Local Inflammatory Markers Research Articles

O-280 Vaginal microbiota transplant (VMT) without antibiotic pretreatment reveals changes in vaginal microbiota and immune profile in women with asymptomatic dysbiosis – a randomized, placebo-controlled trial

Abstract Study question Investigate the safety of VMT and explore its effects on the vaginal microbiomes and local inflammation in healthy, asymptomatic volunteer women screened for vaginal dysbiosis Summary answer We demonstrate safety and engraftment of donor bacteria in 50% of recipients, and a distinct shift in local inflammatory markers following resolution of vaginal dysbiosis What is known already Dominance by a subset of vaginal Lactobacillus species is considered healthy. Dysbiosis characterized by high abundance of anaerobic species is observed in around one third of women, and affects fertility, pregnancy outcomes, risk of STIs and UTIs, and potentially cancer treatment. Current use of antibiotics is often insufficient with high recurrence rates and potentially antibiotic resistance. There is a clear need for alternatives and recently it was shown that VMT following antibiotic treatment resulted in long-lasting improvements in symptoms of bacterial vaginosis in four out of five patients, but RCTs without the use of antibiotics remain to be reported. Study design, size, duration We performed a randomized, placebo-controlled double-blinded trial in 34 recipients randomized in an active:placebo 3:1 ratio. Subjects in the active arm received VMTs from donor cervicovaginal secretions (CVS) while placebo participants received saline. All recipients were dosed on 3 consecutive days with active arm recipients receiving all 3 VMTs originating from the same donor. All recipients were followed for safety and efficacy assessments for 6 menstrual cycles after dosing. Participants/materials, setting, methods Donors: CVS was obtained from healthy subjects screened to be negative for a broad panel of infections and having lactobacilli-dominant vaginal microbiomes. RCT: Healthy, asymptomatic women aged 18 to 45 whose vaginal microbiome had combined lactobacilli relative abundance <10% and combined relative abundance of dysbiosis-associated bacteria >20% were included. VMTs were applied vaginally on three consecutive days. Follow-up visits were performed 1 week after the last VMT and then once every cycle for 6 cycles. Main results and the role of chance Here, we report the successful implementation of a donor program to obtain screened, frozen, and banked donor CVS to be used in the first VMT intervention RCT. In the ITT population, the primary efficacy analysis showed statistically significant increase from baseline in relative abundance of vaginal lactobacilli to follow-up timepoints 1 week and 3 cycles post intervention in the active arm, while no statistical significant change was observed in the placebo arm at any follow-up timepoint. Also, time-resolved metagenomic analysis showed that 50% of women treated with VMT displayed a shift of the vaginal microbiome toward the Lactobacillus-dominated donor CVS sample, confirmed with strain-level engraftment analysis. We observed no serious treatment-associated adverse events and the intervention was well tolerated. Further, the successful “twinning” of the recipient to match the donor microbiome post-VMT also led to a dramatic shift toward an anti-inflammatory environment based on transcriptomic analysis of CVS. Limitations, reasons for caution Recipients were asymptomatic and mainly Caucasian, which limits generalizability to other ethnicities and the symptomatic spectrum of vaginal dysbiosis. Due to COVID-19-related recruitment issues, the cohort size ended below the targeted number of n = 40. Future studies will aim to identify factors driving engraftment and its impact on clinical outcomes. Wider implications of the findings Vaginal microbial interventions including VMT have great potential as the first therapeutic intervention that provides effective and long-term vaginal microbiota restoration, positively impacting a wide variety of diseases and conditions affecting the female reproductive tract. Trial registration number NCT05114031

The interrelation between microbial immunoglobulin coating, vaginal microbiota, ethnicity, and preterm birth

BackgroundVaginal microbiota composition is associated with spontaneous preterm birth (sPTB), depending on ethnicity. Host-microbiota interactions are thought to play an important underlying role in this association between ethnicity, vaginal microbiota and sPTB.MethodsIn a prospective cohort of nulliparous pregnant women, we assessed vaginal microbiota composition, vaginal immunoglobulins (Igs), and local inflammatory markers. We performed a nested case–control study with 19 sPTB cases, matched based on ethnicity and midwifery practice to 19 term controls.ResultsOf the 294 included participants, 23 pregnancies ended in sPTB. We demonstrated that Lactobacillus iners-dominated microbiota, diverse microbiota, and ethnicity were all independently associated with sPTB. Microbial Ig coating was associated with both microbiota composition and ethnicity, but a direct association with sPTB was lacking. Microbial IgA and IgG coating were lowest in diverse microbiota, especially in women of any ethnic minority. When correcting for microbiota composition, increased microbial Ig coating correlated with increased inflammation.ConclusionIn these nulliparous pregnant women, vaginal microbiota composition is strongly associated with sPTB. Our results support that vaginal mucosal Igs might play a pivotal role in microbiota composition, microbiota-related inflammation, and vaginal community disparity within and between ethnicities. This study provides insight in host-microbe interaction, suggesting that vaginal mucosal Igs play an immunomodulatory role similar to that in the intestinal tract.-XF_JK898P7idx1NXSVZ7eVideo

Fecal microbiota composition, serum metabolomics, and markers of inflammation in dogs fed a raw meat-based diet compared to those on a kibble diet.

