Differences In Cytokine Concentrations Research Articles

PSIV-13 Changes in Interferon-γ and Tumor necrosis factor-α in the vaginal environment of cattle during early gestation

Abstract The embryo must create an immuno-tolerant uterine environment via cytokine signaling to prevent rejection and facilitate embryo attachment. Cytokines have a variety of roles in both reproductive and pregnancy-related processes. Cytokines and chemokines are important in inflammatory pathways for uterine remodeling and recruitment of immune cells during pregnancy. Interferon-γ (INFγ) and tumor necrosis factor-a (TNFα) influence T-helper cell immune responses, which can assist in early pregnancy establishment. Therefore, we hypothesized that INFγ and TNFα concentrations following breeding will be differentially expressed based on mating type, time, and pregnancy outcome. To test this, we completed two experiments in which INFγ and TNFα were quantified in vaginal fluid samples on d 0, d 7, d 14, d 21, d 28, and d 35 following mating. In experiment one, Angus heifers (n = 9) were used to determine cytokine concentrations in the vaginal environment between d 0 to d 35, and between artificial insemination (AI) and natural mating. In experiment 2, lactating dairy cows (n = 88) were used to determine if cytokines could be used as a potential biomarker for successful pregnancy. All vaginal flush samples were collected via Foley catheter in 20 mL of sterile saline. Statistical analysis was completed in SAS 9.4 to determine differences in cytokine concentrations based on day of gestation, mating type, and pregnancy outcome. In experiment 1, vaginal concentrations of TNFα were least (P = 0.05) in AI-bred heifers on d 35 (1.15 ± 2.97 pg/mL) and intermediate in non-pregnant heifers (4.39 ± 2.97 pg/mL) compared with natural service-bred heifers on d 7 (18.09 ± 2.97 pg/mL). INFγ was increased (P = 0.05) on d 7 following natural service (3.86 ± 0.76 pg/mL) compared with AI (0.86 ± 0.76 pg/mL). TNFα was increased (P = 0.02) on d 7 following natural service (18.09 ± 2.89 pg/mL) compared with AI (1.93 ± 2.89 pg/mL) and was increased (P = 0.02) by d 7 after a natural mating compared with a pregnancy resulting from AI mating. In experiment two, concentrations of IFNγ were greater (P = 0.04) on d 7 (1.36 ± 0.57 pg/mL) compared with d 8 (0.24 ± 0.57 pg/mL) of gestation. There was also a tendency for TNFα concentrations to decrease (P = 0.09) from d 7 (60.06 ± 26.94 pg/mL) to d 8 (19.67 ± 26.94 pg/mL) of gestation. In conclusion, concentrations of INFγ and TNFα may not be indicative of a viable embryo or pregnancy outcome but do differ based on day of gestation and mating type, which may aid in understanding the role these cytokines play during early gestation.

Cytokine profiles and their roles in development of provoked vulvodynia

Provoked, localized, vulvodynia (PVD) is the main subtype of vulvodynia. Although the etiology of PVD is still a topic of debate, inflammation caused by cytokines responding to a dysregulated microbiome is one of the leading proposed theories. Therefore, the purpose of our study is to further explore the cytokine profiles in the study group with PVD using multiplex immunoassays based on electrochemiluminescence. We compared a panel of 26 distinct cytokines levels in the study group with PVD (n = 23) to the control group (n = 18) and cytokine concentrations were measured using MESO QuickPlex SQ 120 instrument with 5 different multiplex assays. Statistical analysis used the Mann-Whitney U test, two-sided p-values, and a significance level of α = 0.05. Differences in cytokine concentrations are described as negligible, small, medium, or large based on Cliff’s δ. Concentrations of three cytokines were significantly lower in the PVD group: a large difference in IP-10 (p = 0.029*) and medium differences in IL-1RA44Cytokine abbreviations: bFGF; Basic Fibroblast Growth Factor, G-CSF; Granulocyte colony stimulating factor, GM-CSF; Granulocyte-Macrophage Colony-Stimulating Factor, IFN; interferon, IL; Interleukin, IP; Interferon-gamma Inducible Protein, MCP; Monocyte Chemoattractant Protein, MIP; Macrophage Inflammatory Protein, PGDF-BB; Platelet Derived Growth Factor Subunit B, RA; Receptor antagonist, TNF; Tumor Necrosis Factor, VEGF; Vascular Endothelial Growth Factor (p = 0.030*) and IL-12 (p = 0.034*). One cytokine level was significantly higher in the PVD group: a medium difference for IL-6 (p = 0.037*). Due to the lack of consistency in elevation of inflammatory profiles, it is not enough to support persistent inflammation as the etiology behind PVD. However, these findings may indicate there is a possible immune response deficiency in some patients who have PVD. The resemblance of cytokine profile in our study to cytokine profile of people with chronic yeast infection further support this proposed mechanism behind PVD. Future studies involving history and testing for yeast infection are necessary to explore this possibility further.

