Concentrations Of Leukotrienes Research Articles

Mucoactive therapy in the treatment of sinusitis in patients with post-Covid syndrome

Rhinosinusitis, as in the pre-ovoid era, are still the most common diseases in the practice of an otorhinolaryngologist. According to the results of the analysis of medical documentation conducted at the clinic of the St. Petersburg Scientific Research Institute of Ear, Throat, Nose and Speech, more and more patients associate the diseases of the nasopharyngeal space that have appeared in them and the increase in the number of exacerbations of existing chronic diseases of this area with a coronavirus infection. In this regard, we decided to consider approaches to the treatment of rhinosinusitis by summarizing the available data and considering individual clinical cases of patients with sinusitis and COVID-19. Antibiotic resistance associated with uncontrolled administration of antibacterial drugs when infected with SARS-CoV-2 has also become one of the frequent problems at present. Therefore, our article considers the most relevant method in this case, based on the use of phytotherapy. According to the literature, the most effective phytopreparations are preparations containing standardized myrtol, which reduces the viscosity of the mucus of the paranasal sinuses and nasal cavity, increases the frequency of beating of the cilia of the ciliated epithelium, thereby contributing to an increase in the speed of mucociliary transport. Standardized myrtol also has an anti-inflammatory effect by reducing the concentration of leukotrienes and prostaglandin, neutralizing free OH radicals, inhibiting oxidation processes myrtol reduces toxic effects on the nasal mucosa and paranasal sinuses, and also acts on pathogenic flora as a bacteriostatic drug. Clinical recommendations of the Ministry of Health of the Russian Federation from 2021 according to the treatment of acute sinusitis and the European recommendations of the European Position Paper on Rhinosinusitis and Nasal polyps (EPOS 2020), based on the data of controlled studies presented in them, the use of phytopreparations in the treatment of acute and chronic sinusitis is confirmed both in combination with antibacterial therapy and without antibacterial drugs in combination with irrigation therapy and the use of topical decongestants.

Сучасні підходи до вибору базисної терапії легкої персистуючої бронхіальної астми у дітей

Мета роботи: удосконалення лікування легкої персистуючої бронхіальної астми на підставі розроблення диференційної програми протизапальної терапії. Зі 120 дітей сформовані 3 групи залежно від ефективної базисної терапії (антилейкотрієновий препарат, флутиказону пропіонат або їх комбінація), яка дозволила досягти контролю над захворюванням. Результати. Найбільш високі концентрації лейкотрієнів C4/D4/E4 у сироватці крові спостерігалися у дітей 3-ї групи, навіть після лікування вони залишалися вищими, ніж у дітей контрольної групи. При рівні лейкотрієнів від 500 до 1000 пг/мл шанси досягти контролю над бронхіальною астмою за допомогою антилейкотрієнового препарату у 5,9 раза вищі порівняно з інгаляційними глюкокортикостероїдами (95% довірчий інтервал 2,01–18,2). А коли рівень лейкотрієнів (C4/D4/E4) перевищував 1000 пг/мл, комбіноване лікування є ефективнішим, ніж терапія антилейкотрієновим препаратом (відношення шансів (OR) = 6,7; СІ95% 2,5–17,8) та флутиказону пропіонатом (OR = 83,3; СІ95% 9,9– 667,9).Висновки. Антилейкотрієновий препарат як монотерапія показаний дітям 6–7 років з легкою персистуючою бронхіальною астмою при сироваткових рівнях лейкотрієнів від 500 до 1000 пг/мл. При рівні лейкотрієнів нижче від 500 пг/мл доцільно призначати флутиказону пропіонат, вище від 1000 пг/мл — комбінацію антилейкотрієнового препарату та інгаляційного глюкокортикостероїду.

Differential Lipid Mediator Involvement in the Different Forms of Genetic Frontotemporal Dementia: Novel Insights into Neuroinflammation.

