Plasma Levels Of Inflammatory Cytokines Research Articles

Mirogabalin improves cancer-associated pain but increases the risk of malignancy in mice with pancreatic cancer.

Mirogabalin, a selective voltage-gated calcium channel α2δ ligand, improves peripheral neuropathic pain; however, its effects on patients with cancers including pancreatic ductal adenocarcinoma (PDAC) remain unknown. We analyzed the effects of mirogabalin on a KPPC ( LSL-KrasG12D/+; Trp53flox/flox; Pdx-1cre/+ ) mouse model of PDAC. Six-week-old KPPC mice received oral mirogabalin (10 mg/kg/day) (n = 10) or vehicle water (n = 14) until the humane end point. Cancer-associated pain was evaluated using the scores of hunching and mouse grimace scale (MGS). Tumor status and plasma cytokine levels were determined using histopathological analysis and cytokine array, respectively. The effects of mirogabalin on the proliferative ability of PDAC cell lines were determined. The scores of the hunching and MGS improved after mirogabalin administration with a decrease in the plasma levels of inflammatory cytokines, such as tumor necrosis factor-α, interleukin-6, and interferon-γ. Although no significant difference in the survival rate was observed, mirogabalin significantly increased pancreatic tumor size and proliferative index of Ki-67 and cyclins. Local arginase-1 + M2-like tumor-associated macrophages and CD31 + tumor blood vessels increased after mirogabalin administration. By contrast, the number of α-smooth muscle actin + cancer-associated fibroblasts, desmoplastic stroma, and CD8 + T cells decreased. Local myeloperoxidase + tumor-associated neutrophils and CD45R + B cells were unaltered. Mirogabalin enhanced the proliferative ability of PDAC cell lines with the upregulation of cyclins and cyclin-dependent kinases; however, it inhibited the potential of pancreatic stellate cells in vitro. Therefore, our results suggest that mirogabalin improves cancer-associated pain but enhances the proliferative potential of PDAC in vitro and in vivo.

Peripheral inflammation and cognition in older adults with and without cognitive impairment

AbstractBackgroundUnderstanding how different inflammatory markers relate to cognition is important for the development of therapeutic targets in Alzheimer’s disease (AD). However, many studies have relied on a single, broad measure of inflammation or a select handful of markers that have been identified in the AD literature. We examined a large set of plasma inflammatory markers that were statistically grouped via principal component analysis (PCA). Associations between inflammation component scores and cognition were examined.MethodParticipants included 41 cognitively normal (CN) and 64 cognitively impaired (CI; 47 mild cognitive impairment and 17 AD) older adults from the Center for Neurodegeneration and Translational Neuroscience. Composite scores for memory, executive functioning, processing speed, and language were created by averaging age‐adjusted z scores for two tests per domain. Plasma levels of inflammatory cytokines were measured using the Luminex platform and Biorad 48‐plex cytokine assay. Five markers with null values were excluded, and the remaining markers were included in a PCA. Relationships between the inflammation component scores and cognition were examined using linear regression models with inflammatory marker as the predictor and each cognitive composite separately as the dependent variable, with adjustment for education, sex, and clinical diagnosis (CN vs CI).ResultThe PCA produced a 7‐component solution that accounted for 75% of the variance. In the final models, Components 2 (β = 0.260, p = 0.020) and 4 (β = 0.235, p = 0.036) were positively associated with memory, Components 2 (β = 0.231, p = 0.052), 4 (β = 0.260, p = 0.031), and 6 (β = 0.280, p = 0.016) were positively associated with executive functioning, and Components 2 (β = 0.221, p = 0.032) and 4 (β = 0.283, p = 0.007) were positively associated with processing speed. Inflammatory components were not significantly related to language.ConclusionHigher levels of peripheral inflammatory cytokines were associated with better memory, executive functioning, and processing speed in older adults, even when accounting for clinical diagnosis. Acute elevations of certain inflammatory markers may be associated with a protective role in cognitive aging. Longitudinal research examining stability of cytokine levels and association with cognition in AD trajectories is needed.

