Morphologic evaluation and expression of matrix metalloproteinases-2 and 9 and nitric oxide during experimental periodontal disease in rat

The immunopathologic and inflammatory mechanisms involved in periodontal disease (PD) include the participation of host resident, inflammatory cells and chemical mediators. Metalloproteinases (MMPs) and nitric oxide (NO) play essential role in extracellular matrix turnover of periodontal tissue destruction. In this study, by means of RT-PCR through semi-quantitative densitometric scanning methods, the expression of MMPs -2 and -9 and inducible NO synthase (iNOS) was temporally and spatially investigated during the destructive mechanisms of experimentally induced PD in rats. Samples from different periods were microscopically analyzed and compared with the contralateral side (control). Our results showed significant expression of MMP-9 and iNOS in tissues affected by PD, as compared with controls, three days after PD induction, simultaneously with the beginning of alveolar bone loss. At 7 days post induction, only the MMP-9 mRNA presented a significantly higher expression, as compared with the respective controls. Thus, in the rat ligature-induced PD, MMP-9 and iNOS might importantly participate in the early stages of the disease, including inflammatory cell migration, tissue destruction and alveolar bone resorption. Also, we may suggest that the exuberant presence of PMNs may be related to the important expression of iNOS and MMP-9 found at 3 days post induction.

Down-regulation of expression of osteoblast and osteocyte markers in periodontal tissues associated with the spontaneous alveolar bone loss of interleukin-10 knockout mice

The aim of this study was to unravel the mechanisms by which interleukin (IL)-10, a potent pleiotropic cytokine, modulates alveolar bone homeostasis in C57BL/6 wild-type (WT) and IL-10 knockout (IL-10 KO) mice, evaluated at 8, 24, and 48 wk of age. Interleukin-10 KO mice presented significant alveolar bone loss when compared with WT mice, and this was not associated with changes in leukocyte counts or bacterial load. The levels of expression of messenger RNA (mRNA) for tumor necrosis factor-alpha (TNF-alpha), IL-1 beta, IL-6, transforming growth factor-beta (TGF-beta), receptor activator of nuclear factor kappa B ligand (RANKL), osteoprotegerin (OPG), and matrix metalloproteinase 13 (MMP13) were similar between both strains, whereas a significant decrease of tissue inhibitor of metalloproteinase 1 (TIMP1) mRNA expression was found at 48 wk in IL-10 KO mice. The osteoblast markers core binding factor alpha1 (CBFA1) and type I collagen (COL-I) were expressed at similar levels in both strains, whereas the levels of alkaline phosphatase (ALP) and osteocalcin (OCN), and those of the osteocyte markers phosphate-regulating gene endopeptidases (PHEX) and dentin matrix protein 1 (DMP1) were significantly lower in IL-10 KO mice. Our results demonstrate that the alveolar bone loss in the absence of IL-10 was associated with a reduced expression of osteoblast and osteocyte markers, an effect independent of microbial, inflammatory or bone-resorptive pathways.

PPAR-gamma agonist rosiglitazone prevents inflammatory periodontal bone loss by inhibiting osteoclastogenesis

Rosiglitazone (RGZ), an oral anti-hyperglycemic agent used for non-insulin-dependent diabetes mellitus, is a high-affinity synthetic agonist for peroxisome proliferator-activated receptor-gamma (PPAR-gamma). Both in vitro and in vivo experiments have also revealed that RGZ possesses anti-inflammatory properties. Therefore, in the present study, we investigated the anti-inflammatory effects of RGZ in a rat model of periodontal disease induced by ligature placed around the mandible first molars of each animal. Male Wister rats were divided into four groups: 1) animals without ligature placement receiving administration of empty vehicle (control); 2) animals with ligature receiving administration of empty vehicle; 3) animals with ligature receiving administration with oral RGZ (10 mg/kg/day); and 4) animals with ligature receiving administration of subcutaneous RGZ (10 mg/kg/day). Thirty days after induction of periodontal disease, the animals were sacrificed, and mandibles and gingival tissues were removed for further analysis. An in vitro assay was also employed to test the inhibitory effects of RGZ on osteoclastogenesis. Histomorphological and immunohistochemical analyses of periodontal tissue demonstrated that RGZ-treated animals presented decreased bone resorption, along with reduced RANKL expression, compared to those animals with ligature, but treated with empty vehicle. Corresponding to such results obtained from in vivo experiments, RGZ also suppressed in vitro osteoclast differentiation in the presence of RANKL in MOCP-5 osteoclast precursor cells, along with the down-regulation of the expression of RANKL-induced TRAP mRNA. These data indicated that RGZ may suppress the bone resorption by inhibiting RANKL-mediated osteoclastogenesis elicited during the course of experimental periodontitis in rats. (C) 2009 Elsevier B.V. All rights reserved.

