Pleural Transport Physiology: Insights from Biological Marker Measurements in Transudates

Aims: The aim of this study was to evaluate the physicochemical properties of the pleural mesothelial barrier and of the biological markers that facilitate or eliminate the passage of molecules through the pleura. Methods and Material: Pleural fluid samples from sixty-five patients with heart failure were analyzed. The biological markers studied were lactate dehydrogenase (LDH), adenosine deaminase (ADA), interleukin-6 (IL-6), C reactive protein (CRP), tumor necrosis factor-α (TNF-α), carcinoembryonic antigen (CEA), copper/zinc superoxide dismutase (CuZnSOD), matrix metalloproteinase-2 (MMP-2), -3 (MMP-3), -7(MMP-7), -8 (MMP-8) and -9 (MMP-9). Based on the pleural fluid/serum ratio, these molecules were divided into three groups: a) the LDH-like group with a pleural fluid/serum ratio between 0,4 and 0,8 (LDH, CEA, CuZnSOD, ADA, CRP, MMP-8), b) molecules with a pleural fluid/serum ratio less than 0,4 (MMP-7 and MMP-9) and c) molecules with a pleural fluid/serum ratio equal or above 1 (TNF-α, IL-6, MMP-2 and MMP-3). Results: No correlation between the molecular radius and the pleural fluid to serum ratio of the above biological markers was found. Conclusions: The molecular size is not a major determinant for the passage of molecules through the mesothelial barrier. Several other factors may influence the transport of the above molecules to pleural cavity, such as their charge and shape. © Eleni et al

Serum Amyloid Alpha in Parapneumonic Effusions

Study objectives. To assess serum amyloid alpha (SAA) pleural fluid levels in parapneumonic effusion (PPE) and to investigate SAA diagnostic performance in PPE diagnosis and outcome. Methods. We studied prospectively 57 consecutive patients with PPE (empyema (EMP), complicated (CPE), and uncomplicated parapneumonic effusion (UPE)). SAA, CRP, TNF-α, IL-1β, and IL-6 levels were evaluated in serum and pleural fluid at baseline. Patients were followed for 6-months to detect pleural thickening/loculations. Results. Pleural SAA levels (mg/dL) median(IQR) were significantly higher in CPE compared to UPE (P < 0.04); CRP levels were higher in EMP and CPE compared to UPE (P < 0.01). There was no significant difference between IL-1β, IL-6, TNF-α level in different PPE forms. No significant association between SAA levels and 6-month outcome was found. At 6-months, patients with no evidence of loculations/thickening had significantly higher pleural fluid pH, glucose levels (P = 0.03), lower LDH (P = 0.005), IL-1β levels (P = 0.001) compared to patients who presented pleural loculations/thickening. Conclusions. SAA is increased in complicated PPE, and it might be useful as a biomarker for UPE and CPE diagnosis. SAA levels did not demonstrate considerable diagnostic performance in identifying patients who develop pleural thickening/loculations after a PPE

Atopic diseases and inflammation of the brain in the pathogenesis of autism spectrum disorders

Autism spectrum disorders (ASDs) affect as many as 1 in 45 children and are characterized by deficits in sociability and communication, as well as stereotypic movements. Many children also show severe anxiety. The lack of distinct pathogenesis and reliable biomarkers hampers the development of effective treatments. As a result, most children with ASD are prescribed psychopharmacologic agents that do not address the core symptoms of ASD. Autoantibodies against brain epitopes in mothers of children with ASD and many such children strongly correlate with allergic symptoms and indicate an aberrant immune response, as well as disruption of the blood–brain barrier (BBB). Recent epidemiological studies have shown a strong statistical correlation between risk for ASD and either maternal or infantile atopic diseases, such as asthma, eczema, food allergies and food intolerance, all of which involve activation of mast cells (MCs). These unique tissue immune cells are located perivascularly in all tissues, including the thalamus and hypothalamus, which regulate emotions. MC-derived inflammatory and vasoactive mediators increase BBB permeability. Expression of the inflammatory molecules interleukin (IL-1β), IL-6, 1 L-17 and tumor necrosis factor (TNF) is increased in the brain, cerebrospinal fluid and serum of some patients with ASD, while NF-kB is activated in brain samples and stimulated peripheral blood immune cells of other patients; however, these molecules are not specific. Instead the peptide neurotensin is uniquely elevated in the serum of children with ASD, as is corticotropin-releasing hormone, secreted from the hypothalamus under stress. Both peptides trigger MC to release IL-6 and TNF, which in turn, stimulate microglia proliferation and activation, leading to disruption of neuronal connectivity. MC-derived IL-6 and TGFβ induce maturation of Th17 cells and MCs also secrete IL-17, which is increased in ASD. Serum IL-6 and TNF may define an ASD subgroup that benefits most from treatment with the natural flavonoid luteolin. Atopic diseases may create a phenotype susceptible to ASD and formulations targeting focal inflammation of the brain could have great promise in the treatment of ASD

