Systemic Response Research Articles

Abstract Su1207: Influence of Duration of Device-based Fever Prevention on Inflammation, Vasopressor Usage, and Oxygen Consumption after Cardiac Arrest - a Substudy of a Randomized Controlled Trial

Background: In the post-resuscitation phase, cardiac arrest patients develop a systemic inflammatory response and fever is likely if not actively treated. The Blood Pressure and Oxygenation Targets after Out-of-hospital Cardiac Arrest (BOX) trial, randomized patients to a total duration of device-based fever prevention for 36 or 72 hours. The trial found no differences in mortality or neurologic outcome. Use of device-based fever prevention for a shorter or longer duration could however result in clinically relevant differences in the magnitude of inflammation, vasopressor usage, and oxygen consumption. Aim: To assess the effect of a shorter or longer duration of device-based fever prevention on inflammation, vasopressor usage, and oxygen consumption in the intervention period from 36 to 72 hours. Methods: The BOX trail [NCT03141099] randomized comatose resuscitated out-of-hospital cardiac arrest (OHCA) patients to three different treatment targets in the post-resuscitation phase: mean arterial blood pressure target of 63 or 77mmHg; oxygenation targets of 68-75 or 98-105 mmHg; and duration of device-based fever prevention in comatose patients for a total of 36 or 72 hours. Endpoints at 48 and 72 hours: leukocytes; CRP; vasoactive-inotropic score (VIS); systemic oxygen consumption determined by use of Fick’s principle. Analyses were performed both on the total BOX population, and for a sensitivity analysis, selectively on patients still comatose at 72 hours (i.e., not awake or dead) as these had the most exposure to the intervention. Results: The total population consists of 789 patients, with 393 and 396 randomized to a total duration of device-based fever prevention for 36 and 72 hours respectively. For the analysis of patients still comatose at 72 hours, 122 and 148 were available for analysis in the 36-hour and 72-hour group respectively. There were no differences between the 36- and 72-hour group for leukocytes, CRP, VIS, or oxygen consumption during the intervention period from 36 until 72 hours, neither for the total population (Figure 1, all p>0.05) or the cohort of patients still comatose at 72 hours (Figure 2, all p>0.05). Conclusions: There were no group differences for patients randomized to device-based fever prevention for 36 or 72 hours with respect to the magnitude of inflammation, vasopressor usage, or oxygen consumption. These findings were consistent for the cohort as a whole, and when examining those with most exposure to the intervention.

THE EFFECT OF NF-ΚB ON THE INTENSITY OF OXIDATIVE STRESS IN THE CEREBRAL CORTEX OF RATS UNDER THE COMBINED INFLUENCE OF THE LIGHT-DARK CYCLE, SYSTEMIC INFLAMMATORY RESPONSE, AND SODIUM GLUTAMATE ADMINISTRATION

To date, scientists have confirmed the link between the development of oxidative stress and disruptions in the light regime, as well as the systemic inflammatory response. The impact of monosodium glutamate on increasing oxidative damage to rat brain tissue has also been identified. The aim of this study was to investigate the role of the transcription factor NF-κB by examining the effect of its inhibitor, ammonium pyrrolidinedithiocarbamate, on the development of oxidative stress in the cerebral hemispheres of rats, in combination with acute desynchronosis, systemic inflammatory response, and monosodium glutamate administration. The study was conducted on 45 white Wistar rats weighing 150-200 g, divided into three groups: control (n=15), a group subjected to a combination of acute desynchronosis, systemic inflammatory response, and sodium glutamate administration (n=15), and a group exposed to the combination of acute desynchronosis, systemic inflammatory response, and received sodium glutamate and pyrrolidinedithiocarbamate (n=15). To induce acute desynchronosis, the rats were initially kept under a regular light-dark cycle (12 hours of light, 12 hours of darkness) for 3 weeks, followed by a shift in the light-dark phases by 6 hours back over the next 3 days. The systemic inflammatory response was modeled through intraperitoneal administration of Salmonella typhi lipopolysaccharide. During the first week, lipopolysaccharide was administered three times at a dose of 0.4 μg per 1 kg of body weight, and during the following seven weeks, it was given once a week. Sodium glutamate, at a dose of 30 mg/kg, dissolved in 0.5 ml of distilled water, was administered intragastrically for 20 days. The NF-kB activation inhibitor ammonium pyrrolidinedithiocarbamate (Sigma-Aldrich, Inc., USA) was administered at a dose of 76 mg/kg three times a week for 20 days. In a 10% homogenate of the cerebral hemispheres, the following were measured: the rate of superoxide anion radical production, the content of thiobarbituric acid-reactive substances (TBARS), the increase in these parameters, as well as the activity of catalase and superoxide dismutase. Administration of pyrrolidinedithiocarbamate in combination with acute desynchronosis, systemic inflammatory response, and sodium glutamate reduced the rate of basic superoxide anion radical production by 10%, NADPH-induced production by 17.6%, NADH-induced production by 13%, reduced the concentration and growth of TBC-active products by 6.6% and 14.6%, respectively, increased the activity of superoxide dismutase by 35.2%, catalase by 10.5% compared to the group exposed to the combination of acute desynchronosis, systemic inflammatory response, and sodium glutamate administration. Conclusion. The administration of pyrrolidinedithiocarbamate in combination with a systemic inflammatory response, acute desynchronosis and the action of sodium glutamate reduces the production of the superoxide anion radical, the concentration and increase in TBC-active products, enhances antioxidant protection that indicates the possible influence of the nuclear factor NF-κB on the development of oxidative processes in the cerebral hemispheres of rats.

