The aim for the present study was to investigate the effects of olive leaf (Olea europea L.) extract (OLE) on the control of Yersinia ruckeri infection in rainbow (Oncorhynchus mykiss) trout and to assess the impact on the expression of immune-related genes in the spleen and serum biochemical parameters of rainbow trout. Five experimental diets were prepared by adding 0.0%, 0.1%, 0.25%, 0.50% and 1.0% of OLE. Each diet was fed to triplicate groups of fish (mean body weight 51.22 ± 3.04 g) twice a day (at 09:00 and 17:00 h) for 60 days. The dietary supplementation of OLE did not affect growth performance and feed utilization (P>> 0.05). Major changes due to graded levels of OLE in the diets were observed in blood biochemical parameters (P < 0.05). TNFα, IL1-β and IL-8 gene expressions were significanlty up-regulated in OLE 0.1% group compared with others (P < 0.05). Also, diet supplemented with OLE reduced mortality in rainbow trout fed with OLE 0.1% added diet. Present study suggests that OLE especially at 0.1% added feed may effectivelly enhance the serum biochemical parameters, survival rate and immune gene expression in rainbow trout.

Cardio/cerebral-vascular diseases seriously threaten human health and are the leading cause of death. As such, there is great interest in identifying a potential mechanism that controls the development process of cardio/cerebral vascular diseases. Present studies demonstrate that inflammasomes play an important role in the process of ischemic cardio/cerebral vascular diseases (ICCVDs). Among the pathological process of ICCVDs, inflammasomes activated the sterile inflammatory response that accelerated the development of diseases and aggravated the acute lesion of tissue. As the most thoroughly studied inflammasome, the NLRP3 inflammasome has been proven to be a potential therapeutic target for ICCVDs. In this review, we summarized the mechanisms of Chinese herbal medicine which can affect ICCVDs via the regulation of the NLRP3 inflammasome. Our study discovers that active compounds of Chinese medicines have a negative effect on NLRP3 in different ICCVDs models. Astragaloside IV may influence the receptor of the cell membrane to inhibit NLRP3 activation. Resveratrol, colchicinesis, salvianolic acid <em>B</em>, chrysophanol and sulforaphane may directly damage the formation of NLRP3 by inhibiting ASC or Caspase-1. Most of the active natural compounds can negatively regulate the downstream products of NLRP3 inflammasome such as IL-18 and <em>IL1</em> <mml:math><em>β</em></mml:math> . In addition, Chinese medicines such as sinomenine, ruscogenin, resveratrol, arctigenin and cepharanthineas may downregulate NLRP3 inflammasome by inducing autophagy activation. Due to the advantages of multi-target effects, Chinese herbal medicine can be treated as a splendid therapy for ICCVDs by inhibiting NLRP3 inflammasome.

OBJECTIVE

This double-blind, placebo-controlled clinical study compared multiple applications of the antimicrobial photodynamic therapy (aPDT) treatment protocol, to systemic doxycycline as adjuvant to scaling and root planing (SRP) on type 2 diabetic patients on clinical, systemic and immune-inflammatory outcomes.

METHODS

Thirty patients with Hba1c >7% were allocated in two groups, SRP + Doxy (n = 15) using systemic doxycycline 100 mg/day (14 days) and SRP + aPDT (n = 15) with multiple applications (0, 3, 7 and 14 days). Primary outcome was glycated haemoglobin levels (HbA1c). Clinical parameters: plaque score (PS), bleeding on probe, probing depth, suppuration, gingival recession, and clinical attachment level, percentage of pockets with desired clinical endpoint were measured at baseline and 3 months after therapy. Cytokine profile was assessed at 0, 1 and 3 month to measure IL1-β, TNF-α and TGF-β on gingival crevicular fluid.

RESULTS

No significant difference was detected on HbA1c, between treatments. The SRP + aPDT group showed advantage on reducing moderate pockets in single-rooted teeth at 3 months. SRP + aPDT presented better results at 3 months on IL1-β levels. There were no significant differences between TNF-α and TGF-β.