Despite the potential health risks associated with feeding raw and non-traditional diets, the use of these diets in dogs is increasing, yet the health outcomes associated with these diets is not well understood. This study investigates the effect of feeding dogs a kibble or raw meat-based diets on fecal microbiota composition, serum metabolomics and inflammatory markers. Clinically healthy dogs with a history of consuming either kibble (KD, n = 27) or raw meat-based diets (RMBD, n = 28) for more than 1 year were enrolled. Dogs were fed a standardized diet of either a single brand of KD or RMBD for 28 days. Serum and fecal samples were collected for analysis of microbiota, metabolomics, and inflammatory markers. Multiple regression analysis was performed for each of the metabolites and inflammatory markers, with feed group, age and BCS included as independent variables. The fecal microbiota composition differed between the KD and RMBD groups. Beta-diversity and some indices of alpha-diversity (i.e., Shannon and Simpson) were different between the two diet groups. Sixty- three serum metabolites differed between KD and RMBD-fed dogs with the majority reflecting the differences in macronutrient composition of the two diets.Fecal IAP, IgG and IgA were significantly higher in RMBD dogs compared to KD dogs, while systemic markers of inflammation, including serum c-reactive protein (CRP), galectin, secretory receptor of advanced glycation end-products (sRAGE), haptoglobin, and serum IgG were similar in dogs fed either diet. Diet composition significantly affected fecal microbiota composition and metabolome. Although it had a potentially beneficial effect on local inflammatory markers, feeding RMBD had no impact on systemic inflammation. The influence of these changes on long term health outcomes provides an area for future study.

Local Postoperative Graft Inflammation in Pancreas Transplant Patients With Early Graft Thrombosis

Background. Graft thrombosis is the main cause of early graft loss following pancreas transplantation, and is more frequent in pancreas transplant alone (PTA) compared with simultaneous pancreas-kidney (SPK) recipients. Ischemia-reperfusion injury during transplantation triggers a local thromboinflammatory response. We aimed to evaluate local graft inflammation and its potential association with early graft thrombosis. Methods. In this observational study, we monitored 67 pancreas-transplanted patients using microdialysis catheters placed on the pancreatic surface during the first postoperative week. We analyzed 6 cytokines, interleukin-1 receptor antagonist (IL-1ra), IL-6, IL-8, interferon gamma-induced protein 10 (IP-10), macrophage inflammatory protein 1β (MIP-1β), IL-10, and the complement activation product complement activation product 5a (C5a) in microdialysis fluid. We compared the dynamic courses between patients with pancreas graft thrombosis and patients without early complications (event-free) and between PTA and SPK recipients. Levels of the local inflammatory markers, and plasma markers C-reactive protein, pancreas amylase, and lipase were evaluated on the day of thrombosis diagnosis compared with the first week in event-free patients. Results. IL-10 and C5a were not detectable. Patients with no early complications (n = 34) demonstrated high IL-1ra, IL-6, IL-8, IP-10, and MIP-1β concentrations immediately after surgery, which decreased to steady low levels during the first 2 postoperative days (PODs). Patients with early graft thrombosis (n = 17) demonstrated elevated IL-6 (P = 0.003) concentrations from POD 1 and elevated IL-8 (P = 0.027) concentrations from POD 2 and throughout the first postoperative week compared with patients without complications. IL-6 (P < 0.001) and IL-8 (P = 0.003) were higher on the day of thrombosis diagnosis compared with patients without early complications. No differences between PTA (n = 35) and SPK (n = 32) recipients were detected. Conclusions. Local pancreas graft inflammation was increased in patients experiencing graft thrombosis, with elevated postoperative IL-6 and IL-8 concentrations, but did not differ between PTA and SPK recipients. Investigating the relationship between the local cytokine response and the formation of graft thrombosis warrants further research.

Peripheral and local inflammatory markers associated with epicardial fat volume in patients underwent open heart surgery