INFLUENCE OF SODIUM GLUTAMATE ON REACTIVE OXYGEN AND NITROGEN SPECIES PRODUCTION IN KIDNEY TISSUES OF RATS UNDER ACUTE DESYNCHRONIZATION AND LIPOPOLYSACCHARIDE-INDUCED SYSTEMIC INFLAMMATORY RESPONSE

The study investigated the effect of sodium glutamate on the production of reactive oxygen and nitrogen species (ROS/RNS) in kidney tissues of rats under conditions of acute desynchronization (AD) and lipopolysaccharide (LPS)-induced systemic inflammatory response (SIR) and, thus, contributed to a deeper understanding of the mechanisms involved in the development of renal dysfunction under these conditions. The experiment was conducted on 21 male Wistar rats (210-230 g), randomly divided into three groups: intact animals, a group exposed to AD modeling under LPS-induced systemic inflammatory response (Group II), and a group (Group 3) exposed to AD modeling under LPS-induced systemic inflammatory response, who received sodium glutamate in a dose of 20 mg/kg. The administration of sodium glutamate significantly increased the generation rate of the superoxide anion radical by 13.8% through microsomal monooxygenases, by 8.7% through the mitochondrial respiratory chain, and by 7.7% through leukocyte NADPH oxidase compared to Group 2. However, sodium glutamate did not affect the total NO synthase activity or the activity of its constitutive and inducible isozymes in the kidney homogenate compared to Group 2. The uncoupling index of constitutive NO synthases in kidney tissues also remained unchanged under the administration of sodium glutamate, but the peroxynitrites were 20.5% higher, and S-nitrosothiols were 11.6% higher, than in group 2. The findings indicate that sodium glutamate significantly increases oxidative-nitrosative stress in rat kidney tissues under AD and LPS-induced SIR, resulting in excessive ROS/RNS formation.

Association of Systemic Inflammatory Response Syndrome With Cardiovascular Events After Mitral Transcatheter Edge-to-Edge Repair.

Systemic inflammatory response syndrome (SIRS) following cardiovascular interventions is associated with adverse events during hospitalization and follow-up. Mitral transcatheter edge-to-edge repair is increasingly utilized for treatment of mitral regurgitation (MR). We investigated whether SIRS following mitral transcatheter edge-to-edge repair may occur and be associated with adverse clinical outcomes. A total of 158 consecutive patients with severe MR undergoing mitral transcatheter edge-to-edge repair were studied. SIRS was defined by leukocytosis (≥12 × 109/L) and fever (≥38 °C) within 48 hours after intervention. Baseline inflammation was measured by absolute neutrophil and lymphocyte counts and neutrophil-lymphocyte ratio. The primary end point of major cardiovascular events was the composite of nonfatal myocardial infarction, nonfatal stroke, and all-cause death. Recurrent MR at follow-up was also recorded. The mean patient age was 80.8±8.8 years. Forty-four (27.9%) developed SIRS. Neutrophil-lymphocyte ratio correlated with onset of leukocytosis and fever (P=0.04). During a median follow-up of 12.5 (5.4-17.4) months, the primary end point occurred in 27 (17.1%) patients (6 myocardial infarction, 5 strokes, and 16 deaths). Patients with SIRS more often had severe MR (79.5% versus 62.7%, P=0.02) at follow-up. After adjustment for pertinent variables, SIRS (HR 2.73 [95% CI, 1.08-6.86]; P=0.03) was independently associated with major cardiovascular events. SIRS after mitral transcatheter edge-to-edge repair is a strong independent predictor of major cardiovascular events. Closer follow-up is warranted because patients with SIRS have more severe MR at follow-up.

