Primary Response Research Articles

Respiratory Syncytial Virus Vaccines: Analysis of Pre-Marketing Clinical Trials for Immunogenicity in the Population over 50 Years of Age.

Immunosenescence refers to age-related alterations in immune system function affecting both the humoral and cellular arm of immunity. Understanding immunosenescence and its impact on the vaccination of older adults is essential since primary vaccine responses in older individuals can fail to generate complete protection, especially vaccines targeting infections with increased incidence among the elderly, such as the respiratory syncytial virus. Here, we review clinical trials of both candidate and approved vaccines against respiratory syncytial virus (RSV) that include adults aged ≥50 years, with an emphasis on the evaluation of immunogenicity parameters. Currently, there are 10 vaccine candidates and 2 vaccines approved for the prevention of RSV in the older adult population. The number of registered clinical trials for this age group amounts to 42. Our preliminary evaluation of published results and interim analyses of RSV vaccine clinical trials indicates efficacy in older adult participants, demonstrating immunity levels that closely resemble those of younger adult participants.

Abstract 6596: BRMS1 downregulation alters the tumor microenvironment and induces immunotherapy resistance in lung adenocarcinoma

Abstract Background: Lung cancer is the leading cause of cancer-related deaths globally. During cancer progression, tumor cells interact with tumor-infiltrating immune cells (TIICs) creating an immunosuppressive microenvironment. Therefore, it is essential to delineate the communication dynamics between cancer cells and TIICs to identify novel pathways for therapeutic targeting. BRMS1 is a metastasis suppressor gene that is frequently downregulated in lung adenocarcinoma (LUAD). We, and others, have shown that loss of BRMS1 results in increased distant metastatic disease. Given the strong correlation between immune suppression, metastasis, and immunotherapy resistance we sought to elucidate if BRMS1 contributes to these processes. Specifically, we hypothesized that loss of BRMS1 in LUAD cells influences the immune cell composition, shaping an immunosuppressive microenvironment and decreasing tumor response to immunotherapy. Methods: To experimentally address our hypothesis, we generated KrasG12D P53fl/fl Brms1−/− mice inducing spontaneous tumors with Ad-Cre intratracheal inoculation. RNA-seq scRNA-seq on Brms1+/+ and Brms1−/− tumors, coupled with cell type annotations using mMCP-counter and DCATS were used to identify diverse TIIC subsets. To validate our observation, we performed immunofluorescence and flow cytometry to assess the number of TIICs. To explore the impact of Brms1 loss on the efficacy of anti-PD1 immunotherapy, a syngeneic CMT167 mouse model with BRMS1KD cells implanted in the flank was treated with an anti-PD1 antibody. Results: Differential gene expression analysis of RNA-seq data revealed a reduction in CCL9, CCL24, CLEC1B, PYCARD and other proinflammatory molecules in Brms1−/− tumors. Gene ontology and GSEA highlighted a diminished immune response signature in Brms1−/- tumors. In contrast, hallmark gene sets like IFNγ response, IL6 signaling, and inflammation are highly enriched in Brms1+/+ tumors. mMCP-counter suggested a decrease in CD8+ T cells in Brms1−/- tumors. Likewise, scRNA-seq data confirmed a significant decrease in cytotoxic CD8+ T cells in the Brms1−/− mice. Immunofluorescence and flow cytometric analysis showed a reduction in cytotoxic CD8+ T cells with low proliferative potential (Ki67+) in the Brms1−/− mice. Postmortem on our syngeneic mouse tumors after a 30-day anti-PD1 antibody treatment revealed a significantly larger mean tumor size in the BRMS1KD group compared to the WT group (942.4 mm³ vs. 184 mm³; p<0.023). Immunophenotyping on these tumors using flow cytometry also demonstrated reduced cytotoxic CD8+ T cells in the KD group post-treatment. Conclusion: Our data suggest that reduced BRMS1 expression in LUAD alters CD8+ T cell composition and primary tumor response to immunotherapy. Implications of our findings underscore the potential of BRMS1 as a therapeutic target in the context of immunotherapy-resistant LUAD. Citation Format: Manendra B. Lankadasari, Yuan Liu, Brooke Mastrogiacomo, Yingjie Zhu, Elizabeth G. Dunne, Cameron N. Fick, Choudhary Harsha, Jorge S. Reis-Filho, David R. Jones. BRMS1 downregulation alters the tumor microenvironment and induces immunotherapy resistance in lung adenocarcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 6596.

Humoral Immune Response to SARS-CoV-2 Spike Protein Receptor-Binding Motif Linear Epitopes.