Hyaluronic acid levels are increased in complicated parapneumonic pleural effusions

Background and Aim. Hyaluronic acid (HA) is a component of extracellular matrix and may play a role in the pleural inflammation which is implicated in parapneumonic effusions.The aim of the current study was to investigate HA levels in serum and pleura in patients with parapneumonic effusions. Methods. We prospectively studied pleural and serum levels of HA in 58 patients with pleural effusions due to infection (complicated and uncomplicated parapneumonic effusions), malignant effusions and transudative effusions due to congestive heart failure. In addition to HA, TNF-α and IL-1β levels were determined in pleural fluid and serum by ELISA. Results. The median±SD HA levels (pg/ml) in pleural fluid of patients with complicated effusions (39.058±11.208) were significantly increased (p<0.005), compared to those with uncomplicated parapneumonic effusions (11.230±1.969), malignant effusions (10.837±4.803) or congestive heart failure (5.392±3.133). There was no correlation between pleural fluid and serum HA values. Pleural fluid TNF-α levels (146±127 pg/mL) and IL-1β levels (133.4±156 pg/mL) were significantly higher in patients with complicated parapneumonic effusions compared to patients with other types of effusion (p<0.05). No significant association between HA and TNF-α or IL-1β was found. Conclusions. HA may play a significant role in the inflammatory process which characterises exudative infectious pleuritis. Further investigation might reveal whether HA is a useful marker in the management of parapneumonic effusions

Assessment of oxidant and antioxidant markers in the pleural fluid and serum of patients with pleural effusions