Inflammatory Profile of Chronic Rhinosinusitis With Nasal Polyp Patients in Brazil: Multicenter Study.

To determine the inflammatory profile of CRSwNP in Brazil and characterize the subgroups of CRSwNP patients in this population through cluster analysis. Multicenter cross-sectional study involving 15 centers representing different regions of Brazil. Clinical data of 166 patients and 80 controls, aged 18 to 70 years old, number of surgeries for CRS, history of asthma and aspirin sensitivity, and Lund-Mackay scores on CT scans. During nasal endoscopy, we obtained the Lund-Kennedy scores and collected 2 samples of nasal polyps: one for eosinophil and neutrophil tissue counts and one to quantify different cytokines. 79.6% of our patients had 10 or more eosinophils/HPF. CRSwNP groups exhibited significantly lower concentrations of TNF-alpha and significantly higher concentrations of IFN-gamma, CCL11/Eotaxin, CCL24/Eotaxin-2/MPIF-2, and CCL26/Eotaxin-3 versus the control group (Kruskal-Wallis test). Comparison between CRSwNP groups (≥10 vs <10 eosinophils/HPF) showed no difference in cytokine concentration (Mann-Whitney test). Hierarchical clustering and PCA according to cytokine concentrations revealed 2 main Clusters, with a significantly higher concentration of all cytokines in Cluster 1 (n = 35) than in Cluster 2 (n = 121), except IL-6 and IL-33 (Mann-Whitney test). According to ROC curve analysis the best cut-off to differentiate the 2 clusters was 43 eosinophils/HPF. The group with ≥43 presented a higher prevalence of men and a higher Lund-Mackay score (Mann-Whitney test). CRSwNP patients in Brazil present mixed inflammation, with 2 distinct groups (high and low inflammatory pattern) that can be distinguished by tissue eosinophilia of ≥43 eosinophils/HPF cut-off in nasal polyps.

Evaluation of the Levels of Selected Cytokines and Their Possible Influence on the Development of Cardiovascular and Pulmonary Complications in Patients after COVID-19.

Background and Objectives: The aim of this study was to evaluate the levels of selected cytokines and their possible influence on the development of cardiovascular and pulmonary complications in patients hospitalized at the Silesian Centre for Heart Disease in Zabrze after having undergone COVID-19. Materials and methods: The study included 76 randomly selected patients from the SILCOVID-19 database. The median time from symptom onset to the study visit was 102 (86-118) days. The median age of the study group was 53 (44-60) years. Assays of a panel of 30 cytokines were carried out in the serum of patients on a Luminex100 platform using the Milliplex MAP kit from Merck KGaA Germany. Results: There were no statistically significant differences in most of the cytokines analyzed between patients with confirmed or excluded lung lesions or cardiac abnormalities. Additionally, no statistically significant differences in cytokine concentrations according to gender, age, comorbidity of diabetes, renal disease, hypertension, increased risk of thrombotic disease, or psychological disorders were demonstrated. There were high concentrations of cytokines such as platelet-derived growth actor-AA (PDGF-AA), monocyte chemoattractant protein-1 (MCP-1), monokine-induced gamma interferon (MIG), and vascular endothelial growth factor-A (VEGF-A). Conclusions: No direct impact of the dependencies between a panel of cytokines and the incidence of cardiovascular and pulmonary complications in patients hospitalized at the Silesian Centre for Heart Disease in Zabrze after having undergone COVID-19 was demonstrated. The demonstration of high levels of certain cytokines (PDGF-AA, VEGF, MIG, and IP10) that are of significance in the development of many lung diseases, as well as cytokines (MCP-1) that influence the aetiopathogenesis of cardiovascular diseases seems to be highly concerning in COVID-19 survivors. This group of patients should receive further monitoring of these cytokine levels and diagnostic imaging in order to detect more severe abnormalities as early as possible and administer appropriate therapy.

PSII-9 Circulating Cytokine Profiles of Bos Indicus Cattle Prior to Artificial Insemination in Relation to Pregnancy Status

