Avaliação da expressão dos genes cFOS, IL-1b, CYP1a1 e CYP1b1 em Danio rerio expostos a Benzo[a]pireno e tratados com ligantes do receptor P2X7

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Polimorfismos genéticos e expressão de marcadores envolvidos em processos inflamatórios, angiogênicos e de hipóxia na doença falciforme

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Urinary MCP-1 and RBP: Independent predictors of renal outcome in macroalbuminuric diabetic nephropathy

Background: Albuminuria has been considered a sine qua non condition for the diagnosis of diabetic nephropathy (DN) and has been widely used as a surrogate outcome of chronic kidney disease (CKD). However, recent data suggest that albuminuria may fail as a biomarker in a subset of patients, and the search for novel markers is intense. Methods: We analyzed the role of urinary RBP and of serum and urinary cytokines (TGF-beta, MCP-1 and VEGF) as predictors of the risk of dialysis. doubling of serum creatinine or death (primary outcome. PO) in 56 type 2 diabetic patients with macroalbuminuric DN. Results: Mean follow-up time was 30.7 +/- 10 months. Urinary RBP and MCP-1 were significantly higher in patients presenting the PO, whereas no difference was shown for TGF-beta or VEGF. In the Cox regression, urinary RBP. MCP-1 and VEGF were positively associated and serum VEGF was inversely related to the risk of the PO. However, after adjustments for creatinine clearance, proteinuria, and blood pressure only urinary RBP (OR 11.6; 95% CI 2.7-49.2, p = 0.001 for log RBP) and urinary MCP-1 (OR 11.0; 95% CI 1.6-76.4, p = 0.02 for log MCP-1) remained as significant independent predictors of the PO. Conclusion: Urinary RBP and MCP-1 are independently related to the risk of CKD progression in patients with macroalbuminuric DN. Whether these biomarkers have a role in the setting of normoalbuminuria and microalbuminuria in DN should be further investigated. (C) 2012 Elsevier Inc. All rights reserved.

Experimental arthritis exacerbates Aggregatibacter actinomycetemcomitans-induced periodontitis in mice

Aim This study aimed to investigate whether chronic antigen-induced arthritis (AIA) influences infection-induced periodontitis (PD) in mice and whether PD modifies the clinical course of AIA. The contribution of anti-TNF-a therapy was also evaluated. Materials and methods The PD was induced in C57BL/6 mice by oral infection with Aggregatibacter actinomycetemcomitans. AIA was induced after infection. Anti-TNF-a and chlorhexidine therapies were used to investigate the role of TNF-a and oral infection on PD and AIA interaction. Maxillae, knee joints, lymph nodes and serum samples were used for histomorphometric, immunoenzymatic and/or real time-PCR analyses. Results Antigen-induced arthritis exacerbated alveolar bone loss triggered by PD infection. In contrast, PD did not influence AIA in the evaluated time-points. PD exacerbation was associated with enhanced production of IFN-? in maxillae and expression of the Th1 transcription factor tBET in submandibular lymph nodes. Increased serum levels of IL-6 and C-reactive protein were also detected. Anti-TNF-a and antiseptic therapies prevented the development and exacerbation of infectious-PD. Anti-TNF-a therapy also resulted in reduced expression of IFN-?, TNF-a and IL-17 in maxillae. Conclusions Altogether, the current results indicate that the exacerbation of infection-induced PD by arthritis is associated with an alteration in lymphocyte polarization pattern and increased systemic immunoreactivity. This process was ameliorated by anti-TNF-a and antiseptic therapies.

