Unpredictable Chronic Stress Alters Adenosine Metabolism in Zebrafish Brain.

Stress is considered a risk factor for several human disorders. Despite the broad knowledge of stress responses in mammals, data on the relationship between unpredictable chronic stress (UCS) and its effects on purinergic signaling are limited. ATP hydrolysis by ectonucleotidases is an important source of adenosine, and adenosine deaminase (ADA) contributes to the control of the nucleoside concentrations. Considering that some stress models could affect signaling systems, the objective of this study was to investigate whether UCS alters ectonucleotidase and ADA pathway in zebrafish brain. Additionally, we analyzed ATP metabolism as well as ada1, ada2.1, ada2.2, adaL, and adaasi gene expression in zebrafish brain. Our results have demonstrated that UCS did not alter ectonucleotidase and soluble ADA activities. However, ecto-ADA activity was significantly decreased (26.8%) in brain membranes of animals exposed to UCS when compared to the control group. Quantitative reverse transcription PCR (RT-PCR) analysis did not show significant changes on ADA gene expression after the UCS exposure. The brain ATP metabolism showed a marked increase in adenosine levels (ADO) in animals exposed to UCS. These data suggest an increase on extracellular adenosine levels in zebrafish brain. Since this nucleoside has neuromodulatory and anxiolytic effects, changes in adenosine levels could play a role in counteracting the stress, which could be related to a compensatory mechanism in order to restore the homeostasis.

Decrease of serum adenine nucleotide hydrolysis in an irritant contact dermatitis mice model: potential P2X7R involvement.

Extracellular adenosine 5'-triphosphate (ATP) has significant effects on a variety of pathological conditions and it is the main physiological agonist of P2X7 purinergic receptor (P2X7R). It is known that ATP acting via purinergic receptors plays a relevant role on skin inflammation, and P2X7R is required to neutrophil recruitment in a mice model of irritant contact dermatitis (ICD).The present study investigated the effects of chemical irritant croton oil (CrO) upon ATP, ADP, and AMP hydrolysis in mice blood serum, and the potential involvement of P2X7R. The topical application CrO induced a decrease on soluble ATP/ADPase activities (~50 %), and the treatment with the selective P2X7R antagonist, A438079, reversed these effects to control level. Furthermore, we showed that CrO decreased cellular viability (52.6 % ± 3.9) in relation to the control and caused necrosis in keratinocytes (PI positive cells). The necrosis induced by CrO was prevented by the pre-treatment with the selective P2X7R antagonist A438079. The results presented herein suggest that CrO exerts an inhibitory effect on the activity of ATPDase in mouse serum, reinforcing the idea that ICD has a pathogenic mechanism dependent of CD39. Furthermore, it is tempting to suggest that P2X7R may act as a controller of the extracellular levels of ATP.

Spinal blockage of P/Q- or N-type voltage-gated calcium channels modulates functional and symptomatic changes related to haemorrhagic cystitis in mice.

BACKGROUND AND PURPOSE: Spinal voltage-gated calcium channels (VGCCs) are pivotal regulators of painful and inflammatory alterations, representing attractive therapeutic targets. We examined the effects of epidural administration of the P/Q- and N-type VGCC blockers Tx3-3 and Phα1ß, respectively, isolated from the spider Phoneutria nigriventer, on symptomatic, inflammatory and functional changes allied to mouse cyclophosphamide (CPA)-induced haemorrhagic cystitis (HC). The effects of P. nigriventer-derived toxins were compared with those displayed by MVIIC and MVIIA, extracted from the cone snail Conus magus. EXPERIMENTAL APPROACH: HC was induced by a single i.p. injection of CPA (300 mg·kg(-1) ). Dose- and time-related effects of spinally administered P/Q and N-type VGCC blockers were assessed on nociceptive behaviour and macroscopic inflammation elicited by CPA. The effects of toxins were also evaluated on cell migration, cytokine production, oxidative stress, functional cystometry alterations and TRPV1, TRPA1 and NK1 receptor mRNA expression. KEY RESULTS: The spinal blockage of P/Q-type VGCC by Tx3-3 and MVIIC or N-type VGCC by Phα1ß attenuated nociceptive and inflammatory events associated with HC, including bladder oxidative stress and cytokine production. CPA produced a slight increase in bladder TRPV1 and TRPA1 mRNA expression, which was reversed by all the toxins tested. Noteworthy, Phα1ß strongly prevented bladder neutrophil migration, besides HC-related functional alterations, and its effects were potentiated by co-injecting the selective NK1 receptor antagonist CP-96345. CONCLUSIONS AND IMPLICATIONS: Our results shed new light on the role of spinal P/Q and N-type VGCC in bladder dysfunctions, pointing out Phα1ß as a promising alternative for treating complications associated with CPA-induced HC.