Abstract Cytokines and chemokines (cytokines with chemotactic activity) influence inflammatory responses and can upregulate pro- or anti-inflammatory immune responses and influence fertility. The aim of this study was to characterize the circulating cytokines of Bos indicus females prior to fixed-timed artificial insemination (FTAI). Bos indicus females on operations within a 20-mile radius were subjected to the 7-day CO-Synch + CIDR protocol on day (d) -9 and FTAI on d 0. Blood samples were collected on d -9 (CIDR IN), d -2 (CIDR OUT), and d 0 (AI). Cattle were body condition scored (BCS) on d 0. Pregnancy status was determined by transrectal ultrasonography for resulting non-pregnant females (n = 44) and resulting pregnant females (n = 34). Cytokine analyses on blood plasma samples were conducted using the RayBiotech Quantibody Bovine Cytokine Array Q1 kit per manufacturer's instructions. Cytokines and chemokines measured included interferon alpha (IFNa), interferon-gamma (IFNg), interleukin (IL)1a, IL1F5, IL13, IL21, chemokine-ligand (CXCL) 9, CXCL10, macrophage inflammatory protein (MIP)1b, and tumor necrosis factor alpha (TNFa). Data were analyzed using PROC GLM in SAS 9.4 with fixed effects for pregnancy status, ranch, day, and parity. Body condition scores did not significantly differ by pregnancy status (P &amp;gt; 0.05). There were no differences in cytokine concentrations by day of the synchronization protocol or between cows and heifers (P &amp;gt; 0.05). There were differences by location (P &amp;lt; 0.05) for most cytokines except MIP1b and TNFa. Circulating concentrations of the pro-inflammatory cytokines IFNg, IL1a, and TNFa were greater (P &amp;lt; 0.05) in resulting non-pregnant females compared with pregnant females (2,323.5 ± 260.5 vs. 1,502.8 ± 246.8 pg/mL, 894.6 ± 96.8 vs. 598.3 ± 91.7 pg/mL, and 6,052.6 ± 627.4 vs. 4,156.6 ± 594.3 pg/mL, respectively). Circulating concentrations of the chemokines CXCL9, CXCL10, and MIP1b were also greater (P &amp;lt; 0.05) in resulting non-pregnant females compared with pregnant females (4,887.0 ± 538.3 vs. 2,905.3 ± 509.8 pg/mL, 12,167.3 ± 1,496.2 vs. 6,295.4 ± 1,417.2 pg/mL, 586.4 ± 56.8 vs 324.4 ± 53.8 pg/mL, respectively). Interestingly, circulating concentrations of the anti-inflammatory cytokines IL13, IL1F5, and IL21 were greater (P &amp;lt; 0.05) in resulting non-pregnant females compared with pregnant females (11,147.7 ± 1,009.9 vs. 6,752.1 ± 956.6 pg/mL, 1,189.3 ± 111.7 vs. 744.5 ± 105.8 pg/mL, and 1,1153.6 ± 898.0 vs. 8,409.7 ± 850.6 pg/mL, respectively). No differences by pregnancy status were found for concentrations of IFNa (P = 0.13). These results suggest that an increase in circulating cytokines and chemokines prior to FTAI negatively impacts fertility in Bos indicus cattle. Future research is necessary to determine the threshold of circulating cytokine concentrations that subsequently impact pregnancy establishment and maintenance.

Exploring the Involvement of Cytokines in Pediatric Patients Afflicted by Simple Febrile Seizures: A Case-Control Study.

Most children under six with seizures experience febrile seizures (FS), which occur with a temperature of 38°C or higher and no apparent infectious or metabolic causes. FS has a multifaceted etiology, involving genetic and environmental factors. This study aimed to explore the connection between the cytokine system (part of the immune system related to inflammation) and FS to uncover potential relationships. This research was carried out on 50 patients experiencing FS and 25 patients experiencing only fever served as cases and controls, respectively. The patient's blood was obtained under sterile circumstances from the antecubital/femoral vein as soon as feasible following the commencement of seizures. The complete hemogram analysis was done using the Mindray BC-5800 auto hematology analyzer (Mindray Medical International Limited, Shenzhen, China). The cases group had significantly higher interleukin-4 (IL-4) concentrations (292.85 pg/mL) than controls (81.04 pg/mL), indicating a statistically significant difference (p<0.05), respectively. In the current research, case participants had a significantly lower mean level of interleukin-6 (IL-6) than control participants. The average IL-6 concentration in cases was 28.00 pg/mL, whereas in controls was 93.09pg/mL. Patients with FS have an important function for the cytokine network to perform. The findings showed a significant difference in cytokine concentrations between patients with FS and control subjects, highlighting a potential link between the cytokine system and FS. Additionally, lower levels of IL-6 in case participants suggest a complex role of cytokines in FS, emphasizing the importance of the cytokine network in this condition.

Cytokines in equine platelet lysate and related blood products.