CONCLUSIONS

Both treatments improved clinical and systemic outcomes (Hba1c). SRP + aPDT performed better in moderate probing pocket depth on single-rooted teeth, reduced favourably inflammation in short term, and may be an alternative to systemic antibiotics. (Clinicaltrials.org ID NCT01595594).

<A<em>b</em>stractText>To study the putative effects of Advanced Oxidation Protein Products (AOPPs) and Advanced Glycation End Products (AGEs) in the development and progression of cardiovascular disease (CVD).</A<em>b</em>stractText><p><div>(<em>b</em>)Methodology</<em>b</em>)</div>AGEs, AOPPs, e-NOS, lipid profile, circulating stress and inflammatory <em>b</em>iomarkers were evaluated among fifty cardiovascular patients and fifty controls. Independent student's <i>t</i>-test was done for statistical analysis.</p><A<em>b</em>stractText>The malondialdehyde mean level in CVD patients (5.45 nmol/ml) was significantly higher than control (1.36 nmol/ml) (p value = 0.018). Nitric oxide in CVD patients (55.72 ng/ml) was remarka<em>b</em>ly increased as compared to normal su<em>b</em>jects (19.19 ng/ml). A significant change in the mean serum level of AGEs in CVD patients (2.74 ng/ml) and normal individuals (0.85 ng/ml) was recorded (p value = 0.000). The AOPPs also showed significant increased levels in CVD group (132.07 ng/ml) in comparison with normal su<em>b</em>jects (83.05 ng/ml) (p value = 0.011). The mean eNOS serum level in CVD group (15.50 U/L) was higher than control group (11.28 U/L) (p value = 0.004). Cardiovascular disease patients, in comparison with healthy controls, showed increased level of total cholesterol (5.48 mmol/L vs 4.45 mmol/L), triglycerides (2.59 mmol/L vs 1.24 mmol/L), and low density lipoprotein (2.47 mmol/L vs 2.31 mmol/L) along with decrease in high density lipoprotein (1.39 mmol/L vs 1.74 mmol/L). The mean MMP-11 serum levels in CVD group (98.69 ng/ml) was almost dou<em>b</em>le of control group (45.60 ng/ml) (p value = 0.017). The mean serum level of TNF-α and <em>IL1</em>-α were 32.16 pg/ml and 6.64 pg/ml in CVD patient. The significant decreasing trend of SOD (p value = 0.041), CAT (p value = 0.018), GSH (p value = 0.036) and GRx (p value = 0.029) <em>b</em>ut increasing drift of GPx (0.023) level was o<em>b</em>served in CVD patients.</A<em>b</em>stractText><A<em>b</em>stractText>This study provides strong evidence that CVD patients presented with elevated oxidative stress, enhanced inflammation and lipid profile in their serum. Therefore, the study strongly approves that AGEs, AOPPs, inflammatory and lipoxidative <em>b</em>iomarkers hold predictive potential in causing and aggravating the disease, thus <em>b</em>y controlling these factors CVD progression can <em>b</em>e inhi<em>b</em>ited.</A<em>b</em>stractText>

(<em>b</em>)Background:</<em>b</em>) Better understanding of the contri<em>b</em>ution of donor aging and comor<em>b</em>idity factors of expanded criteria donors (ECD) to the clinical outcome of a transplant is a challenge in kidney transplantation. We investigated whether the features of donor-derived stromal vascular fraction of perirenal adipose tissue (PRAT-SVF) could <em>b</em>e indicative of the deleterious impact of the ECD microenvironment on a renal transplant. (<em>b</em>)Methods:</<em>b</em>) A comparative analysis of cellular components, transcriptomic and vasculogenic profiles was performed in PRAT-SVF o<em>b</em>tained from 22 optimal donors and 31 ECD deceased donors. We then investigated whether these parameters could <em>b</em>e associated with donor aging and early allograft dysfunction. (<em>b</em>)Results:</<em>b</em>) When compared with the PRAT-SVF of non-ECD donors, ECD PRAT-SVF displayed a lower proportion of stromal cells, a higher proportion of inflammatory NK cells. The glo<em>b</em>al RNA sequencing approach indicated a differential molecular signature in the PRAT-SVF of ECD donors characterized <em>b</em>y the over-expression of CXCL1 and <em>IL1</em>-<em>β</em> inflammatory transcripts. The vasculogenic activity of PRAT-SVF was highly varia<em>b</em>le <em>b</em>ut was not significantly affected in marginal donors. Periorgan recruitment of monocytes/macrophages and NK cells in PRAT-SVF was associated with donor aging. The presence of NK cell infiltrates was associated with lower PRAT-SVF angiogenic activity and with early allograft dysfunction evaluated on day 7 and at 1 month post-transplant. (<em>b</em>)Conclusions:</<em>b</em>) Our results indicate that human NK cell su<em>b</em>sets are differentially recruited in the periorgan environment of aging kidney transplants. We provide novel evidence that PRAT-SVF represents a non-invasive and timely source of donor material with potential value to assess inflammatory features that impact organ quality and function.