Local and systemic humoral immune responses to Histophilus somni recombinant antigens administered intranasally and subcutaneously to dairy calves

AbstractBovine respiratory disease (BRD) causes significant economic losses in dairy calves. Induction of an early immune response via parenteral vaccination is complicated by the interference of colostral immunity. In this study, we investigated early immunization against selected conserved bacterial antigens. Calves were vaccinated twice intranasally and then subcutaneously with Histophilus somni recombinant proteins (rOMP40 or rHsp60) mixed with one of two adjuvants: CpG ODN2007 or MPLA. The control group (Con) was treated with PBS. The first immunization was done between 24 and 48 h of life and then twice in two weeks intervals. Blood, nasal, and saliva secretion samples were collected directly before vaccination (S1–S3) and then on 42–44 (S4) and 59–61 (S5) day of life. Antibodies (IgG1/IgG2/IgM/IgA in serum; IgG1/IgA in secretions) against both vaccine antigens were quantified in all samples. Intranasal and subcutaneous vaccinations using the described formulas did not increase antibody reactivity against the tested proteins. The reactivity of serum IgG1, IgM, and IgA anti-rOMP40 antibodies was significantly higher in S1 in all groups than that in the other samplings (p˂0.01). Significant differences in the reactivity of serum anti-rOMP40 antibodies between groups were identified in S1 (IgA reactivity was higher in the CpG vs. MPLA group; p < 0.05), S4 (IgM reactivity was higher in Con vs. CpG group; p < 0.05), and S5 (IgG1 reactivity was higher in MPLA vs. Con group; p < 0.05). The lack of consistent changes in antibodies after immunization (S4 and S5) hinders the drawing of conclusions regarding the effect of immunization on antibody reactivity. In the future, establishing a proper immunization window and adjuvants for nasal vaccines against bacterial pathogens causing BRD in calves remains to be determined.

EXTH-57. EXPLORING NOVEL ROUTES FOR NON-INVASIVE DRUG ADMINISTRATION: INTRADERMAL DELIVERY FOR THE TREATMENT OF BRAIN TUMORS

Abstract Effective non-invasive drug delivery to brain tumors remains challenging, despite extensive exploration of these methods, due to the blood-brain barrier (BBB). Intranasal drug delivery has emerged as a promising alternative approach, as it circumvents limitations on tumor access imposed by the BBB when administering therapeutics systemically and limits the occurrence of systemic toxic responses to treatment. The intranasal route of administration allows therapeutic access to brain tumors through pathways involving the olfactory and trigeminal nerves (TNs) that connect the nasal passage to the brain. However, effective nose-to-brain delivery is impeded by rapid mucosal drug clearance in the nasal cavity, drug aspiration, and ingestion. Since the TN arises from the brainstem and its branches innervate the facial skin, in addition to muscles, meninges, and respiratory mucosa, we tested the hypothesis that facial intradermal administration targeting the V2 branch of the TN could be an alternative strategy to harness TN-mediated brain delivery to bypass the BBB. Here, we developed Spherical Nucleic Acids (SNAs) consisting of a 15nm spherical gold nanoparticle core functionalized with radially oriented fluorophore-tagged ssDNA as a model therapeutic. We observed the delivery of the SNAs to the brain starting 4 hours post intradermal injection, with the SNA accumulation in the tumor persisting over time. Using ex vivo fluorescent imaging, comprehensive immunofluorescence microscopy, and flow cytometry we confirmed the SNA accumulation in the brain, dura, TN, and cervical lymph nodes. In particular, we demonstrated that SNAs were enriched in the neuronal cell bodies of the trigeminal ganglia and within the epineurium and perineurium of the TN. The SNAs were also enriched in CD45-positive immune cells. In conclusion, this study credentialed facial intradermal administration of SNAs as a promising non-invasive method for brain drug delivery to target intracranial tumors.

ETIOPATHOGENETIC MECHANISMS OF IMMUNE DYSFUNCTION IN COMBATANTS WITH LOWER LIMB SOFT TISSUE INJURIES UNDER CHRONIC STRESS