In equine medicine, the use of regenerative therapeutics has gained growing attention, but is still a new and complex field with room for improvement. Platelet lysate (PL) can be used as therapeutic agent but is also a promising supplement for the culture of multipotent mesenchymal stromal cells. To enable a targeted use of PL both in clinic and laboratory, it is crucial to learn more details on its effective ingredients. While so far, mainly growth factor components have been analyzed in platelet-based products such as PL, the current study focuses on the content of cytokines in serum, plasma, platelet concentrate and PL. Blood was harvested from 20 clinically healthy horses and subjected to blood count and chemistry analysis, as well as to further processing to PL. Plasma and platelet concentrate were produced by a buffy-coat-based method and PL was produced from the concentrate by freeze-thawing. Samples from each horse were analyzed regarding interleukin (IL)-1β, -4, -6 and -10, interferon-γ and tumor necrosis factor-α concentrations using sandwich ELISAs. Cytokine concentrations in serum, plasma, concentrate and PL were similar and correlated significantly. However, there was a large inter-individual variability in cytokine concentrations between the different donor horses. The samples from some donor animals had overall very high cytokine concentrations, while samples from other donors had no measurable cytokine ingredient. This pattern was observed for all cytokines. There was a noticeable link between high cytokine concentrations in the blood products and abnormal findings in blood chemistry. Cytokine concentrations in samples from horses with abnormal findings were significantly higher than in samples from the remaining horses. The interindividual differences in cytokine concentrations could be highly relevant when using PL for therapy and cell culture, as the mode of action of the PL is likely changed depending on the presence of pro- and anti-inflammatory cytokines. Blood chemistry might be useful to predict cytokine concentrations in blood products.

In vitro detection of suspected drug in maculopapular drug reaction to antibiotics using secreted cytokines from drug-specific activated T cells.

Maculopapular drug eruption (MPE) in reaction to antibiotics is associated with enhanced expression of T-helper (Th)1 cytokines such as interferon gamma (IFN-gamma) or Th2 cytokines such as interleukin (IL)-5. Identifying the culprit drug usually involves rechallenge, which may not be forthcoming. Memory lymphocytes remain responsive to the culprit drug long after the reaction has resolved. On reactivation in vitro, there is increased proliferation and expression of certain markers, which provides us with an opportunity to predict the causal drug. The study aimed to assess drug-specific cytokine production (IL-5 and IFN-gamma) in peripheral blood mononuclear cell (PBMC) culture supernatants to predict the causal antibiotic in cases of MPE. PBMCs of 18 patients who developed MPE to 20 suspected antibiotics (2 patients had 2 suspected antibiotic allergies each), along with 11 drug-matched healthy controls, were incubated for 5 days with the respective drugs at 2 different concentrations. Secreted cytokines were measured in the supernatants, using enzyme-linked immunosorbent assay (ELISA) for IL-5 and IFN-gamma, at 6 h of incubation, then on day 2 and day 5. Drug-specific IL-5 and IFN-gamma production could be demonstrated in 65% and 74% of the cases, respectively. Maximal secretion of IL-5 and IFN-gamma was observed on day 5 and day 2 of incubation, respectively. The cut-off delta values, defined as the difference in cytokine concentration between drug-stimulated and unstimulated samples, were 4 pg mL-1 for IL-5 and 6 pg mL-1 for IFN-gamma. The measurement of drug-specific secretion of IL-5 and IFN-gamma using ELISA is a valuable method for detecting antibiotic-induced MPE.

P346: THE ROLE OF CYTOKINES, TNF, TGFΒ1 AND NEUTROPHIL-TO-LYMPHOCYTE RATIO IN THE COURSE OF THE ACUTE LEUKEMIA

Background: Lymphocytes (Lym) and mature segmental neutrophils (Neu) together with blasts are most commonly detected in the peripheral blood (PC) of patients (pta) with acute leukemia (AL). Besides, Neu, Lym and blast cells are an important source of transforming growth factor β1 (TGFβ1), tumor necrosis factor (TNF) production. Accumulation of TNF, TGFβ1 in the plasma of AL pts is considered an unfavorable sign in AL. However, the role of TNF, TGF β1 in the relationship with neutrophil-to-lymphocyte ratio (NLR) in the course of AL is not definitively established. Aims: Determine the role of cytokines, TNF, TGF β1 in the relationship with NLR to assess the course of AL. Methods: 42 adult AL pts were examined before treatment, who at the time of the examination had blast cells in the peripheral blood (52.54 ± 5.63 x 109 / L). The NLR was calculated from peripheral blood cell counts obtained at the time of diagnosis of AL by dividing the Neu count by the Lym count. The median NLR (0.99) was used to dichotomize patients into high- NLR and low- NLR groups. The concentration of TNF, TGF β1 was determined by biological methods using sensitive cell lines L929 and CCL 64, respectively. Statistical processing of the results was estimated by the value of p ˂ 0, 005. The correlation between the indicators was estimated by the value of r. Results: There were 25 people in the low-NLR group. Median age 66 years (36 - 81 years). Among the pts were 9 people with ALL and 16 people with AML. Number of blasts 55, 83 ± 6.38 x 109 / L; New count was 11.73 ± 2.95 109 / L, Lym count - 21.27 ± 3.64 109 / L. The NLR index is 0.53 ± 0.09. In this group, 88% of pts died in the short term without achieving remission, the remaining 12% of pts had short-term remission. The concentration of TNF in plasma was 1.20 ± 0.13 ng / ml, TGF β1 - 4.02 ± 0.48 ng / ml, which in both cases was higher than in healthy individuals (p˂0.001 and p˂0.05, respectively). TNF levels were positively correlated with Lym (r = 0.30) and blasts (r = 0.40), suggesting a possible role for TNF in neoplastic clone survival. There were 17 people in the high-NLR group, including 10 pts with ALL and 7 people with AML who achieved remission. The median age is 38 years (from 16 to 68 years). The number of blasts was 33.20 ± 9.67 x 109 / L and was significantly lower than in the low-NLR group (p˂0.05); New count was 22.23 ± 0.10 x 109 / L; Lym count - 22.83 ± 3.04 109 / L. The NLR was 1.24 ± 0.16. In this group, 94% of pts had remission. Plasma TNF and TGF concentrations were lower than in the low-NLR group, apparently due to lower blast content. However, no statistical difference in cytokine concentration between the compared groups was found. Summary/Conclusion: More favorable prognostic features in AL could be NLR, the median of which is equal to or greater than 0.99. Under these conditions, a decrease in the number of blast cells, the concentration of cytokines TNF and TGF β1 in the plasma and the achievement of remission were observed. The data obtained may indicate a prognostic role of Neu and Lym blood subpopulations in the course of AL and in the relationship with cytokines, TNF and TGF β1, may be potential targets for the correction of treatment of AL pts.

