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AIM To investigate the presence, localization and the possible correlation of the fibroblast growth factor receptor-2 (FGFR2) with inflammatory resorption of cementum, periodontal ligament and alveolar bone during development of apical periodontitis in mice. METHODOLOGY Apical periodontitis was experimentally induced in mandibular first molars of mice by pulp exposure to the oral environment. Healthy teeth without pulp exposure were used as controls. At 7, 21 and 42 days following pulp exposure, the animals were euthanized and the jaws were prepared for analysis under conventional and fluorescence microscopy, immunohistochemistry (FGFR2), RT-PCR (RNAm levels of RANK, RANKL, OPG, Runx2 and cathepsin K) and enzyme histochemistry (cementoclasts and osteoclasts). Statistical analysis was performed by Kruskal-Wallis tests and Dunn's post hoc tests for multiple comparisons (α = 0.05) using SAS 9.4 software. RESULTS FGFR2-positive cells were not observed in the tissues surrounding healthy teeth but were observed in teeth with periapical lesions from seven days after root canal contamination. At days 21 and 42 after endodontic infection, the increase in periapical lesion size was accompanied by significantly enhanced expression of FGFR2 (P

Background Tooth eruption is a process that is not fully understood. Aim To evaluate whether genetic polymorphisms for RANK/RANKL/OPG are associated with delayed tooth emergence. To evaluate whether the relative expression of this genes is associated with persistent primary teeth. Design To evaluate whether genetic polymorphisms for RANK/RANKL/OPG could be involved in delayed tooth emergence, saliva samples from 160 children, aged 6-13 years old, were analysed. To test if there is correlation between gene expression of RANK/RANKL/OPG in children with delayed tooth emergence and persistent primary teeth, periapical tissue from 15 children with persistent primary teeth and from 15 control subjects were collected for qPCR analysis. Results Fifty-six children with delayed tooth emergence (35%) had at least one permanent tooth with delayed emergence. The T allele in RANKL (rs9594738) increased the risk of delayed tooth emergence (P = 0.02; OR = 1.71, 95%CI 1.09-2.75). The relative gene expression for RANKL and the ratio RANKL/OPG in children with delayed tooth emergence and persistent primary teeth were lower compared to controls (P = 0.02 and P = 0.005, respectively). Conclusions Data suggest that the polymorphism rs9594738 in RANKL is associated with delayed permanent tooth emergence. Moreover, reduced relative gene expression of RANKL in periapical tissue is associated with persistent primary teeth.

Objective To verify the association between 25(OH)D level and polymorphisms in the vitamin D receptor gene (VDR) with the disturbance in the dental development and eruption. Design A total of 183 children from two datasets were evaluated. The first dataset was a case-control (15:15) designed to assess if persistent primary tooth (PPT) is associate with serum 25(OH)D level and with genetic polymorphisms in VDR. The second dataset of genomic DNA samples from 54 children with delayed tooth eruption (DTE) and 99 controls were analysed to verify if genetic polymorphisms in VDR (rs2228570 and rs739837) are associated with DTE. The 25(OH)D and the genotyping/allele distribution were analysed using the T-test and chi-square test, respectively. Results The level of 25(OH)D in the PPT group (24.9 ± 6.4 mg/mL) was significantly lower than the control (30.0 ± 7.0 mg/mL) (p=.047). Our data show that children with 25(OH)D deficiency are more likely to present PPT (OR = 2.36; 95%CI: 1.51, 3.70). The rs739837 and rs2228570 polymorphisms were not associated with DTE (OR = 1.44; 95%CI: 0.87, 2.39 and OR = 0.80; 95%CI: 0.45, 1.44, respectively). Conclusions Vitamin D deficiency is a risk factor for PPT.

