Fluid Cytokine Concentrations Research Articles

Differential Control of T-Cell Subsets by Recombinant Human PLD2 in a Mouse Model of Allergic Asthma

ABSTRACT Background Phospholipase D2 (PLD2) enzymes are expressed on the cytoplasmic membrane of bacteria, fungi, plants, and animals. Recently, extensive research has linked PLD2 to the chronic inflammatory activity of cells. Allergic asthma is a chronic airway inflammation disease. In this context, a recombinant human phospholipase D2 (rhPLD2) was designed and modified from the wild-type PLD2 to study its effects in an ovalbumin (OVA) induced murine model of asthma. Methods Hematoxylin and eosin staining was used for lung histopathology. Cytokine concentrations in bronchoalveolar lavage fluid (BALF) were measured using ELISA kits. The ratio of T-bet and GATA-3 expression level in spleen and lymph nodes following rhPLD2 administration was assessed through RT-PCR. Phenotyping analysis of Treg cells from peripheral blood was performed by flow cytometry. Results It indicated that OVA-induced mice exhibited elevated pulmonary eosinophilia and allergic inflammation in the airways, along with increased expression of IFN-γ, IL-4 in the lung BALF. Administration of rhPLD2 alleviated lung inflammation and significantly reduce the number of eosinophils in peripheral blood and BALF. RhPLD2 also reversed the IFN-γ/IL-4 ratio at the molecular level in BALF and the T-bet/GATA-3 ratio in lymphocytes of the lung, spleen, lymph nodes at the genetic level. Furthermore, FACS analysis demonstrated that rhPLD2 increased the frequency of both IL-10+Treg cells and CD25+ Treg cells. Conclusion From a therapeutic perspective, rhPLD2 alleviates allergic airway inflammation by balancing Th1/Th2 homeostasis and increasing Treg cells. It has been shown to function in immunoregulatory activities in OVA-induced asthma mice.

Immune response and cytokine profiles in post-laminectomy pain syndrome: comparative analysis after treatment with intrathecal opioids, oral opioids, and non-opioid therapies.

This study explores the interaction between cytokines, cell-mediated immunity (T cells, B cells, and NK cells), and prolonged morphine administration in chronic neuropathic pain patients without cancer-related issues. Despite evidence of opioid immunomodulation, few studies have compared these interactions. In a cross-sectional and comparative study, 50 patients with chronic low back radicular pain ("Failed Back Surgery Syndrome") were categorized into intrathecal morphine infusion (IT group, n = 18), oral morphine (PO group, n = 17), and non-opioid treatment (NO group, n = 15). Various parameters, including plasma and cerebrospinal fluid (CSF) cytokine concentrations, lymphocyte immunophenotyping, opioid escalation indices, cumulative morphine dose, and treatment duration, were assessed. CSF IL-8 and IL-1β concentrations exceeded plasma levels in all patients. No differences in T, B, and NK lymphocyte numbers were observed between morphine-treated and non-treated patients. Higher plasma IL-5 and GM-CSF concentrations were noted in IT and PO groups compared to NO. CSF IFNγ concentrations were higher in PO and NO than IT. Positive correlations included CD4 concentrations with opioid escalation indices, and negative correlations involved NK cell concentrations, CSF TNFα concentrations, and opioid escalation indices. Positive correlations were identified between certain cytokines and pain intensity in IT patients, and between NK cells and cumulative morphine dose. Negative correlations were observed between CSF IL-5 concentrations and pain intensity in IT and PO, and between opioid escalation indices and CSF cytokine concentrations in PO and IT. Associations between cytokines, cellular immunity, and prolonged morphine treatment, administered orally and intrathecally were identified.

Use of regenerative medicine in the treatment of endometritis in mares: A systematic review and meta-analysis

Defining the optimal therapy for endometritis remains a significant challenge for clinicians. Given the public health threat posed by antibiotic resistance and the inconclusiveness of traditional therapies, regenerative medicine has been proposed as an alternative. The objective of this study was to conduct a comprehensive systematic review and meta-analysis, to investigate the efficacy of regenerative medicine products in the treatment of both post-breeding persistent and chronic degenerative endometritis (PBIE/CDE) in mares, following the PRISMA guidelines. This research could be a comprehensive scientific reference for determining appropriate treatments and clinical strategies.All studies exploring the use of regenerative medicine therapies (i.e., plasma products, autologous conditioned serum, mesenchymal stem cells MSCs, and MSC derivatives) in the treatment of PBIE/CDE were included, regardless of the specific protocol used, the evaluated outcomes, or the diagnostic method employed. Two authors independently gathered data and evaluated the risk of bias for each study. Treatment effects were assessed using risk ratios for dichotomous data, accompanied by 95 % confidence intervals. Data were aggregated utilizing the fixed-effects model. The quality of evidence for each outcome was evaluated using GRADE criteria.Eighteen studies were included in the systematic review, while fifteen trials were included in the meta-analysis. A sub-meta-analysis was conducted separately on platelet-derived products, as well as on MSCs and their derivatives. The results demonstrated an overall positive effect of regenerative therapies in treating PBIE/CDE, particularly those involving MSCs and their derivatives. The positive outcomes include an anti-inflammatory effect, characterized by reduced intrauterine fluid accumulation, neutrophils, and cytokine concentrations. Additionally, improvements in pregnancy, foaling, and embryo recovery rates have been observed in some cases. Despite the limited number of randomized controlled studies and the high variability among protocols, including the timing of treatment, type, and volume of products used, the use of regenerative products, especially MSCs and their derivatives, has promising results in terms of both efficacy and safety for treating PBIE/CDE in mares.