The pathophysiology of frontotemporal dementia (FTD) is poorly understood but recent studies implicate neuroinflammation as an important factor. However, little is known so far about the role of the resolution pathway, the response to inflammation that allows tissue to return to a homeostatic state. We aimed to measure the concentrations of lipid mediators including specialized proresolving mediators (SPMs) and proinflammatory eicosanoids in the cerebrospinal fluid (CSF) of people with FTD. 15 people with genetic FTD (5 with C9orf72 expansions, 5 with GRN mutations, and 5 with MAPT mutations) were recruited to the study along with 15 age- and sex-matched healthy controls. Targeted liquid chromatography-tandem mass spectrometry techniques were used to measure the CSF concentrations of lipid mediators in the docosahexaenoic acid (DHA), n-3 docosapentaenoic acid, eicosapentaenoic acid, and arachidonic acid (AA) metabolomes. Only the C9orf72 expansion carriers had higher concentrations of SPMs (DHA-derived maresins and DHA-derived resolvins) compared with controls. In contrast, GRN and MAPT mutation carriers had normal concentrations of SPMs but significantly higher concentrations of the proinflammatory AA-derived leukotrienes and AA-derived thromboxane compared with controls. Additionally, the C9orf72 expansion carriers also had significantly higher concentrations of AA-derived leukotrienes. This initial pilot study of lipid mediators provides a window into a novel biological pathway not previously investigated in FTD, showing differential patterns of alterations between those with C9orf72 expansions (where SPMs are higher) and GRN and MAPT mutations (where only proinflammatory eicosanoids are higher).

Leukotrienes in Tumor-Associated Inflammation.

Leukotrienes are biologically active eicosanoid lipid mediators that originate from oxidative metabolism of arachidonic acid. Biosynthesis of leukotrienes involves a set of soluble and membrane-bound enzymes that constitute a machinery complex primarily expressed by cells of myeloid origin. Leukotrienes and their synthetic enzymes are critical immune modulators for leukocyte migration. Increased concentrations of leukotrienes are implicated in a number of inflammatory disorders. More recent work indicates that leukotrienes may also interact with a variety of tissue cells, contributing to the low-grade inflammation of cardiovascular, neurodegenerative, and metabolic conditions, as well as that of cancer. Leukotriene signaling contributes to the active tumor microenvironment, promoting tumor growth and resistance to immunotherapy. This review summarizes recent insights into the intricate roles of leukotrienes in promoting tumor growth and metastasis through shaping the tumor microenvironment. The emerging possibilities for pharmacological targeting of leukotriene signaling in tumor metastasis are considered.

Development and Characterization of an Allergic Asthma Rat Model for Interventional Studies.

Allergic asthma is one of the most common chronic diseases of the airways, however it still remains underdiagnosed and hence undertreated. Therefore, an allergic asthma rat model would be useful to be applied in future therapeutic strategy studies. The aim of the present study was to develop an objective model of allergic asthma in atopic rats that allows the induction and quantification of anaphylactic shock with quantitative variables. Female Brown Norway rats were intraperitoneally sensitized with ovalbumin (OVA), alum and Bordetella pertussis toxin and boosted a week later with OVA in alum. At day 28, all rats received an intranasal challenge with OVA. Anaphylactic response was accurately assessed by changes in motor activity and body temperature. Leukotriene concentration was determined in the bronchoalveolar lavage fluid (BALF), and total and IgE anti-OVA antibodies were quantified in blood and BALF samples. The asthmatic animals’ motility and body temperature were reduced after the shock for at least 20 h. The asthmatic animals developed anti-OVA IgE antibodies both in BALF and in serum. These results show an effective and relatively rapid model of allergic asthma in female Brown Norway rats that allows the quantification of the anaphylactic response.

Modern approaches to the treatment of osteoarthritis

The data given in the article is in accordance with the latest recommendations of ESCEO years 2016, 2019 in regard to possibilities of the sympthom-modifying and structure-modifying action of chondroitin sulfate in the treatment of patients with osteoarthritis. There is the data on the therapy effect when treating patients with osteoarthritis and with periarticular tissues damage with chondroitin sulfate injections (Alflutop, Biotehnos) having strong evidence base, including randomized placebo-controlled studies. In the randomized study it is shown reliably high effectiveness of Alflutop against placebo in regard to main clinical manifestations of the desease as well as possibility to slow down progress of the destructive processes at osteoarthritis of the knee-joint. The data given on estimation of effectiveness and safety of alternate introduction of Alflutop at osteoarthritis 2 ml intramuscularly every other day (10 injections) compared with a standard mode of use of Alflutop 1 ml intramuscularly every day for 20 days. Equal effectiveness of the two modes of Alflutop introduction is demonstrated subject to good tolerance of the drug. Ability of Alflutop to supress the mediators of inflammation, such as interleukins 6 and 8, and degradations at osteoarthritis, as well as to stimulate the proliferation and renewal of chondrocytes is shown according to the results of an international experimental study, explaining its ability to cause a quick analgesic and anti-inflammatory effect. Alflutop inhibits in vitro extracellular release of the pro-inflammatory cytokine interleukin 6 (IL-6) for 16 per cent and chemokine IL-8 for 35 per cent, inhibits pro-angiogenic factor VEGF for 56 per cent, has an antioxidant effect, which contributes to the synthesis of cartilage macromolecules, a decrease in the activity of matrix metalloproteases (MMPs), a decrease in the concentration of prostaglandins and leukotrienes, and a decrease in chondrocyte apoptosis. The data on the effectiveness of the Alflutop injection course in case of damage to the periaticular tissues in the shoulder, elbow, knee joint is given. The data on the gastroprotective effect of the drug, demonstrated in experimental and clinical studies is given.