The assessment of 8-isoprostane and Copper/Zinc Superoxide Dismutase (CuZnSOD) in pleural fluid (PF) and serum of patients with exudative pleural effusion in order to examine their role in the pathogenesis of pleural effusions and their diagnostic value in the differentiation between complicated and uncomplicated parapneumonic effusions. Materials and methods: We studied 214 consecutive patients with pleural effusions were admitted in the Respiratory Department of the University Hospital of Larissa. The effusions were classified as malignant (n=88), tuberculous(n=24), uncomplicated parapneumonic effusions(UPPE)(n=31), complicated parapneumonic effusions(CPPE)(n=20), empyema (n=17), and congestive heart failure(Transudates) (n=34). 8-isoprostane and CuZnSOD levels were measured with ELISA. Results: PF 8-isoprostane levels were higher in patients with PE (55.71 pg/mL) compared to those with malignant (25.10 pg/mL, p35.1pg/mL represent a good marker for the differentiation between CPPE and UPPE.Σκοπός της μελέτης ήταν ο προσδιορισμός του 8-ισοπροστανίου(8-isoprostane) και της υπεροξειδικής δισμουτάσης χαλκού/ψευδαργύρου(CuZnSOD), στο υπεζωκοτικό υγρό(ΥΥ) και στον ορό ασθενών με εξιδρωματική υπεζωκοτική συλλογή, ώστε να εκτιμηθεί ο ρόλος τους στη παθογένεση των υπεζωκοτικών συλλογών και η χρησιμότητά τους στη διάκριση των επιπλεγμένων από τις μη επιπλεγμένες παραπνευμονικές συλλογές. Υλικό & Μέθοδος: Μελετήθηκαν 214 ασθενείς που νοσηλεύτηκαν στην Πνευμονολογική Κλινική του Πανεπιστημιακού Νοσοκομείου Λάρισας, οι συλλογές των οποίων ταξινομήθηκαν στις ακόλουθες ομάδες σύμφωνα με τη τελική διάγνωση. Κακοήθεια(n=88), φυματιώδης πλευρίτιδα(n=24), μη επιπλεγμένη παραπνευμονική συλλογή(ΜΕΠΣ)(n=31), επιπλεγμένη παραπνευμονική συλλογή(ΕΠΣ)(n=20), εμπύημα(n=17) και συμφορητική καρδιακή ανεπάρκεια(Διίδρωμα)(n=34). Το 8-isoprostane και η CuZnSOD προσδιορίστηκαν με ELISA. Αποτελέσματα: Στο ΥΥ τo 8-isoprostane βρέθηκε αυξημένο στις ΠΣ(55.71 pg/mL) συγκριτικά με τις κακοήθεις(25.10 pg/mL, p<0.0001) και φυματιώδεις συλλογές(25.13 pg/mL, p<0.0001). Στο λόγο ΥΥ/ορό, αυξημένες τιμές παρατηρήθηκαν στις ΠΣ(1.42 pg/mL), σε σχέση με τις κακοήθεις(0.77 pg/mL, p=0.0004) και φυματιώδεις συλλογές (0.69 pg/mL, p=0.0004). Η CuZnSOD στο ΥΥ ασθενών με ΠΣ(126.5 ng/mL) βρέθηκε υψηλή συγκριτικά με το διίδρωμα(83.40 ng/mL, p=0.0012). Στο λόγο η CuZnSOD βρέθηκε αυξημένη στην ΠΣ(1.68) σε σχέση με διίδρωμα και κακοήθεια(0.62 και 0.89, p<0.0001 και p<0.05, αντίστοιχα). Επιπλέον, το 8-isoprostane βρέθηκε υψηλό στο εμπύημα(420.2288.4 pg/mL) συγκρινόμενο με τις ΜΕΠΣ(32.32pg/mL, p<0.0001). Η συγκέντρωση του 8-isoprostane βρέθηκε υψηλή στις ΕΠΣ(65.00pg/mL) σε σχέση με τις ΜΕΠΣ(32.32pg/mL, p<0.0001). Η CuZnSOD στο ΥΥ βρέθηκε αυξημένη στο εμπύημα(1301881.2ng/mL) συγκρινόμενη με ΕΠΣ(141.3ng/mL,p<0.0001) και ΜΕΠΣ(83.20ng/mL, p<0.0001). Αυξημένες βρέθηκαν οι συγκεντρώσεις της CuZnSOD στο ΥΥ στις ΕΠΣ(141.3 ng/mL) συγκριτικά με τις ΜΕΠΣ(83.20 ng/mL, p<0.0001). Στο λόγο τα επίπεδα του 8-isoprostane και της CuZnSOD βρέθηκαν υψηλότερα στο εμπύημα(12.91 και 10.84,αντίστοιχα) σε σχέση με τις ΜΕΠΣ (1.0 και 1.6,p<0.0001 και p<0.0001,αντίστοιχα) και ΕΠΣ(2.14 και 0.9, p<0.0001 και p<0.0001,αντίστοιχα). Στο ΥΥ το 8-isoprostane με όριο την τιμή 35.1 pg/mL, παρουσίασε ευαισθησία 100.0% και ειδικότητα 58.1%(AUC=0.848) ως προς τη διάκριση των ΕΠΣ από τις ΜΕΠΣ. Συμπεράσματα: Αυξημένα επίπεδα 8-isoprostane και CuZnSOD παρατηρήθηκαν στις ΠΣ και τα εμπυήματα σε σχέση με τις κακοήθεις και φυματιώδεις υπεζωκοτικές συλλογές. Ο προσδιορισμός των παραπάνω δεικτών πιθανόν μπορεί να βοηθήσει στην διάκριση των ΕΠΣ από τις ΜΕΠΣ

Extracellular vesicles are increased in the serum of children with autism spectrum disorder, contain mitochondrial DNA, and stimulate human microglia to secrete IL-1β