Blockade of Cluster of Differentiation 38 Alleviates Sepsis-Induced Acute Lung Injury in Mice Through Inflammation Suppression via Regulating Toll-Like Receptor 4/Nuclear Factor kappa B p65

This study aimed to investigate the role of CD38 inhibition in sepsis-induced acute lung injury (ALI) using a murine model. ALI, commonly associated with excessive inflammation, was induced by cecal ligation and puncture (CLP). We observed an increase in CD38 expression in lung tissue over time in the ALI mice. To address this, we administered the CD38 inhibitor, 78C, subcutaneously at a dose of 10 mg/kg. Following treatment, we assessed lung function, inflammatory cell levels, and cytokine concentrations in bronchoalveolar lavage fluid. We also examined the activity of the TLR4/NF-κB p65 signaling pathway in lung tissue. Results showed that 78C treatment improved lung function and reduced inflammatory cells and cytokines in ALI mice. Moreover, 78C inhibited the expression of the TLR4/NF-κB p65 signaling pathway in lung tissue. This CD38 blockade effectively mitigated inflammation levels in lung tissue, ameliorated lung function, and alleviated sepsis-induced ALI induced by CLP. These findings suggest that targeting CD38 to modulate the TLR4/NF-κB p65 inflammatory pathway holds promise as a therapeutic strategy for sepsis-induced ALI.

The Relationship Between Intra-articular Fracture Energy and a Patient's Inflammatory Response.

Prior studies have demonstrated elevated inflammatory cytokine concentrations in the synovial fluid of articular fracture patients postinjury. Similarly, CT-based fracture energy measurements have been correlated with posttraumatic osteoarthritis risk after pilon fracture. The purpose of this study was to determine the associations between synovial fluid cytokine levels, fracture energy, and overall trauma to the body in articular fracture patients. Acute tibial plateau, tibial plafond, and rotational ankle fracture patients were prospectively enrolled from December 2011 through January 1, 2019. Synovial fluid concentrations of interleukin-1 beta, interleukin-1 receptor antagonist, IL-6, IL-8, IL-10, matrix metallopeptidase-1, MMP-3, and MMP-13 were quantified. Patient CT scans were used to calculate fracture energy. The Injury Severity Score (ISS) was used to relate cytokine levels to whole-body injury severity. Spearman rho correlation coefficients were calculated to assess the relationship between injury severity metrics and synovial fluid cytokine, chemokine, and matrix metallopeptidase concentrations. Eighty-seven patients were enrolled with 42 had a tibial plateau fractures (OTA/AO 41B1-2, 41B2-14, 41B3-3, 41C1-3, 41C2-4, 41C3-16), 24 patients had a tibial plafond fracture (OTA/AO 43B1-2, 43B2-4, 43B3-5, 43C1-2, 43C2-3, 43C3-8), and 21 had a rotational ankle fracture (OTA/AO 44B1-3, 44B2-3, 44B3-6, 44C1-4, 44C2-5). Fracture energy significantly differed between fracture patterns, with ankle fractures involving substantially less fracture energy (median = 2.92 J) than plafond (10.85 J, P < 0.001) and plateau fractures (13.05 J, P < 0.001). After adjustment for multiple comparisons, MMP-3 was significantly correlated with transformed fracture energy (r = 0.41, 95% confidence interval [CI], 0.22-0.58, P < 0.001), while IL-1β was significantly correlated with the Injury Severity Score (Spearman ρ = 0.31, 95% CI, 0.08-0.49, P = 0.004). Synovial fluid MMP-3 concentration was significantly correlated with CT-quantified fracture energy in intra-articular fracture patients. Given that in clinical practice fracture energy tends to correlate with posttraumatic osteoarthritis risk, MMP-3 may warrant further investigation for its role in posttraumatic osteoarthritis development after articular fracture. Prognostic Level III. See Instructions for Authors for a complete description of levels of evidence.

Human Placental Mesenchymal Stem Cells and Derived Extracellular Vesicles Ameliorate Lung Injury in Acute Respiratory Distress Syndrome Murine Model.

This study investigates the therapeutic potential of human placental mesenchymal stem cells (P-MSCs) and their extracellular vesicles (EVs) in a murine model of acute respiratory distress syndrome (ARDS), a condition with growing relevance due to its association with severe COVID-19. We induced ARDS-like lung injury in mice using intranasal LPS instillation and evaluated histological changes, neutrophil accumulation via immunohistochemistry, bronchoalveolar lavage fluid cell count, total protein, and cytokine concentration, as well as lung gene expression changes at three time points: 24, 72, and 168 h. We found that both P-MSCs and EV treatments reduced the histological evidence of lung injury, decreased neutrophil infiltration, and improved alveolar barrier integrity. Analyses of cytokines and gene expression revealed that both treatments accelerated inflammation resolution in lung tissue. Biodistribution studies indicated negligible cell engraftment, suggesting that intraperitoneal P-MSC therapy functions mostly through soluble factors. Overall, both P-MSC and EV therapy ameliorated LPS-induced lung injury. Notably, at the tested dose, EV therapy was more effective than P-MSCs in reducing most aspects of lung injury.

