Proinflammatory Changes Research Articles

Quantitative proteomics of acutely-isolated mouse microglia identifies novel immune Alzheimer\u2019s disease-related proteins

BackgroundMicroglia are innate immune cells of the brain that perform phagocytic and inflammatory functions in disease conditions. Transcriptomic studies of acutely-isolated microglia have provided novel insights into their molecular and functional diversity in homeostatic and neurodegenerative disease states. State-of-the-art mass spectrometry methods can comprehensively characterize proteomic alterations in microglia in neurodegenerative disorders, potentially providing novel functionally relevant molecular insights that are not provided by transcriptomics. However, comprehensive proteomic profiling of adult primary microglia in neurodegenerative disease conditions has not been performed.MethodsWe performed quantitative mass spectrometry based proteomic analyses of purified CD11b+ acutely-isolated microglia from adult (6 mo) mice in normal, acute neuroinflammatory (LPS-treatment) and chronic neurodegenerative states (5xFAD model of Alzheimer’s disease [AD]). Differential expression analyses were performed to characterize specific microglial proteomic changes in 5xFAD mice and identify overlap with LPS-induced pro-inflammatory changes. Our results were also contrasted with existing proteomic data from wild-type mouse microglia and from existing microglial transcriptomic data from wild-type and 5xFAD mice. Neuropathological validation studies of select proteins were performed in human AD and 5xFAD brains.ResultsOf 4133 proteins identified, 187 microglial proteins were differentially expressed in the 5xFAD mouse model of AD pathology, including proteins with previously known (Apoe, Clu and Htra1) as well as previously unreported relevance to AD biology (Cotl1 and Hexb). Proteins upregulated in 5xFAD microglia shared significant overlap with pro-inflammatory changes observed in LPS-treated mice. Several proteins increased in human AD brain were also upregulated by 5xFAD microglia (Aβ peptide, Apoe, Htra1, Cotl1 and Clu). Cotl1 was identified as a novel microglia-specific marker with increased expression and strong association with AD neuropathology. Apoe protein was also detected within plaque-associated microglia in which Apoe and Aβ were highly co-localized, suggesting a role for Apoe in phagocytic clearance of Aβ.ConclusionsWe report a comprehensive proteomic study of adult mouse microglia derived from acute neuroinflammation and AD models, representing a valuable resource to the neuroscience research community. We highlight shared and unique microglial proteomic changes in acute neuroinflammation aging and AD mouse models and identify novel roles for microglial proteins in human neurodegeneration.

Grape polyphenols corrects ageing-related detriments in neutrophil functionality via modulation of specific molecular targets.

Oxidative stress and inflammation are intricately interlinked as aetiological factors in the context of ageing and chronic disease-related accelerated ageing. Previous research by our group has highlighted the anti-oxidant and anti-inflammatory potential of grape-derived polyphenols in the context of acute inflammation and oxidative stress. The aim here was to add to this by assessing efficacy of the treatment (acutely) to address ageing-associated cumulative pro-oxidant and pro-inflammatory changes in an in vitro model. Blood from young and aged humans was analysed for baseline oxidative stress and inflammatory status. Isolated neutrophils were acutely exposed to the polyphenol treatment in vitro. The chemokinetic capacity of treated and control neutrophils in response to fMLP was subsequently determined in a Dunn chamber, using live cell imaging. Neutrophils were also analysed for the expression of selected molecular markers associated with functional capacity and oxidative stress. Results indicate that the aged population had significantly worse oxidative stress and inflammatory profiles (higher plasma conjugated dienes and MPO) than young controls. Neutrophils isolated from both young and aged individuals had improved chemokinetic accuracy and capacity after in vitro polyphenol treatment. Additionally, increased shedding of CD16 and expression of CD66b suggested sites via which the polyphenol achieved improved neutrophil motility. We conclude that grape seed-derived polyphenols facilitated improved neutrophil functionality by acting on the molecular targets elucidated here.

Suppression of Neuropathy Development in Diabetic Rage-Deficient Mice Is Associated with Absence of M1/M2 Macrophage Skewing in the Sciatic Nerve