The worldwide spread of SARS-CoV-2 has led to a significant economic and social burden on a global scale. Even though the pandemic has concluded, apprehension remains regarding the emergence of highly transmissible variants capable of evading immunity induced by either vaccination or prior infection. The success of viral penetration is due to the specific amino acid residues of the receptor-binding motif (RBM) involved in viral attachment. This region interacts with the cellular receptor ACE2, triggering a neutralizing antibody (nAb) response. In this study, we evaluated serum immunogenicity from individuals who received either a single dose or a combination of different vaccines against the original SARS-CoV-2 strain and a mutated linear RBM. Despite a modest antibody response to wild-type SARS-CoV-2 RBM, the Omicron variants exhibit four mutations in the RBM (S477N, T478K, E484A, and F486V) that result in even lower antibody titers. The primary immune responses observed were directed toward IgA and IgG. While nAbs typically target the RBD, our investigation has unveiled reduced seroreactivity within the RBD's crucial subregion, the RBM. This deficiency may have implications for the generation of protective nAbs. An evaluation of S1WT and S2WT RBM peptides binding to nAbs using microscale thermophoresis revealed a higher affinity (35 nM) for the S2WT sequence (GSTPCNGVEGFNCYF), which includes the FNCY patch. Our findings suggest that the linear RBM of SARS-CoV-2 is not an immunodominant region in vaccinated individuals. Comprehending the intricate dynamics of the humoral response, its interplay with viral evolution, and host genetics is crucial for formulating effective vaccination strategies, targeting not only SARS-CoV-2 but also anticipating potential future coronaviruses.

Age-related changes in responsiveness to non-invasive brain stimulation neuroplasticity paradigms: A systematic review with meta-analysis

ObjectivesWe aimed to summarise and critically appraise the available evidence for the effect of age on responsiveness to non-invasive brain stimulation (NBS) paradigms delivered to the primary motor cortex. MethodsFour databases (Medline, Embase, PsycINFO and Scopus) were searched from inception to February 7, 2023. Studies investigating age group comparisons and associations between age and neuroplasticity induction from NBS paradigms were included. Only studies delivering neuroplasticity paradigms to the primary motor cortex and responses measured via motor-evoked potentials (MEPs) in healthy adults were considered. Results39 studies, encompassing 40 experiments and eight NBS paradigms were included: paired associative stimulation (PAS; n = 12), repetitive transcranial magnetic stimulation (rTMS; n = 2), intermittent theta burst stimulation (iTBS; n = 8), continuous theta burst stimulation (cTBS; n = 7), transcranial direct and alternating current stimulation ((tDCS; n = 7; tACS; n = 2)), quadripulse stimulation (QPS; n = 1) and i-wave periodic transcranial magnetic stimulation (iTMS; n = 1). Pooled findings from PAS paradigms suggested older adults have reduced post-paradigm responses, although there was considerable heterogeneity. Mixed results were observed across all other NBS paradigms and post-paradigm timepoints. Conclusions/significanceWhilst age-dependent reduction in corticospinal excitability is possible, there is extensive inter- and intra-individual variability both within and between studies, making it difficult to draw meaningful conclusions from pooled analyses.

Abstract 894: Radiomics at baseline MRI are associated with tumor shrinkage in patients with unresectable renal cell carcinoma treated with neo-adjuvant Cabozantinib

Abstract Background: Neoadjuvant Cabozantinib targeted therapy (nCabo) has been shown to facilitate tumor size reduction in patients with renal cell carcinoma (RCC). However, there is unmet clinical need to identify pre-treatment markers predictive of response to nCabo to limit financial, dose-related toxicity. We investigated computationally derived radiomic signatures from baseline magnetic resonance imaging (MRI) for their association with response to tumor shrinkage following nCabo.Methods Seventeen patients with non-metastatic, clinical stage ≥T3, biopsy-confirmed clear cell RCC were enrolled in a phase II clinical trial (NCT04022343) examining primary tumor response to nCabo. Patients underwent 3 Tesla MRI at baseline, 6 and 12 weeks. Partial response (PR) or stable disease (SD) was determined per Response Evaluation Criteria in Solid Tumors at 12 weeks. Tumor regions of interest (ROI) were delineated on axial, arterial phase T1-weighted (T1W) series under the guidance of expert radiologist. T1W were normalized with respect to enhancement in cortex region of kidney. 75 radiomic features quantifying texture heterogeneity were derived from tumor ROI on a per-voxel basis at baseline. Wilcoxon ranksum test was used to evaluate significant differences in radiomics between patient groups PR and SD. Results: Following nCabo, 6/17 patients experienced PR. 22 radiomic features from T1W quantifying intensity based heterogeneity showed significant differences between patients with SD and PR after nCabo (p<0.05; top 3 in Table). None of the clinical variables showed significant differences between SD and PR. Conclusion: In this proof-of-concept preliminary study, we observed that radiomics at baseline MRI may allow for identifying RCC patients who would favorably respond to nCabo in terms of tumor shrinkage. Future studies will include analysis on longitudinal data, correlation against survival on large-scale datasets. Association between tumor shrinkage following nCabo and radiomic features, clinical variables Stable Disease (N=6) Partial Response (N=11) Category Feature P-value median IQR median IQR Radiomic Haralick 0.01 1.50 0.51 2.31 0.51 Gradient Filter 0.02 88.06 107.28 289.58 107.28 CoLlAGe 0.03 224.28 10.73 210.50 10.73 Clinical Age 0.12 56.00 14.00 52.50 14.00 BMI 0.48 29.90 8.50 27.30 8.50 modified Glasgow Prognostic Score 0.21 1.00 1.00 0.00 1.00 Baseline Tumor Size (cm) 0.48 9.50 5.15 9.50 5.15 ECOG PS at presentation 0.23 0.00 1.00 0.00 1.00 Charlson Comorbidity Index 0.72 6.00 6.00 4.00 6.00 Citation Format: Liping LI, Edouard Nicaise, Benjamin Schmeusser, Abhishek Midya, Amir Davarpanahfakhr, Anant Madabhushi, Mehmet Asim Bilen, Viraj A. Master, Rakesh Shiradkar. Radiomics at baseline MRI are associated with tumor shrinkage in patients with unresectable renal cell carcinoma treated with neo-adjuvant Cabozantinib [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 894.