Oral Administration of Oleic or Linoleic Acid Accelerates the Inflammatory Phase of Wound Healing

The effects of oral ingestion of oleic (OLA) and linoleic (LNA) acids on wound healing in rats were investigated. LNA increased the influx of inflammatory cells, the concentration of hydrogen peroxide (H(2)O(2)) and cytokine-induced neutrophil chemoattractant-2 alpha beta (CINC-2 alpha beta), and the activation of the transcription factor activator protein-1 (AP-1) in the wound at 1 hour post wounding. LNA decreased the number of inflammatory cells and IL-1, IL-6, and macrophage inflammatory protein-3 (MIP-3) concentrations, as well as NF-kappa B activation in the wound at 24 hours post wounding. LNA accelerated wound closure over a period of 7 days. OLA increased TNF-alpha concentration and NF-kappa B activation at 1 hour post wounding. A reduction of IL-1, IL-6, and MIP-3 alpha concentrations, as well as NF-kappa B activation, was observed 24 hours post wounding in the OLA group. These data suggest that OLA and LNA accelerate the inflammatory phase of wound healing, but that they achieve this through different mechanisms.

Influence of Mid-Trimester Amniotic Fluid on Endogenous and Lipopolysaccharide-Mediated Responses of Mononuclear Lymphoid Cells

Problem We evaluated the influence of amniotic fluid (AF) on immune mediator production by mononuclear leukocytes. Method of study Thirty mid-gestation AFs were incubated with peripheral blood mononuclear cells (PBMCs) in the presence or absence of lipopolysaccharide (LPS). Supernatants were tested for interleukin (IL) -6, 10, 12, 23, tumor necrosis factor-alpha (TNF-alpha) and monocyte chemotactic protein (MCP)-1. Results Endogenous mediator production was minimal or non-detectable. AF stimulated endogenous MCP-1, IL-6 and TNF-alpha release. In the presence of LPS, production of MCP-1 and IL-10 by PBMCs was enhanced eightto ninefold by AF. Release of IL-6 and IL-23 was enhanced less than twofold by the addition of AF while TNF-alpha production was unchanged. AF-stimulated mediator production was similar irrespective of pregnancy outcome. Conclusion Selective AF stimulation of LPS-mediated MCP-1 and IL-10 release may be a mechanism to promote antibody production and the influx of phagocytic cells to engulf pathogens while downregulating the production of pro-inflammatory cytokines.

Spironolactone prevents alterations associated with cardiac hypertrophy produced by isoproterenol in rats: involvement of serum- and glucocorticoid-regulated kinase type 1

Persistent beta-adrenergic receptor stimulation with isoproterenol is associated with cardiac hypertrophy as well as cardiac synthesis of angiotensin II. Serum- and glucocorticoid-regulated kinase type 1 (SGK-1) is a key mediator in structural, functional and molecular cardiac effects of aldosterone in rats. This study was designed to investigate the cardiac effects of the mineralocorticoid receptor antagonist spironolactone on the response to isoproterenol treatment in rats, as well as the involvement of the main mediator of cellular aldosterone action, SGK-1, in the heart. Male Wistar rats received isoproterenol (3 mg kg-1 day-1) or vehicle for 15 days. Half of the animals in each group were simultaneously treated with spironolactone (200 mg kg-1 day-1). Systolic and diastolic blood pressures were not significantly different among groups. Treatment with spironolactone normalized the increased left ventricular end-diastolic pressure observed in isoproterenol-treated rats. Isoproterenol treatment induced cardiac hypertrophy and increased collagen content, both of which were normalized by spironolactone treatment. The mRNA levels of transforming growth factor beta, connective tissue growth factor, matrix metalloprotease 2, matrix metalloprotease inhibitor 2, tumour necrosis factor a, interleukin 1 beta, p22phox and xanthine dehydrogenase were increased (P < 0.05) in isoproterenol-treated rats, and this effect was prevented by spironolactone (P < 0.05). Spironolactone also reduced the elevated SGK-1 expression in isoproterenol-treated rats. The observed reduction of the principal mediator of aldosterone cellular actions, SGK-1, by spironolactone in hearts from isoproterenol-treated rats suggests a role of mineralocorticoids in the cardiac hypertrophy, fibrosis, inflammation, oxidation and diastolic dysfunction induced by isoproterenol treatment in rats.

