IL-1ra Concentrations Research Articles

Cytokine markers of clinical variants of infective endocarditis

Introduction. Infective endocarditis is a bacterial disease. Pathogen is localized mainly on heart valves and endocardium. This condition is accompanied by immunopathological manifestations and potential generation of septic process being unfavorable prognosis for disease outcome. Currently, the causes of death of patients with endocarditis have been increasingly presented as thromboembolic complications, which severity depends on variant of the disease course. An important role in imbalanced immune system in the infectious process is assigned to altered intercellular interaction mediated via cytokine network. Activated immune cells produce cytokines, which investigating is important in terms of interpreting changes in immune system functionality, assessing the severity of diseases and controlling therapeutic effectiveness. Infective endocarditis remains a severe disease associated with high mortality despite current advances in diagnostics and treatment. A timely diagnostic process and early initiation of treatment are major factors for successful patient management necessitating to improve diagnostics of various clinical variants of endocarditis course, taking into account pathogenetically relevant cytokines. Objective was to comparatively evaluate the importance of serum cytokine concentrations in patients with uncomplicated course of infective endocarditis and its thromboembolic complications and determine cytokine markers of various variants of endocarditis course. Materials and methods. An immunological examination of 119 blood serum samples from patients with confirmed diagnosis of infectious endocarditis and 20 samples from apparently healthy persons was carried out. Depending on the clinical disease form, the patients were divided into 4 groups: group 1 primary infective endocarditis (PIE) with thromboembolic complications (n = 24), 2 PIE without thromboembolic complications (n = 34), 3 secondary IE with thromboembolic complications (n = 27), 4 secondary IE without thromboembolic complications (n = 34). The control group consisted of 20 apparently healthy subjects. Immunological studies of serum cytokine concentrations were conducted in all groups. Results. Statistically significant increase in serum concentration of IL-10, IL-6, VEGF-A, IL-18, IL-1Ra and IL-8 was revealed in all clinical groups of patients compared to those in control group (p 0,05). By correlation analysis, we found a significant positive relationship between IL-10 and IL-18 or IL-6. An increase in the concentration of IL-10 leads to increased level of pro-inflammatory IL-18 and IL-6. The marker of secondary endocarditis was observed as increased level of serum concentrations of IFN. A characteristic feature of primary infective endocarditis with thromboembolic complications was revealed as significantly increased serum concentration of IL-8, IL-1Ra and IL-6. Markers of secondary complicated endocarditis were identified as increased level of IL-6, VEGF-A and IL-18.

Articular fragment restoration is critical to mitigate post-traumatic osteoarthritis in a porcine pilon fracture model

ObjectiveDuring articular fracture reconstruction, orthopedic surgeons are frequently faced with the dilemma of retaining small articular fragments versus discarding these fragments. The purpose of this study was to compare post-traumatic osteoarthritis (PTOA) development between tibial plafond fractures and plafond fractures with a missing articular fragment (MF) in a porcine model. DesignHigh-energy tibial plafond fractures in skeletally mature Yucatan mini pigs (n ​= ​12) were created. During surgery, a 3 ​× ​3 mm section of the articular surface was removed in six animals (MF group). Ankle synovial fluid was analyzed for IL-1β, IL-1Ra, IL-6, IL-8, and IL-10 concentrations obtained at initial surgery and 12 weeks post-surgery. Plafond and talus sections were evaluated for subchondral bone porosity and stained with Sanderson's Rapid Bone Stain and blindly evaluated to determine the Osteoarthritis Research Society International (OARSI) grade and vascular invasion. ResultsFractured ankles had greater concentrations of IL-1β, IL-1Ra, IL-6, IL-8, and IL-10 compared to control ankles. There was no difference in cytokine concentrations between fractured and fractured ​+ ​MF ankles. Fractured ankles had significantly greater bone porosity, vascular invasion, and OARSI grade as compared to the control group. In comparing tibial plafonds, the MF group had significantly more bone porosity, more vascular invasion, and a higher average OARSI grade than the anatomically reconstructed group. In comparing the talus, the MF group had higher average OARSI grade and similar bone porosity. ConclusionsArticular fractures with a MF had worse PTOA development as measured by bone porosity, vascular invasion, and OARSI grade than the anatomically reconstructed fractures.

CD4 Tcells are rapidly depleted from tuberculosis granulomas following acute SIV co-infection.

HIV/Mycobacterium tuberculosis (Mtb) co-infected individuals have an increased risk of tuberculosis prior to loss of peripheral CD4 Tcells, raising the possibility that HIV co-infection leads to CD4 Tcell depletion in lung tissue before it is evident in blood. Here, we use rhesus macaques to study the early effects of simian immunodeficiency virus (SIV) co-infection on pulmonary granulomas. Two weeks after SIV inoculation of Mtb-infected macaques, Mtb-specific CD4 Tcells are dramatically depleted from granulomas, before CD4 Tcell loss in blood, airways, and lymph nodes, or increases in bacterial loads or radiographic evidence of disease. Spatially, CD4 Tcells are preferentially depleted from the granuloma core and cuff relative to B cell-rich regions. Moreover, live imaging of granuloma explants show that intralesional CD4 Tcell motility is reduced after SIV co-infection. Thus, granuloma CD4 Tcells may be decimated before many co-infected individuals experience the first symptoms of acute HIV infection.

Comparison of the Impacts of a Dynamic Exercise Program vs. a Mediterranean Diet on Serum Cytokine Concentrations in Women With Rheumatoid Arthritis. A Secondary Analysis of a Randomized Clinical Trial.