Cytokine profile of oral fluid in patients with food allergy associated with atopic dermatitis and psoriasis

The world is experiencing a rapid increase in the prevalence of allergic and autoimmune diseases. It is known that allergic inflammation is most often systemic, involving various organs and systems in the pathological process, such as the skin, respiratory and gastrointestinal tract with the development of dermatorespiratory, dermato-intestinal and other manifestations. The study of the features of the cytokine profile in oral fluid (saliva) deserves special attention, since these characteristics reflect not only local, but also systemic disorders. Of particular relevance is the study of local cytokine regulation of intercellular interactions in food allergies. Our objective was to study the concentration of IL-4, IL-10, IFNγ, secretory IgA in salivary fluid, the concentrations of total immunoglobulin E and eosinophilic cationic protein in blood serum of the patients with atopic dermatitis and psoriasis with concomitant food allergies.The study included patients with atopic dermatitis (AD, group 1, n = 20), psoriasis with concomitant food allergy (PS, group 2, n = 27), psoriasis without concomitant allergies (PS, comparison group 3, n = 23). Quantitative assessment of the cytokine concentrations (IL-4, IL-10, IFNγ, sIgA) in salivary fluid was carried out by enzyme-linked immunosorbent assay. Concentrations of total immunoglobulin E and eosinophilic cationic protein in blood serum were determined by indirect immunofluorescence. The obtained results were processed using the Statistica 8.0 applied software.In groups of patients with atopic dermatitis (Group 1) and psoriasis with concomitant food allergy (Group 2), we have noted a statistically significant increase of salivary IL-4 and IL-10, as well as of total immunoglobulin E concentrations in blood serum as compared with a group of patients with psoriasis without concomitant allergies (group 3), and with control group. When studying concentrations of IFNγ in saliva, no statistically significant intergroup differences were found. The concentration of sIgA in saliva was significantly higher in the groups of patients with atopic dermatitis and psoriasis accompanied by food allergies in comparison with control group and the group of psoriatic patients without food allergies (group 3).The cytokine profile of saliva is characterized by unidirectional changes in food allergy. Skin seems to be the shock organ in this condition, regardless of nosological form of the disease (atopic dermatitis or psoriasis). Salivary fluid is an easily accessible material when assessing the state of mucosal immunity in food allergies.

Better post-operative outcomes at 1-year follow-up are associated with lower levels of pre-operative synovitis and higher levels of IL-6 and VEGFA in unicompartmental knee arthroplasty patients.

Osteoarthritis (OA) is associated with inflammation, and residual inflammation may influence outcomes following knee arthroplasty. This may be more relevant for patients undergoing unicompartmental knee arthroplasty (UKA) due to larger remaining areas of native tissue. This study aimed to: (1) characterise inflammatory profiles for medial UKA patients and (2) investigate whether inflammation markers are associated with post-operative outcomes. This prospective, observational study has national ethics approval. Bloods, synovial fluid, tibial plateaus and synovium were collected from medial UKA patients in between 1 January 2021 and 31 December 2021. Cytokine and chemokine concentrations in serum and synovial fluid (SF) were measured with multiplexed assays. Disease severity of cartilage and synovium was assessed using validated histological scores. Post-operative outcomes were measured with Oxford Knee Score (OKS), Forgotten Joint Score (FJS-12) and pain scores. The study included 35 patients. SF VEGFA was negatively correlated with pre-operative pain at rest (r -0.5, p = 0.007), and FJS-12 at six-week (r 0.44, p = 0.02), six-month (r 0.61, p < 0.01) and one-year follow-up (r 0.63, p = 0.03). Serum and SF IL-6 were positively correlated with OKS at early follow-up (serum 6weeks, r 0.39, p = 0.03; 6months, r 0.48, p < 0.01; SF 6weeks, r 0.35, p = 0.04). At six weeks, increased synovitis was negatively correlated with improvements in pain at rest (r -0.41, p = 0.03) and with mobilisation (r -0.37, p = 0.047). Lower levels of synovitis and higher levels of IL-6 and VEGFA were associated with better post-operative outcomes after UKA, which could be helpful for identifying UKA patients in clinical practice. Level IV case series.

