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Abstract Background/objectives Previous studies have shown that N-acetylcysteine (NAC) supplementation with the simultaneous inclusion of HFD prevents salivary glands from oxidative stress and mitochondrial dysfunction. In this experiment, we examined if NAC supplementation could reverse the harmful effect of HFD on mitochondrial function, reduce the severity of apoptosis, and the activity of pro-oxidative enzymes in the salivary glands of rats with confirmed hyperglycemia. Subjects/methods Wistar rats were fed the standard or high-fat (HFD) diet for 10 weeks. After 6 weeks of the experiment, HFD rats were diagnosed with hyperglycemia and for the next 4 weeks, the animals were given NAC intragastrically. In the mitochondrial fraction of the parotid (PG) and submandibular salivary glands (SMG), we assessed redox status, inflammation, and apoptosis. Results The inclusion of NAC increased the activity of mitochondrial complexes I and II + III as well as decreased the concentration of interleukin-1β, tumor necrosis factor α, and caspase-3, but only in the parotid glands of rats with hyperglycemia compared to the HFD group. However, N-acetylcysteine supplementation did not reduce the activity of caspase-9 or the Bax/Bcl-2 ratio in PG and SMG mitochondria. In both salivary glands we observed reduced activity of cytochrome c oxidase, NADPH oxidase, and xanthine oxidase, as well as hindered production of ROS and lower ADP/ATP radio, but the levels of these parameters were not comparable to the control group. Conclusions We demonstrated that NAC supplementation restores the glutathione ratio only in the mitochondria of the submandibular salivary glands. The supply of NAC did not significantly affect the other measured parameters. Our results indicate that NAC supplementation provides little protection against free radicals, apoptosis, and inflammation in the salivary gland mitochondria of HFD rats. Stimulated salivary secretion in hyperglycaemic rats supplemented with NAC seems to be closely related to mitochondrial respiratory capacity and appropriate ATP level.

Blanka Wolszczak-Biedrzycka,1 Justyna Dorf,2 Marzena Wojewódzka-Żelezniakowicz,3 Małgorzata Żendzian-Piotrowska,4 Violetta Joanna Dymicka-Piekarska,2 Joanna Matowicka-Karna,2 Mateusz Maciejczyk4 1Department of Psychology and Sociology of Health and Public Health, University of Warmia and Mazury in Olsztyn, Olsztyn, 10-900, Poland; 2Department of Clinical Laboratory Diagnostics, Medical University of Bialystok, Bialystok, 15-089, Poland; 3Department of Emergency Medicine and Disasters, Medical University of Bialystok, Bialystok, 15-089, Poland; 4Department of Hygiene, Epidemiology and Ergonomics, Medical University of Bialystok, Bialystok, 15-089, PolandCorrespondence: Blanka Wolszczak-Biedrzycka, Email blanka.wolszczak@uwm.edu.plIntroduction: In coronavirus disease (COVID-19), inflammation takes center stage, with a cascade of cytokines released, contributing to both inflammation and lung damage. The objective of this study is to identify biomarkers for diagnosing and predicting the severity of COVID-19.Materials and Methods: Cytokine levels were determined in the serum from venous blood samples collected from 100 patients with COVID-19 and 50 healthy controls. COVID-19 patients classified based on the Modified Early Warning (MEWS) score. Cytokine concentrations were determined with a multiplex ELISA kit (Bio-Plex Pro™ Human Cytokine Screening Panel).Results: The concentrations of all analyzed cytokines were elevated in the serum of COVID-19 patients relative to the control group, but no significant differences were observed in interleukin-9 (IL-9) and IL-12 p70 levels. In addition, the concentrations of IL-1α, IL-1β, IL-1ra, IL-2Rα, IL-6, IL-12 p40, IL-18, and tumor necrosis factor alpha (TNFα) were significantly higher in symptomatic patients with accompanying pneumonia without respiratory failure (stage 2) than in asymptomatic/mildly symptomatic patients (stage 1).Conclusion: The study revealed that IL-1ra, IL-2Rα, IL-6, IL-8, IL-12 p40, IL-16, and IL-18 levels serve as potential diagnostic biomarkers in COVID-19 patients. Furthermore, elevated IL-1α levels proved to be valuable in assessing the severity of COVID-19.Keywords: cytokines, COVID-19, MEWS score, biomarkers, severity, diagnosis