Abstract Introduction Epicardial fat (EF) is a source of inflammation, and it is associated with cardiovascular diseases. However, the correlation between EF volume (EFV) and local and peripheral biomarkers as well as adipogenesis behavior were not studied yet. Our objective was to a) Find the peripheral and local markers related to EFV independent of age and BMI. b) Study the relationship between the adipogenesis capacity and the EFV in each patient. Materials and Methods We included 33 patients underwent open heart surgery with preoperative cardiac CT imaging. Blood samples of 24 patients were obtained, neutrophils were isolated using polymorphprep and 16 activity markers were analyzed. Twelve biomarkers in plasma associated with metabolism and inflammation were analyzed. The epicardial adipose tissue biopsies were obtained during the surgery. The fat pads were homogenized and RNA was isolated and retrotranscripted to cDNA. Then 17 markers associated with adipogenesis and inflammation were analyzed by real time PCR. Stromal cells from EF of 21 patients were isolated after collagenase activity and cultured for 14 days and then induced to adipogenesis for the next 14 days. Multimarkers on plasma, peripheral neutrophils, EF and primary cultures were analyzed by multiplex assay and real time PCR, respectively. EFV was measured from CT, percentage of EFV in total heart volume was calculated. The association between FV and biomarkers were evaluated with independent sample t-test, linear regression analysis and correlation analysis. Results The main clinical characteristics of studied patients were (69 ± 9 years old, 30 ± 4 kg/m², 76% HT, 67% DLP, 33% CAD, 27% HF, 30% AF, 30% T2DM). EFV (148±55 cm³) was more correlated with age (r=0.4, p=0.023), but not with BMI. Percentage of EFV in total heart volume (12.9±4.9%) was positively correlated with lymphocytes and monocytes markers, CD3G and CXCR2 levels (r=0.5, p=0.008; r=0.49, p=0.028 respectively), and inversely with FABP4 (r=0.46, p=0.043) in EF. Circulating inflammatory markers, plasma C5a and MPO on neutrophils were associated with a greater amount of EFV (r=0.7, p=0.001 for C5a and r=0.75, p=0.001 for MPO) and percentage of EFV in total heart volume (r=0.63, p=0.003 for C5a and r=0.84, p<0.000 for MPO). Neutrophils markers, NGAL (r=0.49, p=0.048), LF (r=0.52, p=0.034), DEFA3 (r=0.57, p=0.023) were also correlated with percentage of EFV to total heart volume. Lineal regression determined that among local markers CD3G was the best EF predictor (r=0.4, p=0.008); regarding peripheral markers, the C5a expression levels on neutrophils was the best EF predictor (r=0.61, p=0.002). There were no association between adipogenesis ability and EFV among studied patients. Conclusion EFV from patients undergoing open heart surgery were associated with local and peripheral inflammatory markers. It might contribute to cardiovascular disease progression. Long term follow-up is needed to clarify this process.

Impact of microbiota and host immunologic response on the efficacy of anticholinergic treatment for urgency urinary incontinence.

Studies within the past decade have suggested associations among composition of the urinary microbiota, local immune responses, and urinary incontinence symptoms. To investigate these relationships, we evaluated the structure of the urinary microbiome, local inflammatory markers, and patient responses prior to and at 6-weeks after treatment with anticholinergic medication for urgency urinary incontinence (UUI). Using a prospective pilot study, we enrolled women who presented with UUI symptoms and were prescribed treatment with anticholinergics. Catheterized urine samples were collected from participants at their baseline and 6-week follow-up visits for microbiological (standard and 16S rRNA gene phylotyping analyses) and cytokine analysis along with the UDI-6 questionnaire and 2-day bladder diary. Patients were Caucasian, post- menopausal, with a median age of 64 and median BMI of 30.1kg/m2. Among the patients, 75% had UUI symptoms for less than 2years, but with a frequency of at least a few times a week or every day. Most women were prescribed 10mg oxybutynin ER daily at enrollment. Patients had varied urinary microbiota by culture and 16S phylotyping, with species of Lactobacillus being the most common, in six samples, in addition to taxa associated with Enterococcus, Staphylococcus, and mixed flora. Cytokine levels showed no differences before and after treatment with anticholinergics, nor correlation with urinary bacteria or microbiome composition. Our pilot study suggests factors in addition to the urinary microbiome and local immune responses may be involved in patients' response to anticholinergics for UUI.

Radiotherapy-induced dynamic changes in the lymphocyte-to-monocyte ratio in patients with laryngeal cancer indicate poor prognosis.

We hypothesized that markers of inflammation correlate with response to radiotherapy in patients with non-metastatic laryngeal cancer (LC). Our aim was to assess peripheral and local markers of inflammation including lymphocyte to monocyte ratio (LMR), neutrophil to lymphocyte ratio (NLR), platelet to lymphocyte ratio (PLR), infiltrating CD8+ lymphocytes (TILsCD8), and programmed death 1 ligand (PD-L1) expression. We performed a retrospective single-center analysis of LC patients administered definitive (R-RT) or postoperative radiotherapy (PORT). The primary endpoint was overall survival (OS) in relation to peripheral and local inflammatory markers and their dynamic changes during RT. Study group included 215 patients (R-RT, n=116; PORT, n=99). The baseline (t0) NLR and LMR were significantly correlated with OS in the R-RT group. In patients with high and low NLR at t0, the five-year OS was 33% and 56% (p=0.010) and in high and low LMR at t0, the five-year OS was 56% and 27% (p=0.003), respectively. The LMR increase during R-RT predicted better prognosis: the five-year OS in high and low LMR was 57% and 31% at t2 (after 2 weeks of RT) (p=0.015), 49% and 26% at t4 (p< 0.001), and 50% and 25% at t6 (p=0.013), respectively. Multivariable analysis shows that the worse performance status (p=0.003), the presence of nodal metastases (p=0.0001), and low baseline LMR (p=0.049) in the R-RT group, and the presence of nodal metastases (p=0.035) and completion treatment on time (p=0.042) in PORT group were associated with poor prognosis. The PD-L1 expression had no significant prognostic value in any of the examined patients. The baseline LMR and its dynamic changes during R-RT and baseline NLR are independent prognostic factors in patients with nonmetastatic LC. PD-L1 expression and number of TILsCD8 have no prognostic value in R-RT and PORT group.