Effects of firocoxib, trimethoprim sulfamethoxazole and altrenogest on inflammation and foal survival after administration to mares with experimentally induced placentitis

Bacterial placentitis is a leading cause of pregnancy loss in mares. Infection, inflammation and prostaglandin production induce preterm delivery leading to poor foal survival. Treatment strategies for placentitis that control infection and inhibit inflammation are vital. The goals of this study were: 1. Assess the anti-inflammatory effects of firocoxib, trimethoprim sulfamethoxazole (TMS) and altrenogest administered to mares with placentitis; 2. Determine the efficacy of firocoxib, TMS and altrenogest for improving foal outcome in mares with placentitis. We hypothesized that this treatment would significantly decrease pro-inflammatory mediators in fetal fluids and tissues when compared to untreated, infected mares and would also improve foal survival. Placentitis was induced in 13 pregnant mares (270-300 days gestation; via intracervical inoculum of Streptococcus zooepidemicus). Mares were randomly assigned to: infected/untreated (UNTREATED; n=6); or infected/treated groups (TREATED; n=7; administered firocoxib, TMS, and altrenogest from the onset of clinical signs of placentitis until abortion/delivery). Fetal fluids, membranes and tissue samples were obtained at abortion/delivery for quantification of TNF-α, IL-1β, IL-10, IL-6, IL-8, PGE2, PGF2α, and matrix metalloproteinases (MMP) -1, -3, -9 using ELISA (fluids) and real-time PCR (tissues). P < 0.05 was considered significant. Gestation length and foal survival were significantly higher in TREATED (mean days post-infection = 40; 7/7 live foals) than UNTREATED (mean days post-infection = 14; 2/6 live foals). Administration of firocoxib, TMS andaltrenogest resulted in significantly lower amniotic fluid concentrations of IL-1β, IL-6, PGE2 and PGF2α in TREATED than UNTREATED. Amniotic fluid IL-10 concentration tended (P < 0.1) to be lower in TREATED than UNTREATED. Cytokine and prostaglandin concentrations in allantoic fluid were not different between groups except PGE2 which tended (P < 0.1) to be lower in TREATED than UNTREATED. The relative quantity of cytokine and MMP mRNA expression in fetal membranes and foal tissues (non-survivors) was log-transformed and estimated marginal means determined. Fetal membranes from TREATED had significantly lower mRNA expression for IL-8 and TNFα compared to membranes from UNTREATED. mRNA expression of IL-1β, IL-6 and MMP3 tended (P < 0.1) to be lower in membranes from TREATED than UNTREATED. mRNA expression of IL-1β, IL-8, IL-10, TNFα, MMP1, 3 and 9 were significantly different between fetal membrane sites. In non-surviving foals, IL-8 mRNA expression was significantly lower in liver than lung while MMP3 expression was significantly higher in liver than lung or spleen. These data provide physiologic evidence that administration of firocoxib, TMS and altrenogest to mares with placentitis has broad anti-inflammatory effects and may improve foal survival. Research funding: Grayson Jockey Club Research Foundation; Merial Animal Health; Kentucky Thoroughbred Breeders Association, University of Florida, University of Georgia.

The intrafollicular concentration of leptin as a potential biomarker to predict oocyte maturity in in-vitro fertilization

Oocyte maturity is critical to the development potential of the embryo and pregnancy outcomes in natural and in-vitro fertilization (IVF). In IVF, oocyte maturity is typically evaluated using morphological criteria, although such assessment remains highly subjective. To identify reliable biomarkers of oocyte maturity, this study investigates the relationship between follicular cytokine concentrations and oocyte maturity in IVF patients with different ovarian reserves. In this prospective study, follicular fluid was collected during oocyte retrieval and the concentrations of cytokines involved in ovarian folliculogenesis were determined. Follicular fluid cytokine concentrations were compared between participants in three groups according to serum anti-Mullerian hormone (AMH) concentration, as follows: low AMH, < 2 ng/mL; normal AMH, 2–5 ng/mL; and high AMH, > 5 ng/mL. Pearson's correlation coefficient analysis showed that the number of mature oocytes correlated positively and strongly with serum AMH level (r = 0.719; p < 0.01). The leptin concentration in follicular fluid was significantly higher in women with normal AMH level than in those with low or high levels. ROC curve analysis showed that the follicular fluid levels of leptin (area under ROC curve, 0.829; 95% confidence interval, 0.659–0.998; p < 0.01) and SCF (area under ROC curve, 0.706; 95% confidence interval, 0.491–0.921; p = 0.087) were the best predictors of oocyte maturity. At an optimal cut-off value of 16 ng/mL, leptin had positive predictive value (sensitivity) up to 70% and negative predictive value (specificity) of 91% for indicating oocyte maturity. The concentration of leptin in follicular fluid is closely related to ovarian reserve and may serve as a biomarker to predict oocyte maturity.

TLR-activated mesenchymal stromal cell therapy and antibiotics to treat multi-drug resistant Staphylococcal septic arthritis in an equine model.