RBC Transfusion to Severe Anemic Murine Neonates Leads to Systemic Inflammation

Background: Transfusions of red blood cells (RBCT) are necessary and life-saving in premature and critically ill infants, who experience severe anemia due to physiologic and iatrogenic factors. Recently, we showed that severe anemia was associated with increased intestinal permeability, demonstrated by identifying endotoxin in the bloodstream. The risks of experiencing severe anemia during critical developmental periods must be balanced with the risks of transfusions, which can lead to the Systemic Inflammatory Response Syndrome (SIRS). In this study, we investigated the role of severe anemia in endotoxin sensitization of the liver, and of RBCT-triggered cytokine storm, in murine neonates. Methods: C57BL/6 (WT) and TLR4-KO mouse pups were studied in 4 groups (n=6 each): (1) naïve controls; (2) non-anemic RBC transfused; (3) anemic (hematocrit 20-24%); and (4) anemic RBC transfused. Pups in anemic groups underwent facial vein phlebotomy on days P2, 4, 6, 8, and 10. Leukoreduced and 7-day refrigerator-stored packed RBCs from allogeneic (FVB) adult donors were transfused intravenously into the retro-orbital plexus (20 mL/kg) of P10 pups. Blood was collected 2, 4, and 6 hours post-transfusion, and plasma was separated for quantifying inflammatory cytokines (IL-1β, TNF-α, CXCL-1, IL-6, IFN-γ) using a Luminex assay. Plasma non-transferrin bound iron (NTBI) levels were measured at 2 hours post-transfusion. To measure leaky gut associated liver endotoxin sensitization, genetically-modified E. coli, expressing green fluorescent protein (GFP; 108 colony-forming units), were administered by gavage; these bacteria were measured in the liver of anemic pups by immunofluorescence using anti-GFP antibody. To investigate the endotoxin-associated monocyte-macrophage system in the liver, single cell suspensions were prepared and flow cytometry was performed by staining with antibodies against CD45, CD11b, F4/80, CD11c, Ly6G and Ly6C. Results: RBC transfusions led to the acute release of inflammatory mediators (IL-1β, TNF-α, CXCL-1, IL-6, IFN-γ) within 2 hours, in pups with a normal hematocrit ("transfusion controls"), as well as in pups rendered anemic. However, inflammatory cytokine levels were significantly higher in RBC-transfused anemic pups as compared to transfusion controls, and remained elevated for several hours. We excluded the inflammatory response at 6 hours post-transfusion, because intestinal inflammation was observed after that time by the release of intestinal fatty acid binding protein (iFABP). No changes were found in plasma NTBI levels between transfusion control and anemic-transfused pups, indicating that the acute inflammatory response in the anemic-transfused groups is independent of liver iron accumulation due to stored RBC clearance. However, severely anemic mouse pups had elevated amounts of orally administered E. coli-GFP in the liver relative to control mice, indicating that anemia allows endotoxin delivery and sensitizes the liver by bacterial translocation across the anemic intestine. The TLR4-KO mice study showed that overall plasma levels of inflammatory cytokines (TNF-α, IL-6, IFN-γ) were reduced, as compared to anemic-transfused WT pups, but remained elevated in the plasma of anemic-transfused TLR4-KO pups, as compared to TLR4-KO transfused-control pups, indicating that this acute inflammatory response does not depend on TLR4. By flow cytometry, anemic (P10) pups showed an increased number of CD11bhiF4/80loLy6C+ hepatic monocytes as compared to control, but no changes in CD11bhiF4/80hi hepatic macrophages (i.e., Kupffer cells); qRT-PCR studies confirm that these anemic hepatic monocytes contained mRNA transcripts of IL-1β, TNF-α, CXCL-1, IL-6, and IFN-γ. In addition, the anemic mouse pups that were treated with recombinant haptoglobin (rHp, 100µg/kg) showed reduced plasma levels of IL-1β, TNF-α, CXCL-1, IL-6, and IFN-γ at 2-6 hours post-transfusion of packed-stored RBCs. Conclusions: In this mouse model, the severe anemia-associated low-grade inflammatory state in the liver leads to increased numbers of hepatic monocytes with inflammatory cytokines mRNA; in this condition, RBC transfusions activate these monocytes and cause systemic inflammation. Pre-treatment of rHp was reduce the RBCT-associated systemic inflammatory response.

Gandouling Mitigates CuSO4-Induced Heart Injury in Rats.

Simple SummaryWilson’s disease (WD) is a rare autosomal recessive inherited disorder of copper (Cu) metabolism, which is one of the few neurogenetic diseases with a cure. Cu deposition in the heart tissue is a mechanism of cardiac involvement. Importantly, Gandouling (GDL) effectively promotes the excretion of intracellular Cu. Therefore, the purpose of this study was to evaluate the protective effects of GDL on heart injuries and the underlying mechanisms in a copper sulfate (CuSO4)-induced animal model. The results showed that the protective effect of GDL on the heart was superior to that of penicillamine.We assessed the protective effects of Gandouling (GDL) on copper sulfate (CuSO4)-induced heart injuries in Sprague–Dawley rats, which were randomly divided into the control, CuSO4, GDL + CuSO4 and penicillamine + CuSO4 groups. The rats received intragastric GDL (400 mg/kg body weight) once per day for 42 consecutive days after 56 days of CuSO4 exposure, and penicillamine was used as a positive control. The levels of plasma inflammatory cytokines (IMA, hFABP, cTn-I and BNP) were determined using the enzyme-linked immunosorbent assay. The histopathological symptoms were evaluated using hematoxylin and eosin staining and transmission electron microscopy. To determine the underlying mechanism, Western blotting was conducted for the detection of the heme oxygenase 1 (HO-1) expression. The results revealed that GDL supplementation alleviated the histopathological symptoms of the rat heart tissue, promoted Cu excretion to attenuate impairment, and significantly decreased inflammatory cytokine levels in the plasma (p < 0.01). In addition, GDL increased the HO-1 expression in the rat hepatic tissue. The protective effect of GDL on the heart was superior to that of penicillamine. Overall, these findings indicate that GDL alleviates hepatic heart injury after a Cu overaccumulation challenge, and GDL supplements can be beneficial for patients with Wilson’s disease.

Calcium-Based Antimicrobial Peptide Compounds Attenuate DNFB-Induced Atopic Dermatitis-Like Skin Lesions via Th-Cells in BALB/c Mice.