The effect of exercise on cytokine concentration in equine autologous conditioned serum.

Autologous conditioned serum (ACS) is a commonly administered intra-articular treatment for the management of osteoarthritis in athletic horses. To investigate the influence of exercise on the concentration of cytokines in a non-commercial method of ACS production. Non-randomised cross over design. Whole blood was obtained from eight healthy Standardbred horses immediately prior to, 1h and 24 h following a single bout of exhaustive exercise. Blood was processed using a non-commercial method of ACS production. Fluorescent microsphere immunoassay (FMIA) analysis was performed to quantify Interleukin 1 receptor antagonist (IL-1Ra), Interleukin-10 (IL-10), Interleukin 1β (IL-1β) and tumour necrosis factor α (TNF-α) concentrations at each time point. Mixed effect repeated measures analysis of variance (ANOVA) was used to compare the pre-exercise and post-exercise cytokine concentrations. Significance was set at P < 0.05. A reduced concentration of IL-1Ra (median 584.4, IQR 81.9-5098 pg/ml, p= 0.004) and an increased concentration of TNF-α (11.92, 9.28-39.75 pg/ml, P= .05) at 1h post-exercise were observed when compared with baseline values (IL-Ra 7349, 1272-10760 pg/ml; TNF TNF-α 11.16, 8.36-32.74 pg/ml). No difference in cytokine concentrations of IL-10 or IL-1β were found between any of the time points. The large biological variability and small sample size represents limitations of this study. These results suggest that a single bout of intense exercise can reduce the concentration of the anti-inflammatory cytokine IL-1Ra and increase the concentration of the pro-inflammatory cytokine TNF-α, reducing the 'anti-inflammatory' cytokine composition of ACS. Our findings suggest that collection of blood for ACS production should be performed no sooner than 24 h following a single episode of intense exercise.

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.

Concentration of vaginal and systemic cytokines obtained early in pregnancy and their impact on preterm birth

Objective A number of factors can lead to a maternal pro-inflammatory response resulting in a spontaneous preterm birth. However, it remains unknown if an upregulation in the maternal immune system early in pregnancy leads to an increase in pro-inflammatory cytokines and ultimately preterm birth. Therefore, we hypothesize an increase in vaginal and systemic pro-inflammatory cytokines early pregnancy is associated with an increased risk of preterm birth. Study design Patients initiating prenatal care prior to 14 weeks gestation were recruited for eligibility. A vaginal swab and serum sample was obtained at the first prenatal visit and these were then stored at −80 C. Patients were then followed for their gestational age at delivery. Five patients delivering preterm (cases) were matched with ten patients delivering at term (controls) based on age, BMI, smoking status and ethnicity. The serum and vaginal swabs from the cases and controls were then analyzed for the following cytokines using a multiplex cytokine assay: GM-CSF, IL-1b, IL-6, TNFα, and Rantes. Results A total of 116 patients were screened for eligibility and 96 of these patients had samples obtained prior to 14 weeks gestation. Of these 96, 5 had a spontaneous preterm birth and these were matched to 10 controls. There was no difference detected in the cytokine concentrations of GM-CSF, IL-1b, IL-6, TNFα, and Rantes in the serum or cervicovaginal fluid between cases and controls. Conclusion This study demonstrates there is no difference in cytokine concentrations of several pro-inflammatory cytokines in the vagina or in the serum prior to 14 weeks gestation in patients delivering preterm. Therefore, the concentration of the cytokines analyzed in this study from the vagina and serum have little predictive value on the risk of preterm birth. Further research is needed to deepen our understanding of the mechanisms leading to preterm birth.