Introduction. Combat injuries, including gunshot, shrapnel, and mine-explosive wounds, affect a significant number of soldiers in modern warfare. Notably, most of these injuries involve damage to the soft tissues of the extremities. Surgeons have expressed concerns regarding the unsatisfactory treatment outcomes in this group of combatants, attributing one of the primary challenges to the limited understanding of immune dysfunction pathogenesis in military trauma cases. This study aims to address this gap by examining immune system dysfunctions in combat-related injuries. The objective of this study is to thoroughly analyze and synthesize the key stages of immune dysfunction occurring over extended periods post-combat trauma, including the subsequent development of traumatic disease and various wound complications. Materials and Methods. The rising prevalence of combat trauma among soldiers has intensified interest in studying this issue, prompting surgeons and traumatologists to address its various medical aspects comprehensively. The literature search focused on recent publications, allowing for a targeted analysis of the immunological aspects relevant to military medical traumatology. Results. In the initial stages of severe or combined injuries affecting various tissues—such as tubular bones, joints, blood vessels, and peripheral nerves—systemic inflammatory response syndrome (SIRS) commonly occurs. This stage is marked by an intense activation of innate antibacterial and immune-protective responses, leading to a significant increase in inflammation. This initial response is soon replaced by a prolonged phase known as compensatory anti-inflammatory response syndrome. During this period, immune-protective responses sharply decrease, certain immunocompetent cells become inhibited, and lymphopenia develops. This phase is often accompanied by infectious contamination of wounds with pathogenic and opportunistic microorganisms, resulting in both local purulent-necrotic processes and potentially severe systemic complications, such as septic shock, sepsis, multiple organ failure, and others. The final stage, known as persistent inflammatory, immunosuppressive, catabolic syndrome, is characterized by the chronic progression of traumatic disease, accompanied by ongoing immune system dysfunction in combatants. Conclusion. In the early period of traumatic injury, the wounded experience sharp inflammatory processes and activation of immune defense mechanisms. At subsequent stages, severe disruptions in the functioning of the immune system, damage to internal organs, and the development of catabolic syndrome are recorded. These changes, especially those resulted from exposure to chronic combat stress preceding the injury, aggravate the processes of infectious decontamination of wounds, regeneration of damaged tissues, and the general process of combatant rehabilitation.

CTIM-34. EFFECTS OF NEOADJUVANT IMMUNE CHECKPOINT INHIBITION ON PEDIATRIC AND ADULT HIGH-GRADE GLIOMAS

Abstract High-grade gliomas (HGG) are lethal tumors with few long-term survivors affecting adult and pediatric patients. Studies investigating immune checkpoint inhibition (ICI) suggest administration of neoadjuvant anti-PD1 blockade (neo-aPD1) enhances local and systemic anti-tumor responses. This data is supported by increased T cell density in the tumor and proportion of cytotoxic T cells in peripheral blood. In a previous randomized clinical trial, neo-aPD1 improved survival in adult with recurrent glioblastoma. In this study, we utilized multiplex immunofluorescence (mIF), and single-cell RNA sequencing (scRNA-seq) to investigate the effects of neoadjuvant ICI on the TME and compare between pediatric and adult populations. Tissue samples from adult patients with recurrent GBM treated with neo-aPD1 (n=22) or adjuvant only therapy (n=5) were obtained from our UCLA off-label neo-PD1 study. Tissue samples were also obtained from pediatric patients with recurrent/progressive HGG treated with neoadjuvant ICI from the ongoing Pediatric Neuro-Oncology Consortium study PNOC19. Samples included diagnostic tissue from two patients and on study surgery tissue from four patients. All tissue samples were subjected to mIF and underwent imaging analysis to assess immune cell infiltration by quantifying lymphoid and myeloid cell densities across the two treatment groups. Our preliminary findings elucidated a trend of higher T cell infiltration in adult neoadjuvant compared to adjuvant-only treated patients (median density 58.11 vs 15.15 T-cells/mm2). Compared to their adult counterparts, neoadjuvant ICI treated pediatric patients had even more robust total T cell infiltration (median density 268.10 vs 58.11 T-cell/mm2, pediatric vs adult respectively). scRNA-seq analysis revealed a greater enrichment of monocytic macrophages in the pediatric population compared to tissue resident microglia in the adults. This early data requires analysis of additional samples to better understand differences in the anti-tumor immune response between adult and pediatric recurrent/progressive HGG patients.

Dissecting Immunological Mechanisms Underlying Influenza Viral Nucleoprotein-induced Mucosal Immunity Against Diverse Viral Strains