Rapid development of antibiotic resistance necessitates advancement of novel therapeutic strategies to treat infection. Mesenchymal stromal cells (MSC) possess antimicrobial and immunomodulatory properties, mediated through antimicrobial peptide secretion and recruitment of innate immune cells including neutrophils and monocytes. TLR-3 activation of human, canine and equine MSC has been shown to enhance bacterial killing and clearance in vitro, in rodent Staphylococcal biofilm infection models and dogs with spontaneous multi-drug-resistant infections. The objective of this study was to determine if intra-articular (IA) TLR-3-activated MSC with antibiotics improved clinical parameters and reduced bacterial counts and inflammatory cytokine concentrations in synovial fluid (SF) of horses with induced septic arthritis. Eight horses were inoculated in one tarsocrural joint with multidrug-resistant Staphylococcus aureus (S. aureus). Bone marrow-derived MSC from three unrelated donors were activated with TLR-3 agonist polyinosinic, polycytidylic acid (pIC). Recipient horses received MSC plus vancomycin (TLR-MSC-VAN), or vancomycin (VAN) alone, on days 1, 4, 7 post-inoculation and systemic gentamicin. Pain scores, quantitative bacterial counts (SF, synovium), SF analyses, complete blood counts, cytokine concentrations (SF, plasma), imaging changes (MRI, ultrasound, radiographs), macroscopic joint scores and histologic changes were assessed. Results were reported as mean ± SEM. Pain scores (d7, P=0.01, 15.2±0.2 vs. 17.9±0.5), ultrasound (d7, P=0.03, 9.0±0.6 vs. 11.8±0.5), quantitative bacterial counts (SF d7, P=0.02, 0±0 vs. 3.4±0.4; synovium P=0.003, 0.4±0.4 vs. 162.7±18.4), systemic neutrophil (d4, P=0.03, 4.6±0.6 vs. 7.8±0.6) and serum amyloid A (SAA) (d4, P=0.01, 1,106.0±659.0 vs. 2,858.8±141.3; d7, P=0.02, 761.8±746.2 vs. 2,357.3±304.3), and SF lactate (d7, P<0.0001, 5.4±0.2 vs. 15.0±0.3), SAA (endterm, P=0.01, 0.0 vs. 2,094.0±601.6), IL-6 (P=0.03, 313.0±119.2 vs. 1,328.2±208.9), and IL-18 (P=0.02, 11.1±0.5 vs. 13.3±3.8) were improved in TLR-MSC-VAN vs. VAN horses. Study limitations include the small horse sample size, short study duration, and lack of additional control groups. Combined TLR-activated MSC with antibiotic therapy may be a promising approach to manage joint infections with drug resistant bacteria.

Differences in Synovial Cytokine Profile Associated with Long-Term Clinical Outcomes in Patients with Knee Osteoarthritis Undergoing Corrective Osteotomy with Platelet-Rich Plasma or Stromal Vascular Fraction Post-Treatments.

Functional outcomes and synovial fluid (SF) cytokine concentrations in response to platelet-rich plasma (PRP) or stromal vascular fraction (SVF) post-treatments following open wedge high tibial osteotomy (HTO) in 20 patients with knee osteoarthritis (OA) were examined. Six weeks after surgery, the knees of 10 patients were injected with autologous PRP (PRP subgroup), while another 10 patients were injected with autologous SVF (SVF subgroup) and monitored for 1.5 years. Pain assessment (VAS score) and functional activity (KOOS, KSS, Outerbridge, and Koshino scores) were applied. PRP subgroup performed better compared with the SVF subgroup according to KOOS, KSS, and VAS scores, while the SVF subgroup demonstrated better results according to Outerbridge and Koshino testing and produced more pronounced cartilage regeneration in the medial condyle and slowed down cartilage destruction in its lateral counterpart. SF was collected before and one week after PRP or SVF injections and tested for concentrations of 41 cytokines (Multiplex Assay). In the PRP subgroup, a significant decrease in IL-6 and CXCL10 synovial concentrations was accompanied by an increase in IL-15, sCD40L, and PDGF-AB/BB amounts. The SVF subgroup demonstrated a significant decrease in synovial TNFα, FLT-3L, MIP-1β, RANTES, and VEGF concentrations while SF concentrations of MCP-1 and FGF2 increased. Both post-treatments have a potential for increased tissue regeneration, presumably due to the downregulation of inflammation and augmentation of synovial growth factor concentrations.

Aquaporin-1 inhibition exacerbates ischemia-reperfusion-induced lung injury in mouse

BackgroundIschemia-reperfusion injury (IRI), which involves severe inflammation and edema, is an inevitable feature of the lung transplantation process and leads to primary graft dysfunction (PGD). The activation of aquaporin 1 (AQP1) modulates fluid transport in the alveolar space. The current study investigated the role of AQP1 in ischemia-reperfusion (IR)-induced lung injury. MethodsA mouse model of lung IR was established by clamping the left lung hilar for 1 h and released for reperfusion for 24 h. The AQP1 inhibitor acetazolamide (AZA) was administered 3 days before lung ischemia with a dose of 100 mg/kg per day via gavage. Lung injury was evaluated using the ratio of wet-to-dry weight, peripheral bronchial epithelial thickness, degree of angioedema, acute lung injury score, neutrophil infiltration, and cytokine concentrations in bronchoalveolar lavage fluid. ResultsCompared with sham treatment, ischemia with no reperfusion (IR 0h) and ischemia with reperfusion for 24 h (IR 24 h) significantly upregulated AQP1 expression, increased the wet/dry weight ratio, angioedema, neutrophil infiltration and cytokine production (interleukin -6 and tumor necrosis factor -α) and thickened the peripheral bronchial epithelium. AZA exacerbated inflammation and pulmonary edema. ConclusionAQP1 may exert a protective effect against IR-induced lung injury, which could be attributed to alleviating pulmonary edema and inflammation. AQP1 upregulation might be a potential application to alleviate lung IRI and decrease the incidence of PGD.