Julita Szulimowska,1 Anna Zalewska,1 Katarzyna Taranta-Janusz,2 Grażyna Marczuk-Kolada,3 Małgorzata Żendzian-Piotrowska,4 Mateusz Maciejczyk4 1Department of Restorative Dentistry, Medical University of Bialystok, Bialystok, Poland; 2Department of Pediatrics and Nephrology, Medical University of Bialystok, Bialystok, Poland; 3Department of Pedodontics, Medical University of Bialystok, Bialystok, Poland; 4Department of Hygiene, Epidemiology and Ergonomics, Medical University of Bialystok, Bialystok, PolandCorrespondence: Mateusz Maciejczyk, Department of Hygiene, Epidemiology and Ergonomics, Medical University of Bialystok, Bialystok, 2c Mickiewicza Street, Bialystok, 15-233, Poland, Email mat.maciejczyk@gmail.comIntroduction: Chronic kidney disease (CKD) is a systemic inflammatory disease that leads to multiple organ complications not only in the kidneys and the cardiovascular system, but also in the oral cavity. CKD children experience reduced saliva secretion (hyposalivation), which leads to increased incidence of dental caries and significant impairment of patients’ quality of life. However, the causes of salivary gland dysfunction in children with CKD are unknown. The present study is the first to evaluate the inflammatory and anti-inflammatory profile in the saliva of children with CKD at different stages of renal failure with normal and reduced salivary gland function.Methods: Thirty children with CKD (age 9– 16) and thirty age- and gender-matched healthy children were classified for the study. Salivary inflammatory and anti-inflammatory profile were assayed using the multiplex ELISA assay.Results: We demonstrated statistically significant changes in salivary pro-inflammatory (↑TNF-α, ↓IL-7), anti-inflammatory (↑IL-10), Th1 (↑INF-γ, ↑IL-15), Th2 (↑IL-4, ↑IL-5, ↑IL-6, ↑IL-9) and Th17 (IL-17) cytokines as well as chemokines (↑MCP-1/CCL-2, ↑MIP-1α/CCL3, ↓MIP-1β/CCL4, ↓EOTAXIN/CCL11) and growth factors (↑G-CSF, ↑FGF) in unstimulated saliva of children with CKD compared to the controls. Although the evaluation of the salivary inflammatory profile does not indicate a particular dominance of any of the branches of the immune system, we observed a statistically significant increase in the concentration of all Th2 cytokines assayed. The multivariate regression analysis showed that the content of salivary cytokines, chemokines and growth factors depends on the secretory function of the salivary glands, ie, salivary flow, total protein concentration and amylase activity in the saliva. Salivary MIP-1α/CCL3 was the most effective to differentiate children with CKD and hyposalivation from patients with normal saliva secretion.Discussion: Inflammation is involved in salivary gland dysfunction in children with CKD, although further studies on in vitro and in vivo models are necessary to confirm this hypothesis.Keywords: chronic kidney disease, saliva, inflammation, salivary gland dysfunction, hyposalivation

Justyna Zi&nacute;czuk,1 Konrad Zar&eogon;ba,2 Joanna Kami&nacute;ska,1 Olga Martyna Koper-Lenkiewicz,1 Violetta Dymicka-Piekarska,1 Anna Pryczynicz,3 Katarzyna Guzi&nacute;ska-Ustymowicz,3 Bogu&lstrok;aw K&eogon;dra,2 Joanna Matowicka-Karna,1 Ma&lstrok;gorzata &Zdot;endzian-Piotrowska,4 Anna Zalewska,5 Mateusz Maciejczyk4 1Department of Clinical Laboratory Diagnostics, Medical University of Bialystok, Bia&lstrok;ystok, 15-269, Poland; 2 2nd Clinical Department of General and Gastroenterological Surgery, Medical University of Bialystok, Bia&lstrok;ystok, 15-276, Poland; 3Department of General Pathomorphology, Medical University of Bialystok, Bia&lstrok;ystok, 15-269, Poland; 4Department of Hygiene, Epidemiology and Ergonomics, Medical University of Bialystok, Bia&lstrok;ystok, 15-222, Poland; 5Independent Laboratory of Experimental Dentistry, Medical University of Bialystok, Bia&lstrok;ystok, 15-276, PolandCorrespondence: Justyna Zi&nacute;czukDepartment of Clinical Laboratory Diagnostics, Medical University of Bialystok, Waszyngtona 15a St., Bia&lstrok;ystok, 15-269, PolandTel +48 85 8 31 87 16Email justyna.zinczuk@umb.edu.plMateusz MaciejczykDepartment of Hygiene, Epidemiology and Ergonomics, Medical University of Bialystok, Mickiewicza 2c St., Bia&lstrok;ystok, 15-222, PolandTel +48 85 748 55 85Email mat.maciejczyk@gmail.comPurpose: In this study, we evaluated the total antioxidant capacity, nitrosative stress, and protein/DNA oxidation and glycoxidation products in patients with colorectal cancer regarding histopathological parameters associated with the tumour microenvironment, such as inflammatory infiltration and tumour budding and compare all determined parameters between tumours located in the right and left side of the colon and normal mucosa.Patients and Methods: Ferric reducing antioxidant power (FRAP), nitrosative stress (myeloperoxidase (MPO), nitrogen oxide (NO), peroxynitrite, and nitrotyrosine), protein oxidation products (protein carbonyls (PC), total thiols, and ischemia modified albumin (IMA)), protein glycooxidation products (tryptophan, kynurenine, N-formylkynurenine, dityrosine, Amadori product, advanced glycation end products (AGE)) and 8-hydroxydeoxyguanosine (8-OHdG) were measured in homogenates from normal and cancerous tissue of 30 patients with colorectal cancer.Results: Levels of FRAP (p=0.0009), IMA (p=0.0002), kynurenine (p< 0.0001), N-formylkynurenine (p< 0.0001), dityrosine (p< 0.0001), Amadori products (p=0.0024), AGE (p< 0.0001), MPO (p< 0.0001), NO (p< 0.0001) and nitrotyrosine (p=0.0011) were increased, whereas PC (p=0.0004), tryptophan (p< 0.0001), 8-OHdG (p< 0.0001) and peroxynitrite (p=0.0003) were decreased in the left-side tumour compared to the right-side tumour and normal mucosa.Conclusion: Our results showed that colorectal cancer is related with disturbances in antioxidant defense and increased oxidative and nitrosative damages to proteins and DNA. These parameters may be useful for evaluation the progression and differentiation of the tumour location. We also demonstrated that redox indicators may depend on the histological type of the tumour and may influence tumour invasion depth, presence of lymph node and distant metastasis, vascular and neural invasion, inflammatory infiltration, and tumour budding, which are part of the tumour microenvironment.Keywords: oxidative stress, nitrosative stress, redox biomarkers, colorectal cancer