CHANGES IN FETAL AND MATERNAL DOPPLER PARAMETERS OBSERVED DURING ACUTE SEVERE HYPERTENSION TREATMENT WITH HYDRALAZINE OR LABETALOL: A RANDOMIZED CONTROLLED TRIAL

We evaluated 16 pregnant women with gestational age between 20 and 32 weeks in acute severe hypertension which were randomly allocated to receive either hydralazine or labetalol. Blood pressure and Doppler ultrasound parameters from maternal uterine and fetal middle cerebral and umbilical arteries were assessed during acute severe hypertension and after treatment. A significant reduction in systolic and diastolic blood pressure was observed in both groups. A significant change in Doppler parameters was observed only in pregnant women who received hydralazine: an increase in uterine arteries resistance index. We concluded that both drugs were highly effective in reducing blood pressure in these women. Despite the observed increase in resistance index of uterine arteries associated with hydralazine, the use of hydralazine and labetalol were not related to any significant changes in fetal Doppler, which is reassuring about the safety of these drugs when treating acute severe hypertension in pregnancy. (E-mail: wpmartins@gmail.com) (C) 2011 World Federation for Ultrasound in Medicine & Biology.

Treatment with cannabidiol reverses oxidative stress parameters, cognitive impairment and mortality in rats submitted to sepsis by cecal ligation and puncture

Oxidative stress plays an important role in the development of cognitive impairment in sepsis. Here we assess the effects of acute and extended administration of cannabidiol (CBD) on oxidative stress parameters in peripheral organs and in the brain, cognitive impairment, and mortality in rats submitted to sepsis by cecal ligation and perforation (CLP). To this aim, male Wistar rats underwent either sham operation or CLP. Rats subjected to CLP were treated by intraperitoneal injection with ""basic support"" and CBD (at 2.5, 5, or 10 mg/kg once or daily for 9 days after CLP) or vehicle. Six hours after CLP (early times), the rats were killed and samples from lung, liver, kidney, heart, spleen, and brain (hippocampus, striatum, and cortex) were obtained and assayed for thiobarbituric acid reactive species (TBARS) formation and protein carbonyls. On the 10th day (late times), the rats were submitted to the inhibitory avoidance task. After the test, the animals were killed and samples from lung, liver, kidney, heart, spleen, and brain (hippocampus) were obtained and assayed for TBARS formation and protein carbonyls. The acute and extended administration of CBD at different doses reduced TBARS and carbonyl levels in some organs and had no effects in others, ameliorated cognitive impairment, and significantly reduced mortality in rats submitted to CLP. Our data provide the first experimental demonstration that CBD reduces the consequences of sepsis induced by CLP in rats, by decreasing oxidative stress in peripheral organs and in the brain, improving impaired cognitive function, and decreasing mortality. (C) 2010 Elsevier B.V. All rights reserved.

Effects of beta-carboline harmine on behavioral and physiological parameters observed in the chronic mild stress model: Further evidence of antidepressant properties

The chronic mild stress (CMS) model has been used as an animal model of depression which induces anhedonic behavior in rodents. The present study was aimed to evaluate the behavioral and physiological effects of administration of P-carboline harmine in rats exposed to CMS Procedure. To this aim, after 40 days of exposure to CMS procedure, rats were treated with harmine (15 mg/kg/day) for 7 days. In this study, sweet food consumption, adrenal gland weight, adrenocorticotrophin hormone (ACTH) levels, and hippocampal brain-derived-neurotrophic factor (BDNF) protein levels were assessed. Our findings demonstrated that chronic stressful situations induced anhedonia, hypertrophy of adrenal gland weight, increase ACTH circulating levels in rats and increase BDNF protein levels. Interestingly, treatment with harmine reversed anhedonia, the increase of adrenal gland weight, normalized ACTH circulating levels and BDNF protein levels. Finally, these findings further support the hypothesis that harmine could be a new pharmacological tool for the treatment of depression. (C) 2009 Elsevier Inc. All rights reserved.