HbA1c, And Blood Glucose, Changes When Treating Periodontal Disease with the Perio Protect Method™

Periodontal disease is a chronic wound resulting in host elevated local and systemic inflammatory markers in response to the microorganisms of the periodontal biofilm. Separately from its importance as an oral ailment, chronic periodontitis has gained relevance since it can develop into a systemic condition characterized by hyper-inflammation, disruption of the innate and adaptive immune system, and other system-wide alterations. There is compelling evidence that inflammation and chronic infection play an essential role in the development of inflammatory markers and of type 2 diabetes. Studies in humans suggest that circulating inflammatory marker levels may predict type 2 diabetes years in advance of the onset of the disease. Local and systemic expression of inflammatory cytokines, such as TNF-alpha and IL-6, increase in individuals with periodontitis. TNF-alpha and IL-6 impair intracellular insulin signaling, which may lead to insulin resistance and are significantly associated with cardiovascular events and stroke. This case study demonstrates treating periodontal disease with a direct medication delivery method (Perio Protect Method™) manages the patient’s periodontal disease and lowers daily blood glucose and HbA1c levels.

Metabolic and inflammatory profiles define phenotypes with clinical relevance in female knee osteoarthritis patients with joint effusion.

Osteoarthritis has been the subject of abundant research in the last years with limited translation to the clinical practice, probably due to the disease's high heterogeneity. In this study, we aimed to identify different phenotypes in knee osteoarthritis (KOA) patients with joint effusion based on their metabolic and inflammatory profiles. A non-supervised strategy based on statistical and machine learning methods was applied to 45 parameters measured on 168 female KOA patients with persistent joint effusion, consecutively recruited at our hospital after a monographic OA outpatient visit. Data comprised anthropometric and metabolic factors and a panel of systemic and local inflammatory markers. The resulting clusters were compared regarding their clinical, radiographic and ultrasound severity at baseline and their radiographic progression at two years. Our analyses identified four KOA inflammatory phenotypes (KOIP): a group characterized by metabolic syndrome, probably driven by body fat and obesity, and by high local and systemic inflammation (KOIP-1); a metabolically healthy phenotype with mild overall inflammation (KOIP-2); a non-metabolic phenotype with high inflammation levels (KOIP-3); and a metabolic phenotype with low inflammation and cardiovascular risk factors not associated with obesity (KOIP-4). Of interest, these groups exhibited differences regarding pain, functional disability and radiographic progression, pointing to a clinical relevance of the uncovered phenotypes. Our results support the existence of different KOA phenotypes with clinical relevance and differing pathways regarding their pathophysiology and disease evolution, which entails implications in patients' stratification, treatment tailoring and the search of novel and personalized therapies.

Abstract P5-03-08: Joint associations of mammographic breast density and obesity on the presence of crown-like structures in the breast adipose tissue of breast cancer patients