Fingolimod (FTY) is known to have multiple effects on the immune system and the central nervous system (CNS) in patients with multiple sclerosis (MS). In this study, we evaluated the immunological and neurobiological effects of FTY in MS. Blood and cerebrospinal fluid (CSF) samples were collected from 15 MS patients before first FTY administration and after 4 months of FTY therapy. Immunophenotyping and evaluation of sphingosine-1-phosphate (S1P), neurofilament light chain (NFL), S-100 and neuron-specific enolase (NSE) levels were conducted. After 4 months of FTY therapy, absolute cell count in CSF was decreased from 6.33 to 2.43 MPt/l, accompanied by decreases of CD3+ (2.22 to 0.65 MPt/l) and of CD4+ counts (1.60 to 0.39 MPt/l). In blood, CD3+ (1.05 to 0.09 GPt/l), CD4+ (0.80 to 0.02 GPt/l), CD8+ (0.23 to 0.04 GPt/l) and CD19+ (0.21 to 0.01GPt/l) cell counts were as well reduced. CD14+ cell count remained stable over the same period (0.24 to 0.26GPt/l). NFL and S1P levels in CSF and blood were reduced over time (NFL: CSF 1759 to 1359 pg/l, blood 8.42 to 7.36 pg/l; S1P: CSF 2.12 to 0.71 nmol/l, blood 392.1 to 312.9 nmol/l). Strong correlations between CSF and blood NFL levels were observed. Neuronal damage markers such as S-100 (1.86 to 1.69 μg/l) and NSE (9.53 to 8.67 μg/l) were reduced to a lesser degree than other markers. FTY exerted significant effects on immunological and neurobiological markers in the central and peripheral compartment. Decreases in levels of neuroinflammatory and neurodegenerative markers were already evident after 4 months of treatment. Four-month serum NFL level appears to be a useful marker for FTY efficacy that correlates well with changes in the CNS compartment. KEY MESSAGES: FTY has important immunological effects in both central and peripheral compartments. Cellular effects of FTY effects are more pronounced in the blood than in the CSF. FTY reduces S1P and NFL levels in CSF and serum. Serum NFL appears to be a useful marker for FTY therapy.

Physical effort can elicit differential adaptive changes in the tissues of trained versus untrained rats. Proteolytic activity in the extracellular matrix could be engaged in such adaptation due to its influence on the elasticity of tissues. The effects were investigated of a single physical effort on the activity of elastase, cathepsin K, and plasmin in the skeletal muscles, heart muscles, and aortas of untrained (UT, n=30) and trained (T, n=30) rats. T rats underwent 6 weeks of endurance training. After the last training session, T and UT rats were divided randomly into 3 subgroups. Ten rats from each group (Tpre, n=10) and (UTpre, n=10) were sacrificed. The other 20 rats from each group performed 60 min. of aerobic exercise and were sacrificed immediately post exercise (T0h, n=10; UT0h, n=10) or 3h later (T3h, n=10; UT3h, n=10). Enzyme activity was measured fluorometrically. Cathepsin K and plasmin activity increased in the soleus muscles of UT0h versus UTpre, plasmin activity increased also in UT3h versus UTpre. Elastase, cathepsin K and plasmin activity increased in the heart muscles of T0h and T3h versus Tpre. No aortic differences were observed. Thus, a single bout of physical effort elicited different responses in tissues of T versus UT rats. Increased proteolytic enzyme levels in muscles could influence tissue remodeling. Unchanged aortic cathepsin K levels may help prevent aortic remodeling and neointima formation.