BackgroundRheumatoid arthritis (RA) is a disease characterized by a chronic inflammatory state. High pro-inflammatory cytokine levels are associated with disease activity. Exercise and the Mediterranean diet (MD) exert anti-inflammatory effects; however, their impacts on inflammation in RA patients remains unknown. This study aimed to compare the effects of six-months of dynamic exercise program (DEP) vs. MD on pro- and anti-inflammatory cytokine serum concentrations.MethodsSecondary analysis of a randomized clinical trial in which 90 women with RA were randomly assigned to the DEP (n = 30), MD (n = 30), or control group (n = 30). All patients received pharmacological treatment. Serum concentrations of pro-inflammatory (TNF-α, TNF-β, IL-1β, IL-6 pg/mL) and anti-inflammatory (IL-10, IL-Ra pg/mL) cytokines were measured at baseline and after 6 months using the Luminex technique.ResultsAfter 6 months of follow-up, we found an improvement of the median percentages changes concentrations of TNF-α (DEP, −12.3; MD, −13.3; control, 73.2; p = 0.01), TNF-β (DEP, −67.4; MD, −54.9; control, 0; p = 0.04), and IL-6 (DEP, −19.9; MD, −37.7; control, 45.5; p = 0.04) in the DEP and MED groups in comparison with control group. IL-1Ra concentrations increased only in the MD group (13.8) compared to levels in the control group (−31.7), p = 0.04. There were no statistically significant differences between DEP and MD groups. Only n = 27 participants in the DEP group, n = 26 in the MD group, and n = 21 in the control group completed the follow-up.ConclusionThe DEP and the MD have potential effects in the concentrations of pro-inflammatory cytokines compared with those in a control group. Only the MD elevated the concentration of IL-Ra.Clinical Trial Registration[ClinicalTrials.gov], identifier [NCT02900898].

Cell saver blood transfusions may be associated with a decrease in inflammation and improved outcome measures in pediatric cardiac surgery patients.

Cardiopulmonary bypass (CPB) is a requisite for correction of congenital heart disease by open-heart surgery and induces a systemic inflammatory response that can lead to complications such as acute lung injury and acute kidney injury. In addition, blood transfusions are commonly required for this type of surgery, and they may further exacerbate this inflammatory response and increase morbidity and mortality. We hypothesized that, in contrast to red blood cells, intraoperative cell saver (CS) blood transfusions attenuate the post-CPB proinflammatory cytokine response. Serum cytokine concentrations of IL-10, IL-1RA, IL-6, IL-8, and TNF-α were measured at four time points (preoperatively and postoperatively on postoperative days 0, 1, and 2). Anti-inflammatory IL-10 levels were significantly lower in the CS group on POD 0 than in the control group (mean 1083.2pg/mL vs 2080.2pg/mL, 95%CI 357.4-1636.6, p = .0026). Of the clinical parameters measured, mean BUN and creatinine levels on POD 2 were significantly lower in the CS group (13.79 vs 21.88, p = .004 and 0.45 vs 0.55, p = .055, respectively). In addition, the duration of milrinone use decreased by 80% in the CS group (0.20, 95%CI 0.04, 0.94; p = .048), the median time to extubation in hours was significantly lower in the CS group (3.5 vs 6.5; 95%CI -38.00, -0.50; p = .026), and hospital length of stay was decreased by 60% in the CS group (p = .003). CS transfusions in children may lower postoperative anti-inflammatory IL-10 levels, possibly due to an overall decrease in proinflammatory state, and may be associated with improvements in renal and pulmonary functions.

Comparing Effects of Polypharmacy on Inflammatory Profiles in Older Adults and Mice: Implications for Translational Aging Research.

Aging and multimorbidity are associated with inflammation. Polypharmacy is common in older people with multimorbidity. Given the potential for interactions between polypharmacy and inflammation, the relationship between inflammation and polypharmacy were studied in older adults with multimorbidity and in healthy aging mice. A cross-sectional analysis of data from the 5-year wave of the Concord Health and Ageing in Men Project, a population-based study of community-dwelling men aged ≥70 years. Serum concentrations of 27 cytokines were measured using a multiplex immunoassay. Associations between polypharmacy (≥5 medications) and cytokines were evaluated using multivariable linear regression adjusting for age, frailty, comorbidities, and individual drug classes. Interaction between polypharmacy and Drug Burden Index (DBI―drugs with anticholinergic and sedative effects) was analyzed. Effects of polypharmacy and DBI on serum levels of 23 cytokines were determined in aging male mice treated with chronic polypharmacy or control. Compared to the nonpolypharmacy group (n = 495), CHAMP participants with polypharmacy (n = 409) had significantly higher concentrations of IL-8, IL-6, CCL3, Eotaxin, IL-1ra, IL-1β, IP-10, and lower concentrations of anti-inflammatory cytokine IL-4. In fully-adjusted multivariable models, polypharmacy was positively associated with concentrations of IL-8 and CCL3. There were no significant differences in inflammatory profiles between control and polypharmacy-treated mice. The relationship was not influenced by DBI in men or in mice. Inflammatory markers associated with polypharmacy in older adults were not seen in healthy aged mice administered polypharmacy, and may be related to underlying diseases. The polypharmacy mouse model provides opportunities for mechanistic investigations in translational research.

Dry Eye Disease in Patients With Schizophrenia: A Case-Control Study.