Abstract Background: Obesity is an established risk factor for postmenopausal breast cancer and is associated with poor outcomes. Accumulating evidence suggests crown-like structures in the breast adipose tissue (CLS-B), a marker of local inflammation, play a role in explaining the obesity-breast cancer association. However, it is unknown whether breast tissue composition (i.e., the amount of fibroglandular tissue in the breast relative to fat) is related to CLS-B. Objective: We evaluated whether breast tissue composition, as reflected by mammographic breast density, is associated with breast adipose tissue inflammation, as indicated by the presence of CLS-B, and whether the combination of breast density and obesity increases the presence of CLS-B among newly diagnosed breast cancer patients. Methods: We examined the presence of CLS-B, detected by CD68 immunohistochemistry, in breast adipose tissue obtained via mastectomy from a quadrant uninvolved by tumor among 254 women with stage I–III breast cancer treated at Emory University Hospitals (2007–2012). Patient characteristics, including mammographic breast density (assessed on a mammogram up to 5 years before breast surgery) and body mass index (BMI) at diagnosis, were abstracted from electronic medical records. Mammographic density was assessed using the Breast Imaging Reporting and Data System (BI-RADS) density classification (1=almost entirely fat; 2=scattered fibroglandular densities; 3=heterogeneously dense; and 4=extremely dense); density was further categorized as fatty (BI-RADS 1-2) and dense (BI-RADS 3-4). Age and multivariable (MV)-adjusted logistic regression models were used to estimate odds ratios (ORs) and 95% confidence intervals (CIs) for the independent and joint associations of breast density (dense vs. fatty) and obesity (BMI ≥30 kg/m2 vs. &amp;lt; 30 kg/m2) on the presence of CLS-B. Multivariable models adjusted for age (continuous, years) and parity (nulliparous, parous) with mutual adjustment of BMI (continuous, kg/m2) or mammographic breast density (dense, fatty) depending on the model. Results: Women with obesity were more likely to have fatty breasts than women without obesity (52% vs. 27%). Obesity was strongly associated with the presence of CLS-B in age-adjusted (OR=3.11, 95% CI: 1.79, 5.48) and multivariable models (MV-OR=3.70, 95% CI: 1.96, 7.15). There was no apparent association between dense breast tissue and presence of CLS-B in the age-adjusted model (OR=1.04, 95% CI: 0.59, 1.85). After additional adjustment for BMI and parity, we noted that women with dense breasts had higher odds of having CLS-B compared to those with fatty breasts (MV-OR=2.13, 95% CI: 1.07, 4.41). MV-ORs from the joint model (common referent: not obese, fatty breasts) were 1.54 (95% CI: 0.62, 4.24) for women without obesity but with dense breasts, 3.18 (95% CI: 1.15, 9.56) for women with obesity and fatty breasts, and 6.24 (95% CI: 2.23, 19.2) for women with obesity and dense breasts. Conclusions: Our findings suggest that dense rather than fatty breast tissue is associated with breast adipose tissue inflammation among women with breast cancer. Results of the joint analyses suggest obesity is more strongly predictive of CLS-B presence than breast density. However, density may be an important risk factor for CLS-B among women without obesity while patients with obesity and dense breast tissue are the most likely to have CLS-B present. Future studies may consider the mechanisms by which density leads to increased presence of CLS-B. Joint Associations Table Table 1. Joint associations between mammographic breast density (MD) and body mass index (BMI) on the presence of crown-like structures in breast adipose tissue (CLS-B) among 254 women diagnosed with invasive breast cancel (stage I-III) at Emory University between 2007 and 2012 Citation Format: Maret L. Maliniak, Rebecca L. Seidel, Kimberly Bertrand, Lauren E. McCullough. Joint associations of mammographic breast density and obesity on the presence of crown-like structures in the breast adipose tissue of breast cancer patients [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P5-03-08.

Abstract 13662: The Role of Pcsk9 in Abdominal Aortic Aneurysm

Introduction: Abdominal aortic aneurysm (AAA) is a local weakening and dilatation of the abdominal aorta ≥50%, is associated with high mortality in the event of rupture and has no current pharmacologic treatment. Proprotein convertase subtilisin/kexin type 9 (Pcsk9), an enzyme mainly known for its production in liver, can cause familial hypercholesterolemia due to gain-of-function mutations. When triggered by pro-inflammatory stimuli, Pcsk9 expression by vascular smooth muscle cells (VSMC) has been found to induce cellular polyploidy, senescence, and apoptosis, hallmarks of AAA. Further, our collaborative consortium identified a relationship between Pcsk9 variants and AAA disease in humans utilizing a genome-wide association stud y (GWAS) in the Million Veteran Program. Lastly, our transcriptional profiling experiments in the porcine pancreatic elastase (PPE) model of murine AAA found significant early (Day 3) upregulation of the Pcsk9 gene in aorta.Monoclonal Pcsk9 antibody therapies are effective for hypercholesterolemic patients who are resistant to conventional statin therapy. However, the role of Pcsk9 in AAA has not been explored in depth. If Pcsk9 were found to be pivotal in disease, Pcsk9-inhibitors could be high priority candidates for investigation in limiting AAA. Methods: Using the PPE model, AAA was induced in wildtype C57Bl/6 mice and Pcsk9 knock-out (KO) mice. Aneurysm growth was monitored via sonography at various time points, and local vascular Pcsk9 and inflammatory marker gene expression were measured by qPCR. In vitro, VSMC were treated with exogenous Pcsk9 and assayed for pro-inflammatory marker expression. Results: Sonographic imaging data showed smaller AAA diameters in Pcsk9 KO mice compared with wildtype C57Bl/6 mice. Inflammatory gene expression was significantly downregulated with Pcsk9 KO. In-vitro VSMCs showed upregulated inflammation after exogenous Pcsk9 treatment. Conclusions: Our data suggest that Pcsk9 plays a crucial role in the development of AAA and could be a viable treatment target to inhibit AAA progression.

PCSK9 inhibitors for anti-inflammation in atherosclerosis: protocol for a systematic review and meta-analysis of randomised controlled trials

IntroductionAtherosclerosis is the leading cause of cardiovascular disease (CVD), which is one of the most common causes of morbidity and mortality worldwide. Lipid accumulation and inflammation play a crucial role...

