Experimental sepsis induces sustained inflammation and acetylcholinesterase activity impairment in the hypothalamus.

Sepsis is one of the leading causes of mortality in intensive care units besides causing profound alterations in the brain. One of the structures notably affected during sepsis is the hypothalamus, resulting in important physiopathological consequences. Recently, we provided evidence that the presence of neuroinflammation, oxidative stress, and apoptosis in the hypothalamus of septic rats, is accompanied by impairment of arginine vasopressin (AVP) secretion. We had also demonstrated that sepsis survivor animals present attenuated AVP secretion after osmotic challenge, suggesting a persistent inflammation in the hypothalamus. However, the long-term course of inflammation in the hypothalamus remains unclear. Thus, we induced sepsis by cecal ligation and puncture (CLP) in Wistar rats and, five days after sepsis induction, the hypothalamus of each animal was collected for analysis. Nonmanipulated animals (naive) were used as controls. We found that CLP-induced morphological alterations in microglial cells are accompanied by an increase in Iba-1 immunoreactivity. Moreover, we observed enhanced expression of NF-κB and CREB transcription factors, which are well known to modulate the immune response. Additionally, we found that phosphorylation of GSK3α/ß (a kinase upstream to the CREB signaling pathway) was increased, as well as COX-2, iNOS, and IL-6 that are canonic inflammatory proteins. Thus, our results indicated the presence of sustained activation of resident glial cells that may result in neuroinflammation and cholinergic neurotransmission disruptions in the hypothalamus.

The impact of compression therapy with Unna's boot on the functional status of VLU patients.

OBJECTIVE: To assess disability in patients with venous leg ulcers treated with compression therapy with Unna's boot. METHOD: A descriptive analytic case control study was conducted from June 2010 to May 2011 in an outpatient wound care clinic in interior Brazil. Fifty patients of both sexes, aged 18 years or above, who had had a venous leg ulcer for more than 1 year and a Doppler ankle-brachial index of 0.8-1.0 were selected for the study. Patients were treated with wound dressings and Unna's boot. Disability was assessed using the 20-item Stanford Health Assessment Disability Scale (HAQ-20). Statistical analysis was performed using the Student's t-test, the Kruskal-Wallis test and the chi-square test of independence, all at a significance level of 0.05 (p < 0.05). RESULTS: The mean overall HAQ score at inclusion (baseline) was 2.98, indicating impaired functional capacity. After 8 and 12 months of compression treatment with Unna's boot, the mean overall HAQ scores were 1.35 and 1.0, respectively, indicating good functional capacity. CONCLUSION: Patients with venous leg ulcer reported severe difficulty or serious disability in their daily functioning at baseline; after 8 months of treatment with Unna's boot, these patients were able to perform activities of daily living.

Celecoxib treatment does not alter recruitment and activation of osteoclasts in the initial phase of experimental tooth movement.

In a previous study, we reported that the short-term treatment with celecoxib, a non-steroidal anti-inflammatory drug (NSAID) attenuates the activation of brain structures related to nociception and does not interfere with orthodontic incisor separation in rats. The conclusion was that celecoxib could possibly be prescribed for pain in orthodontic patients. However, we did not analyze the effects of this drug in periodontium. The aim of this follow-up study was to analyze effects of celecoxib treatment on recruitment and activation of osteoclasts and alveolar bone resorption after inserting an activated orthodontic appliance between the incisors in our rat model. Twenty rats (400-420 g) were pretreated through oral gavage with celecoxib (50 mg/kg) or vehicle (carboxymethylcellulose 0.4%). After 30 min, they received an activated (30 g) orthodontic appliance, set not to cause any palate disjunction. In sham animals, the appliance was immediately removed after introduction. All animals received ground food and, every 12 h, celecoxib or vehicle. After 48 h, they were anesthetized and transcardiacally perfused through the aorta with 4% formaldehyde. Subsequently, maxillae were removed, post-fixed and processed for histomorphometry or immunohistochemical analyses. As expected, incisor distalization induced an inflammatory response with certain histological changes, including an increase in the number of active osteoclasts at the compression side in group treated with vehicle (appliance: 32.2 ± 2.49 vs sham: 4.8 ± 1.79, P<0.05) and celecoxib (appliance: 31.0 ± 1.45 vs sham: 4.6 ± 1.82, P<0.05). The treatment with celecoxib did not modify substantially the histological alterations and the number of active osteoclasts after activation of orthodontic appliance. Moreover, we did not see any difference between the groups with respect to percentage of bone resorption area. Taken together with our previous results we conclude that short-term treatment with celecoxib can indeed be a therapeutic alternative for pain relieve during orthodontic procedures.