ObjectiveTo evaluate the clinical features and inflammatory cytokines of dry eye disease (DED) in patients with schizophrenia.MethodsThis is a case-control study. The modified self-rating depression scale (M-SDS) and the ocular surface disease index (OSDI) were used to evaluate the symptoms of depression and DED, respectively. Lipid layer thickness (LLT), partial blink rate (PBR), meibomian gland loss (MGL), tear break-up time (TBUT), corneal fluorescein staining, Schirmer I-test, and eyelid margin abnormalities were also measured. A multiplex ELISA Quantibody array was used to detect the inflammatory cytokines in the tears of all participants.ResultsForty schizophrenic patients and 20 control subjects were included. The mean age was 45.0 ± 9.5 years (range, 22–63 years) in schizophrenic patients and 45.4 ± 16.2 years (range, 23–76 years) in controls (P = 0.914). The ratio of male to female was 1.1 in schizophrenic patients and 1.0 in controls (P = 0.914). Ten women (52.6%) with schizophrenia and 2 (20%) in the control group (P = 0.096) were menopausal or post-menopausal. The OSDI [0.0 (0.0–4.2) vs. 7.3 (2.1–14.6)] and TBUT [4.5 (3.0–6.0) vs. 10.0 (3.5–11.0)] were significantly lower in patients with schizophrenia than in controls (P = 0.003 and P = 0.009, respectively). The rate of MGL [36.5 (17.5–47.5) vs. 8.5 (0.0–17.5)] increased in schizophrenic patients (P < 0.001). Among pro-inflammatory cytokines, the levels of interleukin (IL)-1α, IL-6, IL-11, IL-12A, IL-15, IL-17A, and granulocyte colony-stimulating factor (G-CSF) in tears were elevated in the schizophrenia group (all P < 0.01). Most of the chemokines examined were at increased levels in the tears of schizophrenics (all P < 0.05). The levels of matrix metalloproteinases-9 (MMP-9) and intercellular adhesion molecule-1 (ICAM-1) were also higher in the schizophrenic patients (all P < 0.001). The concentrations of IL-1Ra, tissue-inhibitor of metalloproteinase-1 (TIMP-1), and TIMP-2 in the schizophrenia group were decreased (all P < 0.001). In schizophrenic patients, the level of CCL2 in tears was positively correlated with OSDI (R = 0.34, P = 0.03). The increasing TIMP-1 and decreasing IL-5 were correlated with increasing LLT (R = 0.33, P = 0.035; R = −0.35, P = 0.027, respectively). The level of ICAM-1 was then positively correlated with partial blink rate (PBR) (R = 0.33, P = 0.035). There was a negative correlation between IL-8 and the Schirmer I-test (R = −0.41, P = 0.009).ConclusionsPatients with schizophrenia were more likely to experience asymptomatic DED, with mild symptoms and obvious signs. The inflammatory cytokines in the tears of schizophrenic patients differed greatly from that of non-schizophrenic patients.

Changes in the Synovial Fluid Cytokine Profile of the Knee Between an Acute Anterior Cruciate Ligament Injury and Surgical Reconstruction

Background: Changes in the intra-articular inflammatory state during the immediate period after an acute anterior cruciate ligament (ACL) rupture are not well defined. Purpose: To evaluate changes in the concentration of select proinflammatory and anti-inflammatory synovial fluid cytokines during the interval between an ACL injury and surgical reconstruction. Study Design: Descriptive laboratory study. Methods: In patients with an acute ACL injury, a synovial fluid sample was obtained from the injured knee during the initial office visit within 2 weeks of the inciting traumatic event. An additional synovial fluid sample was collected at the time of ACL reconstruction just before the surgical incision. Synovial fluid samples from both the acute injury and the surgery time points were processed with a protease inhibitor, and the concentrations of 10 cytokines of interest were measured using a multiplex magnetic bead immunoassay. The primary outcome was the change in cytokine concentrations between time points. Results: A total of 20 patients with a mean age of 30.2 ± 8.3 years were included. The acute injury synovial fluid samples were collected at 6.6 ± 3.8 days after the injury. The surgical synovial fluid samples were collected at 31.6 ± 15.6 days after the acute injury samples. Based on a series of linear mixed-effects models to control for the effect of concomitant meniscal injuries and by-patient variability, there was a statistically significant increase in the concentrations of RANTES and bFGF and a statistically significant decrease in the concentrations of IL-6, MCP-1, MIP-1β, TIMP-1, IL-1Ra, and VEGF between time points. Conclusion: This study demonstrates the ongoing alterations in the intra-articular microenvironment during the initial inflammatory response in the acute postinjury period. We identified 6 synovial fluid cytokines that significantly decreased and 2 that significantly increased between the first clinical presentation shortly after the injury and the time of surgery 1 month later. Clinical Relevance: This study describes the molecular profile of the inflammatory changes between the time of an acute ACL injury and the time of surgical reconstruction 1 month later. A greater understanding of the acute inflammatory response within the knee may be helpful in identifying the optimal timing for a surgical intervention that balances the risk of chondral damage with the likelihood of successful, well-healed reconstruction.