Electromyographic standardized indices in healthy Brazilian young adults and data reproducibility

P>The determination of normal parameters is an important procedure in the evaluation of the stomatognathic system. We used the surface electromyography standardization protocol described by Ferrario et al. (J Oral Rehabil. 2000;27:33-40, 2006;33:341) to determine reference values of the electromyographic standardized indices for the assessment of muscular symmetry (left and right side, percentage overlapping coefficient, POC), potential lateral displacing components (unbalanced contractile activities of contralateral masseter and temporalis muscles, TC), relative activity (most prevalent pair of masticatory muscles, ATTIV) and total activity (integrated areas of the electromyographic potentials over time, IMPACT) in healthy Brazilian young adults, and the relevant data reproducibility. Electromyography of the right and left masseter and temporalis muscles was performed during maximum teeth clenching in 20 healthy subjects (10 women and 10 men, mean age 23 years, s.d. 3), free from periodontal problems, temporomandibular disorders, oro-facial myofunctional disorder, and with full permanent dentition (28 teeth at least). Data reproducibility was computed for 75% of the sample. The values obtained were POC Temporal (88 center dot 11 +/- 1 center dot 45%), POC masseter (87 center dot 11 +/- 1 center dot 60%), TC (8 center dot 79 +/- 1 center dot 20%), ATTIV (-0 center dot 33 +/- 9 center dot 65%) and IMPACT (110 center dot 40 +/- 23 center dot 69 mu V/mu V center dot s %). There were no statistical differences between test and retest values (P > 0 center dot 05). The Technical Errors of Measurement (TEM) for 50% of subjects assessed during the same session were 1 center dot 5, 1 center dot 39, 1 center dot 06, 3 center dot 83 and 10 center dot 04. For 25% of the subjects assessed after a 6-month interval, the TEM were 0 center dot 80, 1 center dot 03, 0 center dot 73, 12 center dot 70 and 19 center dot 10. For all indices, there was good reproducibility. These electromyographic indices could be used in the assessment of patients with stomatognathic dysfunction.

Evidence of the presence of T helper type 17 cells in chronic lesions of human periodontal disease

Periodontal disease is a chronic inflammation of the attachment structures of the teeth, triggered by potentially hazardous microorganisms and the consequent immune-inflammatory responses. In humans, the T helper type 17 (Th17) lineage, characterized by interleukin-17 (IL-17) production, develops under transforming growth factor-beta (TGF-beta), IL-1 beta, and IL-6 signaling, while its pool is maintained by IL-23. Although this subset of cells has been implicated in various autoimmune, inflammatory, and bone-destructive conditions, the exact role of T lymphocytes in chronic periodontitis is still controversial. Therefore, in this study we investigated the presence of Th17 cells in human periodontal disease. Gingival and alveolar bone samples from healthy patients and patients with chronic periodontitis were collected and used for the subsequent assays. The messenger RNA expression for the cytokines IL-17, TGF-beta, IL-1 beta, IL-6, and IL-23 in gingiva or IL-17 and receptor activator for nuclear factor-kappa B ligand in alveolar bone was evaluated by real-time polymerase chain reaction. The production of IL-17, TGF-beta, IL-1 beta, IL-6, and IL-23 proteins was evaluated by immunohistochemistry and the presence of Th17 cells in the inflamed gingiva was confirmed by immunofluorescence confocal microscopy for CD4 and IL-17 colocalization. Our data demonstrated elevated levels of IL-17, TGF-beta, IL-1 beta, IL-6, and IL-23 messenger RNA and protein in diseased tissues as well as the presence of Th17 cells in gingiva from patients with periodontitis. Moreover, IL-17 and the bone resorption factor RANKL were abundantly expressed in the alveolar bone of diseased patients, in contrast to low detection in controls. These results provided strong evidence for the presence of Th17 cells in the sites of chronic inflammation in human periodontal disease.

Histological and histomorphometrical alterations of the periodontal ligament in gerbils submitted to teeth extraction

This study verified the effect of unilateral teeth extraction on the periodontal ligament in gerbils (Meriones unguiculatus). Ten adult male gerbils weighing about 50 g had induced occlusal alterations by upper left molar extractions while the other ten animals, only submitted to surgical stress, were considered as controls. The periodontal ligament was characterized by qualitative and quantitative analysis, histological description and histomorphometric quantification. Significant alterations were observed on the left side of the experimental group (P < 0.05), the hypofunctional region, when it was compared with the contralateral side and the corresponding region of the control group. Two months after occlusal alterations induced by unilateral teeth extraction, atrophic histological alterations and a decrease in the periodontal space on the ipsilateral side characterized the periodontal ligament. In this study it was possible to conclude that the gerbil can be used in experimental models attempting to correlate the periodontium`s biological response to various mechanical stresses, as the periodontal ligament was shown to be highly sensitive to occlusal alterations.