Natriuretic peptide receptor-C mediates the inhibitory effect of atrial natriuretic peptide on neutrophil recruitment to the lung during acute lung injury.

Atrial natriuretic peptide (ANP) protects against acute lung injury (ALI), but the receptor that mediates this effect is not known. Transgenic mice with 0 (knockout), 1 (heterozygote), or 2 (wild-type) functional copies of Npr3, the gene that encodes for natriuretic peptide receptor-C (NPR-C), were treated with intravenous infusion of ANP or saline vehicle before oropharyngeal aspiration of Pseudomonas aeruginosa (PA103) or saline vehicle. Lung injury was assessed 4 h following aspiration by measurement of lung wet/dry (W/D) weight, whole lung leukocyte and cytokine levels, and protein, leukocyte, and cytokine concentration in bronchoalveolar lavage fluid (BALF). PA103 induced acute lung injury as evidenced by increases in lung W/D ratio and protein concentration in BALF. The severity of PA103-induced lung injury did not differ between NPR-C genotypes. Treatment with intravenous ANP infusion reduced PA103-induced increases in lung W/D and BALF protein concentration in all three NPRC genotypes. PA103 increased the percentage of leukocytes that were neutrophils and cytokine levels in whole lung and BALF in NPR-C wild-type and knockout mice. This effect was blunted by ANP in wild-type mice but not in the NPR-C knockout mice. NPR-C does not mediate the protective effect of ANP on endothelial cell permeability in settings of PA103-induced injury but may mediate the effect of ANP on inhibition of the recruitment of neutrophils to the lung and thereby attenuate the release of inflammatory cytokines.

O-191 Endometriosis is associated with a lowered cumulative live birth rate by alters the macrophage polarization and the cytokine concentrations in follicular fluid

Abstract Study question The impact of endometriosis on the outcomes of ART is still ambiguous and the mechanism by which endometriosis impacts fertility has not been fully elucidated Summary answer Endometriosis was associated with low CLBR in IVF, which might be due to the change of follicular microenvironment and compromising the quality of embryos. What is known already Despite the great advantages of IVF treatment in endometriosis-linked infertility, the impact of endometriosis on outcomes of infertility management with ART is controversial and few studies focused on the association between endometriosis and CLBR. Numerous mechanisms have been proposed in an effort to delineate the multifaceted pathophysiology that induces impairment of reproductive dynamics in patients with endometriosis. Reactive oxygen species, dysregulation of the immune system and cellular architectural disruption constitute the crucial mechanisms that detrimentally affect oocyte and embryo developmental potential. Study design, size, duration This study retrospectively included 433 patients with endometriosis and 1299 patients with tuber factor infertility to evaluate the impact of endometriosis on IVF pregnancy outcome between January 2016 and December 2018.The basic study prospectively recruited 30 patients with ovarian endometriosis and 35 controls with tubal factor infertility to analyze polarization stage of macrophages in their follicular fluid, and another 20 females with ovarian endometriosis and 30 controls to detect cytokines in their FF. Participants/materials, setting, methods Data on all women undergoing fresh or frozen IVF treatment cycles were analysed to compare the CLBR between endometriosis and tuber factor infertility patients. A cytometry panel of 4 antibodies (CD45, CD3, CD80, CD163) was designed to enumerate the numbers of MI and MII macrophages from the cell sediment from FF samples. Quantibody® array was utilized to determine the concentration of 10 cytokines in FF, including IFN-γ, IL-1α, IL-1β, IL-10,IL-13,IL-4, IL-6, IL-8, MCP-1 and TNF-α. Main results and the role of chance The results showed that patients with endometriosis were associated with noticeably fewer retrievable oocytes, a lower oocyte maturity rate, decreased numbers of available and high-quality embryos (all p &amp;lt; 0.001) in comparison with the control group. The clinical pregnancy and live birth rate of the endometriosis group were lower in the frozen-thawed embryo transfer cycles (p = 0.028 and p = 0.008, respectively), which leading to a declined cumulative live birth rate (CLBR) (p = 0.001). Logistic regression analysis indicated a close association between endometriosis and low CLBR (p = 0.002). Furthermore, the numbers of type I and type II macrophages in follicular fluid (FF) of patients with ovarian endometriosis were significantly increased compared with the control group (p &amp;lt; 0.001). The expressions of IL-1α, IL-1β, TNF-α, IL-6, IL-13, and IL-10 in FF were higher in endometrioma group(p &amp;lt; 0.05). The increased expression of cytokines was negatively correlated with embryo outcomes, including the numbers of total oocytes retrieved, mature oocytes, and fertilized oocytes, and the numbers of high-quality blastocysts and embryos. Additionally, IL-6 and IL-8 were positively correlated with AMH, rate of available blastocysts, number of blastocysts formed and available blastocysts, but were negatively associated with rate of MII oocytes. Limitations, reasons for caution The present study being of a retrospective and monocentric study, may not come to a very convincing conclusion, and some unknown biases might still exist due to possible underestimation of some confounders. Wider implications of the findings Collectively, our results indicate that endometriosis does adversely affect pregnancy outcomes of ART, as women with endometriosis produce a lower quantity of oocytes and embryos. Moreover, endometriosis might play a role in oocyte or embryo outcomes via regulating the production of cytokines or the number of immune cells in FF. Trial registration number no

