Increase In IL-6 Levels Research Articles

Interleuquina-6 como marcador pronóstico en el fracaso renal agudo y su regulación dependiente de klotho

Background and objectiveIn acute kidney injury (AKI), a strong inflammatory component is activated in response to the renal damage, and one of the main mediators behind this process is the pro-inflammatory interleukin 6 or IL-6. Beside to this phenomenon, there are also alterations in different components of mineral metabolism, such as those dependent on fibroblast growth factor (FGF)23 and the anti-ageing cofactor klotho. The aim of this work was to explore the association between renal function and systemic levels of IL-6, as well as FGF23 and klotho in the early stages of AKI, analysing the predictive capacity of IL-6 in early mortality associated with AKI. Materials and methodsPlasma levels of IL-6, klotho and FGF23 were analysed in samples from 28 patients with AKI and related to renal function on hospital admission, and after 24 and 72hours. In addition, the predictive capacity of IL-6 on AKI-associated mortality was analysed at the three study time points. In an experimental nephrotoxic -AKI mouse model, systemic IL-6 and FGF23 values were also analysed 24 and 72hours after induction of kidney damage, as well as in mice overexpressing the anti-ageing protein, klotho. ResultsSystemic IL-6 levels increased in AKI patients, especially in hospital admission time, and decreased in parallel with improving renal function. At the same time as IL-6 values increased, there was an increase in FGF23 and a decrease in klotho levels, with a significant and positive correlation between IL-6 and FGF23 levels. In addition, we obtained that systemic IL-6 levels were a good predictor of mortality in these patients, with an area under the curve equal to one at 72hours after AKI. In the experimental mouse AKI model, we also observed an increase in plasma levels in both IL-6 and FGF23 after 24hours of kidney damage. Nevertheless, in transgenic mice overexpressing klotho, there was no such increase in any of them. ConclusionsThere is an association between renal damage and increased levels of IL-6 and FGF23 in patients with AKI, especially on hospital admission time. Moreover, IL-6 levels are able to predict mortality in these patients, being a promising prognostic biomarker at any study time with a strong prediction at 72hours after patient admission. Maintaining adequate klotho levels could prevent the IL-6 mediated inflammatory response and therefore also reduce the degree and severity of renal damage after AKI.

Abstract 851: Uncovering a novel role of succinate metabolism in tregulatory cells of aged epithelial ovarian cancer mice

Abstract Epithelial Ovarian Cancer (EOC), the most fatal gynecologic cancer is a disease of older women, with median age of 63 years at diagnosis. Reduced naive T cell generation and a rise in immunosuppressive regulatory T (Treg) cells are hallmarks of aging, which are strongly correlated with advanced cancer stage. Aging also alters host and cellular metabolism, which can influence Treg number and function. Although metabolic alterations are known to impact ant-tumor immune response, the relationship between age-related metabolic dysfunction and immune cells is not fully understood. We utilized young (2 months) and old (18 months) female C57/B6 mice, and were injected intraperitoneally with 5 × 106 mouse epithelial ovarian cancer ID8p53-/− cells. Old mice with EOC showed decreased survival compared to young mice (median survival 49 vs 65 days). Old mice with EOC presented with decreased CD4+ and CD8+ T cells, but a significant increase in CD4+CD25+FoxP3+ Treg cells in tumor micro environment (TME) and in blood. The Tregs from old mice displayed enhanced expression of IL10 and TGFβ, which corelated with their increased suppression of CD4 and CD8 T cells compared to Tregs isolated from young EOC mice. The CD4+ and CD8+ T cells also displayed decreased effector makers including CD4+IFNγ, CD8+IFNγ and CD8+ Granzyme B, compared to young EOC mice in TME and in blood. The tumor promoting role of Treg was validated by depleting Tregs using anti-CD25 antibody which resulted in decreased tumor burden and improved overall survival, of both young and old mice, more significantly in old EOC mice. Tregs from old mice with EOC displayed enhanced metabolic activity, reflected in increased oxidative phosphorylation (OXPHOS) compared to young EOC mice. Further, on inhibition of OXPHOS, the Tregs from old mice showed decreased expression of immunosuppressive markers Foxp3 and IL10. Targeted metabolomics of TCA cycle in the Tregs, revealed a 5-fold increase in succinate levels in old EOC Tregs compared to young. Succinate treatment enhanced the immunosuppressive Treg function along with an increase in levels of IL10 and TGFβ, while inhibition of succinate synthesis reversed the increase in FOXP3, IL10 and TGFβ. We identified fibroblast growth factor 21 (FGF-21), a hormone essential for T cell development, to be significantly decreased in ascites and plasma, of old mice. Supplementing FGF21 decreased Treg survival, number, and suppressive ability, resulting in increased CD4+ and CD8+ cells in aged EOC mice. FGF21 treatment decreased OXPHOS and succinate levels in aged Tregs. Overall, our observations indicate Tregs to be vital in driving aggressive EOC in old mice. The hyper-suppressive Tregs in old mice reprogram succinate metabolism to drive their suppressive ability which may be negatively regulated by FGF21. Thus, Treg-succinate-FGF21 may be a potential actionable pathway in precision targeting of EOC in the older population. Citation Format: Mary Priyanka Udumula, Harshit Singh, Faraz Rashid, Miriana Hijaz, Shailendra Giri, Ramandeep Rattan. Uncovering a novel role of succinate metabolism in tregulatory cells of aged epithelial ovarian cancer mice [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 851.