Infiltrating macrophage (Mφ) into sciatic nerve (SN) is assumed one of the pathogenesis of diabetic polyneuropathy (DPN). In particular, skewing infiltrating Mφ toward pro-inflammatory (M1) Mφ, but not anti-inflammatory (M2) Mφ, possibly contributes to dysfunction of SN in diabetic state. However, the impacts of Mφ into dorsal root ganglia (DRG) in DPN are not yet well explored. For this purpose, male C57BL/6 mice (W) and receptor for AGEs (RAGE) null mice (R) were recruited and rendered diabetic (D) with STZ injection. After 8 weeks, nerve conduction velocity (NCV) and tail flick test were performed. Subsequently, mice were sacrificed to dissect SN and DRG, which were evaluated in the pathology and mRNA expression by qPCR. WD showed significant delay of NCV and increase in threshold for thermal sensation (p<0.01), while those were maintained in RD. Pathological analysis elucidated decrease in the average size of cell bodies in the DRG neuron of WD compared to W (p<0.01), as well as the expression of CGRP in DRG neurons (p<0.01). RD maintained neuronal cell size and CGRP expression. Significant increase in infiltration of Iba-I positive Mφ emerged in SN of WD compared to W (p<0.05). The increase was more robust, composed mostly of M2 in RD than WD (p<0.01). No increase of Mφ infiltration was identified in DRG of diabetic groups. The expression of iNOS (a marker for M1) was increased in SN of WD compared to W (p < 0.01), but not in that of RD. The expression of Arg1 (a marker for M2) was significantly increased in SN of RD compared to WD (p < 0.01). However, no changes of iNOS and Arg1 expression were detected in DRG. Our results suggest that proinflammatory changes in distal nerve elicits systematic sensory neuropathy in diabetes and correction of M1φ/M2φ imbalance may be beneficial for its prevention. Disclosure S. Osonoi: None. H. Mizukami: Research Support; Self; Sanwa Kagaku Kenkyusho Co., Ltd., Mitsubishi Tanabe Pharma Corporation. S. Ogasawara: None. K. Takahashi: None. K. Sango: None. S. Yagihashi: None.

Increase in Galectin-3 Contributes to Glucose Intolerance and Insulin Resistance following Exposure to the Common Food Additive Carrageenan

Exposure to low concentration of the common food additive carrageenan (CGN) leads to impaired glucose tolerance and insulin resistance in C57BL/6J mice, due to CGN’s pro-inflammatory effects and changes in phospho-IRS1 phosphorylation. CGN is a sulfated polysaccharide with α-1,3 and β-1,4 galactosidic bonds. Other studies showed: 1) CGN reduced activity of arylsulfatase B, which removes sulfate groups from N-acetylgalactosamine 4-sulfate residues of chondroitin 4-sulfate (C4S) and dermatan sulfate; and 2) galectin-3 binds less with C4S when ARSB is reduced. Increase in galectin-3, which binds to β-galactosides, was reported to inhibit insulin signaling (Li P et al) by effects on the insulin receptor. This background led to the current assessment of effects of the no-CGN diet on galectin-3 levels in serum and galectin-3 mRNA in circulating mononuclear cells. Blood samples were obtained from adults with prediabetes before and after dietary intervention. Baseline serum galectin-3 values averaged 8.94 ng/ml (range 4.06-12.64). In 7 patients on a no-CGN diet for 12 weeks, the mean level declined to 7.25 ng/ml (p=0.006, paired t-test). In contrast, serum galectin-3 was unchanged in participants on the CGN-containing control diet and increased by over 2 ng/ml in subjects on their regular diet. mRNA expression of galectin-3 (NGALS3) in circulating mononuclear cells of patients on the no-CGN diet declined by 69%, and increased by 36% in the CGN-containing diet group and by 98% in the regular diet group (p=0.0002, 1-way ANOVA). Also, serum galectin-3 levels increased to 1.72 times baseline in C57BL/6J mice given CGN in their water supply for 12 weeks. In HepG2 cells, exogenous galectin-3 (25 µg/ml) inhibited insulin-induced glucose uptake by 39%, and was highly correlated with decline in tyrosine phosphorylation of IRS-1. These findings suggest that CGN contributes to glucose intolerance and insulin resistance, in part, by effects on galectin-3. Disclosure J.K. Tobacman: None. S. Bhattacharyya: None. L. Feferman: None.