Atopic dermatitis (AD) is a chronic and recurrent inflammatory skin disease, characterized by severe itching and recurrent skin lesions. We hypothesized that a novel treatment involving calcium-based antimicrobial peptide compounds (CAPCS), a combination of natural calcium extracted from marine shellfish, and a variety of antimicrobial peptides, may be beneficial for AD. We established a dinitrofluorobenzene (DNFB)-induced AD model in BALB/c mice to test our hypothesis. We observed mouse behavior and conducted histopathological and immunohistochemical analyses on skin lesions before and after CAPCS treatment. We also characterized the changes in the levels of cytokines, inflammatory mediators, and Toll-like receptors (TLRs) in plasma and skin lesions. The results showed that (i) topical application of CAPCS ameliorated AD-like skin lesions and reduced scratching behavior in BALB/c mice; (ii) CAPCS suppressed infiltration of inflammatory cells and inhibited the expression of inflammatory cytokines in AD-like skin lesions; (iii) CAPCS reduced plasma levels of inflammatory cytokines; and (iv) CAPCS inhibited TLR2 and TLR4 protein expression in skin lesions. Topical application of CAPCS exhibits a therapeutic effect on AD by inhibiting inflammatory immune responses via recruiting helper T cells and engaging the TLR2 and TLR4 signaling pathways. Therefore, CAPCS may be useful for the treatment of AD.

The Role of SGLT2 Inhibitor Ipragliflozin on Cardiac Hypertrophy and microRNA Expression Profiles in a Non-diabetic Rat Model of Cardiomyopathy.

Evidence from clinical trials suggests that the cardioprotective effects of sodium-glucose cotransporter 2 (SGLT2) inhibitors may arise through non-glycemic control-related mechanisms. Further, the cardiovascular advantages of SGLT2 inhibitors are likely present among non-diabetic patients with known cardiovascular diseases (CVDs). Here, we studied the impact of ipragliflozin, a selective SGLT2 inhibitor, on cardiac histopathology and microRNA (miRNA) expression profiles in a non-diabetic rat model of cardiomyopathy. Ipragliflozin was added to chow (0.01% (w/w)) and given to male DahlS.Z-Leprfa/Leprfa (DS/obese) rats for 6 weeks. Similarly aged male DahlS.Z-Lepr+/Lepr+ (DS/lean) rats were treated as controls. Measurements of systolic blood pressure (SBP) and heart rate (HR) were taken every other week. Following ipragliflozin treatment for 6 weeks, we conducted echocardiography, histopathological examination, and miRNA expression analysis (microarray). The impact of ipragliflozin on blood parameters was additionally examined. In DS/obese rats, ipragliflozin reduced SBP without affecting HR, reduced interventricular septal thickness in echocardiography and left ventricular (LV) organ weight, and improved hypertrophy of cardiomyocytes according to histopathological experiments. Further, ipragliflozin reduced plasma inflammatory cytokine levels in DS/obese rats. Additionally, ipragliflozin treatment altered the expression profile of miRNAs related to cardiac hypertrophy and heart failure in the LV compared to DS/obese control rats. Ipragliflozin prevented LV hypertrophy and altered related miRNA expression profiles in non-diabetic DS/obese rats. These findings suggest that miRNAs may play a partial role in regulating the structure of the heart and that SGLT2 inhibitors may exert cardio-protective effects by changing miRNA expression profiles in non-diabetic patients with CVDs.

Maternal probiotics supplementation improves immune and antioxidant function in suckling piglets via modifying gut microbiota.

Probiotics could improve the health, growth, and development of host or their foetuses/offspring via regulating gut microbiota. The present study was conducted to determine the effects of maternal probiotics supplementation on gut microbiota and metabolites of sows and their suckling piglets, as well as plasma biochemical parameters, oxidative/anti-oxidative indexes, and inflammatory cytokine levels of suckling piglets. A total of 32 pregnant Bama mini-pigs were selected and randomly divided into two groups. The sows were fed a basal diet (control group) or a basal diet supplemented with probiotics (probiotics group) from mating to day 21 of lactation. Samples from sows were collected on day 105 of pregnancy and day 21 of lactation and from piglets on day 21 of lactation. The results showed that probiotics supplementation increased the faecal abundances of Ruminococcus, Bacteroides, and Anaeroplasma and decreased Tenericutes on day 105 of pregnancy while increased the abundances of Actinobacteria and Anaerostipes and decreased Proteobacteria and Desulfovibrio on day 21 of lactation. In addition, probiotics supplementation decreased the faecal levels of tryptamine, putrescine, and cadaverine on day 105 of pregnancy and isovalerate and skatole on day 21 of lactation while increased butyrate level on day 21 of lactation. Further studies showed that maternal probiotics supplementation decreased the plasma levels of AMM, TC, LDL-C, Ala, Tau, MDA, H2 O2 , IL-1β, IL-2, IL-6, and IFN-α of suckling piglets. Moreover, maternal probiotics supplementation increased the abundances of Deferribacteres, Fusobacteria, and Fusobacterium while decreased Anaerostipes in piglet's colon. Spearman's correlation analysis revealed a potential link between gut microbiota alterations and their metabolites. Dietary probiotics supplementation during pregnancy and lactation periods could improve sow status, alleviate oxidative stress and inflammation response, and improve nutrient metabolism of piglets by altering the gut microbiota. The probiotics alter maternal and offspring's gut microbiota involving in offspring's physiological and metabolic changes, and present a new perspective that the effects of gut microbiota changes induced by probiotics supplementation will help in addressing the growth and development and health problem of their foetuses/offspring.