Background/objectives: Fatty acid handling proteins are involved in the process of accumulation of lipids in different fat tissue depots. Thus, the aim of the study was to estimate the expression of both fatty acid transport and binding proteins in the subcutaneous (SAT) and visceral adipose tissue (VAT) of patients with morbid obesity without metabolic syndrome, as well as the plasma concentrations of these transporters., Subjects/methods: Protein (Western blotting) and mRNA (Real-time PCR) expression of selected fatty acid handling proteins was assessed in the visceral and subcutaneous adipose tissue of 30 patients with morbid obesity. The control group consisted of 10 lean age-matched patients. Plasma levels of fatty acid protein transporters were also evaluated using ELISA method. Moreover, total plasma fatty acid composition and concentration was determined by gas-liquid chromatography (GLC)., Results: Significant increase in fatty acid translocase (FAT/CD36) mRNA (P = 0.03) and plasmalemmal (P = 0.01) expression was observed in VAT of patients with morbid obesity vs. lean subjects together with elevation of lipoprotein lipase (LPL), as well as peroxisome proliferator-activated receptor γ (PPARγ) in both examined compartments of adipose tissue. Moreover, in obese subjects plasma concentration of RBP4 was markedly elevated (P = 0.04) and sCD36 level presented a tendency for an increase (P = 0.08) with concomitant lack of changes in FABP4 concentration (P > 0.05)., Conclusions: Fatty acid transport into adipocytes may be, at least in part, related to the increased expression of FAT/CD36 in the VAT of morbidly obese patients, which is accompanied by augmented expression of LPL, as well as PPARγ. Probably, alternations in plasma concentrations of RBP4 and sCD36 in obese patients are associated with "unhealthy" fat distribution.

Physical exercise causes adaptive changes, mainly in muscles, but it also influences other organs, including liver. Most changes are beneficial; however, strenuous exercise is a strong stressor, and it can result in splanchnic hypoperfusion with subsequent disturbances in liver homeostasis and energy. Cathepsin B is a protease linked to protein turnover and extracellular matrix degradation. It is also involved in autophagy and the activation of proinflammatory and profibrotic pathways. This study investigated the influences of one session of exercise and endurance training on the mRNA, protein level, and activity of cathepsin B in rat liver. Healthy rats were randomly divided into two groups (n = 30, each); one group was untrained and the other received 6-weeks of endurance training with an increasing load. For each group, rats were sacrificed before (controls, n = 10), immediately after (n = 10), and 3 h after (n = 10) an acute bout of intense exercise. Liver gene expression was evaluated with quantitative real-time PCR. Liver protein content was measured with ELISA. Liver enzyme activity was measured fluorometrically. One session of exercise or training did not influence cathepsin B gene expression or protein concentration at any investigated time point. In untrained rats, cathepsin B activity decreased 3 hours after (P = 0.027) one session of exercise. In trained rats, cathepsin B activity increased immediately (P = 0.005) after one session of exercise. Training did not influence baseline cathepsin B activity. In conclusion, one session of exercise differentially influenced cathepsin B activity in the liver, depending on training status.

The diaphragm is a dome-shaped skeletal muscle indispensable for breathing. Its activity contributes up to 70% of the total ventilatory function at rest. In comparison to other skeletal muscles, it is distinguished by an oxidative phenotype and uninterrupted cyclic contraction pattern. Surprisingly, the research regarding diaphragm diabetic phenotype particularly in the light of lipid-induced insulin resistance is virtually nonexistent. Male Wistar rats were randomly allocated into 3 groups: control, streptozotocin-induced (STZ) type-1 diabetes, and rodents fed with high-fat diet (HFD). Additionally, half of the animals from each group were administered with myriocin, a robust, selective inhibitor of ceramide synthesis and, therefore, a potent agent ameliorating insulin resistance. Diaphragm lipid contents were evaluated using chromatography. Fatty acid transporter expression was determined by Western blot. The STZ and HFD rats had increased concentration of lipids, namely, ceramides (CER) and diacylglycerols (DAG). Interestingly, this coincided with an increased concentration of long-chain (C ≥ 16) saturated fatty acid species present in both the aforementioned lipid fractions. The CER/DAG accumulation was accompanied by an elevated fatty acid transporter expression (FATP-1 in HFD and FATP-4 in STZ). Surprisingly, we observed a significantly decreased triacylglycerol content in the diaphragms of STZ-treated rats.