The effect of N-eicosapentaenoyl ethanolamine on the regulation of cytokine synthesis by blood cells of patients with bronchial asthma <I>in vitro</I>

Introduction. Bronchial asthma is characterized by heterogeneity, multiple phenotypes, and varying clinical manifestations. Cytokines play a crucial role in the inflammatory response in asthma. The quantity, as well as the ratio of certain cytokines, determines the mechanism and type of inflammatory response in asthma, upon which the effectiveness of treatment of this disease depends. That is why the development of new methods of treating patients with asthma, aimed at correcting cytokine imbalance, is required. One of the promising substances is N-eicosapentaenoyl-ethanolamine (NAE-EPA), which exhibits anti-inflammatory properties by affecting cytokines, but remains poorly studied.Aim. To study the dose-dependent effect of N-eicosapentaenoyl-ethanolamine on the production of cytokines by peripheral blood cells, in vitro, in subjects with asthma.Materials and methods. The object of the study was whole blood, diluted 1:5 with culture medium of 15 patients with mild to moderate controlled asthma and 16 healthy subjects. The in vitro experiment was carried out in lipopolysaccharide-stimulated (LPS) blood samples (incubation with LPS at 37°C for 30 minutes). Then, the experimental substance N-acylethanolamine eicosapentaenoic acid (NAE EPA) was added in concentrations of 1.0; 5.0, and 10.0 µM and incubated at 37°C for 6 hours with gentle mixing. Cytokine levels (IL-2, IL-4, IL-6, IL-10, IL17A, TNF-α, and INF-γ) were studied by enzyme-linked immunoassay.Results. Analysis of the level of cytokines in patients with asthma showed that an increase in the plasma levels of IL-2, TNF-α, IL-6, and IL-17A is accompanied by a decrease in the level of regulatory IL-10. When NAE EPA was added at a dosage of 1 µM, no statistically significant changes were detected. Exposure to the experimental substance at a dose of 5 µM contributed to a decrease in IL-6 in the blood cells of patients by 19% (p ˂ 0.05). Exposure to NAE EPA at 10 µM produced the greatest number of statistically significant changes in cytokine levels. There was a decrease in IL-17A by 15% (p ˂ 0.05), IL-2 by 14% (p ˂ 0.05), IL-6 by 50% (p ˂ 0.01), and TNF-α by 10% (p ˂ 0.05) relative to values before exposure.Conclusion. N-eicosapentaenoyl ethanolamine shows potential as a regulator of pro- and anti-inflammatory cytokine synthesis in bronchial asthma with a predominant Th-17 type of immune response. The results obtained may contribute to the development of new treatment strategies for patients with asthma.

O124: Investigating the Immuno-modulatory Effect of Adipose-derived Stem Cells in Radiation- Induced Fibrosis

Abstract Background and Introduction Radiation-induced fibrosis (RIF) is a complication of radiotherapy in the treatment of head and neck carcinoma with high patient morbidity. Autologous fat grafting is a viable method to treat RIF, and adipose-derived stem cells (ADSC) were found to be one of the main components to contribute to that effect. Method Experimental group consists of irradiated human-derived fibroblasts (iHDF) in vitro with the control group unexposed to radiation. Both groups were then inoculated with ADSC-controlled media and co-culture with ADSC, whilst levels of key immunomodulators expressed by iHDF cells were obtained and measured via enzyme-linked immunosorbent assay (ELISA) and real-time reverse transcription-polymerase chain reaction (RT-PCR). Results There is marginal decrease in CCL2 concentrations in co-cultured iHDF samples compared to untreated iHDF samples (p = 0.0006). There is a slight increase in CCL3 levels in iHDF co-culture samples compared to untreated iHDF samples (p = 0.0247). iHDF co-culture samples shows a statistically significant increase in IL-8 levels when compared to untreated samples of iHDF (p = 0.027). Col1A1 is reduced in iHDF samples when compared to control groups. Discussion CCL2 is implicated in recruitment of ADSCs to target sites. The increase in IL-8 secretion in ADSC samples is implicated in the pro-angiogenic properties of ADSC secretions to encourage healing in fibrotic tissue. Conclusion ADSC was demonstrated to have anti-fibrotic capabilities by promoting pro- angiogenic factors, however, experimental evidence points to a simultaneous increase in pro-inflammatory factors. Future works required to further investigate in vivo effects of ADSC.