LTB4 and 5-oxo-ETE from extracellular vesicles stimulate neutrophils in granulomatosis with polyangiitis

Activation of neutrophils is an important mechanism in the pathology of granulomatosis with polyangiitis (GPA). In this study, we evaluated whether extracellular vesicles (EVs) circulating in the plasma of GPA patients could contribute to this process. EVs from the plasma of GPA patients in the active stage of the disease (n = 10) and healthy controls (n = 10) were isolated by ultracentrifugation and characterized by flow cytometry (CD63, CD8) and nanoparticle tracking analysis. Targeted oxylipin lipidomics of EVs was performed by HPLC-MS/MS. EV/oxylipin-induced neutrophil extracellular traps (NETs) were analyzed by confocal microscopy, and released double-stranded DNA (dsDNA) was quantified by PicoGreen fluorescent dye. Reactive oxygen species (ROS) production and neutrophils' EV binding/uptake were evaluated by flow cytometry. Brief priming with granulocyte-macrophage colony-stimulating factor was required for EV-mediated ROS production and dsDNA release. It was observed that priming also increased EV binding/uptake by neutrophils only for EVs from GPA patients. EVs from GPA patients had higher concentrations of leukotriene (LT)B4 and 5-oxo-eicosatetraenoic acid (5-oxo-ETE) as compared with EVs from healthy controls. Moreover, neutrophils stimulated with LTB4 or 5-oxo-ETE produced ROS and released dsDNA in a concentration-dependent manner. These results reveal the potential role of EVs containing oxylipin cargo on ROS production and NET formation by activated neutrophils.

CONCENTRATIONS OF CYSTEINYL LEUKOTRIENES IN VARIOUS BIOLOGICAL FLUIDS OF CHILDREN WITH BRONCHIAL ASTHMA, ATOPIC DERMATITIS AND FOOD PROTEIN INDUCED ENTEROCOLITIS

Clinical manifestation of food allergy is characterized by polymorphic cutaneous, respiratory and gastrointestinal syndromes. Leukotrienes occupy a key place in the pathogenesis of a wide range of inflammatory diseases, including bronchial asthma, allergic rhinitis, atopic dermatitis, hives, allergic conjunctivitis, atherosclerotic cardio-vascular lesions system, inflammatory bowel disease, multiple sclerosis, cancer, etc. Better understanding of general pathophysiological mechanisms of allergic realization put the focus on the studying of cysteinyl leukotrienes biological effects in infants with atopic dermatitis and food-protein induced enterocolitis important. Aim. To optimize the diagnosis of allergic lesions of the gastrointestinal tract in children. Methods. The study was conducted in the allergy center and children clinic of the «Institute Pediatrics, Obstetrics and Gynecology named after academician O. Lukyanova of NAMS of Ukraine». Children were included from September 2017 to June 2018. We examined 60 patients aged from 3 months to 3 years old, including 22 patients with atopic dermatitis, 18 children with food-protein induced enterocolitis, 8 patients with bronchial asthma in the stage of aggravation and 12 practically healthy children (control group). Medical examination have been perfomed, general IG E and specific serum IG E were defined by ImmunoCAP (Phadia, 100), as well as concentrations of cysteinyl leukotrienes (LTB4, LTC4, LTE4) in biological liquids (serum, saliva, urine) using immunoenzyme method using the production sets of the "Enzolifescience" (USA) company on the analyzer-photometer Multiskan Plus "Labsystems". The results of the received data were processed statistically. The probability of differences was estimated with Student’s t-test and Tau Kendall rank correlation test. The difference was considered significant at p <0.05. Results. A significant increase in the concentrations of cysteinyl leukotrienes (C4, D4, E4) in the blood, urine and saliva was found in infants with allergic skin diseases, gastro-intestinal and respiratory tract surveyed during manifestation the disease compared with the control group. Comparison of concentrations of leukotrienes in urine and saliva of children with atopic dermatitis (AD), food-protein induced enterocolitis (FPIE) and asthma did not find credible. However, in the serum of patients with asthma, the concentration of cysteinyl leukotrienes was significantly higher (703.9±68.7) pg / ml than in children with enterocolitis induced by dietary proteins (509.3±57.4) pg / ml and significantly did not differ from patients with atopic dermatitis (695.2±46.3) pg / ml. According to the results of Kendall Tau correlation test, no significant Spearman rank correlation was found between the cysteinyl leukotrienes concentration in blood and urine – r=0.14 (p>0.05), blood and saliva r=0.07 (p>0.05), urine and saliva r=–0.52 (p>0.05). Conclusions. Increase in cysteinyl leukotrienes concentrations in serum, urine and saliva of children of early age with allergic skin diseases, respiratory and gastrointestinal tract was found. The absence of significant Spearman rank correlation between concentrations of leukotrienes in blood and urine, blood and saliva, saliva and urine shows that it is possible to select any biological fluid, saliva or urine, as a non-invasive way to determine the leukotriene concentrations for monitoring activity of allergic inflammation.