Exercise induces changes in muscle fibers and the extracellular matrix that may depend on elastin content and the activity of proteolytic enzymes. We investigated the influence of endurance training on the gene expression and protein content and/or activity of elastin, elastase, cathepsin K, and plasmin in skeletal and heart muscles and in the aorta. Healthy rats were randomly divided into untrained (n=10) and trained (n=10; 6 weeks of endurance training with increasing load) groups. Gene expression was evaluated via qRT-PCR. Elastin content was measured via enzyme-linked immunosorbent assay and enzyme activity was measured fluorometrically. Elastin content was significantly higher in skeletal (P=0.0014) and heart muscle (P=0.000022) from trained rats versus untrained rats, but not in the aorta. Although mRNA levels in skeletal muscle did not differ between groups, the activities of elastase (P=0.0434), cathepsin K (P=0.0343) and plasmin (P=0.000046) were higher in trained rats. The levels of cathepsin K (P=0.0288) and plasminogen (P=0.0005) mRNA were higher in heart muscle from trained rats, but enzyme activity was not. Enzyme activity in the aorta did not differ between groups. Increased elastin content in muscles may result in better adaption to exercise, as may remodeling of the extracellular matrix in skeletal muscle., (© 2016. Published by The Company of Biologists Ltd.)

Background: Tick-borne encephalitis (TBE) is a serious acute central nervous system infection that can result in death or long-term neurological dysfunctions. We hypothesize that changes in sphingosine-1-phosphate (S1P) concentration occur during TBE development., Methods: S1P and interleukin-6 (IL-6) concentrations in blood plasma and cerebrospinal fluid (CSF) were measured using HPLC and ELISA, respectively. The effects of S1P on cytoskeletal structure and IL-6 production were assessed using rat astrocyte primary cultures with and without addition of plasma gelsolin and the S1P receptor antagonist fingolimod phosphate (FTY720P)., Results: We report that acute inflammation due to TBE virus infection is associated with elevated levels of S1P and IL-6 in the CSF of infected patients. This elevated concentration is observed even at the earliest neurologic stage of disease, and may be controlled by glucocorticosteroid anti-inflammatory treatment, administered to patients unresponsive to antipyretic drugs and who suffer from a fever above 39°C. In vitro, treatment of confluent rat astrocyte monolayers with a high concentration of S1P (5 μM) results in cytoskeletal actin remodeling that can be prevented by the addition of recombinant plasma gelsolin, FTY720P, or their combination. Additionally, gelsolin and FTY720P significantly decreased S1P-induced release of IL-6., Conclusions: TBE is associated with increased concentration of S1P and IL-6 in CSF, and this increase might promote development of inflammation. The consequences of increased extracellular S1P can be modulated by gelsolin and FTY720P. Therefore, blocking the inflammatory response at sites of infection by agents modulating S1P pathways might aid in developing new strategies for TBE treatment.