Objectives Neutrophil extracellular traps (NETs) play a role in the pathogenesis of periodontitis and rheumatoid arthritis (RA). However, it remains poorly understood whether NETs participate in the cross-talk between periodontitis and RA. Herein, we investigated the production of NETs in individuals with periodontitis and RA and its association with clinical parameters. The impact of periodontal therapy on RA and NET release was also assessed. Methods The concentration of NETs and cytokines was determined in the saliva and plasma of individuals with early RA (n = 24), established RA (n = 64) and individuals without RA (n = 76). The influence of periodontitis on the production of NETs and cytokines was also evaluated. Results Individuals with early RA had a higher concentration of NETs in saliva and plasma than individuals with established RA or without RA. Periodontitis resulted in an increase in the concentration of NETs of groups of individuals without RA and with early RA. The proportion of individuals with high concentrations of IL-6, IL-10 and GM-CSF was higher among individuals with periodontitis than among individuals without periodontitis. The concentrations of TNF-α, IL-6, IL-17/IL-25 and IL-28A were particularly high in individuals with early RA. Worse periodontal clinical parameters, RA onset and RA activity were significantly associated with circulating NETs. Periodontal therapy was associated with a reduction in the concentration of NETs and inflammatory cytokines and amelioration in periodontitis and RA. Conclusion This study reveals that NETs are a possible link between periodontitis and RA, with periodontal therapy resulting in a dramatic switch in circulating NET levels.

Rheumatoid arthritis (RA) is a chronic inflammatory disease characterized by joint destruction and severe morbidity. Cigarette smoking (CS) can exacerbate the incidence and severity of RA. Although Th17 cells and the Aryl hydrocarbon receptor (AhR) have been implicated, the mechanism by which CS induces RA development remains unclear. Here, using transcriptomic analysis, we show that microRNA-132 is specifically induced in Th17 cells in the presence of either AhR agonist or CS-enriched medium. miRNA-132 thus induced is packaged into extracellular vesicles produced by Th17 and acts as a proinflammatory mediator increasing osteoclastogenesis through the down-regulation of COX2. In vivo, articular knockdown of miR-132 in murine arthritis models reduces the number of osteoclasts in the joints. Clinically, RA patients express higher levels of miR-132 than do healthy individuals. This increase is further elevated by cigarette smoking. Together, these results reveal a hitherto unrecognized mechanism by which CS could exacerbate RA and further advance understanding of the impact of environmental factors on the pathogenesis of chronic inflammatory diseases.

Objective To evaluate the role of neutrophil extracellular traps (NETs) in the genesis of joint hyperalgesia using an experimental model of arthritis and transpose the findings to clinical investigation. Methods C57BL/6 mice were subjected to antigen-induced arthritis (AIA) and treated with Pulmozyme (PLZ) to degrade NETs or Cl-amidine to inhibit NET production. Oedema formation, the histopathological score and mechanical hyperalgesia were evaluated. NETs were injected intra-articularly in wild type (WT), Tlr4−/−, Tlr9−/−, Tnfr1−/− and Il1r−/− mice, and the levels of cytokines and Cox2 expression were quantified. NETs were also quantified from human neutrophils isolated from RA patients and individual controls. Results AIA mice had increased NET concentration in joints, accompanied by increased Padi4 gene expression in the joint cells. Treatment of AIA mice with a peptidyl arginine deiminase 4 inhibitor or with PLZ inhibited the joint hyperalgesia. Moreover, the injection of NETs into joints of naïve animals generated a dose-dependent reduction of mechanical threshold, an increase of articular oedema, inflammatory cytokine production and cyclooxygenase-2 expression. In mice deficient for Tnfr1, Il1r, Tlr4 and Tlr9, joint hyperalgesia induced by NETs was prevented. Last, we found that neutrophils from RA patients were more likely to release NETs, and the increase in synovial fluid NET concentration correlated with an increase in joint pain. Conclusion The findings indicate that NETs cause hyperalgesia possibly through Toll-like receptor (TLR)-4 and TLR-9. These data support the idea that NETs contribute to articular pain, and this pathway can be an alternative target for the treatment of pain in RA.