The nucleoprotein (NP) of type A influenza virus (IAV) is highly conserved across all virus strains, making it an attractive candidate antigen for universal vaccines. While various studies have explored NP-induced mucosal immunity, here we interrogated the mechanistic differences between intramuscular (IM) and intranasal (IN) delivery of a recombinant adenovirus carrying NP fused with a bifunctional CD40 ligand. Despite being less effective than IM delivery in inducing systemic cellular immune responses and antibody-dependent cellular cytotoxicity (ADCC), IN immunization elicited superior antigen-specific recall humoral and cellular response in the nasal associated lymphoid tissue (NALT) of the upper respiratory tract, the initial site of immune recognition and elimination of inhaled pathogens. IN vaccination also induced significantly stronger pulmonary T cell responses in the lower respiratory tract than IM vaccination, in particular the CD8 T cells. Moreover, blocking lymphocyte circulation abrogated IM but not IN immunization induced protection, illustrating the critical role of local memory immune response upon viral infection. Notably, the CD40-targeted nasal delivery not only improved the magnitude but also the breadth of protection, including against lethal challenge with a newly isolated highly pathogenic avian H5N1 strain. These findings are informative for the design of universal mucosal vaccines, where the predominant mode of protection is independent of neutralizing antibodies.

Distinct distribution and responses of IgM+, IgT1+ and IgT2+ B cells in common carp

In teleosts, the immunoglobulin classes produced by B cells are IgM, IgD, and IgT/IgZ. IgT was initially described as an immunoglobulin specialized in mucosal responses; accumulating evidence, however, shows that it is also involved in systemic immune responses. Two types of IgT/IgZ (IgT1 and IgT2) were previously described in common carp, but their further characterization was hampered by the lack of specific tool. In the current study, we developed and validated polyclonal antibodies against carp IgT1 and IgT2 and used them in combination with well validated monoclonal antibody against carp IgM (WCI12), to study the distribution of IgM+, IgT1+ and IgT2+ B cells or their secreted immunoglobulins in various mucosal and systemic organs of carp. Finally, we also preliminary assessed the B cell response to infection with the blood-borne parasite Trypanoplasma borreli. Using these tools, we report on the distinct expression of soluble immunoglobulins in systemic and mucosal compartments. IgT1 and IgM were expressed in mucosal as well as systemic organs and responded to systemic parasitic infection, whereas IgT2 was preferentially expressed at mucosal sites and did not respond to systemic infections. By studying the distribution of B cells in different organs, compartmentalization of the three B cell subtypes was observed in gills and gut, whereas splenic B cells appeared as organized clusters around ellipsoids. Our results provide insights into the distribution and to some extent the function of B cells in carp, indicating that our newly developed tools are valuable for future studies aiming at the further characterization of immune responses of carp to infections and vaccination.

Exploring the predictive factors in the gastrointestinal involvement of patients with immunoglobulin A vasculitis

Background. Immunoglobulin A vasculitis (IgAV), the most common systemic vasculitis in children, typically presents with gastrointestinal (GI) symptoms in about half of cases. This study aimed to analyze the clinical and laboratory findings of patients with IgAV regarding GI involvement. Methods. We compared the GI involvement data of the patients diagnosed with IgAV. Results. Of the 210 patients (60.5% female and 39.5% male), 101 had GI involvement, with abdominal pain being the predominant symptom (n=98). White blood cell, neutrophil, monocyte, and platelet counts, C-reactive protein, neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), monocyte-to-lymphocyte ratio (MLR), systemic immune-inflammation index (SII), and systemic inflammation response index (SIRI) were significantly elevated in patients with GI involvement (p<0.001, p<0.001, p=0.01, p=0.005, p=0.002, p<0.001, p=0.03, p=0.001, p<0.001, p<0.001, respectively). The cutoff values for SII (>1035.7), SIRI (>1.65), NLR (>2.73), and MLR (>0.28) were determined, yielding respective sensitivities of 46%, 59%, 47%, and 53%, specificities of 83.1%, 69.1%, 81.3%, and 71.9%. Corresponding areas under the curve were 0.658, 0.668, 0.649, and 0.634, respectively (all p<0.001). Conclusion. Although IgAV is a self-limiting disease, GI involvement can lead to serious consequences. Systemic inflammatory indices such as SII and SIRI may be indicative in identifying patients with GI involvement.