Comparison of two experimental models of pulmonary hypertension

Objective: To compare two models of pulmonary hypertension (monocrotaline and monocrotaline+pneumonectomy) regarding hemodynamic severity, structure of pulmonary arteries, inflammatory markers (IL-1 and PDGF), and 45-day survival. Methods: We used 80 Sprague-Dawley rats in two study protocols: structural analysis; and survival analysis. The rats were divided into four groups: control; monocrotaline (M), pneumonectomy (P), and monocrotaline+pneumonectomy (M+P). In the structural analysis protocol, 40 rats (10/group) were catheterized for the determination of hemodynamic variables, followed by euthanasia for the removal of heart and lung tissue. The right ventricle (RV) was dissected from the interventricular septum (IS), and the ratio between RV weight and the weight of the left ventricle (LV) plus IS (RV/LV+IS) was taken as the index of RV hypertrophy. In lung tissues, we performed histological analyses, as well as using ELISA to determine IL-1 and PDGF levels. In the survival protocol, 40 animals (10/group) were followed for 45 days. Results: The M and M+P rats developed pulmonary hypertension, whereas the control and P rats did not. The RV/LV+IS ratio was significantly higher in M+P rats than in M rats, as well as being significantly higher in M and M+P rats than in control and P rats. There were no significant differences between the M and M+P rats regarding the area of the medial layer of the pulmonary arteries; IL-1 and PDGF levels; or survival. Conclusions: On the basis of our results, we cannot conclude that the monocrotaline+pneumonectomy model is superior to the monocrotaline model.

Effects of overinflation on procollagen type III expression in experimental acute lung injury

Abstract Introduction In acute lung injury (ALI), elevation of procollagen type III (PC III) occurs early and has an adverse impact on outcome. We examined whether different high-inflation strategies of mechanical ventilation (MV) in oleic acid (OA) ALI alter regional expression of PC III. Methods We designed an experimental, randomized, and controlled protocol in which rats were allocated to two control groups (no injury, recruited [alveolar recruitment maneuver after tracheotomy without MV; n = 4 rats] and control [n = 5 rats]) or four injured groups (one exposed to OA only [n = 10 rats] and three OA-injured and ventilated). The three OA-injured groups were ventilated for 1 hour according to the following strategies: LVHP-S (low volume-high positive end-expiratory pressure [PEEP], supine; n = 10 rats, tidal volume [VT] = 8 ml/kg, PEEP = 12 cm H2O), HVLP-S (high volume-low PEEP, supine; n = 10 rats, VT = 20 ml/kg, PEEP = 5 cm H2O), and HVLP-P (high volume-low PEEP, prone; n = 10 rats). Northern blot analysis for PC III and interleukin-1-beta (IL-1β) and polymorphonuclear infiltration index (PMI) counting were performed in nondependent and dependent regions. Regional differences between groups were assessed by two-way analysis of variance after logarithmic transformation and post hoc tests. Results A significant interaction for group and region effects was observed for PC III (p = 0.012) with higher expression in the nondependent region for HVLP-S and LVHP-S, intermediate for OA and HVLP-P, and lower for control (group effect, p < 0.00001, partial η2 = 0.767; region effect, p = 0.0007, partial η2 = 0.091). We found high expression of IL-1β (group effect, p < 0.00001, partial η2 = 0.944) in the OA, HVLP-S, and HVLP-P groups without regional differences (p = 0.16). PMI behaved similarly (group effect, p < 0.00001, partial η2 = 0.832). Conclusion PC III expression is higher in nondependent regions and in ventilatory strategies that caused overdistension. This response was partially attenuated by prone positioning.