The present study examined the hemodynamics [arterial pressure (AP), AP variability (APV), heart rate (HR), and heart rate variability (HRV)], cardiac function (echocardiographycally), and myocardial inflammation in Balb/c mice submitted to Periodontitis, through the ligation of the left first molar, or Sham surgical procedure. The first protocol indicated that the AP was similar (136 ± 2 vs. 132 ± 3 mmHg in Sham), while the HR was higher in mice with Periodontitis (475 ± 20 vs. 412 ± 18 bpm in Sham), compared to their Sham counterparts. The APV was higher in mice with Periodontitis when evaluated in the time domain (4.5 ± 0.3 vs. 3.4 ± 0.2 mmHg in Sham), frequency domain (power of the LF band of systolic AP), or through symbolic analysis (patterns 0V + 1V), indicating a sympathetic overactivity. The HRV was similar in the mice with Periodontitis, as compared to their Sham counterparts. In the second protocol, the mice with Periodontitis showed decreased cardiac output (10 ± 0.8 vs. 15 ± 1.4 mL/min in Sham) and ejection fraction (37 ± 3 vs. 47 ± 2% in Sham) associated with increased myocardial cytokines (Interleukin-17, Interleukin-6, and Interleukin-4). This study shows that experimental Periodontitis caused cardiac dysfunction, increased heart cytokines, and sympathetic overactivity, in line with epidemiological studies indicating an increased risk of cardiovascular events in clinical Periodontitis.

To evaluate the effect and mechanisms of naringenin in TiO2-induced chronic arthritis in mice, a model resembling prosthesis and implant inflammation. Flavonoids are antioxidant and anti-inflammatory molecules with important anti-inflammatory effect. Mice were daily treated with the flavonoid naringenin (16.7–150 mg/kg, orally) for 30 days starting 24 h after intra-articular knee injection of 3 mg of TiO2. TiO2-induced arthritis resembles cases of aseptic inflammation induced by prosthesis and/or implants. Mice were stimulated with 3 mg of TiO2 and after 24 h mice started to be treated with naringenin. The disease phenotype, treatment toxicity, histopathological damage, oxidative stress, cytokine expression and NFκB were evaluated after 30 days of treatment. Naringenin inhibited TiO2-induced mechanical hyperalgesia (96%), edema (77%) and leukocyte recruitment (74%) without inducing toxicity. Naringenin inhibited histopathological index (HE, 49%), cartilage damage (Toluidine blue tibial staining 49%, and proteoglycan 98%), and bone resorption (TRAP-stained 73%). These effects were accompanied by inhibition of oxidative stress (gp91phox 93%, NBT 83%, and TBARS 41%) cytokine mRNA expression (IL-33 82%, TNFα 76%, pro-IL-1β 100%, and IL-6 61%), and NFκB activation (100%). Naringenin ameliorates TiO2-induced chronic arthritis inducing analgesic and anti-inflammatory responses with improvement in the histopathological index, cartilage damage, and bone resorption.

Anthropomorphic measures among type 1 diabetic patients are changing as the obesity epidemic continues. Excess fat mass may impact bone density and ultimately fracture risk. We studied the interaction between bone and adipose tissue in type 1 diabetes subjects submitted to two different clinical managements: (I) conventional insulin therapy or (II) autologous nonmyeloablative hematopoietic stem-cell transplantation (AHST). The study comprised 3 groups matched by age, gender, height and weight: control (C = 24), type 1 diabetes (T1D = 23) and type 1 diabetes treated with AHST (T1D-AHST = 9). Bone mineral density (BMD) and trabecular bone score (TBS) were assessed by dual X-ray absorptiometry (DXA). 1 H Magnetic resonance spectroscopy was used to assess bone marrow adipose tissue (BMAT) in the L3 vertebra, and abdominal magnetic resonance imaging was used to assess intrahepatic lipids (IHL), visceral (VAT) and subcutaneous adipose tissue (SAT). Individuals conventionally treated for T1D were more likely to be overweight (C = 23.8 ± 3.7; T1D=25.3 ± 3.4; T1D-AHST = 22.5 ± 2.2 Kg/m 2 ; p > 0.05), but there was no excessive lipid accumulation in VAT or liver. Areal BMD of the three groups were similar at all sites; lumbar spine TBS (L3) was lower in type 1 diabetes ( p