Genetic regulation of cytokine inflammation in oncohematological diseases

Objective. To analyze the correlations of the polymorphous variants of the genes the modifiers of immune response (IL1-/+3953, IL1RN*VNTR, TNFA*G-308A) with the development of oncohematological diseases (OHD) and the production of pro-and anti-inflammatory cytokines (IL-1, IL-1Ra, TNF-, INF-, IL-2, IL-4, IL-6, IL-8, IL-10, IL-18).&#x0D; Materials and methods. The examination included 100 children (57 (57 %) boys и 43 (43 %) girls, with the mean age 7.50 (2.512.60 years) suffering from malignant blood diseases. The cytokine content (IL-1, IL-4, IL-6, IL-8, IL-18, IL-1Ra и IL-10) was determined using IFA, the genetic typing of the genetic variants of the genes of cytokines IL1-/+3953, IL1RN*VNTR, TNFA*G-308A PCR and RFLP methods.&#x0D; Results. In case of lethal outcome, 14% of cases, the TNF- IL-6, IL-8, IL-18 INF- and IL-10 levels were reliably higher, compared with the survived patients. Renal function disorder detected among 13% of children was accompanied by an increase in IL-1, IL-6, IL-8, IL-18, IL-1Ra and INF- compared to the patients without nephropathy and the control group (p0.05). Eighteen OHD children with high concentration of IL-1, IL-1Ra, IL-6, IL-8, IL-18 and INF- had fractures (р0,05). Against the background of OHD, the carriage of the genotype А2А2 of the polymorphic variant VNTR IL1RN gene was observed 13 times more often, the carriage of the allele A2 2.16 times more often. The carriers of the genotype A2A2 of the genetic variant VNTR IL1RN gene had an increased risk of nephropathy by 20.89 times, the carriers of the allele A2 3.05 times more often. Children with OHD complicated by bacterial infection by 10.77 times more often had the genotype A2A2 and by 2.45 times more often the allele A2 of the genetic variant VNTR IL1RN gene.&#x0D; Conclusions. The carriers of the minor genotype A2A2 of the gene IL1RN*VNTR had a reliably higher production of the antiinflammatory IL-1, IL-6, IL-8, IL-18 и IL-1Ra. The carriers of the genotype GA of the gene TNFA*G-308A had a significantly higher values of IL-1, IL-18, IL-6, IL-8, TNF-.

Cytokine profile in the peripheral blood and the brain in patients with focal drug-resistant epilepsy

Aim. To study markers of blood-brain barrier dysfunction (BBB) in patients with pharmacoresistant epilepsy (PhRE) – the amount of VEGF in endotheliocytes of brain capillaries, TNF-α in brain tissue and cytokine profile in blood serum.Materials and methods. The study included 30 patients with PhRE who underwent anterior temporal bloc resection. Histological samples of the brain were examined to assess the amount of VEGF and TNF-α; the concentration of cytokines in the blood serum was determined.Results. In the PhRE group, the densitometric density of cells expressing VEGF and the amount of TNF-α in the epileptogenic focus were higher than in the control groups (p &lt; 0.001; p &lt; 0.05). Compared with the control, the serum concentrations of IL-2 (0.98 ± 0.28 pg/ml vs. 2.80 ± 0.71 pg/ml; p &lt; 0.001), IL-8 (14.04 ± 1.46 pg/ml vs. 26.13 ± 3.80 pg/ml; p &lt; 0.001) and EGF (43.72 ± 5.63 pg/ml vs. 83.62 ± 24.06 pg/ml; p &lt; 0.05) were statistically significantly lower in the PhRE group, and the amount of TNF-α (33.09 ± 1.23 pg/ml vs. 24.85 ± 1.32 pg/ml, p &lt; 0.05), IL-4 (43.73 ± 2.57 pg/ml vs. 32.37 ± 5.80 pg/ml, p &lt; 0.05), IL-5 (43.73 ± 2.57 pg/ml vs. 32.37 ± 5.80 pg/ml; p &lt; 0.05), IL-7 (16.65 ± 3.07 pg/ml vs. 8.13 ± 1.67 pg/ml; p &lt; 0.05), GRO (growth-regulated protein) (3054.0 ± 200.8 pg/ml vs. 1367.0 ± 187.3 pg/ml; p &lt; 0.001), VEGF (316.10 ± 55.28 pg/ml vs. 95.22 ± 15.78 pg/ml; p &lt; 0.01) are statistically significantly higher. There were no significant differences in the concentration of IL-1β, IL-1RA, IL-10 and IFN-γ between the PhRE group and the control.Conclusion. Based on the studied cytokine profile, there is no systemic inflammation in patients with PhRE. The established overexpression of VEGF in the brain and an increase in its concentration in the blood, combined with a decrease in serum EGF concentrations and an increase in GRO, as well as pro-inflammatory factors, indicates damage to the BBB. A high amount of TNF-α in the epileptic focus indicates neuroinflammation, and an increased concentration of this marker can be found in the blood of patients with BBB dysfunction.