Influence of immunobiological therapy on microwashing the psoriatic process

Objective — make a comparative analysis of immunological and immunohistochemical changes in the skin psoriasis rash in patients with psoriasis after the course of systemic therapy, as well as evaluate the therapeutic effectiveness of the therapy. Materials and methods. Immunohistochemical research of biopsy material, taken from areas of skin psoriasis of patients on psoriasis, was performed. In addition, for comparison of results of immunogistochemikal research, a study of biopsy material of skin of front abdominal wall was undertaken for the persons of corresponding age without psoriasis (5 patients), was executed while the operation of hernioplastick. In order to set the type and prevalence of local cellular immune and inflammatory reactions, immunohistochemical methodologies were used for the exposure of immune inflamation marker. The indices of cellular immunity in the body were assessed by the change in the relative and absolute number of T­(CD3 + ) and B­(CD19 + ) lymphocytes, as well as subpopulations (CD4 + , CD8 + , CD16 + , CD25 + CD71 + , CD30 + , CD95 + ). Results and discussion. The most positive effect on the indices of the cellular part of the body’s immune system was found in patients with psoriasis who received immunotherapy with etanercept in combination with narrow­band (311 nm) phototherapy. After the treatment of patients in this group, due to a significant decrease in production of TNF and its serum concentration in the blood serum, proinflammatory changes in the cellular system of the immune system decreased, the number of lymphocytes with an early activation marker decreased, which initiated a further cytokine cascade of development and prolongation of the inflamation. Conclusions. It was found that, according to the analysis of the results of clinical studies, as well as immunological and morphological studies conducted after the completion of treatment courses for psoriasis patients, the most effective method is a systemic immunobiological therapy in combination with a narrow wave (311 nm) UV B therapy, regarding terms of remission, normalization of the body’s immune status, regression of inflammatory reactions and hyper­plastic processes in the skin, and also elimination of colonies of microorganisms The genus Staphyloccocus under the stratum corneum epidermis.

Oxidized phospholipids are proinflammatory and proatherogenic in hypercholesterolaemic mice.

Oxidized phospholipids (OxPL) are ubiquitous, are formed in many inflammatory tissues, including atherosclerotic lesions, and frequently mediate proinflammatory changes 1 . Because OxPL are mostly the products of non-enzymatic lipid peroxidation, mechanisms to specifically neutralize them are unavailable and their roles in vivo are largely unknown. We previously cloned the IgM natural antibody E06, which binds to the phosphocholine headgroup of OxPL, and blocks the uptake of oxidized low-density lipoprotein (OxLDL) by macrophages and inhibits the proinflammatory properties of OxPL2-4. Here, to determine the role of OxPL in vivo in the context of atherogenesis, we generated transgenic mice in the Ldlr-/- background that expressed a single-chain variable fragment of E06 (E06-scFv) using the Apoe promoter. E06-scFv was secreted into the plasma from the liver and macrophages, and achieved sufficient plasma levels to inhibit in vivo macrophage uptake of OxLDL and to prevent OxPL-induced inflammatory signalling. Compared to Ldlr-/- mice, Ldlr -/- E06-scFv mice had 57-28% less atherosclerosis after 4, 7 and even 12 months of 1% high-cholesterol diet. Echocardiographic and histologic evaluation of the aortic valves demonstrated that E06-scFv ameliorated the development of aortic valve gradients and decreased aortic valve calcification. Both cholesterol accumulation and in vivo uptake of OxLDL were decreased in peritoneal macrophages, and both peritoneal and aortic macrophages had a decreased inflammatory phenotype. Serum amyloid A was decreased by 32%, indicating decreased systemic inflammation, and hepatic steatosis and inflammation were also decreased. Finally, the E06-scFv prolonged life as measured over 15 months. Because the E06-scFv lacks the functional effects of an intact antibody other than the ability to bind OxPL and inhibit OxLDL uptake in macrophages, these data support a major proatherogenic role of OxLDL and demonstrate that OxPL are proinflammatory and proatherogenic, which E06 counteracts in vivo. These studies suggest that therapies inactivating OxPL may be beneficial for reducing generalized inflammation, including the progression of atherosclerosis, aortic stenosis and hepatic steatosis.

Proinflammatory cytokines and endotoxemia level change in patients with type 2 diabetes and nonalcoholic fatty liver disease after hepatoprotective and probiotic therapy usage

Proinflammatory cytokines and endotoxemia level change in patients with type 2 diabetes and nonalcoholic fatty liver disease after hepatoprotective and probiotic therapy usage

Lower plasma trans-4-hydroxyproline and methionine sulfoxide levels are associated with insulin dysregulation in horses