Abstract 1254: Integration of human omics analysis and new syngeneic tumor organoid models reveals that aberrant RAS/PI3K crosstalk triggers refractoriness in high-grade serous carcinoma

Abstract Refractory high-grade serous carcinoma (HGSC) is one of the most severe clinical problems in gynecology, but we have barely grasped the underlying mechanism of aggressiveness and chemo-resistance. Previous study has shown that homologous recombination genes, BRCA1/2-unmutated HGSC patients possess the potential to be in refractory condition, and their primary response to chemotherapy was heterogenous; thus, there can be subtype-specific determinant factors hatching refractoriness in BRCA1/2-unmutated HGSC patients. To explore the determinant factors, we performed gene set variation analysis and comprehensive log-rank tests on TCGA advanced serous ovarian cancer datasets. As a result, regulation of small GTPase signaling prominently correlated with poor prognosis uniquely in BRCA1/2-unmutated HGSC patients (p-value < 0.001), and eventually, pathway clustering analysis by variational Bayesian Gaussian mixture model discovered aberrant RAS/PI3K crosstalk as peculiar poor prognostic signature of BRCA1/2-unmutated HGSC patients. To elucidate the mechanism of poor progression triggered by RAS/PI3K, we utilized our syngeneic HGSC organoid models newly established from murine fallopian tube epithelium by making Myc overexpressing, and knocking out Trp53 and Rb1; as a supplement, the propriety of these genetic alterations to develop HGSC models was backed up by genetic profiles of high-grade serous ovarian cancer patients in AACR Project GENIE. After that, we knocked out Nf1 and Pten whose deletions were detected relatively with high frequency in HGSC to induce aberrant RAS/PI3K crosstalk in the primitive HGSC model. In-vivo evaluation and growth comparison indicated that aberrant RAS/PI3K crosstalk transformed Brca1/2-unmutated HGSC modeling cells into aggressive phenotypes such as poor survival (p-value < 0.01) and high growth rate (p-value < 0.001). Chemo-sensitivity assay also showed that the genetic manipulation made Brca1/2-unmutated HGSC modeling cells more tolerant to anti-tumor agents like carboplatin (p-value < 0.001), paclitaxel (p-value < 0.01), and olaparib (p-value < 0.01). To investigate biological processes that induce the aggressive and chemo-resistant phenotype, we performed gene set enrichment analysis among our HGSC models. The result showed that aberrant RAS/PI3K crosstalk induces cell cycle acceleration and downregulation of apoptosis in the HGSC cells, which backed up aggressive phenotype and chemo-resistance in our HGSC models. In conclusion, aberrant RAS/PI3K crosstalk transforms Brca1/2-unmutated HGSC modeling cells more aggressive and chemo-resistant, which was well recapitulated by our HGSC modeling organoids. The integration of our computational approach and experimental models will lead us to detect peculiar therapeutic targets against refractory HGSC. Citation Format: Tomohiro Tamura, Kenta Masuda, Shimpei Nagai, Keiyo Imaeda, Juntaro Yamasaki, Eiji Sugihara, Hiroyuki Nobusue, Yuji Otsuki, Rui Yamaguchi, Kazuhiro Sakurada, Wataru Yamagami, Hideyuki Saya, Osamu Nagano. Integration of human omics analysis and new syngeneic tumor organoid models reveals that aberrant RAS/PI3K crosstalk triggers refractoriness in high-grade serous carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1254.