Serum cytokine profiles during engraftment syndrome and acute graft-versus-host disease in adult patients after hematopoietic stem cell transplantation

BackgroundThe underlying biology of engraftment syndrome (ES) following allogeneic hematopoietic stem cell transplantation (HSCT) is not fully elucidated, and the extent of its overlap with acute graft-versus-host disease (aGvHD) remains unclear. In order to establish potential indicator to distinguish ES more accurately, we conducted a retrospective analysis of cytokine levels during HSCT. MethodsA total of 121 consecutive adult patients who underwent HSCT were enrolled in this study. Blood samples for interleukin (IL)-2, IL-2R, IL-4, IL-5, IL-6, IL-8, IL-10, IL-1β, IL-12p70, interferon (IFN)-γ, IFN-α, tumor necrosis factor alpha (TNF-α) and C-reactive protein CRP were regularly assessed after transplantation and during transplantation related adverse events. Additionally, the balance of naïve, central memory and effector memory of CD4+ and CD8+ was analyzed around 30 and 60 days after stem cell infusion, respectively. ResultsThirty (24.79 %) and 33 (27.27 %) patients were diagnosed with ES and aGvHD, respectively. ES was characterized by a significant increase in level of IL-5, IL-6, IL-8 and sIL-2R, while aGvHD was associated with a significant upregulation of IL-6, IL-5, IL-10 and sIL-2R in the patients from grade I to grade IV. Notably, patients got much higher levels of IL-6, IL-5 and sIL-2R when developed to ES than to aGvHD. Moreover, a pronounced shift from naïve to memory cells, both in CD4+ and CD8+ subsets, was found in ES patients. ConclusionsThese findings suggest that cytokine profiles could serve as potential indicators for detecting and differentiating ES and aGvHD, enabling timely clinical intervention. Prospective clinical trials involving larger, independent patient cohorts are required to validate these observations.

CHF6297: a novel potent and selective p38 MAPK inhibitor with robust anti-inflammatory activity and suitable for inhaled pulmonary administration as dry powder.

Inhibition of p38 mitogen-activated protein kinase (MAPKs) is a potential therapeutic approach for the treatment of acute and chronic pulmonary inflammatory conditions. Here, we report the in vitro and in vivo characterization of the anti-inflammatory effects of CHF6297, a novel potent and selective p38α inhibitor designed for inhalation delivery as a dry powder formulation. CHF6297 has been proven to inhibit p38α enzymatic activity with sub-nanomolar potency (IC50 = 0.14 ± 0.06nM), with >1,000-fold selectivity against p38γ and p38δ. In human peripheral blood mononuclear cells (PBMCs) stimulated with lipopolysaccharides (LPS), as well as in human bronchial epithelial cells (BEAS2B) stimulated with TNF-α or cigarette smoke extract (CSE), CHF6297 inhibited interleukin (IL)-8 release with low nanomolar potency. CHF6297 administered to rats by using a nose-only inhalation device as a micronized dry powder formulation blended with lactose dose-dependently inhibited the LPS-induced neutrophil influx in the bronchoalveolar lavage fluid (BALF). CHF6297 administered intratracheally to rats dose-dependently counteracted the IL-1β (0.3mg/kg)-induced neutrophil influx (ED50 = 0.22mg/kg) and increase in IL-6 levels (ED50 = 0.82mg/kg) in the BALF. In mice exposed to tobacco smoke (TS), CHF6297, administered intranasally (i.n.) for 4 days at 0.03 or 0.3mg/kg, dose-dependently inhibited the corticosteroid-resistant TS-induced neutrophil influx in the BALF. In a murine house dust mite (HDM) model of asthma exacerbated by influenza virus A (IAV) (H3N3), CHF6297 (0.1mg/kg, i.n.) significantly decreased airway neutrophilia compared to vehicle-treated IAV/HDM-challenged mice. When CHF6297, at a dose ineffective per se (0.03mg/kg), was added to budesonide, it augmented the anti-inflammatory effects of the steroid. Overall, CHF6297 effectively counteracted lung inflammation in experimental models where corticosteroids exhibit limited anti-inflammatory activity, suggesting a potential for the treatment of acute exacerbations associated with chronic obstructive pulmonary disease (COPD) and asthma, acute lung injury (ALI), and viral-induced hyperinflammation.

Size selection of intrahepatic lesions for cryoablation contributes to abscopal effect and long-term survival in patients with liver metastatic melanoma receiving PD-1 blockade therapy