Evaluation of ductus venosus and inferior vena cava by using multiple Doppler ultrasound parameters in healthy fetuses

The objective of this study was to describe multiple Doppler ultrasound parameters of ductus venosus and inferior vena cava in fetuses at gestational ages ranging from 22 to 38 weeks. In this prospective observational study, Doppler ultrasound exams were performed in 45 healthy fetuses at 22, 26, 34, and 38 weeks. Maximum venous velocity, minimum venous velocity, venous pulsatility index, and venous acceleration time (VAT) (defined as the time between minimum and maximum venous velocity) were evaluated at those gestational periods. Increase in the velocities and decrease in the pulsatility index were observed in these vessels throughout gestational age. The VAT increased with gestational age. The VAT, a Doppler parameter still not described elsewhere, increased during pregnancy could be related to a longer period of time needed to fill the atrium in bigger hearts. In this study, we provide reference values for VAT in healthy fetuses.

STANDARDIZATION OF SOME ELECTROCARDIOGRAPHIC PARAMETERS OF CAPTIVE LEOPARD CATS (LEOPARDUS TIGRINUS)

Thirty-three captive leopard cats, Leopardus tigrinus, were anesthetized with xylazine (1-2 mg/kg) and ketamine (10 mg/kg), and electrocardiograph (ECG) tests were recorded in all leads with 1 cm = 1 mV sensibility and 25 mm/sec speed repeating DII lead at 50 mm/sec speed with the same sensibility. Results expressed by mean and standard deviation were: heart rate (HR) = 107 +/- 17 (bpm); P-wave = 0.048 +/- 0.072 (s) x 0.128 +/- 0.048 (mV); PR interval = 0.101 +/- 0.081 (s); QRS compound = 0.053 +/- 0.012 (s) x 1.446 +/- 0.602 (mV); QT interval = 0.231 +/- 0.028 (s); R-wave (CV(6)LL) = 1.574 +/- 0.527 (mV); R-wave (CV(6)LU) = 1.583 +/- 0.818 (mV); heart rhythm: normal sinus rhythm (15.2%), sinus rhythm with wandering pacemaker (WPM) (60.6%), sinus arrhythmia with WPM (24.2%); electric axis: between +30 degrees and +60 degrees (6.1%), +60 (6.1%), between +60 degrees and +90 degrees (57.6%), +90 degrees (9%), between +90 degrees and +120 degrees (21.2%); ST segment: normal (75.7%), elevation (18.2%), depression (6.1%); T-wave polarity (DII): positive (100%); T-wave (V(10)): absent (6.1%), negative (63.6%), positive (18.2%), and with interference (12.1%). Through ECG data comparison with other species, unique features of Leopardus tigrinus` (leopard cat) ECG parameters were detected. Some of the study animals presented with an R-Wave amplitude that was indicative of left ventricle overload according to patterns for normal domestic cats (Felis cati). Echocardiographic exams revealed normal heart cavities` function and morphology. The aim of this study was to establish some electrocardiographic parameters of captive L. tigrinus.

Electrocardiographic parameters of captive lions (Panthera leo) and tigers (Panthera tigris) immobilized with ketamine plus xylazine

Twenty-seven healthy captive lions (Panthera leo) and 13 healthy captive tigers (Panthera tigris) from S to Paulo Zoo (Fundacao Parque Zoologico de Rio Paulo, Sao Paulo, Brazil) collection were selected for this study. They were anesthetized with ketamine (10 mg/kg) combined with xylazine (1-2 mg/kg) for physical examinations. hematologic and serum chemical analysis and electrocardiogram recording. The main aim of this research was to gather initial information about normal electrocardiographic parameters of large felids. Standard P-QRS-T deflections on leads described for domestic carnivores were analyzed, and they did not greatly differ from those of large felids. taking into account the greater weight and corporal mass of large felids. Heart rate of lions ranged frorn 42 to 76 beats per minute (bpm). Heart rate of tigers ranged from 56 to 97 bpm. In both species, the most common rhythm detected was normal sinus rhythm followed by sinus arrhythmia: wandering, pacemaker was also observed with normal sinus rhythm or sinus arrhythmia. Mean electrical axis lay between +60 degrees and +120 degrees. QRS complexes were predominantly positive in leads DI, DII, DIII, and AVF and negative in AVR and AVL. This Study provides insights into normal electrocardiograms of large felids. Wider investigations on the same subject arc necessary to establish criteria for the recognition of abnormalities in these species and should include other anesthetic drug(s) combinations and reports of electrocardiographic features of animals with cardiac disease and electrolytes disturbances.