BackgroundInsulin dysregulation in horses is a metabolic condition defined by high insulin concentrations in the blood and peripheral insulin resistance. This hyperinsulinemia is often associated with severe damage in the hooves, resulting in laminitis. However, we currently lack detailed information regarding the potential involvement of particular metabolic pathways in pathophysiological causes and consequences of equine insulin dysregulation. This study aimed to assess the dynamic metabolic responses given to an oral glucose test (OGT) in insulin-sensitive and insulin-dysregulated horses by a targeted metabolomics approach to identify novel metabolites associated with insulin dysregulation.ResultsOral glucose testing triggered alterations in serum insulin (26.28 ± 4.20 vs. 422.84 ± 88.86 μIU/mL, p < 0.001) and plasma glucose concentrations (5.00 ± 0.08 vs. 9.43 ± 0.44 mmol/L, p < 0.001) comparing basal and stimulated conditions after 180 min. Metabolome analyses indicated OGT-induced changes in short-chain acylcarnitines (6.00 ± 0.53 vs. 3.99 ± 0.23 μmol/L, p < 0.001), long-chain acylcarnitines (0.13 ± 0.004 vs. 0.11 ± 0.002 μmol/L, p < 0.001) and amino acids (2.18 ± 0.11 vs. 1.87 ± 0.08 μmol/L, p < 0.05). Kynurenine concentrations increased (2.88 ± 0.18 vs. 3.50 ± 0.19 μmol/L, p < 0.01), whereas spermidine concentrations decreased during OGT (0.09 ± 0.004 vs. 0.08 ± 0.002 μmol/L, p < 0.01), indicating proinflammatory conditions after oral glucose load. Insulin dysregulation was associated with lower concentrations of trans-4-hydroxyproline (4.41 ± 0.29 vs. 6.37 ± 0.71 μmol/L, p < 0.05) and methionine sulfoxide (0.40 ± 0.06 vs. 0.87 ± 0.13 μmol/L, p < 0.01; mean ± SEM in insulin-dysregulated vs. insulin-sensitive basal samples, respectively), two metabolites which are related to antioxidant defense mechanisms.ConclusionOral glucose application during OGT resulted in profound metabolic and proinflammatory changes in horses. Furthermore, insulin dysregulation was predicted in basal samples (without OGT) by pathways associated with trans-4-hydroxyproline and methionine sulfoxide, suggesting that oxidative stress and oxidant–antioxidant disequilibrium are contributing factors to insulin dysregulation. The present findings provide new hypotheses for future research to better understand the underlying pathophysiology of insulin dysregulation in horses.

Chebulagic acid Chebulinic acid and Gallic acid, the active principles of Triphala, inhibit TNFα induced pro-angiogenic and pro-inflammatory activities in retinal capillary endothelial cells by inhibiting p38, ERK and NFkB phosphorylation

Tumor necrosis factor-α (TNFα) a pleiotropic cytokine induces pro-inflammatory and pro-angiogenic changes in conditions such as diabetic retinopathy (DR) and neovascular age related macular degeneration (NV-AMD). Hence, inhibition of TNFα mediated changes can benefit the management of DR and NV-AMD. Triphala, an ayurvedic herbal preparation is known to have immunomodulatry functions. In this study we evaluated the alcoholic extract of triphala (AlE) and its compounds Chebulagic acid (CA), Chebulinic acid (CI) and Gallic acid (GA) for their anti-TNFα activity. TNFα induced pro-inflammatory and pro-angiogenic changes in the retinal-choroid microvascular endothelial cells (RF/6A). Treatment with CA/CI/GA and the whole Triphala extract showed characteristic inhibition of MMP-9, cell proliferation/migration and tube formation as well the expression of IL-6, IL-8 and MCP-1 without affecting cell viability. This was mediated by inhibition of p38, ERK and NFκB phosphorylation. Ex vivo angiogenesis assay using chick chorioallantoic membrane (CAM) model also showed that TNFα-induced angiogenesis and it was inhibited by AlE and its active principles. Further, in silico studies revealed that CA, CI and GA are capable of binding the TNFα-receptor-1 to mediate anti-TNFα activity. This study explains the immunomodulatory function of Triphala, evaluated in the context of retinal and choroid vasculopathies in vitro and ex vivo; which showed that CA, CI and GA can be a potential pharmacological agents in the management of DR and NV-AMD.

Alkalinization with potassium bicarbonate improves glutathione status and protein kinetics in young volunteers during 21-day bed rest