X, but not Y, Chromosomal Complement Contributes to Stroke Sensitivity in Aged Animals.

Post-menopausal women become vulnerable to stroke and have poorer outcomes and higher mortality than age-matched men, and previous studies suggested that sex chromosomes play a vital role in mediating stroke sensitivity in the aged. It is unknown if this is due to effects of the X or Y chromosome. The present study used the XY* mouse model (with four genotypes: XX and XO gonadal females and XY and XXY gonadal males) to compare the effect of the X vs. Y chromosome compliment in stroke. Aged (18-20months) and gonadectomized young (8-12weeks) mice were subjected to a 60-min middle cerebral artery occlusion. Infarct volume and behavioral deficits were quantified 3days after stroke. Microglial activation and infiltration of peripheral leukocytes in the aged ischemic brain were assessed by flow cytometry. Plasma inflammatory cytokine levels by ELISA, and brain expression of two X chromosome-linked genes, KDM6A and KDM5C by immunochemistry, were also examined. Both aged and young XX and XXY mice had worse stroke outcomes compared to XO and XY mice, respectively; however, the difference between XX vs. XXY and XO vs. XY aged mice was minimal. Mice with two copies of the X chromosome showed more robust microglial activation, higher brain-infiltrating leukocytes, elevated plasma cytokine levels, and enhanced co-localization of KDM6A and KDM5C with Iba1+ cells after stroke than mice with one X chromosome. The number of X chromosomes mediates stroke sensitivity in aged mice, which might be processed through the X chromosome-linked genes and the inflammatory responses.

The association between heightened ADHD symptoms and cytokine and fatty acid concentrations during pregnancy.

ObjectivePrevious research conducted with samples of children suggest that individuals with attention-deficit/hyperactivity disorder (ADHD) have altered fatty acid concentrations and may have increased systemic inflammation. Whether these differences are also apparent in other populations of individuals with heightened ADHD symptoms (e.g., pregnant adults) is unknown. The goal of the current study was to examine whether there are ADHD-associated differences in polyunsaturated fatty acid concentrations or pro-inflammatory cytokine concentrations during pregnancy, a developmental period when fatty acid concentrations and systemic inflammation have implications for the health of both the pregnant person and the developing child. We hypothesized that plasma levels of the ratio of omega-6s to omega-3s (n-6:n-3) and plasma inflammatory cytokine levels would be higher in individuals with heightened ADHD symptoms, consistent with previous findings in children with ADHD.MethodsData (N = 68) came from a prospective study of pregnant community volunteers who were oversampled for ADHD symptoms. During the 3rd trimester, plasma concentrations of fatty acids and the pro-inflammatory cytokines interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) were assessed. Dietary intake was examined in the 3rd trimester using three 24-h recalls conducted by trained dietitians and by examining plasma levels of conjugated linoleic acid (n-6) and α-linolenic acid (n-3), essential fatty acids that must come from dietary intake.ResultsThe group with heightened ADHD symptoms had higher n-6:n-3s (β = 0.30, p < 0.01) and higher TNF-α concentrations (β = 0.35, p < 0.001) relative to controls. There were no group differences in dietary variables, as assessed by self-report and via plasma concentrations of essential fatty acids. IL-6 was not reliably associated with ADHD status in this sample.ConclusionPregnant individuals with ADHD, on average, had higher plasma n-6:n-3s and higher TNF-α concentrations relative to controls. A difference was not detected in their dietary intake of fatty acids or other relevant nutrients. Though these null findings are inconclusive, they are consistent with the hypothesis that ADHD-associated differences in plasma fatty acid concentrations are the result of ADHD-associated differences in fatty acid metabolism, rather than simply differences in dietary intake.

Unpredictable Sound Stress Model Causes Migraine-Like Behaviors in Mice With Sexual Dimorphism.

Migraine represents one of the major causes of disability worldwide and is more prevalent in women; it is also related to anxiety symptoms. Stress, such as sound stress, is a frequently reported trigger in migraine patients, but the underlying mechanisms are not fully understood. However, it is known that patients with migraine have higher levels of plasma inflammatory cytokines and calcitonin gene-related peptide (CGRP). Stress mediated by unpredictable sound is already used as a model of painful sensitization, but migraine-like behaviors and sexual dimorphism have not yet been evaluated. This study characterized nociception and anxiety-related symptoms after the induction of sound stress in mice. C57BL/6 mice (20–30 g) were exposed to unpredictable sound stress for 3 days, nonconsecutive days. We observed enhanced plasma corticosterone levels on day 1 after stress induction. First, 7 days after the last stress session, mice developed hind paw and periorbital mechanical allodynia, grimacing pain behavior, anxiety-like symptoms, and reduced exploratory behavior. The nociceptive and behavioral alterations detected in this model were mostly shown in female stressed mice at day 7 post-stress. In addition, on day 7 post-stress nociception, these behaviors were consistently abolished by the CGRP receptor antagonist olcegepant (BIBN4096BS, 100 mg/kg by intraperitoneal route) in female and male stressed mice. We also demonstrated an increase in interleukine-6 (IL-6), tumor necrosis factor (TNF-α), and CGRP levels in stressed mice plasma, with female mice showing higher levels compared to male mice. This stress paradigm allows further preclinical investigation of mechanisms contributing to migraine-inducing pain.