Effects of Production Method and Repeated Freeze Thaw Cycles on Cytokine Concentrations and Microbial Contamination in Equine Autologous Conditioned Serum.

Autologous conditioned serum (ACS) is a common intra-articular treatment for osteoarthritis in horses. The objective of this study was to investigate the influence of ACS preparation method on product contamination and concentrations of relevant cytokines and the influence of multiple freeze/thaw cycles. Blood was obtained from 10 healthy Thoroughbred horses and processed in parallel using a commercial and a non-commercial method to obtain ACS. Fluorescent microsphere immunoassay (FMIA) analysis was performed to quantify Interleukin 1 receptor antagonist (IL-1Ra), Interleukin-10 (IL-10), Interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) concentrations in ACS obtained by both production methods. Effect of 3, 4 and 5 freeze/thaw cycles on concentrations of IL-1Ra, IL-10, IL-1β and TNF-α were assessed against baseline samples (2 cycles) in commercial ACS products. Standard aerobic and anaerobic culture methods were applied to both ACS products. Mixed effect one-way analyses of variance (ANOVA) were used to compare the two ACS production method for each cytokine. Repeated measures, mixed effect ANOVA were used to assess the effect of freeze/thaw on cytokine concentrations. Significance was set at P < 0.05. There was no difference in cytokine concentration between production methods (IL-1Ra P = 0.067, IL-1β P = 0.752, IL-10 P = 0.211 and TNF-α P = 0.25). Microbial growth was only observed in two samples obtained using the commercial production method. When compared to baseline, IL-1Ra concentration was decreased following the 5th freeze/thaw cycle (P < 0.001). These results suggest that the concentration of important cytokines are not influenced by ACS production method. When storing ACS samples for future use, freeze/thaw cycles associated with standard clinical practice are unlikely to influence cytokine concentrations. However, the lack of outcome measures associated with 1 or 2 freeze/thaw cycles represents a limitation of this study.

Profile of serum cytokine concentrations in patients with gouty arthritis.

ObjectiveThis study aimed to analyze the changes in serum inflammatory cytokines and anti-inflammatory cytokines in patients with gouty arthritis (GA).MethodsThe clinical data and serum samples in patients with gouty arthritis and those in healthy volunteers were collected in China-Japan Friendship Hospital from July 2018 to January 2019. Serum cytokine concentrations in patients with GA and volunteers (controls) were determined by a chemiluminescence method. The differences in cytokine concentrations were compared between the two groups.ResultsConcentrations of serum interleukin-1 beta (IL-1ß), tumor necrosis factor-alpha (TNF-α), IL-6, IL-8, and IL-4 were significantly higher in patients with acute GA than in controls. Serum concentrations of IL-1ß, TNF-α, IL-6, IL-8, and immunoglobulin E in patients with remission of GA were significantly lower, whereas concentrations of IL-10 and interferon-γ were significantly higher, compared with those in patients with acute GA.ConclusionThis study shows that serum concentrations of IL-1ß, TNF-α, IL-6, IL-8, and IL-4 are significantly elevated in patients with GA, and may be involved in the pathogenesis of GA.

Immune Responses after Heavy Alcohol Consumption: Cytokine Concentrations in Hangover-Sensitive and Hangover-Resistant Drinkers.

This study investigated immunological changes during an alcohol hangover, and the possible difference between hangover-resistant and hangover-sensitive drinkers in terms of immune reactivity. Using a semi-naturalistic design, N = 36 healthy social drinkers (18 to 30 years old) provided saliva samples on a control day (after drinking no alcohol) and on a post-alcohol day. Hangover severity was rated directly after saliva collection. Cytokine concentrations, interleukin (IL)-1β, IL-6, IL-8, IL-10 and tumor necrosis factor (TNF)-α, and hangover severity were compared between both test days and between hangover-sensitive and -resistant drinkers. Data from N = 35 drinkers (17 hangover-sensitive and 18 hangover-resistant) were included in the statistical analyses. Relative to the control day, there were significant increases in saliva IL-6 and IL-10 concentrations on the post-alcohol day. No significant differences in cytokine concentrations were found between hangover-sensitive and hangover-resistant drinkers, nor did any change in cytokine concentration correlate significantly with hangover severity. In line with previous controlled studies assessing cytokines in blood, the current naturalistic study using saliva samples also demonstrated that the immune system responds to high-level alcohol intake. However, further research is warranted, as, in contrast to previous findings in blood samples, changes in saliva cytokine concentrations did not differ significantly between hangover-sensitive and hangover-resistant drinkers, nor did they correlate significantly with hangover severity.