ObjectivesIn this study, we aimed to examine parameters of cryoablation, tumor characteristics, and their correlations with distant tumor response and survival of liver metastatic melanoma patients receiving cryoablation and PD-1 blockade (cryo-PD-1) combination treatment.Materials and methodsA retrospective study was conducted among 45 melanoma patients who received combined PD-1 blockade therapy and cryoablation for liver metastasis from 2018 to 2022. Cox regression was utilized to determine the associations between factors and overall survival (OS). Changes in cytokines and immune cell compositions in peripheral blood samples following the combined treatment were investigated, along with their correlations with treatment response.ResultsThe mean cycle of cryo-PD-1 combination treatment was 2.2 (range, 1–6), and the 3-month overall response rate (RECIST 1.1 criteria) was 26.7%. Of the 21 patients who failed previous PD-1 blockade therapy after diagnosis of liver metastasis, 4 (19.0%) achieved response within 3 months since combination treatment. The diameter of ablated lesion ≤ 30 mm, metastatic organs ≤ 2, and pre-treatment LDH level ≤ 300 U/L were independent prognostic factors for favorable OS. Further analysis showed patients with intrahepatic tumor size of 15–45 mm, and ablated lesion size of ≤ 30 mm had significantly higher 3-month response rate (42.9% vs 12.5%; P = 0.022) and survival time (30.5 vs 14.2 months; P = 0.045) than their counterparts. The average increase in NLR among patients with ablated tumor size of ≤ 3 cm and > 3 cm were 3.59 ± 5.01 and 7.21 ± 12.57, respectively. The average increase in serum IL-6 levels among patients with ablated tumor size of ≤ 3 cm and > 3 cm were 8.62 ± 7.95 pg/ml and 15.40 ± 11.43 pg/ml, respectively.ConclusionSize selection of intrahepatic lesions for cryoablation is important in order to achieve abscopal effect and long-term survival among patients with liver metastatic melanoma receiving PD-1 blockade therapy.

IL-6 and IL-10 Levels in Rats Blood Plasma as Immune Responses Post Radioiodine (I-131) Administration.

The purpose of this study was to analyze the effect of oral administration of radioiodine (I-131) on the immune responses (interleukin 6 and 10) as biodosimetry markers and to support clinical trials of I-131 solution. The design of this study was an in vivo experimental study using twenty-seven male rats (Rattus norvegicus strain Sprague-Dawley) given 100 μL of I-131 solution at a dose of 260 μCi. Blood plasma was taken at 0.25, 0.5, 1, 3, 24, 48, 120, and 168 hours post oral I-131 administration, respectively. Rats without radioiodine administration as a control group. The levels of IL-6 and IL-10 were measured using the enzyme-linked immunosorbent assay (ELISA) method. Statistical analysis was carried out with one-way ANOVA using SPSS version 25 software. IL-6 level began to significantly increase at 0.25 hours post administration of I-131 (14.4 pg/mL ± 2.52 pg/mL, p=0.02). During 7 days of observation, IL-6 levels had 2 peaks of highly significant increase at 0.5 hours (43.57 ± 5.28, p<0.001) and 120 hours (24.08 ± 2.69, p<0.001 compared to control (5.44 ± 0.95 pg/mL). IL-10 level began to significantly increase at 0.25 hours (30.32 ± 3.22 pg/mL, p=0.03) compared to controls (20.61 ± 1.59 pg/mL). The highest increase in IL-6 and IL-10 levels occurred respectively in the first 0.5 hours 8 times and in the first 0.25 hours 1.47 times compared to controls. Internal irradiation with radioiodine resulted in a significant increase in immune cells in exposed blood plasma characterized by the production of the cytokines IL-6 and IL-10. This appears to be a response of immune cells to reduce or stop inflammatory reactions through the release of anti-inflammatory cytokines in an effort to prevent excessive inflammatory responses that can damage cells and tissues.

Dual Immunoglobulin Domain-Containing Cell Adhesion Molecule Increases Early in Renal Tubular Cell Injury and Plays Anti-Inflammatory Role.

Dual immunoglobulin domain-containing cell adhesion molecule (DICAM) is a type I transmembrane protein that presents in various cells including renal tubular cells. This study evaluated the expression and protective role of DICAM in renal tubular cell injury. HK-2 cells were incubated and treated with lipopolysaccharide (LPS, 30 μg/mL) or hydrogen peroxide (H2O2, 100 μM) for 24 h. To investigate the effect of the gene silencing of DICAM, small interfering RNA of DICAM was used. Additionally, to explain its role in cellular response to injury, DICAM was overexpressed using an adenoviral vector. DICAM protein expression levels significantly increased following treatment with LPS or H2O2 in HK-2 cells. In response to oxidative stress, DICAM showed an earlier increase (2-4 h following treatment) than neutrophil gelatinase-associated lipocalin (NGAL) (24 h following treatment). DICAM gene silencing increased the protein expression of inflammation-related markers, including IL-1β, TNF-α, NOX4, integrin β1, and integrin β3, in H2O2-induced HK-2 cell injury. Likewise, in the LPS-induced HK-2 cell injury, DICAM knockdown led to a decrease in occludin levels and an increase in integrin β3, IL-1β, and IL-6 levels. Furthermore, DICAM overexpression followed by LPS-induced HK-2 cell injury resulted in an increase in occludin levels and a decrease in integrin β1, integrin β3, TNF-α, IL-1β, and IL-6 levels, suggesting an alleviating effect on inflammatory responses. DICAM was elevated in the early stage of regular tubular cell injury and may protect against renal tubular injury through its anti-inflammatory properties. DICAM has a potential as an early diagnostic marker and therapeutic target for renal cell injury.