Objective: There is no study analyzing the salivary antioxidant profile in the course of the insulin resistance., Materials and Methods: Rats were divided into two groups. One group was fed with a normal diet, another one with a high fat diet for 5 weeks. The analysis included: catalase (CAT), superoxide dismutase, peroxidase activities, uric acid, and total antioxidant status concentrations., Results: The activity of peroxidase in both kind of glands of insulin resistance rats was significantly reduced than in the control rats. The protein concentration, total amount of total antioxidant status in the parotid glands of insulin resistance rats were significantly lower than in the control glands The total amount of superoxide dismutase, CAT, and uric acid in the parotid glands of insulin resistance rats were significantly elevated in comparison with the control rats. The median values of the total amount of superoxide dismutase, CAT, peroxidase, total antioxidant status were significantly higher in the parotid than in the submandibular glands of the insulin resistance and control rats., Conclusion: Parotid and submandibular glands of rats react differently when exposed to insulin resistance condition; however, the parotid glands seem to be more affected. The main source of antioxidants is parotid glands of rats., (© 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Background & Aims: Nonalcoholic fatty liver disease (NAFLD) is an insulin resistance-related hepatic disorder which can transform to cirrhosis. Insulin resistance deregulates hepatic lipid metabolism, leading to accumulation of cytotoxic lipids including ceramide and diacylglycerols. Myriocin, obtained from fungi traditionally used in Chinese medicine in an effort to attain eternal youth, is a potent pharmacological inhibitor of ceramide de novo synthesis. We examined whether inhibition of ceramide de novo synthesis with myriocin ameliorate hepatic lipid accumulation and reverse NAFLD., Methods: The experiment was carried out on male Wistar rats. The animals were divided into four groups: (i) control group, fed standard rodent diet, (ii) group, fed standard diet also treated with myriocin for 7 days, (iii) group, fed high-fat diet for 5 weeks, (iv) group, fed high-fat diet and treated with myriocin. In liver samples sphingolipids: ceramide, sphingosine and sphingosine-1-phosphatate and neutral lipids, such as diacylglycerols and triacylglycerols were measured. In peripheral blood samples, glucose and insulin levels and aminotransferases activities were measured., Results: High-fat diet feeding caused NAFLD, confirmed by histological assessment, with increased hepatic lipids accumulation and whole-body insulin resistance. After treating with inhibitor of ceramide de novo synthesis, decrease in hepatic ceramide and other toxic lipids were noticed. Moreover, histological analysis of liver samples revealed that inhibition of ceramide de novo synthesis reduced hepatic steatosis., Conclusions: Inhibition of ceramide de novo synthesis reduced hepatic lipid accumulation in rats with NAFLD, this led to amelioration of hepatic steatosis., (© 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Irisin induces the browning of adipose tissue. The goal of this study was to investigate the influence of acute exercise in untrained and trained rats and endurance training on FNDC5 mRNA and irisin levels in white and red skeletal muscle and serum. Rats (n=60) were randomly divided into two groups: untrained and trained (subjected to 6-week endurance training with increasing load). Subgroups of rats from each group were sacrificed before (controls), immediately after, or 3 hours following acute exercise with the same work load. Muscle samples (red and white) and serum were collected. FNDC5 mRNA was evaluated using RT-PCR. Irisin levels were measured using an immunoenzymatic method. Muscle FNDC5 mRNA decreased immediately after acute exercise compared with baseline levels, but not in red muscle in trained rats. Atrend toward a return to baseline appeared 3 hours after the exercise, but only in white muscle in untrained group. Irisin protein levels increased after acute exercise in red muscle 3 hours post-exercise compared with samples taken immediately after exercise, and decreased 3 hours post-exercise compared to pre-exercise level in white muscles. FNDC5 mRNA did not change following training, whereas irisin protein levels increased in red muscle and decreased in white muscle. Serum irisin levels remained unchanged following acute exercise and training. We concluded that changes in irisin mRNA and protein levels in rat muscle after acute exercise are limited and depend on training status and the muscle type. Irisin serum levels remained stable after acute exercise or endurance training.

Renalase is a newly discovered amine oxidase and may lower blood pressure by metabolizing catecholamines. We have hypothesized that exercise and training may regulate renalase expression to control blood pressure. In this study, we investigated changes in renalase expression after exercise and training in white and red portion of the gastrocnemius muscle, kidney, and serum in rats. Rats were either untrained or subjected to six weeks of endurance training, which predominantly recruits red fibers. Rats from each group were sacrificed before (n = 10), immediately after (n = 10), or three hours (n = 10) following exercise. Renalase mRNA and protein levels were measured by RT-PCR and ELISA, respectively. There were no significant changes in renalase expression after prolonged training or acute exercise in the serum or red muscle of rats. However, in white muscle, renalase mRNA and protein levels decreased after acute exercise in untrained rats, whereas, in trained rats, its protein level remained unchanged, despite a decrease in mRNA. Thus, exercise influenced renalase expression only in white muscle fibers that are not predominantly recruited during exercise. The reduction of renalase protein in white muscle suggests that renalase may contribute to blood redistribution between contracting and non-contracting fibers during exercise. In the kidney, renalase protein levels decreased after training, while mRNA levels increased. The reduction in renalase protein levels may contribute to functional kidney hypoperfusion, which has been observed after training. In conclusion, exercise differentially regulates renalase expression in skeletal muscle and kidney.

Plasma gelsolin (pGSN) produced by muscle is an abundant protein of extracellular fluids capable of severing actin filaments and eliminating actin from the circulation. Additionally, pGSN modulates the cellular effects of some bioactive lipids. In this study we test the hypothesis that hormonal and metabolic adaptations to exercise are associated with changes in gelsolin concentration in blood. Plasma samples were collected from twenty healthy males recruited from untrained (UT, n=10) and endurance trained (ET, n=10) groups that performed 30-60 minutes of exercise on a cycloergometer at a workload corresponding to 70% of VO2max. Gelsolin concentration was determined by quantitative Western blot analysis with an anti-human gelsolin antibody. The gelsolin concentration in UT and ET subjects before starting exercise ranged from 104 to 330 and 163 to 337 µg · ml(-1) respectively. After 30 minutes of exercise we observed a significant decrease of plasma gelsolin in the UT group (p<0.05) while the gelsolin concentration in the ET group rose on average from 244 to 271 µg · ml(-1). However, this increase did not reach statistical significance. Endurance training might increase the ability of muscle tissue to express plasma gelsolin as part of an adaptive mechanism.