Monitoring plant responses in field-grown peanuts exposed to exogenously applied chitosan under full and limited irrigation levels

In recent decades, numerous studies have examined the effects of climate change on the responses of plants. These studies have primarily examined the effects of solitary stress on plants, neglecting the simultaneous effects of mixed stress, which are anticipated to transpire frequently as a result of the extreme climatic fluctuations. Therefore, this study investigated the impact of applied chitosan on boosting the resistance responses of peanuts to alkali and mixed drought-alkali stresses. Peanuts were grown in mid-alkaline soil and irrigated with full irrigation water requirements (100%IR), represented alkali condition (100% IR × alkali soil) and stress conditions (70% IR × alkali soil—represented mixed drought-alkali conditions). Additionally, the plants were either untreated or treated with foliar chitosan. The study evaluated various plant physio-chemical characteristics, including element contents (leaves and roots), seed yield, and irrigation water use efficiency (IWUE). Plants that experienced solitary alkali stress were found to be more vulnerable. However, chitosan applications were effective for reducing (soil pH and sodium absorption), alongside promoting examined physio-chemical measurements, yield traits, and IWUE. Importantly, when chitosan was applied under alkali conditions, the accumulations of (phosphorus, calcium, iron, manganese, zinc, and copper) in leaves and roots were maximized. Under mixed drought-alkali stresses, the results revealed a reduction in yield, reaching about 5.1 and 5.8% lower than under (100% IR × alkali), in the first and second seasons, respectively. Interestingly, treated plants under mixed drought-alkali stresses with chitosan recorded highest values of relative water content, proline, yield, IWUE, and nutrient uptake of (nitrogen, potassium, and magnesium) as well as the lowest sodium content in leaves and roots. Enhances the accumulation of (N, K, and Mg) instead of (phosphorus, calcium, iron, manganese, zinc, and copper) was the primary plant response to chitosan applications, which averted severe damage caused by mixed drought-alkali conditions, over time. These findings provide a framework of the nutrient homeostasis changes induced by chitosan under mixed stresses. Based on the findings, it is recommended under mixed drought-alkali conditions to treat plants with chitosan. This approach offers a promising perspective for achieving optimal yield with reduced water usage.

Evaluation of immunomodulatory effect of aqueous extract of aloe vera in Wistar albino rat models

Aim of the study wasto evaluate the immunomodulatory effects of Aqueous extracts of Aloe vera in Wistar albino rats using humoral immune response (antibody titre), a cellular immune response, and a neutrophil adhesion test after oral administration. : 24 healthy Wistar albino rats of either sex were divided into 4 different groups containing 6 rats for each immunomodulatory model. Group I ,the control group received gum acacia suspended in normal saline; Group II rats were treated with dexamethasone (1 mg/kg bw)whereas Groups III & IV received AloeVera aqueous extract at a dose of 125 mg/kg and 250 mg/kg, respectively. Each rat was antigenically challenged by injecting 0.1 ml of 0.5 × 10 SRBCs (suspended in normal saline) intraperitoneally. The study of humoral immune response was seen by measurement of antibody titre obtained on 20 day (7 day of Ag challenge) and 27 day (14 day of challenge) with sRBC considered the primary and secondary humoral immune response, respectively. For the cellular immune response, foot pad edema was calculated due to a hypersensitivity reaction after injection of sRBC into the .rat hind paw. A test of neutrophil adhesion is used to evaluate immunomodulatory activity. Aloe vera aqueous extracts at 250 mg/kg significantly improved a significant increase in paw edema volume indicated increased cell-mediated immunity, and elevated Ab titre on the 20th and 27th days served as a marker of increased humoral immune response. Additionally, AVE 250 mg/kg caused an increase in neutrophil adhesion, demonstrating its immunomodulatory effects. Aloe vera aqueous extracts at a dose of 250 mg/kg demonstrated immunomodulatory activity in a model using Wistar albino rats.Aloe vera extract at a dose of 250 mg/kg significantly increased hemagglutination antibody titers compared to the control group, indicating enhanced humoral immunity. This study demonstrates the immunostimulant properties of orally administered Aloe vera aqueous extract in Wistar albino rats. The extract increased antibody production, enhanced cell-mediated immunity, and improved neutrophil function. These findings support the potential of Aloe vera as a natural immunomodulator and warrant further exploration of its therapeutic applications.

MiR-515-5p targeting Toll-like receptor 4 regulates myeloid differentiation primary response gene 88/nuclear factor-kappa B pathway to inhibit apoptosis and inflammatory response of osteoarthritis chondrocytes