The relationship of synonasal symptoms with systemic and local markers of inflammation in children with bronchial asthma and allergic rhinitis

The relationship of synonasal symptoms with systemic and local markers of inflammation in children with bronchial asthma and allergic rhinitis

Resveratrol decreases local inflammatory markers and systemic endotoxin in patients with aggressive periodontitis.

Background:Inflammation is hypothesized to contribute to the pathogenesis of periodontitis. Resveratrol (RV) is known for its anti-inflammatory properties. The purpose of this study was to investigate the inhibitory effect of RV on local inflammatory markers and systemic endotoxin in patients with periodontitis.Methods:A total of 160 patients with periodontitis were enrolled in this study. The selected patients were randomly divided into four groups and received placebo, high-dose (500 mg/d) of RV (HRV, n = 40), middle-dose (250 mg/d) of RV (middle dose RV (MRV), n = 40) and low-dose (125 mg/d) of RV (low dose RV (LRV), n = 40) with orally administration. All patients received an 8-week treatment. The periodontal status of patients with periodontitis was recorded by using clinical attachment level (CAL), bleeding index (BI), oral hygiene index-simplified (OHI-S), and probing pocket depth (PPD). The levels of inflammatory markers in serum and gingival crevicular fluid (GCF), and systemic levels of endotoxin were evaluated using high sensitivity enzyme-linked immuno sorbent assay.Results:Outcomes showed that symptoms of periodontitis determined by CAL, BI OHI-S and PPD were improved by RV compared to placebo. RV treatment decreased inflammatory markers in serum and GCF compared to placebo in patient with periodontitis. Systemic endotoxin declined more in the RV group than the placebo-treated group.Conclusion:In conclusion, data in the current study indicate that RV is an efficient drug for the treatment of patients with periodontitis. The findings of the present study find that RV inhibits systemic local inflammatory markers and systemic endotoxin and suggest that 500 mg/d RV is the ideal dose for patients with periodontitis.

Interleukin-6 and YKL-40 predicted recurrent stroke after ischemic stroke or TIA: analysis of 6 inflammation biomarkers in a prospective cohort study

ObjectiveContribution of individual and combined inflammatory markers in prognosis after stroke was still undefined. We aimed to investigate the association of systemic and local vascular inflammatory markers and recurrent stroke as well as impact on poor functional outcome.MethodsIn this pre-specified substudy of the Third China National Stroke Registry (CNSR-III), 10,472 consecutive acute ischemic stroke or TIA patients with available centralized-measured levels of Interleukin-6 (IL-6), high sensitive C-reactive protein (hsCRP), IL-1 receptor antagonist (IL-1Ra), lipoprotein-associated phospholipase A2 mass (Lp-PLA2) and activity (Lp-PLA2-A), and YKL-40 from 171 sites were enrolled. The primary outcomes consisted of stroke recurrence and poor functional outcome defined as modified Rankin Scale (mRS) score of 2–6 within 1 year.ResultsThere were 1026 (9.8%) and 2395 (23.4%) patients with recurrent stroke and poor functional outcome within 1 year. The highest quartiles of IL-6 (adjusted HR, 1.36; 95% CI 1.13–1.64; P = 0.001), hsCRP (adjusted HR, 1.41; 95% CI 1.17–1.69; P = 0.0003) and YKL-40 (adjusted HR, 1.28; 95% CI 1.06–1.56; P = 0.01) were associated with increased risk of recurrent stroke; and the highest quartiles of IL-6 (adjusted OR 1.93; 95% CI 1.64–2.27; P < 0.0001), IL-1Ra (adjusted OR 1.60; 95% CI 1.37–1.87; P < 0.0001), hsCRP (adjusted OR 1.60; 95% CI 1.37–1.86; P < 0.0001) and YKL-40 (adjusted OR 1.21; 95% CI 1.03–1.42; P = 0.02) were correlated with increased risk of poor functional outcome. In the multivariate stepwise regression analysis including all markers with backward selection, elevated levels of IL-6 or YKL-40 were associated with recurrent stroke (IL6: OR, 1.34; 95% CI 1.19–1.52; P < 0.0001; YKL-40: OR, 1.01; 95% CI 1.01–1.03; P = 0.004) and poor functional outcome (IL6: OR, 1.68; 95% CI 1.46–1.93; P < 0.0001; YKL-40: OR, 1.02; 95% CI 1.01–1.03; P = 0.0001). Adding IL-6 and YKL-40 significantly increased the area under the receiver operating characteristic curves for the prediction models of Essen Stroke Risk Score (0.03, P < 0.0001) and Totaled Health Risks in Vascular Events Score (0.07, P < 0.0001), and yielded continuous net reclassification improvement (19.0%, P < 0.0001; 33.0, P < 0.0001).ConclusionsIn the patients with ischemic stroke or TIA, IL-6 and YKL-40 were independently associated with recurrent stroke and poor functional outcome, and improved risk classification of clinical risk algorithms.