Impairment of the hematological response and interleukin-1β production in protein-energy malnourished mice after endotoxemia with lipopolysaccharide

The objectives of this study were to determine if protein-energy malnutrition (PEM) could affect the hematologic response to lipopolysaccharide (LPS), the interleukin-1β (IL-1β) production, leukocyte migration, and blood leukocyte expression of CD11a/CD18. Two-month-old male Swiss mice were submitted to PEM (N = 30) with a low-protein diet (14 days) containing 4% protein, compared to 20% protein in the control group (N = 30). The total cellularity of blood, bone marrow, spleen, and bronchoalveolar lavage evaluated after the LPS stimulus indicated reduced number of total cells in all compartments studied and different kinetics of migration in malnourished animals. The in vitro migration assay showed reduced capacity of migration after the LPS stimulus in malnourished animals (45.7 ± 17.2 x 10(4) cells/mL) compared to control (69.6 ± 7.1 x 10(4) cells/mL, P ≤ 0.05), but there was no difference in CD11a/CD18 expression on the surface of blood leukocytes. In addition, the production of IL-1β in vivo after the LPS stimulus (180.7 pg·h-1·mL-1), and in vitro by bone marrow and spleen cells (41.6 ± 15.0 and 8.3 ± 4.0 pg/mL) was significantly lower in malnourished animals compared to control (591.1 pg·h-1·mL-1, 67.0 ± 23.0 and 17.5 ± 8.0 pg/mL, respectively, P ≤ 0.05). The reduced expression of IL-1β, together with the lower number of leukocytes in the central and peripheral compartments, different leukocyte kinetics, and reduced leukocyte migration capacity are factors that interfere with the capacity to mount an adequate immune response, being partly responsible for the immunodeficiency observed in PEM.

Il mercato e-MID: intraday e crisi finanziaria

Il lavoro di tesi ha l’obiettivo di dare seguito e profondità alle verifiche già condotte negli anni scorsi da Monticini e Baglioni, da Angelini e da Vento e La Ganga. Nel dettaglio nella serie storica dei tassi scambiati, sul comparto ON, del mercato e-Mid, riscontrano empiricamente che i tassi scambiati nelle prime ore del mattino sono in media superiori rispetto a quelli del pomeriggio. In pratica emerge un costo orario positivo. Il campione è costituito da 196.571 contratti eseguiti tra il 1 gennaio 2007 ed il 31 marzo 2009, sul comparto ON. Le analisi si sono concentrate sull’intensità del reversal patttern, oltre che su base giornaliera anche su base settimanale (day of the week) e su base mensile (month of the year effect). Le medesime analisi sono state condotte sull’ultimo giorno del periodo di mantenimento, e su particolari giorni che hanno scandito la crisi finanziaria. Per osservare la dinamica del reversal pattern prima e durante la crisi, le medesime verifiche sono condotte su tre sub campioni. Il primo considera tutti i contratti dall’inizio e fino all’8 agosto compreso; il secondo da quest’ultimo e fino al 12 settembre 2008 compreso; l’ultimo dal fallimento della Lehman e fino al 31 marzo 2009. I risultati confermano la presenza del reversal, per tutti i giorni e per tutti i mesi. Tuttavia quando il campione è suddiviso, le stime del primo campione, perdono di significatività, in particolare il giovedì ed il mese di febbraio. Sel medesimo periodo di campionamento al fine di verificare se il reversal pattern sia attribuire in parte anche alla liquidità del mercato, si è indagato su tutte le proposte in bid ed in offer, e dei relativi volumi, ovvero sul bid ask spread, il relativo prezzo medio ed i volumi. Anche in questo caso le analisi si sono concentrate oltre che sul campione originario, anche sui tre sub campioni in cui è stato successivamente suddiviso. . Quando infine si analizzano volumi e proposte, si nota innanzitutto che, al mattino, quando i prezzi scambiati sono maggiori il bid ask spread è minimo. Viceversa il pomeriggio quando il BAS è minore. La stessa dinamica si osserva per lo spread effettivo (SE), il quale è pari alla differenza tra prezzo medio delle contrattazioni e prezzo medio delle proposte. Infine i volumi sia delle proposte che delle contrattazioni, sono maggiori al mattino rispetto al pomeriggio e tendono a decrescere con l’intensificarsi della crisi. Inoltre i volumi in bid sono sempre maggiori rispetto a quelli in offer al mattino piuttosto che al pomeriggio.