Central NO-cGMP pathway in thermoregulation and survival rate during polymicrobial sepsis.

Sepsis induces production of inflammatory mediators such as nitric oxide (NO) and causes physiological alterations, including changes in body temperature (Tb). We evaluated the involvement of the central NO-cGMP pathway in thermoregulation during sepsis induced by cecal ligation and puncture (CLP), and analyzed its effect on survival rate. Male Wistar rats with a Tb probe inserted in their abdomen were intracerebroventricularly injected with 1 microL NG-nitro-L-arginine methyl ester (L-NAME, 250 microg), a nonselective NO synthase (NOS) inhibitor; or aminoguanidine (250 microg), an inducible NOS inhibitor; or 1H-[1,2,4]oxadiazolo[4,3,-a]quinoxalin-1-one (ODQ, 0.25 microg), a guanylate cyclase inhibitor. Thirty minutes after injection, sepsis was induced by cecal ligation and puncture (CLP), or the rats were sham operated. The animals were divided into 2 groups for determination of Tb for 24 h and assessment of survival during 3 days. The drop in Tb seen in the CLP group was attenuated by pretreatment with the NOS inhibitors (p < 0.05) and blocked with ODQ. CLP rats pretreated with either of the inhibitors showed higher survival rates than vehicle injected groups (p < 0.05), and were even higher in the ODQ pretreated group. Our results showed that the effect of NOS inhibition on the hypothermic response to CLP is consistent with the role of nitrergic pathways in thermoregulation.

Effects of short-term acetaminophen and celecoxib treatment on orthodontic tooth movement and neuronal activation in rat.

Non-steroidal anti-inflammatory drugs (NSAIDs) have been used for pain relief in orthodontics, but clinical studies reported that they may reduce tooth movement (TM). By other side, TM seems to activate brain structures related to nociception, but the effects of NSAIDs in this activation have not been studied yet. We analyzed the effect of short-term treatment with acetaminophen or celecoxib in the separation of rat upper incisors, as well as in neuronal activation of the spinal trigeminal nucleus, following tooth movement. Thirty rats (400-420 g) were pretreated through oral gavage (1 ml/dose) with acetaminophen (200mg/kg), celecoxib (50mg/kg) or vehicle (carboxymethylcellulose 0.4%). After 30 min, they received an activated (30 g) orthodontic appliance for TM. In controls, this appliance was immediately removed after its introduction. Rats received ground food, and every 12h, one of the drugs or vehicle. After 48 h, they were anesthetized, maxilla was radiographed, and were perfused with 4% paraformaldehyde. Brains were further processed for Fos immunohistochemistry. TM induced incisor distalization (p<0.05) and neuronal activation of the spinal trigeminal nucleus. Treatment with both drugs did not affect tooth movement, but reduced c-fos expression in the caudalis subnucleus. No changes in c-fos expression were seen in the oralis and interpolaris subnuclei. We conclude that neither celecoxib nor acetaminophen seems to affect tooth movement, when used for 2 days, but both drugs are able to reduce the activation of brain structures related to nociception. Short-term treatment with celecoxib, thus, may be a therapeutic alternative to acetaminophen when the latter is contraindicated.

Rapid maxillary expansion causes neuronal activation in brain structures of rats.