Vascular network expansion, integrity of blood–brain interfaces, and cerebrospinal fluid cytokine concentration during postnatal development in the normal and jaundiced rat

BackgroundSevere neonatal jaundice resulting from elevated levels of unconjugated bilirubin in the blood induces dramatic neurological impairment. Central oxidative stress and an inflammatory response have been associated with the pathophysiological mechanism. Cells forming the blood–brain barrier and the choroidal blood–CSF barrier are the first CNS cells exposed to increased plasma levels of unconjugated bilirubin. These barriers are key regulators of brain homeostasis and require active oxidative metabolism to fulfill their protective functions. The choroid plexus-CSF system is involved in neuroinflammatory processes. In this paper, we address the impact of neonatal hyperbilirubinemia on some aspects of brain barriers. We describe physiological changes in the neurovascular network, blood–brain/CSF barriers integrities, and CSF cytokine levels during the postnatal period in normobilirubinemic animals, and analyze these parameters in parallel in Gunn rats that are deficient in bilirubin catabolism and develop postnatal hyperbilirubinemia.MethodsGunn rats bearing a mutation in UGT1a genes were used. The neurovascular network was analyzed by immunofluorescence stereomicroscopy. The integrity of the barriers was evaluated by [14C]-sucrose permeability measurement. CSF cytokine levels were measured by multiplex immunoassay. The choroid plexus-CSF system response to an inflammatory challenge was assessed by enumerating CSF leukocytes.ResultsIn normobilirubinemic animals, the neurovascular network expands postnatally and displays stage-specific regional variations in its complexity. Network expansion is not affected by hyperbilirubinemia. Permeability of the blood–brain and blood–CSF barriers to sucrose decreases between one- and 9-day-old animals, and does not differ between normobilirubinemic and hyperbilirubinemic rats. Cytokine profiles differ between CSF and plasma in all 1-, 9-, and 18-day-old animals. The CSF cytokine profile in 1-day-old animals is markedly different from that established in older animals. Hyperbilirubinemia perturbs these cytokine profiles only to a very limited extent, and reduces CSF immune cell infiltration triggered by systemic exposure to a bacterial lipopeptide.ConclusionThe data highlight developmental specificities of the blood–brain barrier organization and of CSF cytokine content. They also indicate that a direct effect of bilirubin on the vascular system organization, brain barriers morphological integrity, and inflammatory response of the choroid plexus-CSF system is not involved in the alteration of brain functions induced by severe neonatal jaundice.

Supplemental Protein and a Multinutrient Beverage Speed Wound Healing after Acute Sleep Restriction in Healthy Adults

BackgroundPhysiologic and psychologic stress slow healing from experimental wounds by impairing immune function. ObjectivesWe aimed to determine whether supplemental protein and multinutrient supplementation improved wound healing markers after acute stress induced by acute sleep restriction. MethodsIn this single-blind, crossover study in generally healthy young adults (18 males/2 females; mean ± SD age: 19.7 ± 2.30 y), experimental wounds were created by removing the top layer of forearm blisters induced via suction after 48 h of 72-h sleep restriction (2-h nightly sleep), a protocol previously shown to delay wound healing. Skin barrier restoration (measured by transepidermal water loss) assessed wound healing ≤10 d postblistering, and local immune responses were evaluated by serial measurement of cytokine concentrations in fluid collected at wound sites for 48 h postblistering. Participants consumed controlled, isocaloric diets with either 0.900 g · kg−1 · d−1 protein plus placebo (PLA) or 1.50 g · kg−1 · d−1 protein plus multinutrient beverage [l-arginine: 20.0 g/d; l-glutamine: 30.0 g/d; omega-3 (n–3) fatty acids: 1.00 g/d; zinc sulfate: 24.0 mg/d; cholecalciferol: 800 IU/d; and vitamin C: 400 mg/d] (NUT) during sleep restriction and for 4 d afterwards. ResultsSkin barrier restoration (primary outcome) was shorter for NUT (median: 3.98 d; IQR: 1.17 d) than for PLA (median: 5.25 d; IQR: 1.05 d) (P = 0.001). Cytokines from wound fluid (secondary outcome) increased over time (main effect of time P ≤ 0.001), except IL-13 (P = 0.07); however, no effects of treatment were observed. ConclusionsSupplemental nutrition may promote wound healing after sleep restriction in healthy adults including military personnel, the latter of which also have a high incidence of wounds and infection.This trial was registered at clinicaltrials.gov as NCT03525184.

Cytokines in temporomandibular joint synovial fluid and tissue in relation to inflammation.