Rare and complicated forms of infantile hemangiomas: new therapeutic outlooks

Infantile hemangiomas (IHs) are the most common benign neoplastic pathology of childhood; their natural history generally involves three phases: after the onset, which usually occurs in the first weeks of life, there is the proliferation phase where the IH reaches its maximum development and it is followed by the spontaneous involution which leads to the IH regression. The duration and the extent of these phases may vary widely even though in most of the cases the involution process begins around twelve months of life and the regression, complete or partial, is completed around seventh-ninth year of life. The majority of the IHs does not require any treatment. However, 10%-20% is likely to develop serious complications, functional impairments or aesthetic alterations and entail a timely treatment. Although there is no treatment protocol currently shared, therapies usually used in cases with a complication risk consist in: systemic or intralesional steroids as a first choice; interferon α, vincristine and/or bleomicin as second or third choice and/or surgical treatment. Propranolol, a non-selective beta-blocker, has been used for cardiovascular diseases even in childhood for decades. Since 2008 it has been widely used in the IHs treatment, although it is still "off-label". In literature there are hundreds of cases and some clinical studies that show the effectiveness and safety of this drug for this indication. Thanks to a multidisciplinary team (Dermatologists, Cardiologists, Paediatricians, and Radiologists) of S. Orsola-Malpighi Hospital, a clinical study, which has been previously approved by the ethics committee, is carried out in order to evaluate the efficacy and safety of systemic propranolol in the treatment of IHs in paediatric age. At the end of 2012, 78 patients underwent this treatment: the results we have obtained so far show a good efficacy and safety profile in agreement with the data provided by the literature.

Die Beeinflussung einer Th2-Zell-vermittelten Immunantwort durch Interleukin-1-alpha am Beispiel des murinen allergischen Asthmas

Dass durch IL-1α die Th1-vermittelte Immunreaktion der kutanen Leishmaniasis beeinflusst werden kann, konnte unsere Arbeitsgruppe bereits zeigen. Daran anknüpfend war das Ziel meiner Dissertation zu prüfen, ob sich diese Erkenntnisse im Modell des murinen allergischen Asthmas reproduzieren lassen, auch im Sinne eines zukünftigen therapeutischen Nutzens für die Behandlung dieser epidemiologisch hochrelevanten Erkrankung. Daneben sollte die Verwendung der gesamten murinen Lunge als Quelle für Untersuchungsmaterial erschlossen werden. Zu diesem Zweck wurden bei BALB/c Mäusen ein (OVA)/Alum-induziertes allergisches Asthma in Gegenwart oder Abwesenheit von IL-1α generiert. Anschließend wurde eine broncheoalveoläre Lavage (BAL) durchgeführt, bzw. die komplette linke Lunge gewonnen und prozessiert. Der Einfluss von IL-1α auf den Phänotyp der Th2-vermittelten Immunantwort ließ sich auf zytomorphologischer, durchflusszytometrischer und Zytokin-Ebene nachweisen. So zeigte ein Teil unserer Ergebnisse, dass nach der frühen Gabe eine Tendenz zur Verlagerung des Gewichtes der Abwehrreaktion von Th2 in Richtung Th1 besteht. Ebenso fanden sich Hinweise für eine relative Augmentation der Th2-Antwort durch den Einsatz von IL-1α zu einem späteren Zeitpunkt. Eine absolute Verstärkung der Th2-Reaktion auf OVA durch IL-1α konnten wir nicht messen. Hier scheint mit OVA allein schon eine maximale Ausprägung erreicht zu sein. Neben den Th1/Th2-Effekten wurden auch einige gegenläufige Beobachtungen gemacht, welche nicht a priori durch das Th1/Th2-Paradigma zu erklären sind, sondern den Einfluß von IL-1α auf andere Systeme belegen, wie z. B. die Wirkung auf CCL28 und regulatorische T-Zellen. Für die Gewinnung von inflammatorischen Zellen aus der kompletten Lunge und deren weitere Untersuchung konnten wir eine Methode entwickeln und standardisieren, welche relativ einfach in der Durchführung ist und zuverlässig eine im Verhältnis zur BAL hohe Zellzahl liefert, was wiederum ein breites Spektrum an weiteren Untersuchungen erlaubt. Durch den Vorgang der mechanischen und enzymatischen Prozessierung scheinen die funktionellen Eigenschaften der Zellen nicht wesentlich beeinträchtigt zu sein. Der Einsatz von IL-1α resultierte letztendlich in einem Mischbild an hervorgerufenen Veränderungen und es bedarf noch weiterer Studien, um die unterschiedlichen induzierten Mechanismen sauber voneinander zu trennen und den therapeutischen Nutzen von IL-1α im allergischen Asthma zu evaluieren.