Blood Inflammatory Cytokines as Predictors of Depression in Patients With Glioma.

BackgroundDepression commonly develops as a comorbid disorder related to glioma, which affects the patients’ physical function and prognosis. Circulating inflammatory cytokines are potential predictors of depression in disparate cancers. However, less research has specifically investigated this aspect within the context of glioma.Study objectivesThe objective of this study was to investigate the occurrence of depression in patients with glioma and draw a comparison of the ability to predict it through diverse inflammatory cytokines.MethodsA total of 203 patients with stage I–IV glioma were enrolled in this study. Depression was evaluated according to the Hamilton Depression Scale, and the plasma inflammatory cytokines levels were simultaneously measured. We performed the receiver operating characteristic (ROC) analysis to confirm the abilities of identified inflammatory cytokines to predict depression.ResultsAmong the 203 patients with glioma, 135 (66.5%) showed obvious depressive symptoms. Proinflammatory cytokines, including interleukin (IL)-6 (area under the curve (AUC) = 0.76) and tumor necrosis factor (TNF)-α (AUC = 0.75), showed good performance in accurately predicting depression in patients with glioma. These inflammatory cytokines indicated great potential to be depression biomarkers regardless of the patients’ disparate treatment experience.ConclusionWith their relatively simple and time-saving measurement procedures, inflammatory cytokines should be seriously considered effective clinical screening and diagnostic tools, as well as potential biomarkers for depression in patients with glioma.

Altered levels of circulating mitochondrial DNA in elderly people with sarcopenia: Association with mitochondrial impairment.

Age-related chronic inflammatory process is often referred to as "inflammaging", which had been described in several human disorders, including sarcopenia. Recently, mitochondrial DNA (MtDNA) has moved into the spotlight as a "damage-associated molecular pattern" (DAMP) agent that can potentially elicit inflammation. Yet, the roles of this mitochondrial DAMP have never been investigated in sarcopenia. Cross-sectional study. From January 2021 to June 2021, elderly outpatients ≥65years and able to finish a comprehensive geriatric assessment were recruited in our study. Participants were divided into sarcopenia group and non-sarcopenia group according to the DXA scans and grip strength. Genomic DNA was extracted from plasma and peripheral blood mononuclear cells (PBMCs), and changes in MtDNA copies were quantified using qPCR. Plasma levels of inflammatory cytokines were measured using ELISA kits. Loss of mitochondrial membrane potential (Δψm) in PBMCs was analyzed using the fluorescent probe JC-1. Participants with sarcopenia were significantly older, more likely to be physically inactive, and had higher levels of circulating cell-free MtDNA (ccf-MtDNA) (all p<0.05). After adjusting for potential confounders, ccf-MtDNA was independently associated with increased odds of sarcopenia (adjusted odds ratio (AOR), 1.576; p=0.009). Furthermore, ROC curve analysis showed that ccf-MtDNA had an area under the curve (AUC) of 0.726 (95% CI: 0.607-0.844; p<0.05) for distinguishing elderly subjects from sarcopenia. Compared with non-sarcopenia subjects, plasma interleukin (IL)-6 and IL-8 were significantly higher in sarcopenia subjects (both p<0.05). By performing a correlation test, it was found that the level of IL-6 was positively correlated with ccf-MtDNA (r=0.301; p<0.05). Then, PBMCs were used as surrogates for mitochondria-rich cells, and the results showed that the relative amplification of MtDNA in PBMCs was significantly reduced (p<0.05), whereas the depolarization of Δψm was significantly increased in sarcopenia subjects (p<0.05). Taken together, our data suggested that circulating MtDNA might be a novel and important source of inflammatory stimuli potentially relevant for sarcopenia in elderly people, and this would provide an attractive therapeutic target to improve this disease.

MO603: Effects of Intradialytic Bicycle Ergometer Exercise on Inflammation Transcription Factors in Patients with Chronic Kidney Disease