BACKGROUND

Implant-retained maxillofacial prostheses should be biocompatible, regardless of the primers and adhesives used to bond the acrylic resin and facial silicone. The authors are unaware of any study evaluating the influence of these primers and adhesives on the biocompatibility of maxillofacial prostheses.

OBJECTIVE

The purpose of this in vitro study was to evaluate the cytotoxic effect of primers and an adhesive used to bond acrylic resin and facial silicone during the fabrication of implant-retained maxillofacial prostheses.

METHODS

Twenty-eight circular specimens made of resin and silicone were fabricated, either bonded or nonbonded with primer and adhesive. The specimens were divided into 7 groups: resin; silicone; resin+silastic medical adhesive type A+silicone; resin+DC 1205 primer silicone; resin+Sofreliner primer+silicone; resin+DC 1205 primer+silastic medical adhesive type A+silicone; and resin+Sofreliner primer+silastic medical adhesive type A+silicone. Eluates of the materials tested were prepared by setting 4 specimens of each experimental group in Falcon tubes with medium and incubating at 37°C for 24 hours. The eluate cytotoxicity was evaluated by an assay of survival/proliferation ((3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide [MTT] test) in cultures of human keratinocytes. The levels of IL1, IL6, TNFα, and the chemokine MIP-1α were evaluated by enzyme-linked immunosorbent assay. The mRNA expressions for MMP-9, TGF-β, and collagen type IV were analyzed by the real time polymerase chain reaction. Data were submitted to analysis of variance with Bonferroni post hoc tests (α=.05).

RESULTS

An increased cell proliferation was observed for the RAS group, with statistically significant differences (P<.001) compared with the unstimulated group. The RDCpS group showed the highest IL6 concentration values (P<.001). No significant statistical difference was found in the relative quantification of mRNA for collagen type IV, MMP9, or TGFβ between the groups (P>.05).

CONCLUSIONS

The RAS group showed the highest cell proliferation percentage, while the RDCpS group exhibited the highest IL6 concentration values. No detectable levels of IL1β, TNF α, or CCL3/MIP1α were observed. The tested materials showed no toxic effects on the HaCaT cell line.

Chronic obesity has increased worldwide, in conjunction with Type 2 diabetes. Chronic obesity causes systemic inflammation that may result in functional deterioration of the gastrointestinal barrier. However, gastrointestinal conditions associated with chronic obesity have not been comprehensively investigated. The purpose of this study was to evaluate morphological changes in small intestine barrier structures during chronic obesity. A mouse model of chronic obesity induced by monosodium glutamate treatment was established. At postnatal week 15, pathological changes including in small intestinal epithelial cells, were analyzed in chronically obese mice compared with controls. Numerous gaps were identified between small intestinal epithelial cells in chronically obese mice, and levels of both desmosomal and tight junction proteins were significantly lower in their small intestinal epithelial cells. Additionally, numbers of intestinal inflammatory cells, particularly macrophages, were significantly increased in chronically obese mice, as were levels of the inflammatory factors, TNF-alpha and IL1-beta, in blood samples from the mouse model. These findings suggest that functional deterioration of adhesion structures between small intestinal epithelial cells causes gastrointestinal barrier function failure, leading to a rise in intestinal permeability to blood vessels and consequent systemic inflammation, characterized by macrophage infiltration.