Effect of immune drugs to treat acute viral nasopharyngitis

The task in treating acute nasopharyngitis (ANP) deals with reducing the disease symptoms and the risk of complications. The lack of reliable antiviral drugs makes it important to search for appropriate medicines among other pharmacotherapeutic groups.The study involves a comparative analysis of the efficiency and estimates potential: the recombinant interferon α2b and the compound containing fungal β-D-glucans used in treat ANPThe studies involved patients with ANP from 18 to 55 years old. As many as 152 people were examined including the following: 38 were practically healthy people (group 1); and 114 patients wuth ANP: 38 people (group 2) was subject to a standard therapy (vasoconstrictor nasal drops, nasal cavity irrigation using 0.1% Miramistine solution, gargling using the Furacilin solution); forty people (group 3) were administered application of intranasal interferon α2b of 105IU, it was delivered with a spray into each nasal passage twice a day; 36 people (group 4) were administered an immunotropic drug containing β-D-glucans orally twice a day. The duration of drug administration lasted 7 days. Polymerase chain reaction (PCR) was used to identify the ANP etiological factor. Concentrations of cytokines IL-1β, IL-1ra were estimated using enzyme immunoassay (ELISA) technique. Clinical efficiency was assessed through score approach. The following symptoms were taken into account: general malaise, sore throat, character of nasal discharge, and the difficulty of nasal breathing. The results of the study were analyzed using parametric and nonparametric statistical methods. In 60.0% the nasal secretions of patients revealed RV. The distribution of cytokine concentrations in nasal secretions in group 1 indicated that the concentration of IL-1β was in the range of 20.0-25.0 pg/ml, and the concentration of IL-1ra was about 1250.0-2500.0 pg/ml. Developing ANP stimulated an increase in IL-1β concentration up to 30.0-70.0 pg/ml in nasal secretions of patients without affecting IL-1ra concentrations. On day 7 of treatment, the cytokine concentrations among the patients treated using the immunotropic drugs were the same as in the group of healthy individuals. There were no significant changes in cytokine production on day 7 in the group of patients undergoing the standard treatment. Application of proposed immunobiological medicines to ANP does not result in overproduction of proinflammatory cytokine IL-1β in nasal secretion. This confirms that these drugs are promising in the treating strategy including reduction of the risk of developing complications.

Study on the inflammatory response in the contralateral eye after phacoemulsification combined with intraocular lens implantation in the first eye

AIM: To explore whether there is an inflammatory response in the contralateral eye after cataract surgery in the first eye, and try to explore the development of the inflammatory response. METHODS: In this paired-sample study, aqueous humor samples from 37 bilateral age-related cataract patients were collected just before each cataract surgery. Samples collected during the first-eye cataract surgery were marked as Group A, samples collected during the second-eye cataract surgery were marked as Group B. In addition, the samples were classified into five groups according to different operation intervals between the first and the second cataract surgery. Finally, the concentration of 13 selected cytokines (including TNF-α, IL-6, IL-8, CCL2/MCP-1, IL-6 Rα, IL-1 ra, MIP-1α, MIP-1d, CCL4, IL-2, CXCL9, TIMP-1, CCL11) in aqueous humor samples was detected by Luminex-200 (RnD) simultaneously in accordance with the manufacturer’s instructions. RESULTS: Among the 13 selected cytokines, there was statistically difference between the first and the second eyes in concentration of IL-6, MIP-1α, CCL2/MCP-1, TNF-α, MIP-1d, IL-2 and TIMP-1 (P&lt;0.05), and the expression levels of all these seven cytokines in the second eyes were higher than those in the first eyes, and the concentration of IL-6 and MIP-1a in the second eyes was significantly higher than that in the first eyes (P&lt;0.001). Compared with the first eyes, the concentration of CCL4, CCL11, IL-6Rα, CXCL9, and IL-1ra in the second eyes only showed slightly increase, while IL-8 showed a slight decrease, all with no statistically difference (all P&gt;0.05). The concentration differences of IL-6, MCP-1/CCL2, IL-2, MIP-1d between the first eyes and the second eyes were most obvious at the third week after phacoemulsification combined with intraocular lens implantation (PHACO+IOL) in the first eye. CONCLUSION: This study revealed that there may be an inflammatory response in the contralateral eye after cataract surgery in the first eye, and the inflammatory response in the second eye appears to be most significant at the third week after phacoemulsification combined with intraocular lens implantation (PHACO+IOL) in the first eye.

Cytokine Profiles Associated With Worse Prognosis in a Hospitalized Peruvian COVID-19 Cohort.

Cytokines, chemokines and growth factors present different expression profiles related to the prognosis of COVID-19. We analyzed clinical parameters and assessed the expression of these biomarkers in patients with different disease severity in a hospitalized Peruvian cohort to determine those associated with worse prognosis. We measured anti-spike IgG antibodies by ELISA and 30 cytokines by quantitative suspension array technology in 123 sera samples. We analyzed differences between patients with moderate, severe and fatal COVID-19 by logistic regression at baseline and in longitudinal samples. Significant differences were found among the clinical parameters: hemoglobin, neutrophils, lymphocytes and C-reactive protein (CRP), creatinine and D-dimer levels. Higher anti-spike IgG antibody concentrations were associated to fatal patient outcomes. At hospitalization, IL-10, IL-6, MIP-1α, GM-CSF, MCP-1, IL-15, IL-5, IL1RA, TNFα and IL-8 levels were already increased in fatal patients´ group. Meanwhile, multivariable analysis revealed that increased GM-CSF, MCP-1, IL-15, and IL-8 values were associated with fatal outcomes. Moreover, longitudinal analysis identified IL-6 and MCP-1 as the main risk factors related to mortality in hospitalized COVID-19 patients. In this Peruvian cohort we identified and validated biomarkers related to COVID-19 outcomes. Further studies are needed to identify novel criteria for stratification of SARS-CoV-2 infected patients at hospital entry. BackgroundIn the most severe forms of SARS-CoV-2 infection, large numbers of innate and adaptive immune cells become activated and begin to produce pro-inflammatory cytokines, establishing an exacerbated feedback loop of inflammation.MethodsA total of 55 patients with laboratory-confirmed COVID-19 admitted to the Hospital Nacional Guillermo Almenara Irigoyen in Lima, Peru were enrolled during August-October 2020. Of these, 21 had moderate disease, 24 severe diseases and 10 died. We measured 30 cytokines and chemokines by quantitative suspension array technology and anti-spike IgG antibodies using a commercial ELISA. We evaluated these parameters in peripheral blood every 2-5 days until patient discharge or death. Patient information and clinical parameters related were obtained from the respective clinical histories.ResultsThe frequency of obesity differed among the 3 groups, being most frequent in patients who died. There were also significant differences in clinical parameters: hemoglobin, segmented neutrophils, lymphocytes,C-reactive protein, creatinine and D-dimer levels. Greater anti-spike IgG antibody concentrations were associated to fatal outcomes. In univariate analyses, higher baseline concentrations of IL-6, MIP-1α, GM-CSF, MCP-1, IL-15, IL-5, IL1RA, TNFα, IL-8 and IL-12p70 correlated with severity, while multivariable analysis showed that increased concentrations in 4 biomarkers (GM-CSF, MCP-1, IL-15, IL-8) were associated with fatal outcomes. Longitudinal analysis showed IL-6 (hazard ratio [HR] 6.81, 95% confidence interval [CI] 1.6-28.7) and MCP-1 (HR 4.61, 95%CI 1.1-19.1) to be related to mortality in hospitalized COVID-19 patients.ConclusionsCytokine, chemokine and growth factor profiles were identified and validated related to severity and outcomes of COVID-19. Our findings may be useful to identify novel criteria for COVID-19 patient stratification at hospital entry.