Abstract BACKGROUND AND AIMS Oxidative stress and inflammation are involved in the pathophysiological mechanisms in patients with chronic kidney disease (CKD) on hemodialysis (HD). In this context, the therapeutic potential of targeting the gene transcription factors associated with these mechanisms has been studied; for instance, nuclear factor erythroid 2-related factor 2 (Nrf2), the master regulator of antioxidant responses and, nuclear factor κappa B (NF-κB) that promotes the inflammatory response. However, the effects of physical exercise on the modulation of these factors are under investigation in CKD patients. Therefore, the aim of the present study was to evaluate the effects of an intradialytic aerobic exercise training program on the expression of transcription factors (Nrf2 and NF-κB) in patients with CKD on HD. METHODS This is a longitudinal, randomized clinical trial with a washout period and crossover performed with 18 patients randomized into two groups: Exercise (individualized intradialytic aerobic exercise on an adapted stationary exercise bike) three times per week for 3 months and control (without exercise). After the washout period (1 month), the exercise group became the control, and the other group performed the exercises for another 3 months. Peripheral blood mononuclear cells (PBMCs) were isolated, and quantitative real-time PCR analysis was performed to evaluate Nrf2, NF-kB and NAD(P)H: quinone oxidoreductase-1 (NQO1) mRNA expression. The plasma levels of inflammatory cytokines, tumour necrosis factor-α (TNF-α), C-reactive protein (CRP) and interleukin-6 (IL-6) were evaluated by ELISA. RESULTS Eighteen patients [11 men, 44.1 ± 8.4 years, 17.3 (6.6–124) months on HD] completed all the study. The obtained data revealed that the intervention did not affect the mRNA expression of Nrf2, NQO1 and NF-κB (Table 1). Also, TNF-α levels were not changed. However, IL-6 showed a tendency to decrease after the exercise intervention (P = 0.054). CONCLUSION Three months of intradialytic aerobic exercise were insufficient for modulating the transcription factors associated with antioxidant activity (Nrf2 and NQO1) and inflammation (NF-κB) in patients with CKD.

Anti-atherosclerotic effects of geraniin through the gut microbiota-dependent trimethylamine N-oxide (TMAO) pathway in mice

BackgroundCardiovascular disease is a leading cause of death, which signifies the urgent need for effective anti-atherosclerotic strategies. Gut microbiota-dependent trimethylamine-N-oxide (TMAO) is associated with atherosclerosis, and geraniin, a natural polyphenol with various biological activities, might play key role in this process. PurposeWe aimed to investigate the pharmacological activity of geraniin in atherosclerosis through remodeling the gut microbiota. MethodsC57BL/6J ApoE−/− mice were administrated geraniin for 12 weeks. The colon contents were analyzed via 16S rRNA sequencing. Pathological staining was performed to evaluate the atherosclerotic characteristics. Cytokine assays detected the levels of plasma inflammatory cytokines. RAW264.7 cells were cultured in vitro and treated with TMAO. Tandem Mass Tag quantitative proteomics analysis and western blot were performed to investigate the effect of TMAO in macrophages. ResultsThe plasma TMAO level in mice significantly decreased after geraniin intervention. The predominant intestinal microflora from geraniin-treated mice were Bacteroides (65.3%) and Firmicutes (30.6%). Pathological staining demonstrated that administration of geraniin attenuated atherosclerotic characteristics. After geraniin treatment, plasma levels of IL-1β, IL-6, and TNF-α in mice were significantly reduced, and IL-10 levels were significantly increased. Proteomics analysis demonstrated the number of differentially expressed proteins after TMAO administration. In vitro study suggested that the atherogenic effect of TMAO could be attributed to changes in CD36, transmembrane protein 106a, apolipoprotein C1, macrophage scavenger receptor types I and II, and alpha-2-macroglobulin. ConclusionGeraniin might be an effective prospective drug against cardiovascular diseases, and the gut microbiota is a potential target to reduce the risk of atherosclerotic disease.

Presence of Tumor Necrosis Factor-Alpha in Urine Samples of Patients With Chronic Low Back Pain Undergoing Chiropractic Care: Preliminary Findings From a Prospective Cohort Study.

Background and aimsLow back pain is the leading cause of years lived with disability worldwide. Chiropractors employ different interventions to treat low back pain, including spinal manipulative therapy, although the mechanisms through which chiropractic care improves low back pain are still unclear. Clinical research and animal models suggest that spinal manipulation might modulate plasma levels of inflammatory cytokines, which have been involved in different stages of low back pain. More specifically, serum levels of Tumor Necrosis Factor-alpha (TNF-α) have been found to be elevated in patients with chronic low back pain. We aimed to investigate whether urine from chronic low back pain patients could be an appropriate medium to measure concentrations of TNF-α and to examine possible changes in its levels associated to chiropractic care.MethodsUrine samples were collected from 24 patients with chronic low back pain and TNF-α levels were analyzed by ELISA before and after 4–6 weeks of care compared to a reference value obtained from 5 healthy control subjects, by means of a Welch’s t-test. Simultaneously, pain intensity and disability were also evaluated before and after care. Paired t-tests were used to compare mean pre and post urinary concentrations of TNF-α and clinical outcomes.ResultsSignificantly higher baseline levels of urinary TNF-α were observed in chronic low back pain patients when compared to our reference value (p < 0.001), which were significantly lower after the period of chiropractic treatment (p = 0.03). Moreover, these changes were accompanied by a significant reduction in pain and disability (both p < 0.001). However, levels of urinary TNF-α were not correlated with pain intensity nor disability.ConclusionThese results suggest that urine could be a good milieu to assess TNF-α changes, with potential clinical implications for the management of chronic low back pain.

Altered Levels of Plasma Inflammatory Cytokines and White Matter Integrity in Bipolar Disorder Patients With Suicide Attempts.