Reactive carbonyl compounds, carbonyl stress, and neuroinflammation in methyl alcohol intoxication

Methyl alcohol intoxications are characterized by high lethality and high prevalence of serious visual and brain damage in survivors. The mechanisms of toxic brain damage are complex and the role of carbonyl stress has not been studied yet. We measured the acute and follow-up concentrations of reactive carbonyl compounds in patients with acute methyl alcohol intoxication. Blood samples were collected from 28 subjects hospitalized with confirmed methyl alcohol intoxication and from 36 subjects who survived poisoning 2 years after discharge. Serum concentrations of C6–12 reactive aldehydes were measured by liquid chromatography–electrospray ionization–tandem mass spectrometry. The acute concentrations of all measured reactive aldehydes were higher than the follow-up concentrations: 36.4 ± 4.8 vs. 21.6 ± 5.2 ng cm−3 for C6; 38.9 ± 5 vs. 17.0 ± 2.0 ng cm−3 for C7; 18.8 ± 3.9 vs. 4 ± 0 cm−3 for C8; 36.5 ± 3.9 vs. 19.0 ± 3.0 ng cm−3 for C9; 6.1 ± 0.4 vs. 4.0 ± 0.5 ng cm−3 for C10; 13.6 ± 3.0 vs. 3.7 ± 0.6 ng cm−3 for C11; and 7.8 ± 0.4 vs. 4.7 ± 0.4 ng cm−3 for C12 (all p < 0.001). The patients who survived the intoxication had higher concentration of reactive carbonyl compounds than those who died: 38.6 ± 5.9 vs. 28.3 ± 1.7 ng cm−3 for C6 (p = 0.002); 20.7 ± 4.7 vs. 11.8 ± 1.2 ng cm−3 for C8 (p = 0.001); 37.7 ± 4.8 vs. 31.8 ± 3.8 ng cm−3 for C9 (p = 0.042); and 7.9 ± 0.6 vs. 7.3 ± 0.5 ng cm−3 for C12 (p = 0.022). A significant association was present between severity of metabolic acidosis, anion gap, and the acute concentration of measured biomarkers: r = − 0.39; p = 0.046 for C6; r = − 0.42; p = 0.035 for C7; r = − 0.48; p = 0.012 for C8; r = − 0.39; p = 0.046 for C9; and r = − 0.47; p = 0.015 for C11. The acute concentration of C6–C12 reactive aldehydes positively correlated with the acute serum concentration of leukotrienes (all p < 0.05). Acute elevation of serum concentration of reactive carbonyl compounds suggests that carbonyl stress is involved in the mechanisms of leukotriene-mediated neuroinflammatory response to methyl alcohol-induced toxic brain damage.