Physical inactivity is associated with lean body mass wasting, oxidative stress and pro-inflammatory changes of cell membrane lipids. Alkalinization may potentially counteract these alterations. We evaluated the effects of potassium bicarbonate supplementation on protein kinetics, glutathione status and pro- and anti-inflammatory polyunsaturated fatty acids (PUFA) in erythrocyte membranes in humans, during experimental bed rest. Healthy, young, male volunteers were investigated at the end of two 21-day bed rest periods, one with, and the other without, daily potassium bicarbonate supplementation (90mmol×d-1), according to a cross-over design. Oxidative stress in erythrocytes was evaluated by determining the ratio between reduced (GSH) and oxidized glutathione (GSSG). Glutathione turnover and phenylalanine kinetics, a marker of whole body protein metabolism, were determined by stable isotope infusions. Erythrocyte membranes PUFA composition was analyzed by gas-chromatography. At the end of the two study periods, urinary pH was 10±3% greater in subjects receiving potassium bicarbonate supplementation (7.23±0.15 vs. 6.68±0.11, p<0.001). Alkalinization increased total glutathione concentrations by 5±2% (p<0.05) and decreased its rate of clearance by 38±13% (p<0.05), without significantly changing GSH-to-GSSG ratio. After alkalinization, net protein balance in the postabsorptive state improved significantly by 17±5% (p<0.05) as well as the sum of n-3 PUFA and the n-3-to-n-6 PUFA ratio in erythrocyte membranes (p<0.05). Alkalinization during long-term inactivity is associated with improved glutathione status, anti-inflammatory lipid pattern in cell membranes and reduction in protein catabolism at whole body level. This study suggests that, in clinical conditions characterized by inactivity, oxidative stress and inflammation, alkalinization could be a useful adjuvant therapeutic strategy.

Early synovial B-cell infiltration as a candidate mechanism of pathogenesis in the OA of obesity

Purpose: Obesity is a major risk factor for the development of osteoarthritis (OA), the greatest cause of disability in the US. With the incidence of both obesity and OA on the rise, the comorbid disease burden is only worsening, yet there are no disease modifying therapies approved for the use in OA to date. Recent studies, from our lab and others, have discovered that synovial inflammation and immune cell infiltration are associated the accelerated progression of OA in obese mice and humans. These findings led us to hypothesize that obesity-induced pathobiological change in the synovium is the seminal driver of accelerated OA degeneration of joints. To test this hypothesis, we performed a transcriptomic analysis of synovial tissue from lean and obese mice, at a time point well before any histologic joint degeneration is evident. Methods: Male C57BL/6J mice were fed a low fat or high fat diet (HF) for 16 weeks, at which point the synovial membrane was removed for RNA isolation. RNA samples were then sequenced using Illumina’s HiSeq2500v4, and the next generation sequencing (NGS) data were processed and aligned. Following alignment, the significantly differentially expressed genes (DEG) were evaluated for cell type enrichment using CTen, as well as enriched canonical pathways and diseases and functions using Ingenuity Pathway Analysis (IPA). To analyze the various cell populations in the synovial membrane, we performed flow cytometry on cells isolated from lean and obese synovium. Results: Before any histologic evidence of cartilage degeneration, significant pro-inflammatory changes were observed in the obese synovium, suggesting these changes may be crucial to disease pathogenesis. Cell type enrichment analysis identified the macrophage as a significant mediator of early synovial pathogenic change (Fig. 1a). Correlating with this, pathway analysis of RNAseq data revealed significant induction of numerous inflammatory pathways including activation of NF-κB, stimulation of macrophages, and inflammation of the joint (Fig. 1b). Additionally, we discovered a potentially novel mediator of the early pathogenesis of the OA of obesity; the B-cell. Cell type enrichment analysis identified a significant B-cell signature in the obese synovial membranes (Fig. 1a). IPA analysis provides further evidence for the role of the B-cell, as pathways related to the B-cell including B-cell receptor signaling, quantity of B lymphocytes (Fig. 1c, d), and PI3K signaling in B lymphocytes were significantly activated. To confirm the presence of the B-cell in the synovial membrane, we performed flow cytometry on cells isolated from lean and obese synovium. This analysis identified an increased population of B-cells residing in the synovial membrane of obese mice, providing further evidence for the transcriptome analysis (Fig. 1 e, f). Conclusions: Significantly, this study is the first direct evidence that synovial disease occurs long before discernable joint degeneration, supporting the concept that the OA of obesity is preceded by a disease state in the synovial membrane. Transcriptome analysis identified a significant inflammatory change in the synovium of obese mice, with the macrophage being recognized as a prominent mediator of disease. Additionally, both cell type enrichment analysis and pathway analysis identified the B-cell as a potentially novel mediator of the OA of obesity. These findings warrant deeper study of the B-cell in the OA of obesity, where we hypothesize it plays a role as an antigen presenting cell for activation of other immune cells which in aggregate are responsible for supporting a proinflammatory environment leading to joint degeneration.