ObjectiveBipolar disorder (BD) has a higher lifetime rate of suicide attempts (SA) than other psychiatric disorders. Furthermore, BD patients with SA (BD + S) are prone to a worse quality of life. However, the pathophysiology of BD + S is poorly understood. To further reveal the potential mechanisms of BD + S, abnormalities in peripheral plasma inflammatory cytokines and brain white matter (WM) in BD + S, as well as the correlation between them are investigated.MethodsWe tested the levels of TNF-α, IL-1β, and IL-6 in peripheral plasma and collected the diffusion tensor imaging (DTI) data from 14 BD + S, 24 BD patients without SA (BD-S), and 26 healthy controls (HCs). The three groups were matched by age and gender. The levels of TNF-α, IL-1β, and IL-6 were detected by Luminex multifactor detection technology, and the fractional anisotropy (FA) values were employed to depict the alterations of WM. Partial correlation analyses were conducted to detect correlations between levels of TNF-α, IL-1β, and IL-6 and changes of WM, and the relationships between severity of clinical symptoms, including scores of HAMD-17 and YMRS, and cytokine levels or FA values in all groups.ResultsFor plasma inflammatory cytokines, there was no significant difference in their levels except for IL-6 among the three groups. Post-hoc analyses revealed that increased IL-6 level was only detected in BD + S (p < 0.05, Bonferroni correction). For DTI, BD + S showed specifically decreased FA in the bilateral middle cerebellar peduncle and the left superior corona radiata compared to BD-S and HCs (p < 0.05, Bonferroni correction). Additionally, both BD + S and BD-S groups revealed decreased FA in the bilateral body and genu of corpus callosum (CC) compared to HCs (p < 0.05, Bonferroni correction). No significant correlation between plasma inflammatory cytokines and WM integrity was found. In the BD + S group, we found negative correlation between the scores of YMRS and FA values of the left middle cerebellar peduncle (r = −0.74, p = 0.035).ConclusionThe inflammation and impaired WM integrity may provide a scientific basis to understand the potential mechanisms of BD + S.

Effects of an intravenous ketamine infusion on inflammatory cytokine levels in male and female Sprague\u2013Dawley rats

BackgroundKetamine, a multimodal dissociative anesthetic drug, is widely used as an analgesic following traumatic injury. Although ketamine may produce anti-inflammatory effects when administered after injury, the immunomodulatory properties of intravenous (IV) ketamine in a non-inflammatory condition are unclear. In addition, most preclinical studies use an intraperitoneal (IP) injection of ketamine, which limits its clinical translation as patients usually receive an IV ketamine infusion after injury.MethodsHere, we administered sub-anesthetic doses of a single IV ketamine infusion (0, 10, or 40 mg/kg) to male and female Sprague–Dawley rats over a 2-h period. We collected blood samples at 2- and 4-h post-ketamine infusion to determine plasma inflammatory cytokine levels using multiplex immunoassays.ResultsThe 10 mg/kg ketamine infusion reduced spontaneous locomotor activity in male and female rats, while the 40 mg/kg infusion stimulated activity in female, but not male, rats. The IV ketamine infusion produced dose-dependent and sex-specific effects on plasma inflammatory cytokine levels. A ketamine infusion reduced KC/GRO and tumor necrosis factor alpha (TNF-α) levels in both male and female rats, interleukin-6 (IL-6) levels in female rats, and interleukin-10 (IL-10) levels in male rats. However, most cytokine levels returned to control levels at 4-h post-infusion, except for IL-6 levels in male rats and TNF-α levels in female rats, indicating a different trajectory of certain cytokine changes over time following ketamine administration.ConclusionsThe current findings suggest that sub-anesthetic doses of an IV ketamine infusion may produce sex-related differences in the effects on peripheral inflammatory markers in rodents, and further research is warranted to determine potential therapeutic effects of an IV ketamine infusion in an inflammatory condition.

Elevated midtrimester maternal plasma cytokines and preterm birth in patients with cerclage

Cerclage is used for the prevention of spontaneous preterm birth; however, many patients at high risk of spontaneous preterm birth who have a cerclage in place eventually deliver before term. Although inflammation, measured by biomarkers (eg, cytokines), is a known risk factor for preterm delivery, evaluation of inflammation to determine pregnancy outcomes among patients with cerclage is poorly understood. We sought to examine levels of maternal plasma inflammatory cytokines in the midtrimester among asymptomatic patients with a cervical cerclage (placed for any indication, including history, ultrasound, and examination indications) to evaluate the association between cytokine levels and preterm birth. This was a prospective cohort study of singleton, nonanomalous pregnancies who had a cerclage placed at <24 weeks of gestation from 2015 to 2018 at a single tertiary institution. Maternal plasma was collected perioperatively whenever possible. A custom magnetic bead Luminex cytokine assay was used to measure plasma inflammatory cytokine levels from these stored samples. The primary outcome was preterm birth at <37 weeks of gestation. A statistical cut point was calculated for each cytokine level to assess its optimal sensitivity and specificity for spontaneous preterm birth prediction. Patients were classified as having a "high" or "low" result for each cytokine based on this cut point. Receiver operating characteristic curve analysis was performed to estimate sensitivity, specificity, and positive and negative predictive values for spontaneous preterm birth prediction. Cox proportional-hazards regression modeled the association between the number of "high" inflammatory cytokines and gestational age at delivery, adjusting for confounders. Additional analyses were performed on the subgroup of patients with history-indicated cerclage and those with an ultrasound- or examination-indicated cerclage. A total of 43 patients participated in this study: 20 (46.5%) had spontaneous preterm birth (median, 30.9 weeks of gestation; interquartile range, 28.4-35.0). Plasma samples were collected at a median of 0 (interquartile range, -2 to 17) days concerning cerclage placement and a median of 18 (interquartile range, 13-21) weeks of gestation. Based on the statistical cut point for each cytokine level, 7% of patients had zero, 20.9% had 1, 18.6% had 2, 20.9% had 3, and 32.6% had ≥4 "high" cytokine results. Each additional "high" cytokine level was associated with earlier delivery (hazard ratio, 1.51; 95% confidence interval, 1.25-1.81) even after controlling for ultrasound- or examination-indication for cerclage (hazard ratio, 1.73; 95% confidence interval, 0.95-3.15). The presence of ≥4 "high" cytokine levels was 70% sensitive and 74% specific for predicting spontaneous preterm birth (area under the curve, 0.846; 95% confidence interval, 0.728-0.964; positive predictive value, 70%; negative predictive value, 73.9%). Among patients with a cervical cerclage, elevated midtrimester maternal plasma cytokine profiles were associated with subsequent preterm birth and can estimate the probability of preterm birth. Confirmation and refinement of this noninvasive panel may provide insight into improved selection of individuals who may benefit from cerclage placement and investigation of therapeutic strategies to mitigate midpregnancy inflammation.