CSIG-13. TARGETING GENETIC DEPENDENCIES EXPOSES DIFFUSE MIDLINE GLIOMA CELL OF ORIGIN VULNERABILITIES

Abstract Diffuse midline glioma (DMG), including diffuse intrinsic pontine glioma (DIPG), are uniformly fatal brain cancers affecting children, adolescents, and young adults. These tumors are characterized by ‘oncohistone’ mutations in histone H3 genes (H3F3A, HIST1H3B, HIST1H3C), resulting in the substitution of lysine 27 to methionine (H3K27M), resulting in dominant negative hypomethylation at lysine 27 (H3K27) and a global loss of gene silencing. CRISPR-Cas9 loss-of-function gene deletion screens identified mutation-independent dependencies on PIK3CA and MTOR genes for tumor growth and proliferation, highlighting a targetable molecular dependency across DMG patient-derived models (n=38). However, systemic PI3K/mTOR inhibition increased blood glucose and insulin levels, promoting hyperinsulinemia/hyperglycemia and reduced efficacy in vivo. To exploit genetic dependencies while maintaining compliance and therapeutic benefit, we optimized combinations of clinically relevant PI3K inhibitors (paxalisib, GCT007, everolimus) with the antiglycemic drug metformin to restore glucose homeostasis and decrease DMG insulin receptor activity in vivo, extending the survival of DIPG xenograft models. Phosphoproteomic profiling of DIPG models treated with PI3K/mTOR inhibitors promoted calcium-activated PKC signaling. The brain-penetrant PKC inhibitor enzastaurin in combination with paxalisib, synergistically extended the survival of DIPG xenograft models, including those mimicking disease progression, with further benefits potentiated using metformin in a multimodality approach. Combined PI3K-PKC inhibition in vivo promoted differentiation of OPC-like DMG to more OC-like cells, enhancing PDGFRA-JAK/STAT proinflammatory signaling, showing features of demyelination and MHC-II+ antigen expression, including PD1/PDL1. In parallel, combined PI3K/mTOR and PDGFRA inhibition using paxalisib and avapritinib synergistically extended the survival of patient-derived xenograft models, showing the preclinical relevance of simultaneously targeting these vulnerabilities. Together, we reveal that therapeutic inhibition of PI3K/mTOR and compensatory PKC signaling, while mitigating side effects with metformin, altered the chromatin architecture, leading to cellular differentiation and opening the door for clinically relevant checkpoint inhibitors. This combination strategy addresses DMG genetic dependencies, cellular and systemic responses to precision therapies, while harnessing the immune system for potential clinical translation.

Extracellular DNA and Deoxyribonuclease Activity as Prognostic Markers in Sepsis

Background/Objectives: Sepsis is characterized by a dysregulated immune response to infection and is associated with high lethality. Extracellular DNA (ecDNA) has drawn significant interest as a damage-associated molecular pattern because of its potential involvement in the pathophysiology of sepsis. Methods: In this study, we examined the ecDNA concentration in 27 adult patients admitted to the intensive care unit. Fluorometry and quantitative PCR were used for the assessment of ecDNA. In addition, deoxyribonuclease activity was measured as a potential modulator of ecDNA. Results: Our findings reveal nearly 5-fold higher concentrations of ecDNA in non-survivors, suggesting its potential as a prognostic indicator for sepsis outcomes on day 7. Interestingly, the subcellular origin of ecDNA was similar between patients diagnosed with systemic inflammatory response syndrome, sepsis, and septic shock. Deoxyribonuclease activity, implicated in the cleavage of ecDNA, was comparable across all patient groups. Conclusions: To establish the prognostic value of ecDNA as a biomarker, further investigations within a larger patient cohort are needed. Nevertheless, our results suggest that high ecDNA in sepsis patients represents a negative prognostic biomarker.

Oral Administration of Zinc Sulfate with Intramuscular Foot-and-Mouth Disease Vaccine Enhances Mucosal and Systemic Immunity

Background/Objectives: Foot-and-mouth disease (FMD) remains a significant global threat to livestock farming. Current commercial FMD vaccines present several challenges, including the risk of infection and adverse injection site reactions due to oil-based adjuvants. The complex immune environment of the gut-associated lymphoid tissue has the potential to induce broad and diverse immune responses. Therefore, we aimed to explore the potential of zinc sulfate as an oral adjuvant to enhance intestinal mucosal immunity and complement the effects of intramuscular (IM) FMD vaccination. Methods: We conducted serological analyses on mice and pigs, measuring secretory IgA (sIgA) levels and evaluating the expression of mucosal immunity-related genes in pigs. These assessments were used to investigate the systemic and mucosal immune responses induced by oral zinc sulfate administration in combination with an IM FMD vaccine. Results: This combination strategy significantly increased structural protein antibody titers and virus neutralization titers in experimental animals (mice) and target animals (pigs) across early, mid-, and long-term periods. Additionally, this approach enhanced the expression of key cytokines associated with mucosal immunity and increased sIgA levels, which are critical markers of mucosal immunity. Conclusions: Oral zinc sulfate administration may synergize with inactivated FMD vaccines, leading to sustained and enhanced long-term immune responses. This novel strategy could reduce the frequency of required vaccinations or allow for a lower antigen dose in vaccines, effectively stimulating the mucosal immune system and boosting systemic immunity. This approach has the potential to improve the overall efficacy of commercial FMD vaccines.