Time Course of Regional Sweat Cytokine Concentrations During Steady State Exercise

Recently, there has been significant interest in the use of sweat as a non-invasive diagnostic biofluid. Research to date has focused primarily on electrolytes, micronutrients, or metabolites as markers of hydration status or metabolic stress, while fewer studies have measured messenger molecules like cytokines and chemokines. The purpose of this study was to determine time course and regional differences in sweat EGF, IFN-γ, IL-1β, IL-1α, IL-1RA, TNF-α, IL-8, IL-10, and IL-6 concentrations during steady state exercise. Eight moderately trained subjects (34±7 y, 80.2±10.2 kg, 52.2±6.4 ml/kg/min VO2max) completed 90 min of cycling at 82±6 %HRmax in the heat (32°C, 50% RH). Preceding exercise, right dorsal forearm (RDF), right triceps (RT), right scapula (RS) and forehead (FH) were cleaned with alcohol and deionized water. Absorbent patches (10 cm2 absorbent pad, 3M Tegaderm™ + Pad) were applied at 0, 30, and 60 minutes into exercise. The patches were removed at 25, 55, and 85 minutes into exercise. Sweat cytokines were measured using Multiplex (EMD Millipore, MagPix) customized kits. Kruskal-Wallis was used to test differences among groups. Dunn's test for multiple comparisons was used for post-hoc analysis. Cytokine measurements are shown as median ± IQR. There were no differences in IL-1β or IL-10 across sites or across time. The majority of samples (62%) were below detection for IFNγ and TNFα. IL-1α (3413±4564 pg/mL; p<0.0001), IL-1RA (343.4±717.2 pg/mL; p=0.0001), IL-6 (0.12±0.03 pg/mL; p=0015), and IL-8 (0.21±2.1 pg/mL; p<0.001) varied across sites. There were specific differences between IL-1α FH (1007±1465 pg/mL) compared to RDF (3785±2354 pg/mL; p=0.0001), RS (5860±9491 pg/mL; p<0.0001), and RT (4236±9877 pg/mL; p<0.0001). There were also specific differences when FH IL-1RA (876.6±1146.6 pg/mL) was compared to RDF (182.1±257.3 pg/mL; p=0.0013) and RS (91.6±385.6 pg/mL; p=0.0004). FH IL-6 (0.11±0.03 pg/mL) showed significant differences when compared to RDF (p=0.03), RS (p=0.02), and RT (p=0.002). FH IL-8 (4.9±9.9 pg/mL) also showed significant differences when compared to RDF (p<0.0001), RS (p=0.0001), and RT (p=0.0001). FH IL-1α varied across time (2274±2191, 864.9±1418, 589.5±1014 pg/mL at 25, 55, and 85 min respectively; p=0.046), with specific differences between 25 and 85 min (p=0.049). EGF varied across time for all skin regions (FH p=0.0001; RDF p=0.01; RS=0.003; RT p=0.009) with significant differences between 25 and 85 min for all sites (FH p<0.0001; RDF p=0.009; RS p=0.003; RT p=0.01). RS EGF also showed significant differences between 25 and 55 min (p=0.04). In conclusion, EGF decreased significantly from 25 to 85 min for all sites. There was significant regional variation in IL-1α, IL-1 RA, IL-6, and IL-8, especially when FH was compared to RDF, RS, and RT. This study suggests that cytokine concentrations vary across skin regions and sample collection timing which could impact the usage of sweat as a diagnostic tool. Disclosure Statement: All authors are employed by the Gatorade Sports Science Institute, a division of PepsiCo, Inc. The views expressed in this abstract are those of the authors and do not necessarily reflect the position or policy of PepsiCo, Inc.