Heat stress impairs performance parameters, induces intestinal injury, and decreases macrophage activity in broiler chickens

Studies on environmental consequences of stress on animal production have grown substantially in the last few years for economic and animal welfare reasons. Physiological, hormonal, and immunological deficits as well as increases in animals` susceptibility to diseases have been reported after different stressors in broiler chickens. The aim of the current experiment is to describe the effects of 2 different heat stressors (31 +/- 1 and 36 +/- 1 degrees C/10 h per d) applied to broiler chickens from d 35 to 42 of life on the corticosterone serum levels, performance parameters, intestinal histology, and peritoneal macrophage activity, correlating and discussing the obtained data under a neuroimmune perspective. In our study, we demonstrated that heat stress (31 +/- 1 and 36 +/- 1 degrees C) increased the corticosterone serum levels and decreased BW gain and food intake. Only chickens submitted to 36 +/- 1 degrees C, however, presented a decrease in feed conversion and increased mortality. We also showed a decrease of bursa of Fabricius (31 +/- 1 and 36 +/- 1 degrees C), thymus (36 +/- 1 degrees C), and spleen (36 +/- 1 degrees C) relative weights and of macrophage basal (31 +/- 1 and 36 +/- 1 degrees C) and Staphylococcus aureus-induced oxidative burst (31 +/- 1 degrees C). Finally, mild multifocal acute enteritis characterized by an increased presence of lymphocytes and plasmocytes within the jejunum`s lamina propria was also observed. The stress-induced hypothalamic-pituitary-adrenal axis activation was taken as responsible for the negative effects observed on the chickens` performance and immune function and also the changes of the intestinal mucosa. The present obtained data corroborate with others in the field of neuroimmunomodulation and open new avenues for the improvement of broiler chicken welfare and production performance.

Laser phototherapy effect on protein metabolism parameters of rat salivary glands

The aim of this study was to evaluate the effects of infrared diode laser phototherapy (LP) on tissues of the submandibular gland (SMG) and parotid gland (PG). Wistar rats were randomly divided into experimental (A and B) and control (C) groups. A diode laser, 808 nm wavelength, in continuous wave mode, was applied to the PG, SMG and sublingual gland in the experimental groups on two consecutive days. The doses were 4 J/cm(2) and 8 J/cm(2), and total energy was 7 J and 14 J, respectively. The power output (500 mW) and power density (277 mW/cm(2)) were the same for both experimental groups. In order to visualize the area irradiated by the infrared laser, we used a red pilot beam (650 nm) with 3 mW maximum power for the experimental groups. For the control group, the red pilot beam was the only device used. The SMG and PG were removed after 1 week of the first irradiation. Total protein concentration, amylase, peroxidase, catalase and lactate dehydrogenase assays were performed, as well as histological analysis. Statistical tests revealed significant increase in the total protein concentration for groups A and B in the parotid glands (P < 0.05). Based on the results of this study, LP altered the total protein concentration in rats` parotid glands.

CO(2) Laser (10.6 mu m) Parameters for Caries Prevention in Dental Enamel

Although CO(2) laser irradiation can decrease enamel demineralisation, it has still not been clarified which laser wavelength and which irradiation conditions represent the optimum parameters for application as preventive treatment. The aim of the present explorative study was to find low-fluence CO(2) laser (lambda = 10.6 mu m) parameters resulting in a maximum caries-preventive effect with the least thermal damage. Different laser parameters were systematically evaluated in 3 steps. In the first experiment, 5 fluences of 0.1, 0.3, 0.4, 0.5 and 0.6 J/cm(2), combined with high repetition rates and 10 mu s pulse duration, were chosen for the experiments. In a second experiment, the influence of different pulse durations (5, 10, 20, 30 and 50 mu s) on the demineralisation of dental enamel was assessed. Finally, 3 different irradiation times (2, 5 and 9 s) were tested in a third experiment. In total, 276 bovine enamel blocks were used for the experiments. An 8-day pH-cycling regime was performed after the laser treatment. Demineralisation was assessed by lesion depth measurements with a polarised light microscope, and morphological changes were assessed with a scanning electron microscope. Irradiation with 0.3 J/cm(2), 5 mu s, 226 Hz for 9 s (2,036 overlapping pulses) increased caries resistance by up to 81% compared to the control and was even significantly better than fluoride application (25%, p < 0.0001). Scanning electron microscopy examination did not reveal any obvious damage caused by the laser irradiation. Copyright (C) 2009 S. Karger AG, Basel