Meal‐Induced Pro‐Inflammatory Responses in Guatemalan Adults Are Associated with Body Mass Index And Are More Pronounced in Women

BackgroundPopulations in low‐ and middle‐income countries (LMICs) often experience a combination of undernutrition in childhood and an obesogenic environment in adulthood. Research to address their emerging cardiometabolic disease is sparse, especially their response to stressors such as a meal challenge.ObjectiveOur objective was to describe pro‐inflammatory changes in response to a standardized meal challenge.MethodsParticipants are from a longitudinal cohort study established in 1969 in Guatemala. In 2015–17 (n=1131), mean age 45 y (range 37–54 y), pre‐ and two‐hour post‐meal challenge (600kcal standardized mixed meal, 30% fat, 50% carbohydrates, and 20% protein) plasma samples were obtained. Measurements included lipid profile, glucose (GLU), insulin (INS), alanine aminotransferase (ALT), aspartate aminotransferase (AST), and pro‐ and anti‐inflammation cytokines, including interleukin‐6 and 10 (IL‐6, IL‐10), adiponectin, leptin, soluble TNF receptor II (TNFsR), and resistin. Data from an initial random subset (n=119, 58% women) is presented. The impact of the meal challenge was assessed as percent changes (denoted as %D), which were related to body mass index (BMI) values. Analyses were stratified by sex.ResultsMean (SD) BMI was 29.4 (5.3) for women and 27.0 (4.1) for men. Women had higher fasting HDL‐cholesterol, apoA‐I, insulin, hsCRP, adiponectin, leptin, and resistin levels than men (all p&lt;0.05). While most biomarkers were within the ‘normal’ fasting range as observed in U.S. and other published reference populations, mean triglyceride (TG), glucose and non‐esterified fatty acids (NEFA) were higher for both genders. Mean hsCRP was high among women and not in men; and mean ALT was high among men.Meal‐induced changes were associated with BMI, and differed by sex. %D‐GLU (mean change 20.4% vs. 5.9%, p&lt;0.001) and %D‐INS (297% vs. 210%, p&lt;0.015) was higher in women than men. %D‐GLU increased with BMI for both genders, while %D‐INS increased with BMI only in women. %D‐leptin (−17.8% women; −35.4% men) and %D‐TNFsR (‐19.3% women; −11.3% men) were negative. The association between %D‐leptin with BMI was positive in women and negative in men (p&lt;0.001 for heterogeneity by sex). %D‐TNFsR was inversely associated with BMI in women and positively associated in men (p&lt; 0.03 by sex).DiscussionIn this sample, meal‐induced pro‐inflammatory responses were associated with BMI. The changes in key biomarkers, especially in TNFsR, suggest that women tend to have greater meal‐induced pro‐inflammatory responses than men. The gender‐dependent difference in postprandial leptin suggests a more complex role of leptin than a simple pro‐inflammatory marker. These results warrant further investigation.ConclusionMeal‐induced pro‐inflammatory responses were associated with BMI, and differed between men and women in this study.Support or Funding InformationThis work is supported by the National Institute of Health, USA (Grant Number: 5R01HD075784).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

F166. LOW-GRADE INFLAMMATION IN FIRST-EPISODE PSYCHOSIS IS DETERMINED BY WAIST CIRCUMFERENCE INCREASE

BackgroundThere is evidence of low-grade inflammation in psychosis, as measured by the high-sensitivity C-reactive protein (hs-CRP). Significant weight gain is common during the first months of antipsychotic treatment. In the general population, overweight and obesity often lead to systemic low-grade inflammation. Lifestyle factors, such as smoking, can contribute to the pro-inflammatory changes. The metabolic changes in people with first-episode psychosis (FEP) taking place after the onset of psychosis can be especially harmful as these individuals are typically young and without major somatic illnesses. We aimed to study how the low-grade inflammation, measured by hs-CRP, develops in FEP and to clarify the effect of waist circumference increase in the inflammation.MethodsThe Helsinki Early Psychosis Study recruited FEP patients (age 18 to 40 years) attaining their first treatment for psychosis from the catchment area of the Helsinki University Hospital. We recruited 95 FEP patients and 62 controls. The inclusion criterion for the study was receiving a score of at least 4 in Unusual thought content or Hallucinations in the Brief Psychiatric Rating Scale - Extended (BPRS-E). Diagnoses of psychotic disorders according to the DSM-IV criteria were later verified using the Structured Diagnostic Interview for DSM-IV and reviewing all medical records. Substance-induced psychotic disorders and psychotic disorders due to a general medical condition were excluded. We measured the changes in hs-CRP, weight, waist circumference, glucose metabolism and lipids at baseline and at follow-ups of 2 and 12 months. We used linear mixed effects models to analyze the relationship between hs-CRP and waist circumference. In the model, we included a random intercept for each patient and, as fixed effects, we entered sex, time (days from baseline measurement), waist circumference and antipsychotic use at each assessment point, and baseline cigarette smoking.ResultsAt baseline, FEP patients (mean age 26.1 years) had higher insulin resistance, total and LDL cholesterol, apolipoprotein B and triglyceride levels than controls. However, baseline weight and waist circumference, hs-CRP, fasting glucose and HDL cholesterol were similar between patients and controls. A robust change in anthropometric measures and inflammation was evident among patients by 12 months. Hs-CRP was significantly higher in patients at 12-month follow up than at baseline (baseline hs-CRP 0.67 mg/l, IQR 0.33–2.54; 12-month 1.73 mg/l, IQR 0.49–4.21; Wilcoxon signed-rank p = 0.007). When at the baseline the prevalence of overweight or obesity was 30% (28/94) in patients with FEP, by 12 months the prevalence was 59% (35/59) (McNemar′s test p < 0.001). The proportion of patients gaining ≥ 7 % of baseline weight was 68 % (40/59). The median weight gain among patients was 9.6 kg (IQR 1.5–13.6 kg), and the waist circumference increase 6.0 cm (IQR 2.0–13.0 cm). In the mixed effects model waist circumference (p < 0.0001) and sex (p = 0.014) were significantly associated with hs-CRP level.DiscussionWe detected a significant elevation in hs-CRP in people with FEP during the first treatment year. The rise in hs-CRP was determined by waist circumference increase. Patients with FEP are in a marked risk of developing abdominal obesity and subsequent low-grade inflammation during the first year of treatment. Prevention of the early metabolic changes in first-episode psychosis is important, as abdominal obesity and inflammation are associated with increased risk of cardiovascular events and mortality.