Ultrasound-triggered nano delivery of lenvatinib for selective immunotherapy treatment against hepatocellular carcinoma

Although there have been remarkable advances in the treatment of hepatocellular carcinoma (HCC), the prognosis remains poor in those with advanced stage and more effective therapeutic options are urgently needed. Lenvatinib (Len), a multiple receptor tyrosine kinase inhibitor, is an emerging molecular targeted agent for HCC, whose immunomodulatory activities have been investigated. However, Len utilization is limited because of its low metabolic stability, poor bioavailability and dose-dependent toxicity, rendering its direct use insufficient for immune modulation. Stimuli-responsive nanoparticles, which are drug-targeted delivery platforms for on-demand drug release, facilitate deeper and uniform tumour penetration, providing alternative solutions to overcome current limitations. Ultrasound (US) exhibits superior tissue penetration abilities and can produce reactive oxygen species (ROS) at the tumour site to treat deeper tumours. In addition, US serves as an excellent and selective drug delivery mediator for tumour treatment. Herein, we designed US-triggered lenvatinib nanoparticles (Len-RNPs) for selective drug delivery that utilize US-triggered ROS to induce in situ oxidation reactions, resulting in nanoparticle disintegration. Len-RNPs mitigate the toxicity of Len and effectively trigger robust systemic anti-tumour immune responses in a H22 tumour model, resulting in a tumour suppression rate of 95.7%, with 60% of mice being cured. Our study elucidates a novel and precise strategy of combining Len and US therapy for enhanced HCC immunotherapy.

Survival of Patients with Alcohol-Related Liver Disease Cirrhosis—Usefulness of the New Liver Mortality Inpatients Prognostic Score

Background/Objectives: Alcohol can directly damage the liver, causing steatosis, steatohepatitis, cirrhosis, and hepatocellular cancer. The aim of this study was to examine 28-day survival in hospitalized patients with alcohol-related liver disease (ALD) cirrhosis, as well as to develop and validate a new survival prediction model. Methods: A total of 145 patients with ALD cirrhosis were included; 107 were diagnosed with acute decompensation (AD) and 38 with acute-on-chronic liver failure (ACLF). The new liver mortality inpatients (LIV-IN) score was calculated using the following variables: hepatic encephalopathy (HE), hepatorenal syndrome (HRS), ascites, systemic inflammatory response syndrome (SIRS), community-acquired infection (CAI), and fibrinogen. The diagnostic accuracy of the LIV-IN score was tested, along with the model for end-stage liver disease (MELD), model for end-stage liver disease-sodium (MELD-Na), albumin-bilirubin (ALBI), neutrophil-to-lymphocyte ratio (NLR), chronic liver failure consortium-C acute decompensation (CLIF-C AD), and chronic liver failure consortium-acute-on-chronic liver failure (CLIF-C ACLF). Results: Lethal outcome occurred in 46 (31.7%) patients. The mortality rate was higher in the ACLF group (n = 22, 57.9%) compared to the AD group (n = 24, 22.4%) (p < 0.01). The highest predictive power for short-term mortality was observed for the LIV-IN score (AUC 73.4%, p < 0.01). In patients with AD, the diagnostic accuracy of the CLIF-C AD score was better than for the LIV-IN score (AUC 0.699; p = 0.004, AUC 0.686; p = 0.007, respectively). In patients with ACLF, only the LIV-IN score had statistically significant discriminative power in predicting 28-day survival. Conclusions: The liver mortality inpatients prognostic score is a new, reliable prognostic model in predicting 28-day mortality.

Effect and mechanisms of shikonin on breast cancer cells in vitro and in vivo

BackgroundBreast cancer seriously affects physical and mental health of women. Despite advances in the clinical use of different treatments, breast cancer remains a major cause of mortality. Therefore, it is imperative to identify promising treatment options. In the present study, we investigated the effects of shikonin on 4T1 breast cancer cells and its potential mechanisms of action.MethodsBALB/c-derived mouse breast cancer 4T1 is very close to human breast cancer in growth characteristics and systemic response, so 4T1 cells were selected for further experiments. Cell viability, apoptosis, intracellular reactive oxygen species (ROS), mitochondrial activity, and cellular calreticulin (CRT) exposure were assessed to evaluate the antitumor effects and mechanisms of shikonin in vitro. Orthotopic tumor growth inhibition and splenic immune cell regulation by shikonin were evaluated in 4T1 breast cancer orthotopic mice in vivo.ResultsIn vitro, shikonin could inhibit cell proliferation, cause apoptosis, disrupt mitochondrial activity, and induce ROS production and CRT exposure. In vivo, shikonin inhibited tumor growth, increased the proportion of CD8+ T cells, and reduced the proportion of regulatory cells (CD25+ Foxp3+ T cells) in the spleen.ConclusionsShikonin inhibits the growth of 4T1 breast cancer cells by disrupting mitochondrial activity, promoting oxidative stress, and regulating immune function.