Thyroid hormones (T3, T4) are well known modulators of different cellular signals including the sphingomyelin pathway. However, studies regarding downstream effects of T3 on sphingolipid metabolism in skeletal muscle are scarce. In the present work we sought to investigate the effects of hyperthyroidism on the activity of the key enzymes of ceramide metabolism as well as the content of fundamental sphingolipids. Based on fiber/metabolic differences, we chose three different skeletal muscles, with diverse fiber compositions: soleus (slow-twitch oxidative), red (fast-twitch oxidative-glycolytic) and white (fast-twitch glycolytic) section of gastrocnemius. We demonstrated that T3 induced accumulation of sphinganine, ceramide, sphingosine, as well as sphingomyelin, mostly in soleus and in red, but not white section of gastrocnemius. Concomitantly, the activity of serine palmitoyltransferase and acid/neutral ceramidase was increased in more oxidative muscles. In conclusion, hyperthyroidism induced fiber specific changes in the content of sphingolipids that were relatively more related to de novo synthesis of ceramide rather than to its generation via hydrolysis of sphingomyelin.

Objectives: We aim to develop antibacterial peptide mimics resistant to protease degradation, with broad-spectrum activity at sites of infection., Methods: The bactericidal activities of LL-37, ceragenins CSA-13, CSA-90 and CSA-92 and the spermine-conjugated dexamethasone derivative D2S were evaluated using MIC and MBC measurements. Gingival fibroblast counting, interleukin-8 (IL-8) and lactate dehydrogenase (LDH) release from keratinocytes (HaCat) were used to determine effects on cell growth, pro-inflammatory response and toxicity., Results: All tested cationic lipids showed stronger bactericidal activity than LL-37. Incubation of Staphylococcus aureus with half the MIC of LL-37 led to the appearance of bacteria resistant to its bactericidal effects, but identical incubations with CSA-13 or D2S did not produce resistant bacteria. Cathelicidin LL-37 significantly increased the total number of gingival fibroblasts, but ceragenins and D2S did not alter gingival fibroblast growth. Cationic lipids showed no toxicity to HaCat cells at concentrations resulting in bacterial killing., Conclusions: These data suggest that cationic lipids such as ceragenins warrant further testing as potential novel antibacterial agents.

Sphingolipids in digestive system are responsible for numerous important physiological and pathological processes. In the membrane of gut epithelial cells, sphingolipids provide structural integrity, regulate absorption of some nutrients, and act as receptors for many microbial antigens and their toxins. Moreover, bioactive sphingolipids such as ceramide or sphingosine-1-phosphate regulate cellular growth, differentiation, and programmed cell death-apoptosis. Although it is well established that sphingolipids have clinical implications in gastrointestinal tumorigenesis or inflammation, further studies are needed to fully explore the role of sphingolipids in neoplastic and inflammatory diseases in gastrointestinal tract. Pharmacological agents which regulate metabolism of sphingolipids can be potentially used in the management of colorectal cancer or inflammatory bowel diseases. The aim of this work is to critically the review physiological and pathological roles of sphingolipids in the gastrointestinal tract.

Present in the digestive system, sphingolipids are responsible for multiple important physiological and pathological processes. On the membrane of intestinal epithelial cells sphingolipids contribute to structural integrity, regulate absorption of nutrients and may act as receptors for some microorganisms and their toxins. Moreover, bioactive lipid messengers such as ceramide and sphingosine-1-phosphate influence cellular growth, differentiation and programmed cell death, apoptosis. Further studies are needed to fully explore the clinical implications of sphingolipids in neoplastic and inflammatory diseases in the gastrointestinal tract. Pharmacological compounds which regulate metabolism of sphingolipids can be potentially useful in treatment of colon cancer, inflammatory bowel diseases or nonalcoholic fatty liver disease. The aim of this work is to present a critical review of the physiological and pathological role of sphingolipids in the digestive system.

Three bioactive sphingolipids, namely sphingosine-1-phosphate (S1P), ceramide (CER) and sphingosine (SPH) were shown to be involved in ischemia/reperfusion injury of the heart. S1P is a powerful cardioprotectant, CER activates apoptosis and SPH in a low dose is cardioprotective whereas in a high dose is cardiotoxic. The aim of the present study was to examine effects of experimental myocardial infarction on the level of selected sphingolipids in plasma, erythrocytes and platelets in the rat. Myocardial infarction was produced in male Wistar rats by ligation of the left coronary artery. Blood was taken from the abdominal aorta at 1, 6 and 24 h after the ligation. Plasma, erythrocytes and platelets were isolated and S1P, dihydrosphingosine-1-phosphate (DHS1P), SPH, dihydrosphingosine (DHS) and CER were quantified by means of an Agilent 6460 triple quadrupole mass spectrometer using positive ion electrospray ionization source with multiple reaction monitoring. The infarction reduced the plasma level of S1P, DHS1P, SPH and DHS but increased the level of total CER. In erythrocytes, there was a sharp elevation in the level of SPH and DHS early after the infarction and a reduction after 24 h whereas the level of S1P, DHS1P and total CER gradually increased. In platelets, the level of each of the examined compounds profoundly decreased 1 and 6 h after the infarction and partially normalized in 24 h. The results obtained clearly show that experimental heart infarction in rats produces deep changes in metabolism of sphingolipids in the plasma, platelets and erythrocytes.