To explore the molecular mechanism of miR-515-5p in inhibiting chondrocyte apoptosis and alleviating inflammatory response in osteoarthritis (OA). Human cartilage cell line C28/I2 was cultured in vitro and treated with 10 ng/mL interleukin 1β (IL-1β) for 24 hours to construct an in vitro OA model. C28/I2 cells were transfected with miR mimics, mimics negative control (NC), over expression (oe)-NC, and oe-Toll-like receptor 4 (TLR4), respectively, and then treated with 10 ng/mL IL-1β for 24 hours to establish OA model. Cell proliferation capacity was detected by cell counting kit 8 and 5-Ethynyl-2'-deoxyuridine, cell apoptosis and cell cycle were detected by flow cytometry, and B-cell lymphoma 2 protion (Bcl-2), Bcl-2-associated X protein (Bax), cleaved-Caspase-3, TLR4, myeloid differentiation primary response gene 88 (MyD88), p65 and phosphorylated p65 (p-p65) protein expression levels were detected by Western blot. Real-time fluorescence quantitative PCR was used to detect mRNA expression levels of miR-515-5p and TLR4, and ELISA was used to detect pro-inflammatory factor prostaglandin E2 (PGE2), tumor necrosis factor α (TNF -α), and IL-6 levels in cell supernatant. The potential binding sites between miR-515-5p and TLR4 were predicted by BiBiServ2 database, and the targeting relationship between miR-515-5p and TLR4 was verified by dual luciferase reporting assay. After the treatment of C28/I2 cells with IL-1β, the expressions of miR-515-5p and Bcl-2 protein and the proliferation ability of C28/I2 cells significantly reduced. The expression levels of Bax and cleaved-Caspase-3 protein, the levels of pro-inflammatory factors (PGE2, TNF-α, IL-6) in the supernatant of C28/I2 cells, and the apoptosis of C28/I2 cells significantly increased. In addition, the proportion of the cells at S phase and G 2 phase decreased significantly, and the proportion of cells at G 1 phase increased significantly, suggesting that the cell cycle was blocked after IL-1β treatment. After transfection with miR mimics, the expression level of miR-515-5p in the cells significantly up-regulated, partially reversing the apoptosis of OA chondrocytes induced by IL-1β, and alleviating the cycle arrest and inflammatory response of OA chondrocytes. After treating C28/I2 cells with IL-1β, the mRNA and protein levels of TLR4 significantly increased. Overexpression of miR-515-5p targeted inhibition of TLR4 expression and blocked activation of MyD88/nuclear factor κB (NF-κB) pathway. Overexpression of TLR4 could partially reverse the effect of miR mimics on IL-1β-induced apoptosis and inflammation of OA chondrocytes. miR-515-5p negatively regulates the expression of TLR4, inhibits the activation of MyD88/NF-κB pathway and apoptosis of OA chondrocytes, and effectively alleviates the inflammatory response of the cells.

MyD88-mediated signaling is critical for the generation of seizure responses and cognitive impairment in a model of anti-N-methyl-D-aspartate receptor encephalitis.

We previously demonstrated that interleukin-1 receptor-mediated immune activation contributes to seizure severity and memory loss in anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis. In the present study, we assessed the role of the myeloid differentiation primary response gene 88 (MyD88), an adaptor protein in Toll-like receptor signaling, in the key phenotypic characteristics of anti-NMDAR encephalitis. Monoclonal anti-NMDAR antibodies or control antibodies were infused into the lateral ventricle of MyD88 knockout mice (MyD88-/-) and control C56BL/6J mice (wild type [WT]) via osmotic minipumps for 2 weeks. Seizure responses were measured by electroencephalography. Upon completion of the infusion, the motor, anxiety, and memory functions of the mice were assessed. Astrocytic (glial fibrillary acidic protein [GFAP]) and microglial (ionized calcium-binding adaptor molecule 1 [Iba-1]) activation and transcriptional activation for the principal inflammatory mediators involved in seizures were determined using immunohistochemistry and quantitative real-time polymerase chain reaction, respectively. As shown before, 80% of WT mice infused with anti-NMDAR antibodies (n = 10) developed seizures (median = 11, interquartile range [IQR] = 3-25 in 2 weeks). In contrast, only three of 14 MyD88-/- mice (21.4%) had seizures (0, IQR = 0-.25, p = .01). The WT mice treated with antibodies also developed memory loss in the novel object recognition test, whereas such memory deficits were not apparent in MyD88-/- mice treated with anti-NMDAR antibodies (p = .03) or control antibodies (p = .04). Furthermore, in contrast to the WT mice exposed to anti-NMDAR antibodies, the MyD88-/- mice had a significantly lower induction of chemokine (C-C motif) ligand 2 (CCL2) in the hippocampus (p = .0001, Sidak tests). There were no significant changes in the expression of GFAP and Iba-1 in the MyD88-/- mice treated with anti-NMDAR or control antibodies. These findings suggest that MyD88-mediated signaling contributes to the seizure and memory phenotype in anti-NMDAR encephalitis and that CCL2 activation may participate in the expression of these features. The removal of MyD88 inflammation may be protective and therapeutically relevant.

Retention rate of biologic and targeted synthetic anti-rheumatic drugs in elderly rheumatoid arthritis patients: data from GISEA registry.