Evaluating the Diagnostic and Prognostic Value of Interleukin-6 (IL-6) and Soluble Triggering Receptor Expressed on Myeloid Cells-1 (sTREM-1) in Systemic Inflammatory Response Syndrome (SIRS) and Sepsis in Adults

Evaluating the Diagnostic and Prognostic Value of Interleukin-6 (IL-6) and Soluble Triggering Receptor Expressed on Myeloid Cells-1 (sTREM-1) in Systemic Inflammatory Response Syndrome (SIRS) and Sepsis in Adults

Systemic immune inflammatory response index (SIIRI) in acute myocardial infarction.

Different treatment approaches exist for non-ST elevation acute coronary syndrome (ACS) patients. This study assessed the systemic immune inflammatory response index (SIIRI) for its prognostic value and incremental clinical utility in determining optimal timing for percutaneous coronary intervention (PCI) in non-ST elevation myocardial infarction (NSTEMI) patients, particularly when troponin levels are initially negative. This study included 1270 ACS patients: 437 STEMI, 422 NSTEMI, and 411 unstable angina. Patients were stratified by SIIRI levels measured at admission, and coronary artery disease severity was evaluated using the SYNTAX score. The primary endpoint was major adverse cardiovascular events (MACE), including cardiovascular death, non-fatal myocardial infarction, stroke, and revascularization. Secondary endpoints encompassed individual MACE components and heart failure hospitalisations. The mean age was 54.93 years (83% male). SIIRI levels were significantly higher in STEMI patients (6.83 ± 6.43 × 105) compared to NSTEMI (4.5 ± 5.39 × 105) and unstable angina (3.48 ± 2.83 × 105) (P < 0.001). Area under the curve for SIIRI distinguished NSTEMI and unstable angina from STEMI (0.81 and 0.80), with optimal cut-off points of 4.80 × 105 and 4.25 × 105. In NSTEMI, 24.6% presented within 2 h of symptom onset, were troponin-negative, yet had elevated SIIRI. Post-PCI, SIIRI > 4.93 × 105 correlated with increased MACE at 1 year (17.2% vs 5%). NSTEMI and unstable angina patients with SIIRI values >4.80 × 105 and 4.25 × 105 respectively, may require urgent intervention (<2 h). SIIRI can be of significant utility in patients of NSTEMI who present earlier with negative troponins. SIIRI can also aid in identifying high-risk individuals post-PCI, providing a valuable tool for early and accurate assessment.

TNF-alpha gene promoter's hypomethylation mediates a pro-inflammatory phenotype in peripheral blood monocytes from apical periodontitis individuals.

Epigenetic regulation of the key inflammatory genes plays a crucial role in controlling monocyte/macrophage-mediated local and systemic responses to bacterial challenges. However, it has not been addressed in apical periodontitis (AP). We aimed to explore the methylation pattern of the TNF-α gene promoter and its association with the inflammatory phenotype of peripheral blood monocytes from individuals with AP and controls. A cross-sectional study was conducted, including otherwise healthy individuals with AP (n = 25) and controls (n = 29). Monocytes were isolated from the volunteer's blood samples using a Ficoll gradient followed by negative immunoselection. RNA and DNA were extracted. The DNA methylation profiles of the TNF-α gene promoter region were analyzed using bisulfite sequencing PCR. The mRNA expression levels of DNA methyltransferases 3a (DNMT3a) and Ten Eleven Translocation enzymes 1(TET1) were assessed by qPCR. A fraction of primary monocytes was also cultured for 24 h, and the supernatant was collected to measure cytokine levels through a Luminex assay. Generalized structural equation models (GSEM) evaluated the association between AP, DNA methylation, and TNF-α protein expression controlled for potential covariates. Models included the effect of the methylation of TNF-α gene promoter as a mediator of the association between AP and TNF-α protein expression levels. Monocytes from AP individuals exhibited a heightened secretion of TNF-α and IL-1β and hypomethylation of the TNF gene promoter (p < .05). AP diagnosis was associated with the TNF-α gene promoter´s hypomethylated profile and enhanced pro-inflammatory cytokine levels, while lower methylation of the gene promoter region and -163 CpG single site mediated TNF-α overexpression (p < .05). DNA hypomethylation at the TNF-α gene mediates a proinflammatory phenotype in monocytes from AP patients, supporting a role in the systemic response.