OBJECTIVE

Small studies suggest that fetal sex alters maternal inflammation. We examined the association between fetal sex, preeclampsia and circulating maternal immune markers.

METHODS

This was a secondary data analysis within a nested case-control study of 216 preeclamptic women and 432 randomly selected normotensive controls from the Collaborative Perinatal Project. All women had singleton, primiparous pregnancies without chronic health conditions. Logistic regression was used to calculate odds ratios (OR) and 95% confidence intervals (CI) for associations between female fetal sex and preeclampsia. Outcomes included preeclampsia, preterm preeclampsia (<37 and <34 weeks), and normotensive preterm birth <37 weeks. Associations between female fetal sex and immune markers [interleukin (IL)-6, IL4, IL5, IL1IL1IL1-beta, interferon (IFN)-gamma, tumor necrosis factor (TNF)-beta, and transforming growth factor-beta] were examined using a statistical method developed for large proportions of censored biomarker data. Models were adjusted for maternal age, race, body mass index, and smoking.

RESULTS

Women with early preterm preeclampsia (<34 weeks) had higher odds of having a female fetus (ORadj. 3.2, 95% CI 1.1-9.6) and women with normotensive preterm birth had lower odds (ORadj. 0.5, 95% CI 0.3-0.9). Female fetal sex was associated with lower first trimester pro-inflammatory IFNγ and IL-12 but higher second trimester pro-inflammatory <em>IL1</em>β and TNFβ, anti-inflammatory IL4r, and regulatory cytokines IL5 and <em>IL1</em>0. Female fetal sex was associated with higher postpartum <em>IL1</em>0 in preeclamptic women only.

CONCLUSIONS

We identified sexual dimorphism in maternal inflammation. Longitudinal studies are needed to determine if fetal sex impacts the maternal immune milieu across pregnancy.

Rationale and objectives: Adiposity and physical fitness levels are major drivers of cardiometabolic risk, but these relationships have not been well-characterized in chronic kidney disease (CKD). We examined the associations of visceral adipose tissue (VAT), subcutaneous adipose tissue (SAT), intrahepatic fat and physical function with inflammation, insulin resistance, and adipokines in patients with CKD STUDY DESIGN: Prospective cohort study Setting and Participants: Participants with Stage 3-5 CKD not receiving chronic dialysis, followed at one of 8 clinical sites in the Chronic Renal Insufficiency Cohort study, and who underwent MRI of the abdomen at an annual CRIC Study visit (n=419) PREDICTORS: VAT volume, SAT volume, intrahepatic fat, body mass index (BMI), waist circumference (WC), and time taken to complete the 400 m walk test (physical function) OUTCOMES: Markers of inflammation (IL1-beta, IL-6, TNF-R1, and TNF-R2), insulin resistance (HOMA-IR), and adipokines (adiponectin- total and high molecular weight [HMW], resistin, and leptin).

Analytical approach: Multivariable linear regression of VAT and SAT volume, intrahepatic fat and physical function, with individual markers (log-transformed values) adjusting for relevant covariates RESULTS: Mean age of the study population was 64.3 years; 41% were females, and the mean eGFR was 53.2 (+/- 14.6) ml/min/1.73 m². Over 85% were overweight or obese, and 40% had diabetes. Higher VAT volume, SAT volume, and liver proton density fat fraction, were associated with lower levels of total and HMW adiponectin, higher levels of leptin and insulin resistance, lower HDL cholesterol and higher serum triglycerides. A slower 400m walk time was associated only with higher levels of leptin, total adiponectin, plasma IL-6 and TNFR-1 and did not modify the associations between fat measures and cardiometabolic risk factors LIMITATIONS: Lack of longitudinal data and dietary details CONCLUSIONS: Various measures of adiposity are associated with cardiometabolic risk factors. Physical function was also associated with the cardiometabolic risk factors studied and does not modify the associations between fat measures and cardiometabolic risk factors. Longitudinal studies of the relationship between body fat and aerobic fitness with cardiovascular and kidney disease progression are warranted.

Keywords: Obesity; cardiometabolic risk factor; fat; kidney disease; physical function.