Cyclosporine Preconditioning During Ex Vivo Lung Perfusion Improves the Quality of Post Transplant Lung Grafts

Lung grafts on ex vivo lung perfusion (EVLP) exhibit prominent proinflammatory changes and compromised metabolic profiles, leading to deleterious effects on the quality of lung grafts. In this study, we investigated the preconditioning effects of cyclosporine on mitochondrial protection in lung grafts during EVLP as well as following subsequent transplantation.

Canonical NF-κB signaling in myeloid cells promotes lung metastasis in a mouse breast cancer model.

An inflammatory tumor microenvironment is a common characteristic of solid tumors. It is the result of a complex interplay between tumor cells, tumor infiltrating immune cells and other stromal cells. Myeloid cells in the tumor microenvironment are considered major drivers of tumor progression and metastasis and increased numbers of these cells are associated with poor prognosis in various cancer patients. The transcription factor NF-κB is considered the master regulator of inflammatory gene expression and immune cell function. Its activation in various cells of the tumor microenvironment contributes essentially to tumorigenesis. In the present study, the role of canonical NF-κB signaling in myeloid cells in metastatic breast cancer was addressed by myeloid-specific deletion of Ikkβ in the MMTV polyoma middle T (PyMT) mouse model. Ikkβ deletion in myeloid cells did not affect primary mammary tumor growth but significantly reduced lung metastasis. While dissemination from the primary tumor was unaltered, myeloid-specific Ikkβ loss resulted in a strong up-regulation of pro-inflammatory genes and changes in immune cell populations in the lung, creating a tumor-suppressive microenvironment at the distant site. Thus, canonical NF-κB signaling in myeloid cells creates a permissive lung microenvironment that supports breast to lung metastasis.

Differences in Proinflammatory Cytokines and Monocyte Subtypes in Older as Compared With Younger Kidney Transplant Recipients.

The number of elderly patients with end-stage kidney disease requiring kidney transplantation continues to grow. Evaluation of healthy older adults has revealed proinflammatory changes in the immune system, which are posited to contribute to age-associated illnesses via "inflamm-aging." Immunologic dysfunction is also associated with impaired control of infections. Whether these immunologic changes are found in older kidney transplant recipients is not currently known, but may have important implications for risk for adverse clinical outcomes. Three months after transplant, innate immune phenotype was evaluated by flow cytometry from 60 kidney transplant recipients (22 older [≥60 years] and 38 younger [<60 years old]). Multiplex cytokine testing was used to evaluate plasma cytokine levels. Younger patients were matched to older patients based on transplant type and induction immune suppression. Older kidney transplant recipients demonstrated decreased frequency of intermediate monocytes (CD14++CD16+) compared with younger patients (1.2% vs 3.3%, P = 0.007), and a trend toward increased frequency of proinflammatory classical monocytes (CD14++CD16-) (94.5% vs 92.1%) (P = 0.065). Increased levels of interferon-gamma (IFN-γ) were seen in older patients. In this pilot study of kidney transplant recipients, we identified differences in the innate immune system in older as compared with younger patients, including increased levels of IFN-γ. This suggests that age-associated nonspecific inflammation persists despite immune suppression. The ability to apply noninvasive testing to transplant recipients will provide tools for patient risk stratification and individualization of immune suppression regimens to improve outcomes after transplantation.