Оптимізація лікування вірусіндукованих загострень бронхіальної астми у дітей із використанням лізатів молочнокислих бактерій Lactobacillus rhamnosus V

Objective. To study the effectiveness of lactic acid lactic acid bacteria Lactobacillus rhamnosus V in children with virus-induced exacerbations of bronchial asthma. Materials and methods. Were examined 35 patients aged 3 to 16 years old with exacerbation of bronchial asthma in the background of acute respiratory infections. Twenty five children of the main group were in the complex therapy of virus induced asthma exacerbation and were treated by lysate of the Lactic acid bacteria Lactobacillus rhamnosus V ( Del-Immune V ® ) at a dose of 25 mg 1 capsule 2 times a day 30 minutes before meals for 20 days. Ten patients were in the comparison group and obtained standard therapy. General clinical, immunological, allergic and statistical studies were used. Results. Use of Del-Immune V ® in the complex treatment of children with virus-nduced exacerbation of asthma, has led to a significant decrease in the frequency and severity of bronchial obstruction syndrome, which increases the effectiveness of treatment. In the main group, there was a significant increase in the concentration of IFN-gamma and a decrease in the concentration of cysteine leukotrienes in serum, and an increase in the concentration of a saliva IgA in the dynamics of treatment compared with the children in the comparison group. Conclusions. Observed integrated therapeutic effect of Del-Immune V ® use, no side effects and allergic reactions, justifying the appropriateness of Del-Immune V ® inclusion in the complex therapy of virus-induced asthma exacerbations.

Leukotriene mechanisms of allergic enterocolitis pathogenesis in young children

Purpose of the study was to determine the parameters of lipid mediators of allergic inflammation (cysteinyl leukotrienes C4, D4, E4) by enzyme immunoassay in serum, saliva, urine of young children with allergic enterocolitis. Materials and methods. 40 patients aged from 3 months to 3 years (mean age - (19.9 ± 2.7) months) were examined, among them 20 children with allergic enterocolitis, 10 patients with asthma at the acute stage and 10 healthy babies. Clinical, allergological, endoscopic examination and concentrations of cysteinyl leukotrienes (LTB4, LTC4, LTE4) determination in the biological fluids (blood serum, saliva, urine) using the enzyme immunoassay kits (Enzo Life Science, USA) on a Multiskan photometer Plus “Labsystems” were conducted. The obtained data results were processed statistically. Significance of differences was assessed using Student's t-test and Kendall's tau rank correlation analysis. Difference with a p value 0.05), blood and saliva τ = 0.07 (p>0.05), urine and saliva τ = -0.52 (P > 0.05). Conclusions. An increase in cysteinyl leukotrienes concentration in blood serum, urine, saliva was found in young children with allergic enterocolitis in comparison with the control group. The absence of a significant rank correlation between leukotrienes concentration in the blood and urine, blood and saliva, saliva and urine indicates that any biological fluid, in particular saliva or urine, could be selected as a non-invasive method for determination of leukotrienes concentration to monitor the activity of allergic inflammation.

PGE2 increases inflammatory damage in Escherichia coli-infected bovine endometrial tissue in vitro via the EP4-PKA signaling pathway.

Endometritis is the most common bovine uterine disease following parturition. The role of prostaglandin E2 (PGE2) in the regulation of endometrial inflammation and repair is well understood. Excess PGE2 is also generated in multiple inflammatory diseases, including endometritis. However, it remains unclear whether PGE2 is associated with pathogen-induced inflammatory damage to the endometrium. To clarify the role of PGE2 in pathogen-induced inflammatory damage, this study evaluated the production of PGE2, inflammatory factors, and damage-associated molecular patterns (DAMPs) in cultured Escherichia coli-infected bovine endometrial tissue. PGE2 production was significantly higher in E. coli-infected tissue, and in E. coli-infected tissue treated with 15-prostaglandin dehydrogenase (15-PGDH) inhibitors, as compared to uninfected tissue. Phospholipase A2 (PLA2), cyclooxygenase-2 (COX-2), and microsomal prostaglandin E synthase-1 (mPGES-1) were also upregulated in E. coli-infected tissue, while concentrations of arachidonic acid (AA), leukotrienes, DAMPs, and other proinflammatory factors increased. The accumulation of PGE2 clearly damaged the cultured tissue. Treatment with the COX-2, mPGES-1, EP4, and protein kinase A (PKA) inhibitors decreased the production of PGE2, inflammatory factors, and DAMPs, simultaneously alleviating the E. coli-induced endometrial tissue damage. Therefore, the PGE2 that was generated by COX-2 and mPGES-1 accumulated, and this pathogenic PGE2 increased inflammatory damage by upregulating inflammatory factors and DAMPs in E. coli-infected bovine endometrial tissue. This upregulation of inflammatory factors and DAMPs might be regulated by the EP4-PKA signaling pathway.