P-175 Local inflammatory biomarkers and their association with systemic inflammation as well as overall survival in primary metastatic gastroesophageal cancer patients

Immunotherapy has recently led to a major breakthrough in the treatment of metastatic gastroesophageal cancer patients. Yet, further analyses of large trials investigating checkpoint inhibitor monotherapy or combination with standard chemotherapy suggest, that only a subgroup of patients might benefit from addition of immunotherapy to chemotherapy. Despite these advances, prognosis remains poor, especially in patients with Caucasian ethnicity. Thus, novel prognostic as well as predictive biomarkers are desperately needed to improve patient management. Further knowledge on the complex local inflammatory mechanisms might help to improve our understanding on the efficacy of immunotherapy. Thus, the aim of this study was to further characterize local inflammatory processes in patients with advanced gastroesophageal cancer and investigate their association with systemic inflammation as well as the overall survival (OS). We analyzed local inflammatory biomarkers for T-cells (CD3, CD8), macrophages (CD68) and immune checkpoint inhibition (Lymphocyte-activation gene 3 (LAG3)) in previously untreated, primary tumor tissue samples of advanced gastroesophageal cancer patients treated at the Medical University of Vienna between 2003 and 2016. FFPE tissue was stained using an automated immunohistochemistry slide staining system (Roche Ventana Medical Systems Inc., Tucson, AZ, USA). Analyses of the immunohistochemical staining was performed by Definiens Tissue Studio 4.0 software. Systemic serum inflammatory parameters including leucocyte levels (WBC), C-reactive protein levels (CRP) and albumin (Alb) as well as the OS was evaluated retrospectively by hospital chart review. Local inflammatory parameters were then associated with systemic parameters and the OS using log-rank test and Kruskal-Wallis-test. Primary tumor tissue samples (localization: 29% esopheageal, 25% gastroesophageal junction, 46% stomach; histological subtype: 77% adenocarcinoma, 23% squamous cell carcinoma) of 48 patients (65% male) were analyzed. CD3 was positive in 5.6%, CD8 in 1.8%, CD68 in 2.4% and LAG3 in 0.24% of analyzed cells. Slides were then divided by the median expression into samples with more positive and less positive cells. The median OS of the cohort was 7.6 months (95%CI 5.3-9.9). Local inflammatory biomarker could not be statistically significantly associated with the OS (CD3: p=0.317; CD8: p=0.905, CD68: p=0.369; LAG3: p=0.428). In addition, no clinically significant association with systemic inflammatory parameters could be identified (CD3: CRP p=0.168, WBC p=0.706, Alb p=0.849; CD8: CRP p=0.214, WBC p=0.275, Alb p=0.340; CD68: CRP p=0.499, WBC p=0.992, Alb p=0.181; LAG3: CRP p=0.766, WBC p=0.384), only Alb was associated with LAG3 expression (p=0.048). Local inflammation might play an important role as a prognostic and predictive tool. However, the understanding of local inflammation in gastroesophageal cancer is still scarce and, thus, further research is warranted to identify promising novel biomarkers. Additional staining for further local inflammatory markers as well as expansion of sample size are underway to further characterize the local inflammatory microenvironment in advanced gastroesophageal cancer patients.

Adverse Effect in Patients with Psoriasis Treated with Interleukin 17A Inhibitor- Secukinumab

Abstract Secukinumab is fully human monoclonal antibody, IgG-1κ, which selectively attaches to IL-17A and inhibits its effects, which subsequently leads to a decrease of local inflammatory markers. In 2015 it was approved for treatment of patients suffering from psoriasis. We can say that in comparison with other biologic medicine, such as IL-12/23 inhibitors and TNF-α inhibitors, the incidence rate of serious adverse effects related to use of secukinumab is notably lower. Serious adverse effects reported in relation to use of secukinumab were development of mucocutaneous candidiasis, neutropenia and development or aggravation of the inflammatory bowel disease conditions. In this review study we focused on frequent adverse effects and adverse effects of special interest during the secukinumab therapy in treating psoriasis patients. Available data on long-term safety and effects on comorbidities are relatively few. A more extensive and longer term research is needed, as well as critical reevaluation of the criteria for participation in clinical trials in order to obtain data which would be of relevance in clinical practice. A better understanding of adverse effects leads to an improved individual therapeutic approach, increases patient’s satisfaction and results in minimizing these effects.

