative sympathetic activation between mastectomy and breast-conserving surgery may influence, and potentially contribute to, postoperative immune homeostasis to improve survival seen after breast-conserving surgery.