An increased number of elderly individuals affected by rheumatoid arthritis (RA) has been reported, including both patients with RA onset in advanced age and patients aged with the disease. In this registry-based study, we aimed to analyze the retention rate and cause of discontinuation of biologic (b) and targeted synthetic (ts)-disease-modifying anti-rheumatic drugs (DMARDs) in RA patients over 65 year old. RA patients enrolled in the Italian GISEA registry and starting a b- or a ts-DMARD over 65 years of age were included. Demographic, clinical, serologic, and therapeutic features were collected. A total of 1,221 elderly RA patients were analyzed (mean age 71.6 ± 5.2 years). RA was diagnosed before 65 years in 72.5% of cases, a 60.6% of patients experienced a previous b- or ts-DMARD. In patients older than 65 initiating a new b- or ts-DMARDS, tumor necrosis factor alpha inhibitors (TNFi) were prescribed in 29.6% of patients, abatacept in 24.8%, anti-interleukin 6 receptor antagonists (anti-IL6R) in 16.3%, Janus kinases inhibitors (JAKi) in 24.9%, and rituximab in 4.4%. The main causes of discontinuation were primary or secondary inadequate responses (66.1%). The median retention rate for all treatments was 181.3 weeks. A statistically higher retention rate was observed for abatacept when compared to TNFi (p = 0.02), JAKi (p < 0.001), and anti-IL6R (p < 0.001), and for TNFi vs. JAKi (p = 0.013). We described, in a real-life setting, elderly RA patients treated with a biologic or a ts-DMARD in Italy. Loss of efficacy was the main cause of discontinuation, and the DMARD safety profile suggests that age does not contraindicate their use. Our study reinforced that the control of disease activity is mandatory.

Ultrasensitive cortisol electrochemical immunosensor amplifying by Au single-atom nanozymes and HRP enzymes

Cortisol, a corticosteroid hormone as a primary stress hormone response to internal and external stress, has been regarded as a gold standard reliable biomarker to evaluate human mental stress. The double enzymes strategy, using nanozyme and enzyme amplifying the electrochemical signal, has been widely used to improve the performance of electrochemical biosensors. An ultra-sensitive electrochemical cortisol sensor based on Au single-atom nanozymes had been fabricated through HRP labeled anti-cortisol antibody binding with Au by Au-S bond. Based on the high catalytic activity of Au single-atom nanozymes and the high selectivity of HRP-labeled anti-cortisol antibodies, the cortisol electrochemical sensor-based Au single-atom nanozymes had an excellent response to cortisol, such as high electrochemical activity, high sensitivity, high selectivity, and wide linear range (0.15–300 ng mL−1) and low detection (0.48 pg mL−1) through the four-parameter logistic model with 95% confidence. The electrochemical cortisol sensor was used to determine the cortisol concentration of human saliva at different times.

Functional mechanism of baicalein in alleviating severe acute pancreatitis-acute lung injury by blocking the TLR4/MyD88/TRIF signaling pathway.

Severe acute pancreatitis-acute lung injury (SAP-ALI) is a disease with high mortality. This study aims to explore the mechanism of baicalein on SAP-ALI in rats by blocking toll-like receptor-4 (TLR4)/myeloid differentiation primary response gene 88 (MyD88)/TIR-domain-containing adapter-inducing interferon-β (TRIF) signal pathway. The SAP-ALI rat model was established by intraperitoneal injection of 3% pentobarbital sodium (30 mg/kg), with pancreas and intestines turned over, injected with 3.5% sodium taurocholate backward into the bile-pancreatic duct at 0.1 mL/100 g for 12h, and treated with baicalein, lipopolysaccharide (LPS), miR-182 agomir, or miR-182 antagomir. The TLR4/MyD88/TRIF pathway was activated using LPS in SAP-ALI rats after baicalein treatment. Baicalein attenuated inflammatory cell infiltration, alveolar wall edema, decreased W/D ratio and levels of TLR4, MyD88, and TRIF in the lung tissues, reduced levels of inflammatory factors in pancreatic and lung tissues and BALF, diminished ROS, and elevated GSH, SOD and CAT in pancreatic and lung tissues of SAP-ALI rats. Activation of the TLR4/MyD88/TRIF pathway partly abrogated baicalein-mediated improvements in inflammation and oxidative stress in SAP-ALI rats. miR-182 targeted TLR4. miR-182 suppressed inflammation and oxidative stress in SAP-ALI rats by targeting TLR4. Inhibition of miR-182 partly nullified baicalein-mediated attenuation on inflammation and oxidative stress in SAP-ALI rats. In conclusion, baicalein can inhibit the TLR4/MyD88/TRIF pathway and alleviate inflammatory response and oxidative stress in SAP-ALI rats by upregulating miR-182 and suppressing TLR4, thus ameliorating SAP-ALI.

Notoginsenoside R1 alleviates cerebral ischemia/reperfusion injury by inhibiting the TLR4/MyD88/NF-κB signaling pathway through microbiota-gut-brain axis