Association of fracture configuration and callus formation with a concentration of proinflammatory cytokines in children with long bone fractures

Background/Aim. The inflammatory response is of utmost importance in bone healing, but the precise role of cells and cytokines remains unclear. In our study, we examined the association between interleukin-1? (IL-1?), tumor necrosis factors alpha (TNF-?), monocyte chemoattractant protein-1 (MCP-1) and interleukin-8 (IL-8) concentrations, fracture configuration, and callus formation. Methods. Serum cytokine concentrations were determined in 78 non-obese children with long bone fractures (group 1), 10 children with finger fractures (group 2), and 10 healthy controls (group 3). Blood samples were taken immediately after fracture upon hospital admission for groups 1 and 2. Differences in cytokine concentrations were analyzed among groups and catego-rized according to fracture configuration and callus formation. Results. IL-1? and TNF-? levels were lower in patients that went on to produce incomplete callus compared with patients that formed complete callus. Surprisingly, the average IL-1? concentration was highest in the healthy control group. The only significant correlation between IL-1? and TNF-? was in the group with moderate callus formation. MCP-1 level was slightly increased in all patient groups compared to controls, with no mutual difference. An average IL-8 level showed a clear decrease tendency from the group with incompletely formed callus toward the group with completely formed callus compared to controls, without significant difference. Children with epiphysiolysis had the lowest concentrations of cytokines compared with all other fracture types including transverse, oblique, and spiral. There were significantly lower concentrations of IL-1? and MCP-1 in patients with less fragment displacement compared with patients with greater fragment displacement. Conclusion. The systemic inflammatory response is important in physiological bone healing. High early production of IL-1?, TNF-?, and MCP-1 is associated with greater callus formation and better healing outcome, while increased IL-8 level is associated with poor callus formation and worse healing outcome. Our results indicate that epiphysiolysis and larger fragment displacement are associated with delayed fracture healing.

Repeated allergic asthma in early versus late pregnancy differentially impacts offspring brain and behavior development

BackgroundStress during pregnancy and maternal inflammation are two common prenatal factors that impact offspring development. Asthma is the leading chronic condition complicating pregnancy and a common source of prenatal stress and inflammation. ObjectiveThe goal of this study was to characterize the developmental impact of repeated allergic asthma inflammation during pregnancy on offspring behavioral outcomes and brain inflammation. MethodsPregnant female C57BL/6 mice were sensitized with ovalbumin (OVA) or PBS vehicle control and then randomly assigned to receive daily aerosol exposures to the same OVA or PBS treatment during early, gestational days (GD) 2-GD9, or late pregnancy, GD10-GD17. Maternal sera were collected after the first and last aerosol induction regimen and measured for concentrations of corticosterone, anti-OVA IgE, and cytokine profiles. Juvenile male and female offspring were assessed for locomotor and social behaviors and later as adults assessed for anxiety-like, and marble burying behaviors using a series of behavioral tasks. Offspring brains were evaluated for region-specific differences in cytokine concentrations. ResultsIn early gestation, both PBS and OVA-exposed dams had similar serum corticosterone concentration at the start (GD2) and end (GD9) of daily aerosol inductions. Only OVA-exposed dams showed elevations in cytokines that imply a diverse and robust T helper cell-mediated immune response. Male offspring of early OVA-exposed dams showed decreases in open-arm exploration in the elevated plus maze and increased marble burying without concomitant changes in locomotor activity or social interactions. These behavioral deficits in early OVA-exposed male offspring were associated with lower concentrations of G-CSF, IL-4, IL-7, IFNγ, and TNFα in the hypothalamus. In late gestation, both PBS and OVA-exposed dams had increased corticosterone levels at the end of daily aerosol inductions (GD17) compared to at the start of inductions (GD10). Male offspring from both PBS and OVA-exposed dams in late gestation showed similar decreases in open arm exploration on the elevated plus maze compared to OVA male offspring exposed in early gestation. No behavioral differences were present in female offspring across all treatment groups. However, females of dams exposed to OVA during early gestation displayed similar reductions as males in hypothalamic G-CSF, IL-7, IL-4, and IFNγ. DiscussionThe inflammatory responses from maternal allergic asthma in early gestation and resulting increases in anxiety-like behavior in males support a link between the timing of prenatal insults and sex-specific developmental outcomes. Moreover, the heightened stress responses in late gestation and concomitant dampened inflammatory response to allergic asthma suggest that interactions between the maternal immune and stress-response systems shape early life fetal programming.