Sonographic Tophi and Inflammation Are Associated With Carotid Atheroma Plaques in Gout

Objective: Gout and cardiovascular disease are closely related, but the mechanism connecting them remains unknown. This study aims to explore whether urate crystal deposits and inflammation (assessed by ultrasound) are associated with carotid atherosclerosis.Methods: We included consecutive patients with crystal-proven gout newly presenting to a tertiary rheumatology unit. Patients under urate-lowering treatment were excluded. Ultrasound assessment was performed during intercritical periods. Musculoskeletal scans evaluated six joints and four tendons for urate crystal deposits (double contour, aggregates, and tophi), and power Doppler (PD) signal (graded 0–3) as a marker of local inflammation. The sum of locations showing deposits or a positive PD signal (≥1) was registered. Carotids were scanned for increased intima-media thickness (IMT) and atheroma plaques, according to the Mannheim consensus. Associations were analyzed using logistic regression.Results: The study included 103 patients showing sonographic crystal deposits at the examined locations (mean sum 9.9, minimum 2); tophi were the most frequent. Two-thirds of participants presented a positive PD signal (30.1% grade 2–3). In the carotid scans, 59.2% of participants showed atheroma plaques, and 33.0% increased IMT. Tophi (odds ratio [OR] 1.24; 95% confidence interval [CI] 1.03–1.50) and a positive PD signal (OR 1.67; 95% CI 1.09–2.56) were significantly associated with atheroma plaques, while an increased IMT showed no sonographic association.Conclusion: Sonographic crystal deposits and subclinical inflammation were consistently observed in patients with intercritical gout. Tophi and a positive PD signal were linked to carotid atherosclerosis. Our findings may contribute to understanding the complex relationship between gout and atherosclerosis.

Inhibition of the Interaction of TREM-1 and eCIRP Attenuates Inflammation and Improves Survival in Hepatic Ischemia/Reperfusion.

Triggering receptor expressed on myeloid cells-1 (TREM-1) has important implications in sepsis and inflammation and is a novel receptor for extracellular cold-inducible RNA-binding protein (eCIRP). We hypothesize that the inhibition of TREM-1 via its interaction with eCIRP by novel peptide inhibitor M3 or knockout gene will attenuate the inflammation and injury associated with severe hepatic ischemia/reperfusion (I/R). Wild-type (WT) C57BL/6 and TREM-1-/- mice underwent 60 min of 70% hepatic ischemia, with 24 h of reperfusion. Additionally, WT mice underwent hepatic I/R and were treated with M3 (10 mg/kg body weight) or vehicle (normal saline) at the start of reperfusion. Blood and ischemic liver tissues were collected, and analysis was performed using enzymatic assays, enzyme-linked immunosorbent assay, reverse-transcription quantitative polymerase chain reaction, and pathohistology techniques. For survival surgery, mice additionally underwent resection of non-ischemic lobes of the liver and survival was monitored for 10 days. There was an increase in serum levels of tissue markers including aspartate aminotransferase, alanine aminotransferase, and lactate dehydrogenase as well as cytokine levels (IL-6) and histological scoring of hematoxylin and eosin sections in WT I/R mice. These markers decreased substantially in TREM-1-/- mice. Additionally, neutrophil infiltration markers and markers of local inflammation (myeloperoxidase, macrophage inflammatory protein-2, cyclooxygenase-2) were attenuated in TREM-1-/- mice. Similarly, we show a significant decrease in injury and inflammation markers with M3 treatment. Additionally, we demonstrate decreased apoptosis with TREM-1 inhibition. Finally, M3 treatment improved the survival rate from 42% to 75% after hepatic I/R. TREM-1 is an important eCIRP receptor in the inflammatory response of hepatic I/R, and deficiency of TREM-1 via knockout gene or peptide inhibition attenuated liver injury and inflammation, and improved survival. Inhibition of the TREM-1 and eCIRP interaction in hepatic I/R may have important therapeutic potential.

Dehydrated Human Amniotic-Chorionic Membrane Reduces Incisional Hernia Formation in an Animal Model

Currently there are no standard of care treatment strategies for IH prevention (IHP). Dehydrated human amnion-chorion (dHACM) is a healing adjunct that elutes growth factors including several that have reduced IH in animal models. We therefore performed a double-blinded, prospective randomized controlled trial (RCT) to test the hypothesis that dHACM significantly reduces IH formation in a well-studied animal model of acute IH. Forty 16-week-old male Sprague-Dawley rats were randomized to one of four groups: No Treatment vs. dHACM Sheet (Group A), and Saline vs. dHACM Injection (Group B). Each animal underwent a 5-cm midline laparotomy which was incompletely closed with 5-0 plain gut sutures; this was performed by a surgeon blinded to treatment group (first blind). After 28 days, the primary endpoints of IH formation and hernia size were determined by study staff blinded to treatment (second blind). Secondary endpoints included healed fascia tensile strength as determined by tensiometry, systemic and local inflammatory markers as measured by ELISA, and fascial scar collagen I/III ratios per Western blotting. In Group A, No Treatment developed IH at 87.5% vs. 62.5% for Sheet (P=0.28). Hernias that formed in the Sheet group were significantly smaller (P=0.036).In Group B, Injection and Saline yielded identical IH rates of 77.8%. Molecular characterization of fascial scar demonstrated non-inferior tensile strength, collagen I/III ratios, and inflammatory markers in dHACM-treated animals. dHACM sheets significantly reduced the size of IH following laparotomy when compared to no treatment.