BackgroundIschemic stroke (IS) ranks as the second common cause of death worldwide. However, a narrow thrombolysis timeframe and ischemia-reperfusion (I/R) injury limits patient recovery. Moreover, anticoagulation and antithrombotic drugs do not meet the clinical requirements. Studies have demonstrated close communication between the brain and gut microbiota in IS. Notoginsenoside R1 (NG-R1), a significant component of the total saponins from Panax notoginseng, has been demonstrated to be effective against cerebral I/R injury. Total saponins have been used to treat IS in Chinese pharmacopoeia. Furthermore, previous research has indicated that the absorption of NG-R1 was controlled by gut microbiota. Study designThis study aimed to access the impact of NG-R1 treatment on neuroinflammation and investigate the microbiota-related mechanisms. ResultsNG-R1 significantly reduced neuronal death and neuroinflammation in middle cerebral artery occlusion/reperfusion (MCAO/R) models. 16S rRNA sequencing revealed that NG-R1 treatment displayed the reversal of microbiota related with MCAO/R models. Additionally, NG-R1 administration attenuated intestinal inflammation, gut barrier destruction, and systemic inflammation. Furthermore, microbiota transplantation from NG-R1 exhibited a similar effect in the MCAO/R models. ConclusionIn summary, NG-R1 treatment resulted in the restoration of the structure of the blood-brain barrier (BBB) and reduction in neuroinflammation via suppressing the stimulation of astrocytes and microglia in the cerebral ischemic area. Mechanistic research demonstrated that NG-R1 treatment suppressed the toll-like receptor 4/myeloid differentiation primary response 88/nuclear factor kappa B (TLR4/MyD88/NF-κB) signaling pathway in both the ischemic brain and colon. NG-R1 treatment enhanced microbiota dysbiosis by inhibiting the TLR4 signaling pathway to protect MCAO/R models. These findings elucidate the mechanisms by which NG-R1 improve stroke outcomes and provide some basis for Panax notoginseng saponins in clinical treatment.

Fast Frequency Response for Future Low-Carbon Indian Power System: Challenges and Solutions

India has transitioned from being an energy-deficient nation to an energy-surplus nation. As of July 2023, India’s power system has seen a rapid transformation, with renewable energy, including large hydro, accounting for 40% of the total installed capacity at 177 GW. The ongoing energy transition has not only changed the country’s energy mix but also brought new challenges for system operators. The primary challenge is frequency instability due to the reduction of system inertia and governor-based primary frequency response (PFR) availability by large-scale integration of uncertain renewable energy resources (RES) such as wind and solar photovoltaic generators. Furthermore, the absence of frequency control ancillary services market, inadequate penetration of electric vehicles (EVs) and uncertainty of RES generation enhance the complexity of the frequency response services in the Indian power system. In this context, this paper presents a comprehensive review of the frequency response challenges of the future low-carbon Indian power system and underlines the new potential fast frequency response (FFR) services from advanced energy storage and renewable generation technologies. Such FFR services can be swiftly deployed in low-inertia grids through the integration of energy storage systems and inverter-based resources in a shorter span. Modeling of FFR services in security constraint unit commitment (SCUC) can enhance grid resiliency and facilitate high penetration of RES in the Indian power system without loss of frequency stability.

TRIF-IFN-I pathway in Helicobacter-induced gastric cancer in an accelerated murine disease model and patient biopsies

Helicobacter pylori (H.pylori) infection is a known cause of many digestive diseases, including gastritis, peptic ulcers, and gastric cancer. However, the underlying mechanisms by which H.pylori infection triggers these disorders are still not clearly understood. Gastric cancer is a slow progressing disease, which makes it difficult to study. We have developed an accelerated disease progression mouse model, which leverages mice deficient in the myeloid differentiation primary response 88 gene (Myd88-/-) infected with Helicobacter felis (H.felis). Using this model and gastric biopsy samples from patients, we report that activation of the Toll/interleukin-1 receptor (TIR)-domain-containing adaptor inducing interferon-β (TRIF)-type I interferon (IFN-I) signaling pathway promotes Helicobacter-induced disease progression toward severe gastric pathology and gastric cancer development. Further, results implicated downstream targets of this pathway in disease pathogenesis. These findings may facilitate stratification of Helicobacter-infected patients and thus enable treatment prioritization of patients.

Treatment response of advanced HNSCC towards immune checkpoint inhibition is associated with an activated effector memory T cell phenotype.

Locally advanced or metastatic head and neck squamous cell carcinoma (HNSCC) is associated with a poor prognosis. The introduction of PD-1 inhibitors has led to a significant improvement in survival, but only a subpopulation of patients responds to therapy. Current biomarkers cannot reliably identify these patients. The identification of biomarkers for the prediction and monitoring of immunotherapy is therefore of great importance. In this study, we characterized lymphocyte subsets in the peripheral blood of HNSCC patients under PD-1 inhibition. Patients with primary response (n=11) to PD-1 inhibition showed an increase of the CD3+ effector memory (CD3/EM) population and an elevated expression of the activation marker CD69 in CD3+ T cells, particularly in the CD3/EM subpopulation at 3 months when treatment response was assessed. In contrast, patients with primary treatment failure and progressive disease (n=9) despite PD-1 inhibition had lower absolute lymphocyte counts and an increased expression of CTLA-4 in CD3+ T cells at the time of treatment failure compared with baseline, particularly in CD4+ and CD8+ effector memory populations. Our results demonstrate that HNSCC patients' response to immune checkpoint inhibition shows a distinct immune signature in peripheral blood, which could help identify refractory patients earlier. Furthermore, strategies to overcome primary therapy failure by inducing a beneficial T cell phenotype or adding alternative immune checkpoint inhibitors could improve response rates and survival of HNSCC patients.