The effects of fructose-1,6-bisphosphate and dexamethasone on acute inflammation and T-cell proliferation.

OBJECTIVE AND DESIGN: Chronic glucocorticoid treatment is associated with pharmacological resistance. We investigated the auxiliary effects of fructose-1,6-bisphosphate (FBP) on dexamethasone (DEX)-related modulation of inflammation and T-cell proliferation. METHODS: Acute inflammation (pleurisy) was induced by injection of carrageenan into the pleural cavity of rats that were treated in vivo with DEX s. c. and FBP i. p. Peripheral blood mononuclear cells were isolated and T-cell sensitivity to FBP and DEX was evaluated in vitro. RESULTS: FBP and DEX reduced the exudate volume, protein concentration and neutrophils in the pleural cavity. However no synergistic effects were observed when these compounds were tested simultaneously. In contrast, both compounds dose-dependently and synergistically suppressed T-cell proliferation. CONCLUSION: These data suggest that FBP may be beneficial as auxiliary drug for the treatment of patients with acquired glucocorticoid resistance.

Extract of the bristles of Dirphia sp. increases nitric oxide in a rat pleurisy model.

OBJECTIVES: In the present study we assessed the inflammatory potential of venom obtained from caterpillar genus Dirphia in an acute model of lung injury. MATERIAL AND METHODS: Injection of extract from the bristles of Dirphia sp. (EBD) into the pleural cavity of rats elicited an acute inflammation response characterized by fluid accumulation which contained a large number of polymorphonuclear neutrophils (PMNs). RESULTS: The results show that EBD induces an inflammatory response, with a significant increase in PMNs, exudate and nitric oxide within 4 h after a 0.04 mg/kg dose. The administration of anti-inflammatory drugs (fructose-1,6-bisphosphate, dexamethasone, rofecoxib, sodium diclofenac and pyrilamine) significantly reduced the inflammatory effect of EBD. CONCLUSIONS: EBD causes an inflammatory reaction in the pleural cavity of rats involving a variety of inflammatory mediators, its action mechanism probably involving cellular injury and the exacerbated induction of cytokines and nitric oxide.

[Influence of sodium-restricted diet and food allergy in the treatment of epilepsy]. / Influência da dieta hipossódica e da alergia alimentar no tratamento da epilepsia.

An appreciation of the methods on epilepsy treatment has been done since the beginning of the century such as fasting, dehydrating diet and the restriction of sodium chloride. The author says that the latter may be of a beating out factor of epileptic crisis. He presents eight cases treated on restrictive diets on sodium chloride so that a progress is shown in the convulsive board of all patients. Surprising facts were the simultaneous progress on behavior in general, excitability and headaches, told by patients and their relatives. At last, the importance of alimentary allergy is emphasized as cause of epileptic crisis.

Influencia da dieta hipossodica e da alergia alimentar no tratamento da epilepsia. / Influence of sodium-restricted diet and food allergy in the treatment of epilepsy

E feita uma apreciacao dos metodos de tratamento da epilepsia no inicio do seculo como o jejum, a dieta desidratante e a restricao de cloreto de sodio. O autor se fixa neste ultimo, julgando que ele possa ser um fator desencadeante de crises epilepticas. Apresenta oito casos tratados com dietas restritas em cloreto de sodio, havendo melhora do quadro convulsivo em todos os pacientes. Fato surpreendente foi a melhora concomitante do comportamento em geral, da excitabilidade e de dores de cabeca, referidos pelos doentes e seus familiares. Por fim, e realcada a importancia da alergia alimentar como causa de crises epilepticas