Interactive effects of dietary lipids and vitamin E level on performance, blood eicosanoids, and response to mitogen stimulation in broiler chickens of different ages

The effects of the dietary polyunsaturated fatty acids (PUFA) n-6:n-3 ratio and vitamin E (vE) on the levels of pro-inflammatory eicosanoids, the incorporation of docosahexaenoic acid (DHA) and arachidonic acid (AA) into immune tissues, and changes in leukocyte population after phytohemagglutinin (PHA) challenge were investigated in broiler chickens of different ages. One-day-old female broilers (48 per treatment) were fed 4 different wheat-soybean-corn-based diets containing corn oil with a high PUFA n-6:n-3 ratio (HR) or a mixture of linseed and fish oils with a low PUFA n-6:n-3 ratio (LR). Diets contained either 50 mg vE kg−1 of diet (basal vE) or 300 mg vE kg−1 of diet (increased vE). At d 14 and d 34, 8 chickens per treatment were challenged with PHA, and wing web swelling (WWS) was measured. The blood concentration of leukotriene (LTB4), prostaglandin (PGE2), and thromboxane (TBX2) in 17-day-old and 43-day-old chickens was determined. The pattern of AA and DHA incorporation into bursa, spleen, and brain lipids reflected the level of their precursors in the diet. WWS was the highest in chickens fed a LR diet and in 14-day-old chickens (P < 0.01). Leukocyte proportions varied with dietary PUFA n-6:n-3 ratio and with age. The heterophil:lymphocyte ratio was the highest at 6 h post PHA challenge, and was higher in 34-day-old chickens (P < 0.001). TBX2 and PGE2 concentrations were higher in chickens fed HR diet, whereas TBX2 and LTB4 concentrations were lower at high vE level. Lower PGE2 and LTB4, but higher TBX2 concentrations were measured in younger birds (P < 0.001). The results indicated that LR increased the phagocytic cell proportion in the blood; HR promoted the incorporation of AA into the immune tissues, which increased the levels of more pro-inflammatory eicosanoids in the blood; and vE counteracts these effects to some extent. Owing to the immaturity of the immune system, dietary interventions might be promising at the early stage of chicken growth.

Eosinophil transendothelial migration induced by the bronchoalveolar lavage fluid of acute eosinophilic pneumonia.

Acute eosinophilic pneumonia (AEP) is characterized by a massive pulmonary infiltration of eosinophils. Mechanisms regulating the selective accumulation of eosinophils in AEP have not been fully established. The objective of this study was to evaluate the mechanisms of eosinophil accumulation in alveolar spaces through examination of bronchoalveolar lavage fluid (BALF) from AEP patients (AEP-BALF). Eosinophils were isolated from the blood of healthy subjects and were placed on a human pulmonary microvascular endothelial cell monolayer cultured on Transwell filters (Coster, Cambridge, MA, USA). A saline control solution or BALF from patients with AEP, sarcoidosis or hypersensitivity pneumonitis was applied to the lower compartment, and the transendothelial migration of the eosinophils was evaluated. The concentrations of cytokines and chemokines in BALF were also measured. Transmigration of eosinophils across endothelial cells was only induced by the AEP-BALF. This transmigration was blocked by anti-β2 integrin mAb. The concentrations of eotaxin-2 and monocyte chemotactic protein (MCP)-4, which are CC chemokine receptor (CCR) 3 ligands, were elevated in the AEP-BALF, and anti-CCR3 mAb or anti-MCP-4 mAb inhibited the AEP-BALF-induced transmigration of eosinophils. Furthermore, the concentration of leukotriene (LT) B4 was increased in the AEP-BALF, and an LTB4 receptor antagonist partially suppressed the AEP-BALF-induced transmigration of eosinophils. These findings suggest that CCR3 ligands including eotaxin-2 and MCP-4, and LTB4 play a role in the accumulation of eosinophils in AEP.