VAERS Vasculitis Adverse Events Retrospective Study: Etiology Model of Immune Complexes Activating Fc Receptors in Kawasaki Disease and Multisystem Inflammatory Syndromes.

Vasculitis diseases include Kawasaki disease (KD), Kawasaki disease shock syndrome (KDSS), Multisystem Inflammatory Syndrome (MIS), Henoch-Schönlein purpura (HS), or IgA vasculitis, and additional vasculitis diseases. These diseases are often preceded by infections or immunizations. Disease incidence rates are higher in children than in adults. These diseases have been extensively studied, but understanding of the disease etiology remains to be established. Many studies have failed to demonstrate an association between vasculitis diseases and vaccination; this study examines possible associations. Herein, the Vaccine Adverse Event Reporting System (VAERS) database is retrospectively examined for associations between vasculitis diseases and immunizations. For some vaccines, the number of rare cases of KD, MIS, and HS are higher than the background rates. These rare cases are predicted to occur in individuals with (1) genetic risk factors with (2) antibody titer levels above the primary immune response level. Herein, the model of humoral immune response antibodies bound to antigens (pathogen or vaccine) creating immune complexes is proposed. These immune complexes are proposed to bind Fc receptors on immune cells and platelets, resulting in cell activation and the release of inflammatory molecules including histamine and serotonin. Immune complexes and inflammatory molecules including serotonin and histamine likely trigger vasculitis. Elevated serotonin and possibly histamine drive initial vasoconstrictions, disrupting blood flow. Increased blood flow pressure from cardiac capillary vasoconstrictions is predicted to trigger coronary artery aneurysms (CAA) or lesions (CAL) in some patients. For KDSS and MIS patients, these cardiac capillary vasoconstrictions are predicted to result in ischemia followed by ventricular dysfunction. Ongoing ischemia can result in long-term cardiac damage. Cases associated with pathogens are likely to have persistent infections triggering disease onset. The proposed model of immune complexes driving disease initial disease etiology by Fc receptor activation of immune cells and platelets, resulting in elevated histamine and serotonin levels, is testable and is consistent with disease symptoms and current treatments.

Whole proteome screening to develop a potent epitope-based vaccine against Coxiella burnetii: a reverse vaccinology approach

Coxiellosis is known as a threat to human health. This study aimed to develop an epitope-based vaccine against coxiellosis using a whole proteome investigation. In this case, the whole proteome of Coxiella burnetii was collected from the database, then different assessments were performed to select immunogenic proteins. The selected proteins were used for epitopes prediction. The epitope-based vaccine was made using the best-selected epitopes and HBHA protein. The physical and chemical features, as well as secondary and tertiary structures of the developed vaccine were analyzed. The interaction between the developed vaccine and TLR4/MD2 receptor was examined using molecular docking and molecular dynamic simulation. Finally, in silico cloning, codon optimization, and immune response simulation for the developed vaccine were performed. The findings supported a stable, hydrophilic, antigenic and non-allergenic vaccine with a molecular weight equal to 59.261 kDa and 542 amino acid residues in length. The findings showed that the developed vaccine not only could dock to TRL4/MD2 receptor with an affinity of −20.9 kcal/mol and 15 hydrogen bonds, but also the protein-protein complex was stable during molecular dynamic simulation with the binding free energy of −57.9 ± 6.9 kcal/mol. Furthermore, the optimized sequence of the developed vaccine with a CAI value of 0.97, could be cloned into the pET-21a (+) vector. Finally, The results confirmed that the developed vaccine could strongly trigger primary and secondary immune responses. Evidently, the developed vaccine can be an interesting candidate to apply. Communicated by Ramaswamy H. Sarma

The transcription factor NF-κB orchestrates nucleosome remodeling during the primary response to Toll-like receptor 4 signaling

Inducible nucleosome remodeling at hundreds of latent enhancers and several promoters shapes the transcriptional response to Toll-like receptor 4 (TLR4) signaling in macrophages. We aimed to define the identities of the transcription factors that promote TLR-induced remodeling. An analysis strategy based on ATAC-seq and single-cell ATAC-seq that enriched for genomic regions most likely to undergo remodeling revealed that the transcription factor nuclear factor κB (NF-κB) bound to all high-confidence peaks marking remodeling during the primary response to the TLR4 ligand, lipid A. Deletion of NF-κB subunits RelA and c-Rel resulted in the loss of remodeling at high-confidence ATAC-seq peaks, and CRISPR-Cas9 mutagenesis of NF-κB-binding motifs impaired remodeling. Remodeling selectivity at defined regionswas conferred by collaboration with other inducible factors, including IRF3- and MAP-kinase-induced factors. Thus, NF-κB is unique among TLR4-activated transcription factors in its broad contribution to inducible nucleosome remodeling, alongside its ability to activate poised enhancers and promoters assembled into open chromatin.