Immuno-Enhancing Effects of Galium aparine L. in Cyclophosphamide-Induced Immunosuppressed Animal Models.

This study investigates the immunomodulatory potential of Galium aparine L. (GAE) in immunodeficient animals. In this study, animals were categorized into five groups: the normal group, CYP group (cyclophosphamide intraperitoneal injection), GA5 group (cyclophosphamide + 5 μg GAE), GA50 group (cyclophosphamide + 50 μg GAE), and GA500 group (cyclophosphamide + 500 μg GAE). The CYP group exhibited significantly reduced spleen weights compared to the normal group, while the groups obtaining GAE displayed a dose-dependent increase in spleen weight. Furthermore, the GAE demonstrated dose-dependent enhancement of splenocyte proliferating activity, with significant increases observed in both LPS and ConA-induced assays. NK cell activity significantly increased in the GA50 and GA500 groups compared to the CYP group. Cytokine analysis revealed a significant increase in IL-6, TNF-α, and IFN-γ levels in ConA-induced splenocytes treated with GAE. Gene expression analysis identified 2434 DEG genes in the extract groups. Notable genes, such as Entpd1, Pgf, Thdb, Syt7, Sqor, and Rsc1al, displayed substantial differences in individual gene expression levels, suggesting their potential as target genes for immune enhancement. In conclusion, Galium aparine L. extract exhibits immunomodulatory properties. The observed gene expression changes further support the potential of Galium aparine L. extract as a natural agent for immune augmentation.

Evaluation of the effect of cannabidiol on the THLE-2 liver cell line exposed to lead

Environmental and soil pollution increase the likelihood of human exposure to toxic metals. Therefore, there is a need for new methods and substances to protect individuals against the harmful effects caused by toxic metals. The study is the first to aim at determining the protective effect of cannabidiol (CBD) against oxidative stress and inflammation induced by toxic metal exposure in Transformed Human Liver Epithelial-2 (THLE-2) cell lines representing healthy liver cells. The IC50 value was determined by exposing THLE-2 human liver healthy cell line to different molarities of lead (Pb) using the XTT kit. The protective efficacy of CBD was assessed by adding 5 μM CBD in addition to the Pb doses determined at IC50 levels to the Pb groups created in cell lines. The levels of GSH, MDA, MPO, CAT, TNF-α, IL-1β, and IL-6 in cell lines were determined using ELISA kits. The inhibition of toxic metal entry into the cells by CBD was assessed through ICP-MS analysis. The IC50 value for Pb was determined as 10 μM in 2D cell lines and 25 μM in 3D cell lines. It was observed that the application of 5 μM concentration of CBD, along with the determined IC50 doses for Pb, increased the cell proliferation rate. Furthermore, the decrease in GSH and CAT levels and the increase in MDA, MPO, TNF-α, IL-1β, and IL-6 levels observed in cell lines treated only with Pb were reversed with the application of CBD. The ICP-MS analysis revealed that CBD reduced the cellular uptake of Pb. The reversal of oxidative stress and inflammation induced by Pb, the increase in cell proliferation, and the reduction in the cellular uptake of toxic metals by CBD can be considered as strong evidence for the protective use of CBD in Pb exposures.

High-fat diet disrupts the gut microbiome, leading to inflammation, damage to tight junctions, and apoptosis and necrosis in Nyctereutes procyonoides intestines.

Given the burgeoning Nyctereutes procyonoides breeding industry and its growing scale, it is imperative to investigate the impact of high-fat diets on the health of these animals. This study involved 30 male Nyctereutes procyonoides of comparable weights (3 kg ±0.5), randomly assigned to either a control group or a high-fat diet group (n = 15 each). The latter group was fed a mixture of lard and basal diet in a 2:5 ratio, establishing a high-fat diet model in Nyctereutes procyonoides. This diet induced diarrhea and histopathological changes in the Nyctereutes procyonoides. Analysis of the small intestine contents using 16S rRNA sequencing revealed a high-fat diet-induced disruption in the gut microbiota. Specifically, Escherichia-Shigella emerged as the biomarker in the high-fat diet group (P = 0.049), while Vagococcus was prevalent in the control group (P = 0.049), indicating a significant increase in harmful bacteria in the high-fat diet group. Furthermore, this disrupted gut flora correlated with inflammation and oxidative stress, as evidenced by marked increases in TNF-α (P < 0.01), IL-1β (P < 0.05), and IL-6 (P < 0.05) levels, measured via q-PCR, Western blot, and oxidative stress assays. In addition, q-PCR analysis revealed significant upregulation of apoptosis and necrosis markers, including Bax, Caspase3, Caspase9, Caspase12, RIPK3, and RIPK1 (P < 0.01 to P < 0.001), and a concurrent downregulation of the anti-apoptotic gene Bcl-2 (P < 0.01) in the high-fat diet group, consistent with protein expression trends. These findings suggest that a high-fat diet alters the gut microbiome toward a more harmful bacterial composition, escalating inflammatory responses and intestinal tissue permeability, culminating in intestinal cell apoptosis and necrosis.IMPORTANCEThis study examines the impact of high-fat diets on Nyctereutes procyonoides. Our research established a Nyctereutes procyonoides model on a high-fat diet, revealing significant health impacts, such as diarrhea, histological anomalies, and alterations in the gut microbiota. These findings emphasize the importance of preventing health issues and promoting sustainable industry growth. They highlight the significant impact of diet on gut microbiota and overall animal health.