181 Uterine and vaginal bacteria and cytokine profiles prior to artificial insemination between resulting pregnant and open postpartum beef cows

Abstract Numerous factors impact reproductive success in beef cattle; however, the presence and activity of bacteria in the reproductive tract and its effects on fertility is relatively unknown. The objective of this study was to evaluate the relationship between reproductive tract bacterial communities and cytokine profiles prior to timed-AI (TAI) and compare between resulting pregnancy status at d30. Sixty-eight beef cows, 80 ± 2.6 d postpartum at TAI, were synchronized and subjected to TAI on d0. Pregnancy was diagnosed via transrectal ultrasonography on d30. Uterine and vaginal flushes were collected on d -21 and -2 for bacterial DNA extraction to sequence the V1-V3 hypervariable regions of the 16S rRNA gene and to measure pro-inflammatory [interlukin-6 (IL-6)] and anti-inflammatory [IL-10 and transforming growth factor-beta (TGFβ)] cytokine concentrations using validated commercial ELISAs. Concentration data were analyzed using repeated measures in PROC MIXED of SAS and correlations were performed in SAS using Pearson correlation. There were no differences in cytokine concentrations by day or status x day. There were no differences in uterine IL-6 or IL-10 concentrations (P &amp;gt; 0.05). Uterine TGFβ concentrations were overall elevated in pregnant cows compared to open cows (72.6 vs. 13.7 ± 12.1 pg/mL, respectively; P &amp;lt; 0.05). On d -2, a positive correlation exists between TGFβ and the relative abundance of Actinobacteria in the uterus of pregnant cows (r=0.93, P = 0.02). There were no differences in vaginal IL-6 or TGFβ concentrations (P &amp;gt; 0.05). On d -2, vaginal IL-10 concentrations were elevated in open cows compared to pregnant cows (741.6 vs. 602.1 ± 47.0 pg/mL, respectively; P &amp;lt; 0.05) and a positive correlation exists between IL-10 and the relative abundance of Lentisphaerae in the vagina of pregnant cows (r=0.89, P = 0.04). These data suggest a possible relationship between bacterial communities and cytokines concentrations within the reproductive tract of beef cattle which may affect fertility.

Adjunctive Antiseptic Irrigation of Periodontal Pockets: Effects on Microbial and Cytokine Profiles.

To evaluate the effect of adjunctive antiseptic irrigation of periodontal pockets on microbial and cytokine profiles. Fifty-nine patients with severe periodontitis were allocated to one of three groups for scaling and root planing facilitated with different adjunctive antiseptics: 1% polyhexamethyleneguanidine phosphate (PHMG-P) (n = 19), 0.2% chlorhexidine (CHX) (n = 21) or distilled water (n = 19). Gingival crevicular fluid and subgingival bacterial samples were collected at baseline, and at 2 weeks, and 1 and 4 months. The levels of interleukin (IL)-1β, IL-8, IL-10, and IL-17A, matrix metalloproteinase (MMP)-8, Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola, Fusobacterium nucleatum, Aggregatibacter actinomycetemcomitans, and Prevotella intermedia were determined. There were no intergroup differences in cytokine concentrations and bacterial counts at any follow-up, however, varying patterns were observed. In the PHMG-P and water groups IL-1β expression peaked at 2 weeks and then gradually declined. In all three groups, the dynamics of MMP-8 concentration were non-linear, increasing by 2 weeks and then declining to below baseline (p > 0.05). P. gingivalis and T. forsythia declined within the first month and increased thereafter, not regaining the baseline level. Adjunctive antiseptic treatment was associated with changes in biomarkers and bacterial counts in the course of the study. The effects of adjunctive antiseptic irrigation were limited in the applied protocol.

Effects of neural mobilization on individuals with chronic low back pain

ABSTRACT BACKGROUND AND OBJECTIVES: The objective of this study was to evaluate the effect of neural mobilization technique on individuals with chronic low back pain, as well as analyze possible changes in pain, motor behavior and on cytokine quantification before and after treatment. METHODS: Sixteen individuals with mean age of 30.45±10.32 years old were evaluated. The technique consisted of a total of 10 interventions, with a total duration of 10 minutes, divided into four series of 2 minutes each, with a 30-second interval between each series. The following outcomes were evaluated: pain intensity using the visual analog scale; mobility of the lumbar spine with the third finger to the ground test and hip goniometry; concentration of cytokines in serum with the multiplex technique; and quality of life with the Oswestry Disability Index, the Roland-Morris Disability Questionnaire and the World Health Organization Quality of Life Questionnaire-bref. RESULTS: Results showed a reduction in pain intensity of approximately 70% between the assessments (p<0.001). Improvements in lumbar mobility of approximately 25.5% on the third finger to the ground test (p<0.04) and improvement in the hip joint angles (p<0.04) were also observed. A difference in cytokine concentrations, both pro-inflammatory (p<0.009) and anti-inflammatory (p<0.03), was also observed between the assessments. CONCLUSION: The technique was able to reduce pain intensity and consequently increase the mobility of the volunteers. Additionally, a difference in cytokine concentration at the end of the treatment and an improvement in the individuals quality of life were observed.