Relation between Ocular Comfort, Arachidonic Acid Mediators, and Histamine

ABSTRACTPurpose: Contact lenses are associated with discomfort during wear. This may be the result of stimulation of the ocular surface and production of pro-inflammatory mediators which are then released into the tears. This study examined changes in the concentration in tears of arachidonic acid metabolites (AAM) prostaglandins, cysteinyl leukotrienes, and resolvin-D1, as well as histamine in a general contact lens population in the morning and evening.Method: Tears were collected twice a day (morning and evening) for up to 10 days on two different occasions (with and without contact lens wear) from 30 experienced contact lens wearers for analysis of AAM and a separate group (N = 33) for analysis of histamine. Ocular comfort was rated subjectively on an ordinal scale at each time of tear collection. Tears were analyzed using commercial immunoassay-based kits. Statistical analysis was performed using linear mixed model test.Results: Ocular comfort decreased from morning to evening with and without contact lenses (p = 0.001), and the difference in comfort in the evening was greater with contact lens wear (80.9 ± 16.2 vs. 75.5 ± 16.8; p = 0.008). The total concentration of PGs (10.7 ± 10.8 ng/ml), cysteinyl leukotrienes (8.7 ± 0.38 ng/ml), resolvin-D1 (1.6 ± 0.5 ng/ml), or histamine 13.8 ± 10.4 ng/ml) did not change during the day or during contact lens wear (p > 0.05).Conclusions: Prostaglandins, cysteinyl leukotrienes, resolvin-D1, or histamine concentrations did not alter in relation to changes in comfort of the eye during the day or during contact lens wear. These results suggest that release of these mediators is not responsible for contact lens discomfort.

Protolichesterinic Acid, Isolated from the Lichen Cetraria islandica, Reduces LRRC8A Expression and Volume-Sensitive Release of Organic Osmolytes in Human Lung Epithelial Cancer Cells.

We have tested the effect of protolichesterinic acid (PA) on the activity of the volume-sensitive release pathway for the organic osmolyte taurine (VSOAC) and the expression of the leucine-rich-repeat-channel 8A (LRRC8A) protein, which constitutes an essential VSOAC component. Exposing human lung cancer cells (A549) to PA (20 µg/mL, 24 h) reduces LRRC8A protein expression by 25% and taurine release following osmotic cell swelling (320 → 200 mOsm) by 60%. C75 (20 µg/mL, 24 h), a γ-lactone with a C8 carbon fatty acid chain, reduces VSOAC activity by 30%, i.e. less than PA. Stearic acid (20 µg/mL, 24 h) has no effect on VSOAC. Hence, length of PA's fatty acid chain adds to γ-lactone's inhibitory action. 5-Lipoxygenase (5-LO) activity is essential for swelling-induced activation of VSOAC. PA has no effect on cellular concentration of leukotrienes (5-HETE/LTB4 ) under hypotonic conditions, excluding that PA mediated inhibition of VSOAC involves 5-LO inhibition. A549 cells exposed to the chemotherapeutic drug cisplatin (10 μM, 24 h) reveal signs of apoptosis, i.e. 25% reduction in cell viability as well as 1.3-, 1.5- and 3.3-fold increase in the expression of LRRC8A, Bax (regulator of apoptosis) and p21 (regulator of cell cycle progression), respectively. PA reduces cell viability by 30% but has no effect on p21/Bax expression. This excludes PA as a pro-apoptotic drug in A549 cells.

Increased PUFA Content and 5-Lipoxygenase Pathway Expression Are Associated with Subcutaneous Adipose Tissue Inflammation in Obese Women with Type 2 Diabetes

Obese women with type 2 diabetes mellitus (T2DM) have more inflammation in their subcutaneous white adipose tissue (sWAT) than age-and-BMI similar obese women with normal glucose tolerance (NGT). We aimed to investigate whether WAT fatty acids and/or oxylipins are associated with the enhanced inflammatory state in WAT of the T2DM women. Fatty acid profiles were measured in both subcutaneous and visceral adipose tissue (vWAT) of 19 obese women with NGT and 16 age-and-BMI similar women with T2DM. Oxylipin levels were measured in sWAT of all women. Arachidonic acid (AA) and docosahexaenoic acid (DHA) percentages were higher in sWAT, but not vWAT of the T2DM women, and AA correlated positively to the gene expression of macrophage marker CD68. We found tendencies for higher oxylipin concentrations of the 5-LOX leukotrienes in sWAT of T2DM women. Gene expression of the 5-LOX leukotriene biosynthesis pathway was significantly higher in sWAT of T2DM women. In conclusion, AA and DHA content were higher in sWAT of T2DM women and AA correlated to the increased inflammatory state in sWAT. Increased AA content was accompanied by an upregulation of the 5-LOX pathway and seems to have led to an increase in the conversion of AA into proinflammatory leukotrienes in sWAT.