Metodi perturbativi dipendenti dal tempo

Sono rari i problemi risolubili in maniera esatta in meccanica quantistica. Ciò è legato alle difficoltà matemtiche insite nella teoria dei quanti. Inoltre i problemi risolti, pur essendo in alcuni casi delle ottime approssimazioni, sono spesso delle astrazioni delle situazioni reali. Si pensi ad esempio al caso di una particella quantistica di un problema unidimensionale. Questi sistemi sono talmente astratti da violare un principio fondamentale, il principio di indetermi- nazione. Infatti le componenti dell’impulso e della posizione perpendicolari al moto sono nulle e quindi sono note con certezza ad ogni istante di tempo. Per poter ottenere una descrizione dei modelli che tendono alla realtà è necessario ricorrere alle tecniche di approssimazione. In questa tesi sono stati considerati i fenomeni che coinvolgono una interazione variabile nel tempo. In particolare nella prima parte è stata sviluppata la teoria delle rappresentazioni della meccanica quantistica necessaria per la definizione della serie di Dyson. Questa serie oper- atoriale dovrebbe convergere (il problema della convergenza non è banale) verso l’operatore di evoluzione temporale, grazie al quale è possibile conoscere come un sistema evolve nel tempo. Quindi riuscire a determinare la serie di Dyson fino ad un certo ordine costituisce una soluzione approssimata del problema in esame. Supponiamo che sia possibile scomporre l’hamiltoniana di un sistema fisico nella somma di due termini del tipo: H = H0 + V (t), dove V (t) è una piccola perturbazione dipendente dal tempo di un problema risolubile esattamente caratterizzato dall’hamiltoniana H0 . In tal caso sono applicabili i metodi della teoria perturbativa dipendente dal tempo. Sono stati considerati due casi limite, ovvero il caso in cui lo spettro dell’hamiltoniana imperturbata sia discreto e non degenere ed il caso in cui lo spettro sia continuo. La soluzione al primo ordine del caso discreto è stata applicata per poter formu- lare il principio di indeterminazione energia-tempo e per determinare le regole di selezione in approssimazione di dipolo elettrico. Il secondo caso è servito per spiegare il decadimento beta, rimanendo nel campo della teoria quantistica classica (per una trattazione profonda del problema sarebbe necessaria la teoria dei campi).

Dose-response effect of interleukin (IL)-1beta, tumour necrosis factor (TNF)-alpha, and interferon-y on the in vitro production of epithelial neutrophil activating peptide-78 (ENA-78), IL-8, and IL-6 by human endometrial stromal cells

The production of epithelial neutrophil activating peptide-78 (NA-78) and the interleukins IL-8 and IL-6 by endometrial stromal cells is stimulated by pro-inflammatory interleukin-1 (IL-1) and tumour necrosis factor-α (TNF-α). IL-8 is suggested to play a role in the pathogenesis of endometriosis, and in these women the peritoneal fluid concentrations of ENA-78 and IL-8 are increased. TNF-α has been tested together with interferon-γ because of their cooperative stimulation of IL-6. The release of IL-8, however, is inhibited with increasing interferon levels. The aim of the study was the analysis of the production of ENA-78, IL-6 and IL-8 by cultured human endometrial stromal cells in the presence of varying concentrations of IL-1β, TNF-α, and interferon-γ.