Abstract Background Autism spectrum disorder (ASD) has been associated with brain inflammation as indicated by the activation of microglia, but the triggers are not known. Extracellular vesicles (EVs) are secreted from many cells in the blood and other biological fluids and carry molecules that could influence the function of target cells. EVs have been recently implicated in several diseases, but their presence or function in ASD has not been studied. Methods EVs were isolated from the serum of children with ASD (n = 20, 16 males and 4 females, 4–12 years old) and unrelated age and sex-matched normotypic controls (n = 8, 6 males and 2 females, 4–12 years old) using the exoEasy Qiagen kit. EVs were characterized by determining the CD9 and CD81 membrane-associated markers with Western blot analysis, while their morphology and size were assessed by transmission electron microscopy (TEM). Human microglia SV40 were cultured for 24 h and then stimulated with EVs (1 or 5 μg/mL), quantitated as total EV-associated protein, for 24 or 48 h. IL-1β secretion was measured by ELISA. The results were analyzed using the Mann-Whitney U non-parametric test, and all statistical analyses were performed using Graph Pad Prism 5. Results EVs were isolated and shown to be spherical structures (about 100 nm) surrounded by a membrane. Total EV-associated protein was found to be significantly increased (p = 0.02) in patients as compared to normotypic controls. EVs (5 μg/mL) isolated from the serum of patients with ASD stimulated cultured human microglia to secrete significantly more of the pro-inflammatory cytokine interleukin IL-1β (163.5 ± 13.34 pg/mL) as compared to the control (117.7 ± 3.96 pg/mL, p < 0.0001). The amount of mitochondrial DNA (mtDNA7S) contained in EVs from children with ASD was found to be increased (p = 0.046) compared to the normotypic controls. Conclusions These findings provide novel information that may help explain what triggers inflammation in the brain of children with ASD and could lead to novel effective treatments

Mast Cells, Stress, Fear and Autism Spectrum Disorder

Autism Spectrum Disorder (ASD) is a developmental condition characterized by impaired communication and obsessive behavior that affects 1 in 59 children. ASD is expected to affect 1 in about 40 children by 2020, but there is still no distinct pathogenesis or effective treatments. Prenatal stress has been associated with higher risk of developing ASD in the offspring. Moreover, children with ASD cannot handle anxiety and respond disproportionately even to otherwise benign triggers. Stress and environmental stimuli trigger the unique immune cells, mast cells, which could then trigger microglia leading to abnormal synaptic pruning and dysfunctional neuronal connectivity. This process could alter the &ldquo;fear threshold&rdquo; in the amygdala and lead to an exaggerated &ldquo;fight-or-flight&rdquo; reaction. The combination of corticotropin-releasing hormone (CRH), secreted under stress, together with environmental stimuli could be major contributors to the pathogenesis of ASD. Recognizing these associations and preventing stimulation of mast cells and/or microglia could greatly benefit ASD patients

Children with autism spectrum disorders, who improved with a luteolin-containing dietary formulation, show reduced serum levels of TNF and IL-6

Autism spectrum disorders (ASDs) have been associated with brain inflammation as indicated by microglia activation, as well as brain expression and increased plasma levels of interleukin-6 (IL-6) and tumor necrosis factor (TNF). Here we report that serum levels of IL-6 and TNF were elevated (61.95±94.76 pg ml-1 313.8 ±444.3 pg ml-1, respectively) in the same cohort of patients with elevated serum levels of corticotropin-releasing hormone (CRH) and neurotensin (NT), while IL-9, IL-31 and IL-33 were not different from controls. The elevated CRH and NT levels did not change after treatment with a luteolin-containing dietary formulation. However, the mean serum IL-6 and TNF levels decreased significantly (P=0.036 and P=0.015, respectively) at the end of the treatment period (26 weeks) as compared with levels at the beginning; these decreases were strongly associated with children whose behavior improved the most after luteolin formulation treatment. Our results indicate that there are distinct subgroups of children within the ASDs that may be identifiable through serum levels of IL-6 and TNF and that these cytokines may constitute distinct prognostic markers for at least the beneficial effect of luteolin formulation

The mast cell – Neurofibromatosis connection

[No abstract available