Background/objectives: Dysregulated inflammatory profile play an important role in COVID-19 pathogenesis. Moreover, depletion of lymphocytes is typically associated with an unfavorable disease course. We studied the role and impact of p53 and deacetylase Sirtuin 1 (SIRT1) on lymph-monocyte homeostasis and their possible effect on T and B cell signalling.

Methods: Gene expression analysis and flow cytometry were performed on peripheral blood mononuclear cells of 35 COVID-19 patients and 10 healthy donors (HD). Inflammatory cytokines, the frequency of Annexin + cells among CD3 + T cells and CD19 + B cell subsets were quantified.

Results: PBMC from COVID-19 patients had a higher p53 expression, and a higher concentrations of plasma proinflammatory cytokines (IL1β, TNF-α, IL8, IL6) than HD. Deacetylase Sirtuin 1 (SIRT1) expression was significantly decreased in COVID-19 patients, and was negatively correlated with p53 (p = 0.003 r=-0.48). A lower expression of IL-7R and B Cell linker (BLNK), key genes for lymphocyte homeostasis and function, was observed in COVID-19 compared with HD. Reduction of IgK and IgL chains was seen in lymphopenic COVID-19 patients. A significant increase of both apoptotic B and T cells were observed. Inflammatory cytokines correlated positively with p53 (IL-1β: r = 0.5, p = 0.05;IL-8: r = 0.5, p = 0.05) and negatively with SIRT1 (IL1-β: r=-0.5,p = 0.04;TNF-α: r=-0.4, p = 0.04).

Conclusions: Collectively, our data indicate that the inflammatory environment, the dysregulated p53/SIRT1 axis and low expression of IL7R and BLNK may impact cell survival, B cell signalling and antibody production in COVID-19 patients. Further studies are required to define the functional impact of low BLNK/IL7R expression during SARS-COV2 infection.

Keywords: BLNK, Inflammation; COVID-19; IL-7R; p53/SIRT1.

BACKGROUND

More than 90% of malignant tumors of the head and neck are oral squamous cell carcinomas. Early detection of oral squamous cell carcinoma using salivary biomarkers could prevent malignant transformations and enhance patient survival.

METHODS

A systematic search in MEDLINE and the Central Register of Controlled Trials and meta-analysis were undertaken to identify the screening potential of six salivary biomarkers for early oral squamous cell carcinoma detection: interleukins IL-8 and IL1-β, DUSP-1 and S100P messenger RNAs, and miR125a and miR200a microRNAs.

RESULTS

The sensitivities of IL-8 (0.41; 95%CI 0.19-0.99), IL1-β (0.26; 95%CI 0.19-0.99), DUSP-1 (0.61; 95%CI 0.01-0.98), and S100P (0.67; 95%CI 0.32-0.99) were calculated. Specificities of the biomarkers analyzed were found to be IL-8 (0.69; 95%CI 0.66-0.99), IL1-β (0.47; 95%CI 0.46-0.90), DUSP-1 (0.75; 95%CI 0.33-1), and S100P (0.73; 95%CI 0.18-0.99).

CONCLUSIONS

Early detection of oral squamous cell carcinoma was best achieved by screening for salivary messenger RNA DUSP-1 and S100P. Further investigation is required into miRNAs as novel biomarkers.

BACKGROUND

Polymorphisms of genes controlling cytokine production have not been studied in the genetic susceptibility to cutaneous adverse drug reactions (CADR).

OBJECTIVE

The objective was to determine whether polymorphisms in nine cytokine genes were associated to the occurrence of drug reaction with eosinophilia and systemic symptoms (DRESS) compared to drug-induced maculopapular eruption or urticaria and to controls without drug intolerance.

METHODS

Results from 118 patients with a well-defined CADR were compared to 236 controls without drug intolerance living in the same area of France. We assessed nine polymorphisms: interleukin (IL)1-alpha-889C>T (rs 1800587), IL1-beta-511C>T (rs 16944), IL1-RN intron-2-VNTR (rs2234663), IL2-330T>G (rs 2069762), IL4-33C>T (rs 2070874), IL5-745C>T (rs 2069812), IL1A (rs 1800872), IL1C (rs 4778889) and tumour necrosis factor-alpha-308G>A (rs 1800629).