ISSLS PRIZE IN BASIC SCIENCE 2018: Growth differentiation factor-6 attenuated pro-inflammatory molecular changes in the rabbit anular-puncture model and degenerated disc-induced pain generation in the rat xenograft radiculopathy model.

To elucidate the effects of growth differentiation factor-6 (GDF6) on: (i) gene expression of inflammatory/pain-related molecules and structural integrity in the rabbit intervertebral disc (IVD) degeneration model, and (ii) sensory dysfunction and changes in pain-marker expression in dorsal nerve ganglia (DRGs) in the rat xenograft radiculopathy model. Forty-six adolescent rabbits received anular-puncture in two non-consecutive lumbar IVDs. Four weeks later, phosphate-buffered saline (PBS) or GDF6 (1, 10 or 100µg) was injected into the nucleus pulposus (NP) of punctured discs and followed for 4weeks for gene expression analysis and 12weeks for structural analyses. For pain assessment, eight rabbits were sacrificed at 4weeks post-injection and NP tissues of injected discs were transplanted onto L5 DRGs of 16 nude rats to examine mechanical allodynia. The rat DRGs were analyzed immunohistochemically. In GDF6-treated rabbit NPs, gene expressions of interleukin-6, tumor necrosis factor-α, vascular endothelial growth factor, prostaglandin-endoperoxide synthase 2, and nerve growth factor were significantly lower than those in the PBS group. GDF6 injections resulted in partial restoration of disc height and improvement of MRI disc degeneration grades with statistical significance in rabbit structural analyses. Allodynia induced by xenograft transplantation of rabbit degenerated NPs onto rat DRGs was significantly reduced by GDF6 injection. Staining intensities for ionized calcium-binding adaptor molecule-1 and calcitonin gene-related peptide in rat DRGs of the GDF6 group were significantly lower than those of the PBS group. GDF6 injection may change the pathological status of degenerative discs and attenuate degenerated IVD-induced pain.

Age-related pro-inflammatory and pro-angiogenic changes in human aqueous humor.

To reveal age-related aqueous cytokine changes in human aqueous humor. Aqueous humor was collected from 12 young children (3-6.5 years old) and 71 healthy adults (22-106 years old) with cataract but without other systemic or ocular disorders. Levels of 22 cytokines, chemokines and vascular endothelial growth factor (VEGF) were measured and analyzed. The following proteins showed significant increase from childhood to adult: interferon-gamma (IFN-γ), interleukin (IL)-13, IL-6, IL-12(p70), IL-10, CCL2, CCL3, CCL4, CXCL8, CXCL9, CXCL10, IFN-α2 and VEGF (all P<0.05). IFN-γ, IL-13, IL-12(p70), IL-10, CCL3, CXCL9 and VEGF also showed moderate strength age-related increase in the adult group (r>0.5). The strength of correlation between aging and CCL4 were fair (r=0.398). The concentrations of IL-2, IL-4, IL-5, IL-1β and TNF-α were low in both groups. From childhood to adult, the immunological milieu of the anterior chamber become more pro-inflammatory and pro-angiogenic. Such changes may represent the parainflammation state of the human eye.

Src kinase inhibition reduces inflammatory and cytoskeletal changes in ΔF508 human cholangiocytes and improves cystic fibrosis transmembrane conductance regulator correctors efficacy.

Our findings have strong translational potential and indicate that targeting Src kinase and decreasing inflammation may increase the efficacy of pharmacological therapies aimed at correcting the basic ΔF508 defect in CF liver patients. These studies also demonstrate the promise of applying iPSC technology in modeling human cholangiopathies. (Hepatology 2018;67:972-988).

Эпикардиальный жир и фибрилляция предсердий: роль профиброгенных медиаторов

Epicardial adipose tissue (EAT) has certain paracrine functions, which could be associated with proinflammatory and fibrotic changes in myocardium even in patients without structural heart disease but with atrial fibrillation (AF). to evaluate interrelationship between clinical data, parameters of tissue Doppler imaging (TDI), EAT thickness estimated by magnetic resonance imaging (MRI), and biochemical markers of fibrosis and inflammation in patients with nonvalvular AF without concomitant ischemic heart disease (IHD). We included in this study 39 AF patients without valvular pathology or IHD with normal or moderately increased left atrial (LA) dimension (mean age 50.8±13.9 years, 49 % men). The following groups were distinguished for analysis: with idiopathic AF (iAFgroup, n=21); with AF and arterial hypertension (AF+AH group, n=18); with normal (.