Sex-specific effects of ketogenic diet on anxiety-like behavior and neuroimmune response in C57Bl/6J mice

The ketogenic diet (KD) has been shown to reduce anxiety and enhance cognitive functions in neurological diseases. However, the sex-specific effects of KD on anxiety-like behavior in healthy individuals and the underlying molecular mechanisms contributing to these effects, including neuroinflammation, are unelucidated. This study investigated the sex-specific effects of KD on anxiety-like behavior and the neuroimmune response in the prefrontal cortex (PFC) and hippocampus of healthy C57BL/6J male and female mice. Animals were fed either a control diet (CD- 17% fat, 65% carb, 18% protein) or a KD (80% fat, 5% carb, 15% protein) for 4 weeks. KD increased the levels of circulating β-hydroxybutyrate (BHB) both in males and females. However, PFC BHB levels were found to be elevated only in KD males. Moreover, KD did not affect the behavior of females but improved motor abilities and reduced anxiety levels in males. KD suppressed the mRNA expression of the pan microglial markers (Cd68, P2ry12) and induced morphological changes in the male PFC microglia. A sex-specific decrease in IL1β and an increase in IL-10 levels was found in the PFC of KD males. A similar trend was observed in the hippocampus of males where KD reduced the mRNA expression of P2ry12, Il1β, and cFos. Additionally, BHB increased the production of IL-10 whereas it decreased the production of IL1β from human microglia in in-vitro conditions. In summary, these results demonstrate that the anxiolytic and motor function enhancement abilities of KD are male-specific. Reduced pro-inflammatory and improved anti-inflammatory factors in the male PFC and hippocampus may underlie these effects.

The CSF1-CSF1R pathway in the trigeminal ganglion mediates trigeminal neuralgia via inflammatory responses in mice.

Trigeminal neuralgia (TN) is the most severe type of neuropathic pain. The trigeminal ganglion (TG) is a crucial target for the pathogenesis and treatment of TN. The colony-stimulating factor 1 (CSF1) - colony-stimulating factor 1 receptor (CSF1R) pathway regulates lower limb pain development. However, the effect and mechanism of the CSF1-CSF1R pathway in TG on TN are unclear. Partial transection of the infraorbital nerve (pT-ION) model was used to generate a mouse TN model. Mechanical and cold allodynia were used to measure pain behaviors. Pro-inflammatory factors (IL-6, TNF-a) were used to measure inflammatory responses in TG. PLX3397, an inhibitor of CSF1R, was applied to inhibit the CSF1-CSF1R pathway in TG. This pathway was activated in naïve mice by stereotactic injection of CSF1 into the TG. The TN model activated the CSF1-CSF1R pathway in the TG, leading to exacerbated mechanical and cold allodynia. TN activated inflammatory responses in the TG manifested as a significant increase in IL-6 and TNF-a levels. After using PLX3397 to inhibit CSF1R, CSF1R expression in the TG declined significantly. Inhibiting the CSF1-CSF1R pathway in the TG downregulated the expression of IL-6 and TNF-α to reduce allodynia-related behaviors. Finally, mechanical allodynia behaviors were exacerbated in naïve mice after activating the CSF1-CSF1R pathway in the TG. The CSF1-CSF1R pathway in the TG modulates TN by regulating neuroimmune responses. Our findings provide a theoretical basis for the development of treatments for TN in the TG.

Changes in interleukin-2, -4, -6 and -8 expression in the postovulatory sow endometrium after artificial insemination based on conceived or failed to conceive.

The establishment and maintenance of a pregnancy requires proper interaction between the endocrine and immune systems in the uterus. Therefore, it is crucial to understand how changes in endometrial cytokine levels facilitate reproduction. This study aimed to investigate how representative cytokines sequentially changed in the endometrium and whether conception could be attributed to these changes. In this study, artificial insemination was performed twice in 160 sows and ovulation was examined every 3 h using transrectal ultrasonography. Uterine endometrial tissues were obtained via repeated biopsies at 2, 4, 6, 8, 12, 16, and 20 h after ovulation and interleukin (IL)-2, IL-4, IL-6, and IL-8 expression was examined using real-time polymerase chain reaction. The conception rate was 91.9%. The IL-2 levels showed no differences in conception or time. The expression peaks of IL-4 and IL-6 were delayed in sows that failed to conceive within 4-6 h and 2 h, respectively, compared to those that did conceive. In sows that conceived, IL-8 was highest after 2 h, and no difference was observed at other time point, regardless of conception. In sows that failed to conceive, the increase in IL-8 levels might have been cancelled or terminated before the first sampling time. These results highlight the importance of timely increases and subsequent declines in the levels of some cytokines for the establishment of pregnancy. Differences in uterine capacity start just after ovulation; detection and correction of these deviations can improve the reproductive efficiency of sows.