Level of Fatty Acid Binding Protein 5 (FABP5) Is Increased in Sputum of Allergic Asthmatics and Links to Airway Remodeling and Inflammation

BackgroundThe inflammatory processes in the upper and lower airways in allergic rhinitis and asthma are similar. Induced sputum and nasal lavage fluid provide a non-invasive way to examine proteins involved in airway inflammation in these conditions.ObjectivesWe conducted proteomic analyses of sputum and nasal lavage fluid samples to reveal differences in protein abundances and compositions between the asthma and rhinitis patients and to investigate potential underlying mechanisms.MethodsInduced sputum and nasal lavage fluid samples were collected from 172 subjects with 1) allergic rhinitis, 2) asthma combined with allergic rhinitis, 3) nonallergic rhinitis and 4) healthy controls. Proteome changes in 21 sputum samples were analysed with two-dimensional difference gel electrophoresis (2D-DIGE), and the found differentially regulated proteins identified with mass spectrometry. Immunological validation of identified proteins in the sputum and nasal lavage fluid samples was performed with Western blot and ELISA.ResultsAltogether 31 different proteins were identified in the sputum proteome analysis, most of these were found also in the nasal lavage fluid. Fatty acid binding protein 5 (FABP5) was up-regulated in the sputum of asthmatics. Immunological validation in the whole study population confirmed the higher abundance levels of FABP5 in asthmatic subjects in both the sputum and nasal lavage fluid samples. In addition, the vascular endothelial growth factor (VEGF) level was increased in the nasal lavage fluid of asthmatics and there were positive correlations between FABP5 and VEGF levels (r=0.660, p<0.001) and concentrations of FABP5 and cysteinyl leukotriene (CysLT) (r=0.535, p<0.001) in the nasal lavage fluid.ConclusionsFABP5 may contribute to the airway remodeling and inflammation in asthma by fine-tuning the levels of CysLTs, which induce VEGF production.

Bradykinin and cysteinyl leukotriene concentrations in cell-salvaged blood before and after passage through negatively charged filters during clinical use in cancer patients: a pilot study.

It has been suggested that giving cell-salvaged blood through a leucocyte depletion filter can cause hypotension due to bradykinin released when factor XII and platelets are activated by the negatively charged surface of the filter. We measured the concentration of bradykinin and cysteinyl leukotrienes in cell-salvaged blood sampled before and after passage through a negatively charged leucodepletion filter in 24 consecutive patients with gynaecological or bowel cancer undergoing elective surgery with cell salvage. In no case was an increase in bradykinin concentration observed after passage through the filter; in 23 patients the post-filtration bradykinin concentration was zero (p=0.007). The change in the concentration of cysteinyl leukotrienes detected during passage across the filter was not statistically significant (p=0.1). Our findings do not support the suggestion that either bradykinin or cysteinyl leukotrienes are generated in cell-salvaged blood during passage through leucodepletion filters.

Effects of a Mediterranean Diet Intervention on Anti- and Pro-Inflammatory Eicosanoids, Epithelial Proliferation, and Nuclear Morphology in Biopsies of Normal Colon Tissue

This randomized trial evaluated the effects of intervention with either a Healthy Eating or a Mediterranean diet on colon biomarkers in 120 healthy individuals at increased colon cancer risk. The hypothesis was that eicosanoids and markers of proliferation would be favorably affected by the Mediterranean diet. Colon epithelial biopsy tissues and blood samples were obtained at baseline and after 6 mo of intervention. Colonic eicosanoid concentrations were evaluated by HPLC-MS-MS, and measures of epithelial proliferation and nuclear morphology were evaluated by image analysis of biopsy sections. There was little change in proinflammatory eicosanoids and in plasma cytokine concentrations with either dietary intervention. There was, however, a 50% increase in colonic prostaglandin E3 (PGE3), which is formed from eicosapentanoic acid, in the Mediterranean arm. Unlike PGE2, PGE3, was not significantly affected by regular use of non-steroidal anti-inflammatory drugs at baseline, and normal weight subjects had significantly higher colon PGE3 than overweight or obese subjects. Increased proliferation in the colon at baseline, by Ki67 labeling, was associated with morphological features that defined smaller nuclei in the epithelial cells, lower colon leukotriene concentrations and higher plasma cytokine concentrations. Dietary intervention had little effect on measures of epithelial proliferation or of nuclear morphology. The increase in PGE3 with a Mediterranean diet indicates that in normal colon, diet might affect protective pathways to a greater extent than proinflammatory and proliferative pathways. Hence, biomarkers from cancer models might not be relevant in a true prevention setting.