BackgroundSynovial tissue is known to be the origin of inflammation in joint disease. Despite this, synovial fluid is the main biological specimen of choice in temporomandibular joint (TMJ) inflammation and pathology biomarker research. No comparison of TMJ protein content between synovial fluid and synovial tissue has been made.ObjectivesThe aim of this study was to investigate whether cytokine concentrations in synovial fluid can be related to cytokine concentrations in synovial tissue and to analyse correlation of clinical parameters reflecting local inflammation to cytokine concentrations.MethodsSynovial tissue and fluid samples were obtained during the same surgical procedure from a cohort of 101 patients with TMJ disorders. Interleukin (IL) 1β, IL‐6, IL‐8, IL‐10 and tumour necrosis factor α (TNF‐α) were analysed in the samples and an intraindividual correlation made. Various patient‐specific factors related to TMJ inflammation were associated with the cytokine concentrations in synovial fluid and tissue.ResultsNo correlation between cytokine concentration in synovial fluid and synovial tissue was found, except for IL‐8 (ρ = .284, p = .024). Synovial tissue cytokines correlated strongly to inflammation‐related factors: diagnosis (IL‐1β, p = .001; TNF‐α, p = .000; IL‐10, p = .000), TMJ palpation pain (IL‐1β, p = .024; TNF‐α, p = .025), synovitis score (IL‐1β, p = .015) and subjective TMJ pain (TNF‐α, p = .016). Synovial fluid cytokines showed no significant relations to inflammation.ConclusionsThe investigated cytokine concentrations showed weak correlations between synovial fluid and synovial tissue, besides IL‐8. Synovial tissue appeared to reflect inflammation to a higher extent than synovial fluid. Thus, suggesting that synovial tissue research should complement synovial fluid in future explorations of TMJ pathology and inflammation.

Synovial Fluid Cytokine Profile at the Time of Arthroscopy Explains Intermediate-Term Functional Outcomes

Background: The intra-articular immune response after ligamentous, meniscal, or focal chondral knee injuries likely plays a role in intra-articular healing and the onset and progression of posttraumatic osteoarthritis. Purpose: To evaluate the association of synovial fluid cytokine concentrations measured at the time of knee arthroscopy with intermediate-term functional outcomes after knee arthroscopy based on the Lysholm score. Study Design: Cohort study; Level of evidence, 2. Methods: This was a prospective cohort study of patients undergoing arthroscopic knee surgery. Synovial fluid was aspirated from the injured knee immediately before surgical incision, and the concentrations of 10 cytokines were analyzed using immunoassay. Principal component regression was used to create a model to predict patient-reported Lysholm score at a minimum of 5 years postoperatively. Hierarchical clustering was performed to identify groups of patients with similar synovial fluid inflammatory phenotypes. Lysholm scores and cytokine concentrations were compared between clusters. Results: A total of 26 patients (mean age, 40.33 ± 16.40 years) were included in the analysis. The mean duration between surgery and follow-up was 6.69 ± 0.72 years. A model consisting of 2 principal components (PC1, PC2) explained 62.48% of the variance in the cytokine data and 52.03% of the variance in intermediate-term Lysholm score. Hierarchical clustering resulted in 3 patient clusters based on the principal components used in the regression model. Despite no baseline differences in Lysholm score, cluster 3 demonstrated significantly greater intermediate-term Lysholm score compared with cluster 2 (94.33 vs 76.09, respectively; 95% CI, 5.96-30.52; P = .006) and cluster 1 (94.33 vs 52.33, respectively; 95% CI, 24.09-59.91; P = .003). Cluster 3, when compared with the overall means, was characterized by greater PC1 value (1.01 vs 0.00, respectively; P = .030) and greater PC2 value (0.86 vs 0.00, respectively; P = .002). Conclusion: The concentrations of select synovial fluid cytokines assessed at the time of knee arthroscopy can be used to explain more than half of the variance in intermediate-term functional outcomes.

Comparative analyses in transcriptome of human granulosa cells and follicular fluid micro-environment between poor ovarian responders with conventional controlled ovarian or mild ovarian stimulations

BackgroundBoth mild and conventional controlled ovarian stimulation are the frequently used protocols for poor ovarian responders. However, there are some debates about which treatment is better. Moreover, little is known about the follicular physiology after the two ovarian stimulation protocols. This study was intended to investigate the features in granulosa cells and follicular fluid micro-environment after the two different ovarian stimulation protocols in poor responders.MethodsGranulosa cells RNA were sequenced using Illumina Hiseq technology. Specific differently expressed genes and proteins were verified by real-time quantitative PCR and Western blot analysis. Moreover, hormone and cytokine concentrations in the follicular fluid were measured by electrochemiluminescence immunoassay and enzyme-linked immunoabsorbent assay. The correlation between the results of molecular experiments and the laboratory outcomes were analyzed by Spearman correlation analysis.ResultsThe differentially expressed genes between the two groups were involved in 4 signaling pathways related to the follicular development; three proteins pertinent to the TGF-β signaling pathway were expressed differently in granulosa cells between the two, and the constituents in the follicular fluid were also different. Further, a correlation between the TGF-β signaling pathway and the good-quality embryo was observed.ConclusionsThe present study made a comparison for the first time in the transcriptome of human granulosa cells and the follicular fluid micro-environment between poor responders with the conventional controlled ovarian stimulation or the mild ovarian stimulation, showing that the TGF-β signaling pathway may correlate with the good-quality of embryos in the mild group, which may be instrumental to the choice of optimal management for IVF patients.