Neutrophil membrane-mimicking nanodecoys with intrinsic anti-inflammatory properties alleviate sepsis-induced acute liver injury and lethality in a mouse endotoxemia model.

Sepsis-induced acute liver injury often develops in the early stages of sepsis and can exacerbate the pathology by contributing to multiple organ dysfunction and increasing lethality. No specific therapies for sepsis-induced liver injury are currently available; therefore, effective countermeasures are urgently needed. Considering the crucial role of neutrophils in sepsis-induced liver injury, herein, neutrophil membrane-mimicking nanodecoys (NM) were explored as a biomimetic nanomedicine for the treatment of sepsis-associated liver injury. NM administration exhibited excellent biocompatibility and dramatically decreased the plasma levels of inflammatory cytokines and liver injury biomarkers, including aspartate aminotransferase, alanine aminotransferase, and direct bilirubin, in a sepsis mouse model. NM treatment also reduced hepatic malondialdehyde content, myeloperoxidase activity, and histological injury, and ultimately improved survival in the septic mice. Further in vitro studies showed that NM treatment neutralized the neutrophil chemokines and inflammatory mediators and directly mitigated neutrophil chemotaxis and adhesion. Additionally, NM also markedly weakened lipopolysaccharide-induced reactive oxygen species generation, cyclooxygenase-2 expression, nitric oxide secretion, and subsequent hepatocyte injury. Thus, this study provides a promising therapeutic strategy for the management of sepsis-induced acute liver injury.

Abstract 82: The Second X Chromosome Contributes to Stroke Sensitivity in the Absence of Activational Hormonal Effects

Introduction: Stroke is a sexually dimorphic disease. Stroke sensitivity in the elderly is altered by the decline of estrogen levels in females. Our previous studies suggested a X chromosomal effect on stroke sensitivity in aged mice; however, these mice were gonadal intact and the activational effect of hormones cannot be ruled out. In this study, we utilized the XY* mice model in which mice of four genotypes were produced (XX, XO, XY, and XXY) to dissociate the effect of X chromosome from that of Y. To rule out the activational effect of hormones, we gonadectomized all mice at 3 weeks old, and let them survive for 6-8 weeks to achieve “surgical post-menopause”. We hypothesized that the second X chromosome mediates the stroke sensitivity in the absence of activational sex hormones. Two X escapee genes ( Kdm6A and Kdm5C ) that escape from the X chromosome inactivation and involved in microglial activation, were also examined to explore the potential mechanisms. Methods: “Surgical postmenopausal” XY* mice were subjected to a 60-min middle cerebral artery occlusion (MCAO) (n=6-8/gp for stroke and = 4-5/gp for sham). Infarct volumes and behavior deficits were quantified 3 days after MCAO. Plasma inflammatory cytokine (TNFα, IL-1β, IL-4, iNOS, TGF-β1 and IL-10) levels were determined by ELISA. KDM6A and KDM5C expression in microglia was examined by IHC. Results: XX mice had significantly larger infarct size than XO mice in striatum (55.02±12.38% vs. 36.06±9.25%, P=0.0387) and the ipsilateral hemisphere (45.33±10.77% vs. 26.55±6.92%, p=0.0413); the same pattern was seen in XXY vs. XY group in striatum (58.47±14.26% vs. 34.22±15.24%, P=0.0027). Neurological deficit scores were higher in XX vs. XO mice. Mice with two copies of the X chromosome had significantly higher plasma levels of pro-inflammatory cytokines (TNFα, IL-1β) than the mice with one X. Significantly higher levels of KDM6A or KDM5C in Iba-1 positive microglial cells were also found in mice with two vs. one copy of X. Conclusion: The second X chromosome contributes to stroke sensitivity in the absence of activational sex hormones. Our future goal is to use Kdm6A and Kdm5C microglial conditional knockout mice to explore the molecular mechanisms underlying chromosomal factors in stroke sensitivity.