Plasma cytokine levels as markers of pathogenesis and treatment response in patients with non-tuberculous mycobacterial pulmonary disease.

We investigated the value of plasma cytokine levels as markers of pathogenesis and treatment response in patients with non-tuberculous mycobacteria (NTM) pulmonary disease. Plasma cytokine levels were measured and compared among patients with NTM pulmonary disease (n=111), tuberculosis (TB) patients (n=50), and healthy individuals (n=40). Changes during treatment were monitored at 3 and 6 months after treatment. According to the treatment response, NTM patients were classified as 'resistance' or 'sensitivity' responders. The results revealed that five out of twelve cytokines exhibited significantly higher levels in NTM patients compared to controls. Among these, interleukin (IL)-6 demonstrated the strongest discriminating capacity for NTM. Furthermore, when combined with IL-1β, they efficiently distinguished between NTM drug-resistant and drug-sensitive patients, as well as between NTM and TB groups. Additionally, IL-6 levels initially rose and then decreased in the NTM drug-resistant group during the six months of treatment, similar to the behavior of IL-1β in the NTM drug-sensitive group. Subgroup analyses of the sensitive group with differential treatment responses revealed an increase in IL-10 levels in the six-month treatment responders. A high IL-6/IL-10 ratio was associated with increased disease severity of NTM and TB. Collectively, combinations of various plasma cytokines, specifically IL-1β, IL-6, and IL-10, effectively distinguished NTM patients with varying mycobacterial burdens, with IL-6 and IL-10 emerging as potential biomarkers for early treatment response. The combination of IL-6 and IL-1β demonstrated the highest discriminatory value for distinguishing between NTM-resistant and NTM-sensitive groups as well as between NTM and TB groups.

Effect of carboxymethyl cellulose, icodextrin and hyaluronic acid solutions on postoperative intraabdominal adhesion formation in rabbits and the role of cytokines in intraabdominal adhesion formation

In this study, the aim was to investigate the effects of 1% CMC, 4% ICO and 0.4% HA solutions in preventing postoperative IAs and their contribution to peritoneal wound healing, as well as the relationship of cytokines with the formation of postoperative IAs in rabbits. The material of the study consisted of 32 healthy rabbits. Rabbits underwent median laparotomy following general anesthesia. Serosal abraded was created at the antimesenteric border of the cecum. Next, a 3 × 2 cm peritoneum was excised on the right abdominal wall and the defect was closed using with a 2/0 silk suture. The rabbits were randomly assigned to either of the following four treatment groups: CNT (0.9% NaCl), ICO (4% icodextrin), HA (0.4% hyaluronic acid) and CMC (1% carboxymethyl cellulose). Both cecum and peritoneal surfaces were treated with 20 ml each of treatment solutions. On the 7th postoperative day, the rabbits were euthanized with a lethal dose of sodium pentobarbital and the degree of adhesion was evaluated. Samples taken from the peritoneal defect were examined histopathologically and tissue hydroxyproline levels were measured. Serum TNF-α, IL-1 and IL-6 levels were measured from blood samples taken before the surgery and at 1, 6, 24 and 48 hours after the surgery. It was observed that the adhesion grade in the HA group (p &lt; 0.05) and CMC group (p &lt; 0.01) was significantly lower than the control group. Although peritoneal tissue hydroxyproline levels were lower in the other groups compared to the control group, they were not statistically significant (p &gt; 0.05). In rabbits with adhesion formation, it was determined that TNF-α levels increased at the 6th postoperative hour (p &lt; 0.05), and IL-6 levels increased at the 6th, 24th and 48th postoperative hours (p &lt; 0.001) compared to preoperative. In this study, treating tissues with 0.4% HA and 1% CMC solutions suppressed peritoneal inflammation, and this resulted in an increase in the levels of proinflammatory cytokines TNF-α and IL-6. It has been determined that the application of these solutions reduces postoperative adhesion formation. It was concluded that TNF-α and especially IL-6, which are proinflammatory cytokines, can be a non-invasive biomarker in determining postoperative IA formation and evaluating the adhesive process.

Molecular characteristics and pathogenic mechanisms of KPC-3 producing hypervirulent carbapenem-resistant Klebsiella pneumoniae (ST23-K1).