RESULTS

Three polymorphisms exhibited a significant association with CADR (P < 0.05). The combination of the <em>IL1</em>-RN-A2 and <em>IL1</em>-<em>beta</em>-511C alleles was statistically different between cases and controls (P = 0.007) and the A2C haplotype was associated with susceptibility to CADR, particularly in drug reaction with eosinophilia and systemic symptoms (DRESS) patients (odds ratio = 3.22; 95% confidence interval = 1.23-8.41; P = 0.016). The frequency of the <em>IL1</em>0-592A allele was higher in DRESS patients than in controls (dominant model CC vs. CA + AA: P = 0.035). These abnormalities were not evident in maculopapular eruptions or urticaria.

CONCLUSIONS

This is the first study showing that <em>IL1</em>-cluster polymorphisms and haplotypes and the <em>IL1</em>0-592A allele (a low <em>IL1</em>0 producer) are associated with DRESS. These gene variants may decrease drug tolerance and promote herpes virus reactivation.

Background: Current therapeutic options for autoimmune hepatitis (AIH) are limited by adverse events associated with corticosteroids and thiopurines and the limited evidence base for second- and third-line treatment options. Furthermore, current treatment approaches require long-term exposure of patients to pharmacological agents. There have been significant advances in the understanding of the mechanisms underpinning autoimmunity and an expansion in the available therapeutic agents for suppressing autoimmune responses or potentially restoring self-tolerance.

Aim: To review the mechanisms and evidence for experimental therapies that are being actively explored in the management of AIH.

Methods: We have reviewed the literature relating to a range of novel therapeutic immunomodulatory treatment strategies and drugs.

Results: Drugs which block B cell-activating factor of the tumour necrosis factor family (BAFF) and tumour necrosis factor α are currently in clinical trials for the treatment of AIH. Experimental therapies and technologies to increase immune tolerance, such as pre-implantation factor and regulatory T cell therapies, are undergoing development for application in autoimmune disorders. There is also evidence for targeting inflammatory pathways to control other autoimmune conditions, such as blockade of IL1 and IL6 and Janus-associated kinase (JAK) inhibitors.

Conclusions: With the range of tools available to clinicians and patients increasing, it is likely that the therapeutic landscape of AIH will change over the coming years and treatment approaches offering lower corticosteroid use and aiming to restore immune self-tolerance should be sought.

Clostridium difficile (CD) is considered a major health care problem both in developing and developed countries; frequently reported to be associated with enterocolitis and diarrhea in horses and other animals. In this study, we examined acute phase response (APR), cytokines response, neopterin (NP) procalcitonin (PCT) production and oxidative stress condition in horses and foals with C. difficile-induced enterocolitis (CDIE) and evaluated the effectiveness of these parameters as biomarkers for the disease. A total of 407 Arabian horses in 35 stables were examined between January 2017 to December 2018. Only 24 out of 407 horses showed two or more signs of CDIE. The blood level of serum amyloid A (SAA), haptoglobin (HP), proinflammatory cytokines (TNF-α, IL-6 and IL1-β), serum malondialdehyde (MDA), PCT and NPT in horses with CDIE were higher than in healthy horses. Nevertheless, the levels of nitric oxide (NO), superoxide dismutase (SOD) and total antioxidant concentration (TAC) were considerably lower in diseased horses compared to those that were healthy. The ROC curves for eleven selected blood parameters, both in healthy horses and horses with CDIE demonstrated that all examined blood markers had significant levels of differentiation between CDIE cases and healthy controls (AUC > 87.5). The data in this study suggest that the evaluation of acute-phase proteins, cytokines, PCT, NPT, and oxidative stress biomarkers may well be used as a tool for diagnosis and assessment of CDIE and in disease pathogenesis in Arabian horses.

Keywords: Cytokines; Haptoglobin; Neopterin; Oxidative stress; Procalcitonin; Serum amyloid A.