Amnion and Umbilical Cord–Derived Products in Sports Medicine: From Basic Science to Clinical Application

Background: Birth tissue products from amnion, chorion, umbilical cord, amniotic fluid, or cord blood are frequently marketed as viable sources of stem cells and growth factors. It can be difficult for health care professionals to differentiate implied from explicit conclusions in reported product analyses. Purpose: To provide an educational platform for health care professionals to interpret data presented in the promotion of birth tissue products. Study Design: Descriptive laboratory study and expert opinion; Level of evidence, 5. Methods: A cord blood product was analyzed by 3 methods for cell viability, 2 methods for assessment of cell morphology and cell type, multicolor flow cytometry to identify stem cells, and enzyme-linked immunosorbent assay (ELISA) plus Western blot for analysis of interleukin 1 receptor antagonist protein (IL-1ra). These data were compared with analyses reported by the manufacturer. Results: Cell viability in the cord blood product was less than reported by the manufacturer, the cells were primarily leukocytes, no stem cells were present, and the concentration of IL-1ra was falsely increased due to nonspecific antibody binding in the sample. Conclusion: To assess birth tissue products, health care professionals should consider the following: (1) Understanding fluorescent dyes is important for assessing cell viability data—green does not always mean alive. (2) The report of “cells” in the product does not necessarily mean “stem cells”; microscopic images of at least ×20 or a hemogram should be evaluated to determine cell type (leukocyte, red blood cells, etc). (3) There is no single cluster of differentiation (CD) marker on flow cytometry to identify stem cells. (4) Biological tissues are complex substances, and inaccurately increased measurements of growth factors could be present in ELISA results because most ELISAs are not designed or validated for use in biologics. Furthermore, the reported measurement of growth factors should be considered relative to concentrations in native biological tissues and plasma. Clinical Relevance: Health care professionals should be able to interpret cell viability, cell morphology, stem cell analysis using CD markers, and growth factor analysis when considering use of a birth tissue product in patients.

Innate immunity impacts social-cognitive functioning in people with multiple sclerosis and healthy individuals: Implications for IL-1ra and urinary immune markers

Social-cognitive difficulties can negatively impact interpersonal communication, shared social experience, and meaningful relationships. This pilot investigation examined the relationship between social-cognitive functioning and inflammatory markers in people with multiple sclerosis (MS) and demographically-matched healthy individuals. Additionally, we compared the immune marker profile in serum and urine-matched samples. Social cognitive functioning was objectively assessed using The Awareness of Social Inference Test – Short (TASIT-S) and subjectively assessed using self-reports of abilities in emotion recognition, emotional empathy, and cognitive theory of mind. In people with MS and healthy individuals, there were moderate-to-large negative relationships between pro-inflammatory biomarkers (serum IL-1β, IL-17, TNF-α, IP-10, MIP-1α, and urine IP-10, MIP-1β) of the innate immune system and social-cognitive functioning. In MS, a higher serum concentration of the anti-inflammatory marker IL-1ra was associated with better social-cognitive functioning (i.e., self-reported emotional empathy and TASIT-S sarcasm detection performance). However, there were mixed findings for anti-inflammatory serum markers IL-4 and IL-10. Overall, our findings indicate a relationship between pro-inflammatory cytokines and social-cognitive abilities. Future studies may provide greater insight into biologically-derived inflammatory processes, sickness behaviour, and their connection with social cognition.

Surface characterization and biological assessment of corrosion-resistant a-C:H:SiOx PACVD coating for Ti-6Al-4V alloy

The paper focuses on the SiOx-doped amorphous hydrocarbon (a-C:H:SiOx) coating on the titanium (Ti-6Al-4V) alloy substrate obtained by plasma-assisted chemical vapor deposition (PACVD) in a mixture of argon gas and polyphenylmethylsiloxane vapor using a bipolar substrate bias. It is shown that the a-C:H:SiOx coating deposition results in the formation of a negative surface potential important for application of this coating for medical implants. The a-C:H:SiOx coatings improve the corrosion resistance of Ti alloy to 0.5 M NaCl solution and phosphate-buffered saline. In particular, the corrosion current density of the a-C:H:SiOx-coated sample in a 0.5 M NaCl solution at 22 °C decreases from 1∙10−8 to 1.7∙10−10 A/cm2, that reduces the corrosion rate from 9∙10−5 to 15∙10−7 mm/year. The a-C:H:SiOx coating facilitates the surface endothelization of an implant located in the thoracic aorta of a mini pig, and reduces the risk of thrombosis and implant failure. This effect can be explained by the ability of the a-C:H:SiOx coating ability to reduce in vitro a 24-hour secretion of pro-inflammatory interleukins (IL-6, IL-12(p70), IL-15, and IL-17) and cytokines (IFN-g and TNF-a) by blood mononuclear cells (MNCs) and elevates the concentration of anti-inflammatory interleukin IL-1Ra. In vitro analysis shows no cytotoxicity of the a-C:H:SiOx coating for the human blood MNCs, suggesting a promising PACVD on Ti alloys for cardiovascular implants, including pumps for mechanical heart support systems.

Short-term Western-style diet negatively impacts reproductive outcomes in primates.