It is known that the ratio, the level of sphingosine-1-phosphate (S1P)/the level of ceramide (CER) determines survival of the cells. The aim of the present study was to examine the effect of myocardial infarction on the level of different sphingolipids in the uninfarcted area. The experiments were carried out on male Wistar rats: 1, control; 2, after ligation of the left coronary artery (infarct) and 3, sham operated. Samples of the uninfarcted area of the left ventricle were taken in 1, 6 and 24 h after the surgery. The level of sphingolipids, S1P, CER, sphingosine (SPH), sphinganine-1-phosphate (SPA1P) and sphinganine (SPA) was determined. The control values were (ng/mg), S1P-0.33 ± 0.03, SPH-1.02 ± 0.13, SPA1P-0.11 ± 0.01, SPA-0.28 ± 0.04, total CER-20.3 ± 1.8. In infarct, the level of S1P in the uninfarcted area was reduced by ~3 times in 1 and 6 h and decreased further in 24 h. The level of SPH decreased in 1 h and returned to the control thereafter. The total level of CER decreased in 6 h after the infarction. Sham surgery also produced changes in the level of certain sphingolipids. The ratio, the level of S1P/the level of CER was markedly reduced at each time point after the infarction. It is concluded that the reduction in the S1P/CER ratio could be responsible for increased apoptosis in the uninfarcted area after the myocardial infarction in the rat.

Triiodothyronine (T(3)) can influence lipid metabolism via multiple mechanisms, which generally result in an increase of fatty acids (FAs) oxidation. Consequently, we hypothesize that hyperthyroidism may influence intramuscular lipids accumulation. This increased intramuscular lipid turn-over is possibly accompanied by an increase in fatty acid transporters expression (FAT/CD36, FABPpm, FATP-1,4). In the present study we examined the lipid content and fatty acid saturation status of free fatty acids (FFA), triacylglycerols (TAG), diacylglycerols (DAG) and phospholipids (PL) in skeletal muscle of hyperthyroid rats (n=8). We measured also fatty acid transporters as well as AMP-activated protein kinase (pAMPK/AMPK), peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α), acetyl-CoA carboxylase (pACC/ACC), carnitine palmitoyltransferase I (CPT I) and citrate synthase (CS) protein expression in these muscles. In vivo T3 administration, decreased the content of FFA, particularly in the red gastrocnemius and the TAG fraction, in both the red and white portions of the gastrocnemius muscle. Concomitantly, saturated/unsaturated fatty acids (SFA/UFA) ratio was also decreased, but only in the FFA fraction, irrespectively of muscle's fiber composition. In contrast, T(3) treatment had no effect on the lipid content and saturation status in PL fraction. Triiodothyronine induced also modest activation of AMPK/ACC axis with subsequent increased expression of mitochondrial proteins: CPT I and CS. This was accompanied by increased content of FAT/CD36, but only in the red part of gastrocnemius muscle. These findings support the conclusion that hyperthyroidism increases lipid metabolism, especially in skeletal muscles with high capacity for fatty acid oxidation.

Diabetes is associated with disturbances of brain activity and cognitive impairment. We hypothesize that ceramides may constitute an important contribution to diabetes-linked neuro-dysfunction. In our study we used rats injected with streptozotocin (STZ) as a model of severe hyperglycemia. Using the gas-liquid chromatography technique we found a significant increase of ceramide content in brains and a decrease in plasma of diabetic rats. The inhibitor of serine palmitoyltransferase, myriocin, reduced ceramide generation in hyperglycemic brains, although injected alone it exerted a paradoxical effect of ceramide upregulation. Myriocin had no impact on ceramide concentration in the plasma of either control or diabetic rats. The level of ceramide saturated fatty acids was elevated whereas the level of ceramide poly-unsaturated fatty acids was downregulated in brains of all experimental groups. The concentration of ceramide mono-unsaturated fatty acids remained unchanged. The pattern of individual ceramide species was altered depending on treatment. We noted an STZ-evoked increase of brain ceramide C16:0, C18:0 and C20:0 and a strong decline in ceramide C18:2 fatty acid levels. Some changes of brain ceramide pattern were modified by myriocin. We found a decreased amount of total ceramide-ω-6 fatty acids in STZ-treated rat brains and no changes in ceramide-ω-3 concentration. We conclude that ceramides may be important mediators of diabetes-accompanied brain dysfunction., (© 2012 The Authors Journal compilation © 2012 FEBS.)