Edwardsiella tarda phage (ETP-1) was isolated from marine fish farm water to characterize its effect against pathogenic multidrug-resistant E. tarda. According to transmission electron microscopy results, ETP-1 is classified as a member of the family Podoviridae. ETP-1 showed MOI dependent E. tarda growth inhibition, a latent period of 60 min, and burst size of 100 PFU per infected cells. In host range tests, five out of eight E. tarda strains were sensitive to ETP-1 which had efficiency of plating index in the range 1-1.28. ETP-1 was stable over a broad range of pH and temperature. The size of the ETP-1 genome was predicted to be approximately 40 kb. Zebrafish exposed to ETP-1 showed no adverse gene responses to the inflammatory mediator cytokines, il1-β, tnf-α, il-6, and il-10, the chemokine, cxcl-8a, and reactive oxygen species, sod-1. When zebrafish were bath exposed to ETP-1 for 12 days and simultaneously challenged with E. tarda (1.08 × 10⁵ CFU fish^-1), the survival rate was higher in phage exposed fish (68%) compared to that of the control (18%) until 4 days post challenge. Our results suggest that ETP-1 can be used as a potential bio-therapeutic candidate to control multi-drug resistant E. tarda infection in aquaculture.

Multisystem inflammatory syndrome in children (MIS-C) is a life-threatening disease occurring several weeks after severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Deep immune profiling showed acute MIS-C patients had highly activated neutrophils, classical monocytes and memory CD8+ T-cells; increased frequencies of B-cell plasmablasts and double-negative B-cells. Post treatment samples from the same patients, taken during symptom resolution, identified recovery-associated immune features including increased monocyte CD163 levels, emergence of a new population of immature neutrophils and, in some patients, transiently increased plasma arginase. Plasma profiling identified multiple features shared by MIS-C, Kawasaki Disease and COVID-19 and that therapeutic inhibition of IL6 may be preferable to IL1 or TNF-α . We identified several potential mechanisms of action for IVIG, the most commonly used drug to treat MIS-C. Finally, we showed systemic complement activation with high plasma C5b-9 levels is common in MIS-C suggesting complement inhibitors could be used to treat the disease.

OBJECTIVE

Hydrogen sulfide (H(2) S) is one of two volatile sulfur compounds that are known to be the main cause of oral malodor; the other is methyl mercaptan. Other known volatiles existing in mouth air do not contribute significantly to oral malodor originating in the oral cavity. Hydrogen sulfide is also known to be an etiological factor in periodontal disease. However, the effects of H(2) S on alveolar bone remain unclear. The objectives of this study were to determine the apoptotic effects of H(2) S on osteoblasts and to verify the apoptotic molecular pathways.

METHODS

A clonal murine calvaria cell line was incubated with 50 ng/mL of H(2) S. To detect apoptosis, the cells were analysed by flow cytometry and ELISA. Mitochondrial membrane depolarization was assessed using flow cytometry as well. ELISA was used to evaluate the release of cytochrome c into the cytosol and to assess Fas ligand, p53, tumor necrosis factor α, interleukin IL1-α IL-β, IL-2, IL-4, IL-10, interferon-γ, granulocyte-colony stimulating factor and granulocyte-macrophage colony stimulating factor. Caspase-3, -8 and -9 activities were estimated. Expression of BAX and Bcl-2 was assessed by real-time quantitative RT-PCR. DNA fragmentation was detected by single-cell gel electrophoresis. Fas receptors were evaluated by western blotting.

RESULTS

After H(2) S incubation, apoptotic levels increased significantly in a time-dependent manner. Mitochondrial membrane depolarization, the release of cytochrome c, p53 and caspase-3, -8 and -9 and DNA fragmentation were all significantly greater. BAX gene activity was upregulated, whereas Bcl-2 remained low. Fas ligand/Fas receptor, tumor necrosis factor α and other cytokines were not increased to a significant degree.

CONCLUSIONS

At less-than-pathological concentrations in gingival crevicular fluid, H(2) S induces apoptosis in osteoblasts. The molecular mechanisms underlying the apoptotic process include p53, a mitochondrial pathway and caspase-8 activation.