A correlation between pain sensation and neuronal c-fos expression has been analyzed following experimental rapid maxillar expansion (RME). Adult male Wistar rats were anaesthetized and divided into three groups: animals that received an orthodontic apparatus, which was immediately removed after the insertion (control), animals that received an inactivated orthodontic apparatus (without force), and animals that received an orthodontic apparatus previously activated (140 g force). After 6, 24, 48, or 72 h, the animals were re-anaesthetized, and perfused with 4% paraformaldehyde. The brains were removed, fixed, and sections containing brain structures related to nociception were processed for Fos protein immunohistochemistry (IHC). The insertion of the orthodontic apparatus with 140 g was able to cause RME that could be seen by radiography. The IHC results showed that the number of activated neurons in the different nuclei changed according to the duration of appliance insertion and followed a temporal pattern similar to that of sensations described in clinics. The animals that received the orthodontic apparatus without force did not show RME but a smaller c-fos expression in the same brain structures. In conclusion, we demonstrate that orthodontic force used for palate disjunction activates brain structures that are related to nociception, and that this activation is related to the pain sensation described during orthodontic treatment.

Efficacy of intracoronal bleaching techniques with different light activation sources.

AIM: To evaluate ex vivo the efficacy of 35% hydrogen peroxide for intracoronal bleaching when activated by LEDs, halogen lamp or by the walking bleach technique. METHODOLOGY: Forty extracted human maxillary central incisors had their crowns resected 1 mm below the amelo-cemental junction and were submitted to artificial staining in centrifuged rat haemolysed blood. A 2-mm thick glass ionomer cervical plug was placed inside the canal, at the level of the amelo-cemental junction. Samples were divided randomly into five groups: group I received 35% hydrogen peroxide gel activated by LEDs. Group II received 35% hydrogen peroxide gel activated by a halogen lamp-based light curing unit. Group III received 35% hydrogen peroxide gel followed by the walking bleach technique. Group IV was neither artificially stained nor bleached (positive control) and group V was stained, but not bleached (negative control). The shade of the teeth was assessed visually by three independent and calibrated evaluators, before and after bleaching. The results were analysed using Kruskal-Wallis one-way analysis of variance and Dunn's post-test. RESULTS: No statistical differences regarding sample shades were found amongst groups for the tested internal bleaching techniques (P > 0.05). CONCLUSIONS: Hydrogen peroxide for intracoronal bleaching when activated either by LEDs, halogen lamp or by the walking bleach technique presented similar efficacy.

C-fos expression in rat brain nuclei following incisor tooth movement.

In the rat experimental model, molar tooth movement induced by Waldo's method is known to cause a temporally and spatially defined pattern of brain neuronal activation. Since orthodontic correction usually involves the entire dental arch, we used a spring-activated appliance to extend the investigation to incisors, and we included brain regions related to antinociception. Adjustment of the non-activated appliance on incisors resulted in c-fos expression in the dorsal raphe, peri-aqueductal gray matter, and the locus coeruleus, in addition to trigeminal sensory subnuclei and the parabrachial nucleus, where neuronal activation has already been detected in previous studies on molar tooth movement. Appliance activation with a 70-g force resulted in a further increase in Fos-immunoreactive neurons in the trigeminal sensory subnucleus caudalis and in the dorsal raphe. This result suggests that there is a recruitment of neurons related to nociception and to antinociception when tooth movement is increased.

Effect of cholinergic stimulation of the medial septal area on the secretory properties of atrial myocardial fibers

Adult male Wistar rats weighing 240-260 g were implanted with stainless steel guide cannulae into the medial septal area (MSA). Cholinergic stimulation of the MSA increased natriuresis (344.6 + or - 13.8 vs 22.2 + or - 2.1 micronEq for the controls), the number of atrial specific granules (61.0 + or - 6.7 vs 43,8 + or - 3.5 granules/100 micron m**2 sarcoplasma for the controls), and the number of electron-dense vesicles near the sarcolemma or appearing to undergo exocytotic extrusion (50.0 + or - 2.3 vs 21.4 + or - 5.7 vesicles/100 micronm sarcolemma for the controls) It is not yet clear how cholinergic stimulation of the MSA changes the secretory characteristics of atrial myocardial fibers. However, the present study provides evidence that release of an atrial natriuretic factor may be controlled by the central nervous system (CNS). This may occur through the sympathetic and parasympathetic innervation of the heart or through the release of some substance produced by the CNS or produced at another site whose release is controlled by the CNS