This study aimed to comprehensively investigate hypervirulent carbapenem-resistant Klebsiella pneumoniae (CR-hvKP) in the Ningbo region. Importantly, we sought to elucidate its molecular characteristics and pathogenic mechanisms. This information will provide evidence-based insights for preventing and controlling nosocomial infections and facilitate improved clinical diagnosis and treatment in this region. 96 carbapenem-resistant Klebsiella pneumoniae strains were collected from the Ningbo region between January 2021 and December 2022. Whole genome sequencing and bioinformatic methods were employed to identify and characterize CR-hvKP strains at the molecular level. The minimum inhibitory concentrations (MICs) of common clinical antibiotics were determined using the VITEK-2 Compact automatic microbiological analyzer. Plasmid conjugation experiments evaluated the transferability of resistance plasmids. Finally, mouse virulence assays were conducted to explore the pathogenic mechanisms. Among the 96 strains, a single CR-hvKP strain, designated CR-hvKP57, was identified, with an isolation frequency of 1.04%. Whole-genome sequencing revealed the strain to be ST23 serotype with a K1 capsule. This strain harbored three plasmids. Plasmid 1, a pLVPK-like virulence plasmid, carried multiple virulence genes, including rmpA, rmpA2, iroB, iucA, and terB. Plasmid 2 contained transposable element sequences such as IS15 and IS26. Plasmid 3, classified as a resistance plasmid, harbored the bla KPC-3 carbapenem resistance gene. Mouse virulence assays demonstrated a high mortality rate associated with CR-hvKP57 infection. Additionally, there was a significant increase in IL-1β, IL-6, and TNF-α levels in response to CR-hvKP57 infection, indicating varying degrees of inflammatory response. Western blot experiments further suggested that the pathogenic mechanism involves activation of the NF-κB signaling pathway. This study confirms the emergence of hypervirulent CR-hvKP in the Ningbo region, which likely resulted from the acquisition of a pLVPK-like virulence plasmid and a bla KPC-3 resistance plasmid by the ST23-K1 type Klebsiella pneumoniae. Our findings highlight the urgent need for more judicious use of antibiotics to limit the emergence of resistance. Additionally, strengthening infection prevention and control measures is crucial to minimize the spread of virulence and resistance plasmids.

Dysregulation of Peripheral Blood Mononuclear Cells and Immune-Related Proteins during the Early Post-Operative Immune Response in Ovarian Cancer Patients.

Surgical treatment is a cornerstone of ovarian cancer (OC) therapy and exerts a substantial influence on the immune system. Immune responses also play a pivotal and intricate role in OC progression. The aim of this study was to investigate the dynamics of immune-related protein expression and the activity of peripheral blood mononuclear cells (PBMCs) in OC patients, both before surgery and during the early postoperative phase. The study cohort comprised 23 OC patients and 20 non-cancer controls. A comprehensive analysis of PBMCs revealed significant pre-operative downregulation in the mRNA expression of multiple immune-related proteins, including interleukins, PD-1, PD-L1, and HO-1. This was followed by further dysregulation during the first 5 post-operative days. Although most serum interleukin concentrations showed only minor changes, a distinct increase in IL-6 and HO-1 levels was observed post-operatively. Reduced metabolic and phagocytic activity and increased production of reactive oxygen species (ROS) were observed on day 1 post-surgery. These findings suggest a shift towards immune tolerance during the early post-operative phase of OC, potentially creating a window for treatment. Further research into post-operative PBMC activity could lead to the development of new or improved treatment strategies for OC.

Bioactive composition and protective properties of Talium triangulare in dextran sodium sulphate-induced ulcerative colitis in rats

BackgroundsThe prevalence of ulcerative colitis (UC) is gradually becoming a global health challenge. Talinum triangulare, commonly called "waterleaf" (Tu ren shen in Chinese) is a plant that is cultivated in Asia, western Africa, and other tropical rainforest regions as a food and medicine plant. Due to its medicinal significance, Talium triangulare has been used in Chinese traditional medicines to treat various human diseases. The purpose of this study was to evaluate the possible protective effect of aqueous leaf extract of Talium triangulare (TT) on dextran sodium sulphate (DSS)-induced colitis. MethodsExposure of rats to 5 % DSS in drinking water induced colitis, and the effect of 200 mg/kg TT was studied over a 10-day period via oral administration. The effect of TT on anti-inflammatory, redox assays, clinical manifestations (disease activity index), and colonic mucosa histological changes was determined. ResultsDSS induced colonic inflammation in rats with a significant increase in NO, MPO, TNF-α, IL-6, and IL-1β levels compared to the untreated group. Oxidative stress was markedly increased in the DSS group with concomitant suppression of antioxidant enzymes activities. Exposure of rats to DSS causes lipid peroxidation with a marked increase in colonic level of malondialdehyde. Nevertheless, the administration of TT significantly suppressed inflammation in the colon by downregulating expression of the inflammatory indicators. TT suppressed the levels of MDA and H2O2 thus mitigating lipid peroxidation chain reaction. Furthermore, TT attenuated DSS-facilitated oxidative damage by improving the enzymatic activities of the protective enzymes (CAT, GST, and SOD) and increasing GSH level. ConclusionTT enhanced the antioxidant enzymes activities, attenuated oxidative stress, suppressed lipid peroxidation, inhibited inflammation, and reduced DAI score. These beneficial activities of TT could be attributed to its phytoconstituents depicted in GCMS results. The results of this research indicated that TT could contain therapeutic agents for the treatment of inflammatory colitis.