A bioassay for bovine interleukin-1 (IL1) activity is described. The assay is based on the IL1-stimulated proliferation of a mouse T-lymphocyte cell line, D10(N4)M. Bovine mononuclear cells stimulated with lipopolysaccharide produce an interleukin-1-like activity which stimulated the growth of the D10(N4)M cell line in a dose-dependent manner. The stimulatory activity was neutralised by a combination of both anti-human IL1 alpha and anti-human IL1 beta sera. The quantity of IL1-like activity released from the mononuclear cells increased asymptotically with increasing lipopolysaccharide dose.

OBJECTIVE

To investigate the correlation of gut epithelial cell apoptosis with ICE, IL1 beta gene expression in septic mice.

METHODS

Sepsis was induced in mice by cecal ligation and puncture(CLP). Sham-operation group underwent the same manipulation but without CLP. 1, 3, 6 hours after CLP, gut epithelial cells, were isolated. IL1 beta, ICE gene expression was detected quantitatively by RT-PCR. Epithelial cell apoptosis was assessed by flow cytometric method and biochemically by DNA electrophoresis.

RESULTS

The survival rate of CLP mice was 1/10 as compared to 10/10 of sham-operation mice. IL1 beta, ICE mRNA expression in CLP mice was significantly higher than that in the sham-operation group(P < 0.01); IL1 beta mRNA expression was parallel to ICE mRNA expression. The number of epithelial cell apoptosis was correlated excellently to the level of ICE, IL1 beta mRNA expression. Epithelial cell apoptosis could not be detected in sham-operation group at the indicated time points.

CONCLUSIONS

ICE, IL1 beta gene overexpression may be involved in the vulnerability of epithelial cell apoptosis in septic mice.

Disturbance of capillary perfusions due to leukocyte adhesion, disseminated intravascular coagulation, tissue edema is critical components in the pathophysiology of sepsis. Alterations in brain microcirculation during sepsis are not clearly understood. The aim of this study is to gain an improved understanding of alterations through direct visualization of brain microcirculations in an experimental endotoxemia using intravital microscopy (IVM). Endotoxemia was induced in Lewis rats with Lipopolysaccharide (LPS, 15 mg/kg i.v.). The dura mater was removed via a cranial window to expose the pial vessels on the brain surface. Using fluorescence dyes, plasma extravasation of pial venous vessels and leukocyte-endothelial interaction were visualized by intravital microscopy 4 h after LPS administration. Plasma cytokine levels of IL1-beta, IL-6, IFN-gamma, TNF-alpha and KC/GRO were evaluated after IVM. A significant plasma extravasation of the pial venous vessels was found in endotoxemia rats compared to control animals. In addition, a significantly increased number of leukocytes adherent to the pial venous endothelium was observed in septic animals. Endotoxemia also induced a significant elevation of plasma cytokine levels of IL1-beta, IL-6, IFN-gamma, TNF-alpha and KC/GRO. Endotoxemia increased permeability in the brain pial vessels accompanied by an increase of leukocyte-endothelium interactions and an increase of inflammatory cytokines in the plasma.

Interleukin 1 beta (IL1 beta) is an inducible polypeptide with many roles in host defence and homoeostasis. It has also been implicated as a mediator of infectious, inflammatory and autoimmune diseases, and the kinetics of its production are relevant to an understanding of the pathogenesis of these conditions. We report here the time-course of IL1 beta production in human adherent monocytes. Both IL1 beta protein and mRNA were measured following cell activation with bacterial endotoxin (lipopolysaccharide; LPS), and pro-inflammatory crystals of monosodium urate (MSU), which cause arthritis and kidney disease. We also tested other crystal types associated with arthritis, namely hydroxylapatite and calcium pyrophosphate dihydrate. IL1 was absent from unstimulated cells, but IL1 beta mRNA accumulated rapidly after LPS or MSU stimulation and was associated with the later appearance of intracellular IL1 beta protein which was subsequently released from the cells (60% at 9 h). The other crystals failed to induce significant IL1 production. Our findings support the view that production of IL1 beta in human mononuclear cells is based on rapid translation of an inducible pool of mRNA and that no pre-formed mRNA or intracellular protein exists in normal blood monocytes. Further, although IL1 beta is translated without a conventional leader sequence, it is translocated extracellularly with the kinetics of a secretory protein.