A maternal Western-style diet (WSD) is associated with poor reproductive outcomes, but whether this is from the diet itself or underlying metabolic dysfunction is unknown. Here, we performed a longitudinal study using regularly cycling female rhesus macaques (n = 10) that underwent 2 consecutive in vitro fertilization (IVF) cycles, one while consuming a low-fat diet and another 6–8 months after consuming a high-fat WSD. Metabolic data were collected from the females prior to each IVF cycle. Follicular fluid (FF) and oocytes were assessed for cytokine/steroid levels and IVF potential, respectively. Although transition to a WSD led to weight gain and increased body fat, no difference in insulin levels was observed. A significant decrease in IL-1RA concentration and the ratio of cortisol/cortisone was detected in FF after WSD intake. Despite an increased probability of isolating mature oocytes, a 44% reduction in blastocyst number was observed with WSD consumption, and time-lapse imaging revealed delayed mitotic timing and multipolar divisions. RNA sequencing of blastocysts demonstrated dysregulation of genes involved in RNA binding, protein channel activity, mitochondrial function and pluripotency versus cell differentiation after WSD consumption. Thus, short-term WSD consumption promotes a proinflammatory intrafollicular microenvironment that is associated with impaired preimplantation development in the absence of large-scale metabolic changes.

Probiotics, Anticipation Stress, and the Acute Immune Response to Night Shift.

IntroductionSleep disturbance and sleep disruption are associated with chronic, low grade inflammation and may underpin a range of chronic diseases in night shift workers. Through modulation of the intestinal microbiota, probiotic supplements may moderate the effects of sleep disruption on the immune system. The aim of this study was to examine 14 days of daily probiotic supplementation on the acute response of acute phase proteins and immune markers to sleep disruption associated with night shift work (Australia and New Zealand Clinical Trials Registry: 12617001552370).MethodsIndividuals (mean age 41 ± 11 yrs; 74% female) performing routine night shift were randomly assigned to a probiotic group (1 × 1010 colony forming units (CFU) Lactobacillus acidophilus DDS-1 or 1 × 1010 CFU Bifidobacterium animalis subsp. lactis UABla-12) or placebo (n= 29 per group). Participants undertook a 14-day supplementation period that coincided with a period of no night shifts followed by two consecutive night shifts. Blood samples were collected prior to the start of supplementation (V1), prior to commencing the first night shift (V2), after the first night shift (V3) and after the second night shift (V4). Serum was assessed for markers of stress (cortisol), acute phase response (C reactive protein (CRP), erythrocyte sedimentation rate, pentraxin), adhesion markers (serum E-selectin, mucosal vascular addressin cell adhesion molecule 1 (MAdCAM-1), and serum cytokines (interleukin (IL)-1ra, IL-1β, IL-6, tumor necrosis factor (TNF)-α, IL-10). Sleep quality was assessed with the Pittsburgh Sleep Quality Index (PSQI) and a Fitbit activity tracker.ResultsThe groups were well balanced on key markers and the probiotic strains were well tolerated. The 14-day supplementation period that coincided with typical night-day sleep-wake cycles leading up to night shift (V1 to V2) was associated with significant changes in the placebo group in the concentration of serum cortisol (p = 0.01), pentraxin (p = 0.001), MAdCAM-1 (p = 0.001), and IL-1ra (p=0.03). In contrast, probiotic supplementation moderated changes in these serum markers from V1 to V2. No significant interaction effects (time by group) were observed for the serum markers prior to and after night shift work following probiotic supplementation due to the substantial changes in the serum markers that occurred during the normal sleep period from V1 to V2.ConclusionsProbiotics may moderate the effects of anticipatory stress on the immune system in the lead up to night shift.

IL-1 Receptor Antagonist Protects the Osteogenesis Capability of Gingival-Derived Stem/Progenitor Cells under Inflammatory Microenvironment Induced by Porphyromonas gingivalis Lipopolysaccharides.

Mesenchymal stem cells (MSCs) have been considered to be a future treatment option for periodontitis due to their excellent regenerative capability. However, it is still a challenge to protect MSCs' biological properties from multiple bacterial toxins in local inflammatory environment. The present study is aimed at investigating the treatment effect of interleukin-1 receptor antagonist (IL-1ra) on cell proliferation, migration, and osteogenic differentiation of gingival-derived mesenchymal stem cells (GMSCs) under an inflammatory microenvironment induced by Porphyromonas gingivalis lipopolysaccharides (P. gingivalis-LPS). GMSCs derived from Sprague-Dawley (SD) rats' free gingival tissues were treated with P. gingivalis-LPS (10 μg/mL) to create in vitro inflammatory environment. Different concentrations of IL-1ra (0.01-1 μg/mL) were used to antagonize the negative effect of LPS. Cell behaviors including proliferation, cloning formation unit (CFU), cell migration, osteogenic differentiation, mineral deposition, and cytokine production were assessed to investigate the protection effect of IL-1ra on GMSCs under inflammation. The toll-like receptor 4 (TLR4)/nuclear factor kappa B (NF-κB) pathway activated by LPS was evaluated by real-time quantitative polymerase chain reaction (RT-PCR) and western blot. In response to P. gingivalis-LPS treatment, cell numbers, cloning formation rate, cell migration rate, proinflammatory cytokine production, and osteogenic differentiation-associated protein/mRNA expressions as well as mineralized nodules were suppressed in a time-dependent manner. These negative effects were effectively attenuated by IL-1ra administration in a time- and dose-dependent manner. In addition, mRNA expressions of TLR4 and IkBα decreased dramatically when IL-1ra was added into LPS-induced medium. IL-1ra also reversed the LPS-induced TLR4/NF-κB activation as indicated by western blot. The present study revealed that IL-1ra decreased inflammatory cytokine production in a supernatant, so as to protect GMSCs' osteogenesis capacity and other biological properties under P. gingivalis-LPS-induced inflammatory environment. This might be explained by IL-1ra downregulating TLR4-mediated NF-κB signaling pathway activation.