Objectives: Miniscrew has been frequently used, considering that anchorage control is a critical point in orthodontic treatment, and its failure, the main adverse problem. Using two groups of stable (successful) and unstable (failed) mini-implants, this in vivo study aimed to quantify proinflammatory cytokines IL-1 α, IL-6, IL-17, and TNF-α and osteoclastogenesis marker RANK, RANKL, and OPG in gingival tissue, using the real-time polymerase chain reaction technique. Methodology: Thirteen patients of both sexes (11-49 years old) under orthodontic treatment were selected, obtaining 11 successful and 7 failed mini-implants. The mini-implants were placed and removed by the same surgeon, in both jaws. The mean time of permanence in the mouth was 29.4 months for successful and 7.6 months for failed mini-implants. At removal time, peri-mini-implant gingival tissue samples were collected and processed for quantification of the proinflammatory cytokines and osteoclastogenesis markers. Nonparametric Wilcoxon rank-sum test considering the clusters and Kruskal-Wallis test were used for statistical analysis (α=0.05). Results: No significant difference (p>0.05) was observed between the groups for either quantification of cytokines or osteoclastogenesis markers, except for IL-6 (p

Titanium dioxide (TiO2) is a common component of orthopedic prosthesis. However, prosthesis wear releases TiO2, which induces inflammation and osteolysis in peri-prosthetic tissues. Quercetin is a flavonoid widely present in human diet, which presents biological activities such as antinociceptive, anti-inflammatory and antioxidant effects. Therefore, the effect of intraperitoneal treatment with quercetin in TiO2-induced arthritis model was evaluated. In the first set of experiments, mice received injection of TiO2 (0.1-3 mg/knee joint) and articular mechanical hyperalgesia, edema and histopathology analysis were performed in a 30 days protocol. The dose of 3 mg of TiO2 showed the most harmful effect, and was chosen to the following experiments. Subsequently, mice received 3 mg of TiO2 followed by post-treatment with quercetin during 30 days. Quercetin (10-100 mg/kg) inhibited in a dose-dependent manner TiO2-induced knee joint mechanical hyperalgesia, edema and leukocyte recruitment and did not induce damage in major organs such as liver, kidney and stomach. The dose of 30 mg/kg was chosen for the subsequent analysis, and reduced histopathological changes such as leukocyte infiltration, vascular proliferation and synovial hyperplasia (pannus formation) on day 30 after TiO2 challenge. The protective analgesic and anti-inflammatory mechanisms of quercetin included the inhibition of TiO2-induced neutrophil and macrophage recruitment, proteoglycan degradation, oxidative stress, cytokine production (TNF-α, IL-1β, IL-6, and IL-10), COX-2 mRNA expression, and bone resorption as well as activation of Nrf2/HO-1 signaling pathway. These results demonstrate the potential therapeutic applicability of the dietary flavonoid quercetin to reduce pain and inflammatory damages associated with prosthesis wear process-induced arthritis.

OBJECTIVES: The present study was designed to evaluate the bone phenotypes and mechanisms involved in bone disorders associated with hepatic osteodystrophy. Hepatocellular disease was induced by carbon tetrachloride (CCl4). In addition, the effects of disodium pamidronate on bone tissue were evaluated. METHODS: The study included 4 groups of 15 mice: a) C = mice subjected to vehicle injections; b) C+P = mice subjected to vehicle and pamidronate injections; c) CCl4+V = mice subjected to CCl4 and vehicle injections; and d) CCl4+P = mice subjected to CCl4 and pamidronate injections. CCl4 or vehicle was administered for 8 weeks, while pamidronate or vehicle was injected at the end of the fourth week. Bone histomorphometry and biomechanical analysis were performed in tibiae, while femora were used for micro-computed tomography and gene expression. RESULTS: CCl4 mice exhibited decreased bone volume/trabecular volume and trabecular numbers, as well as increased trabecular separation, as determined by bone histomorphometry and micro-computed tomography, but these changes were not detected in the group treated with pamidronate. CCl4 mice showed increased numbers of osteoclasts and resorption surface. High serum levels of receptor activator of nuclear factor-κB ligand and the increased expression of tartrate-resistant acid phosphatase in the bones of CCl4 mice supported the enhancement of bone resorption in these mice. CONCLUSION: Taken together, these results suggest that bone resorption is the main mechanism of bone loss in chronic hepatocellular disease in mice.

Interleukin (IL)-33, a member of the IL-1 cytokine family, is an important modulator of the immune system associated with several immune-mediated disorders. High levels of IL-33 are expressed by the central nervous system (CNS) suggesting a potential role of IL-33 in autoimmune CNS diseases. We have investigated the expression and function of IL-33 in the development of experimental autoimmune encephalomyelitis (EAE) in mice. We report here that IL-33 and its receptor ST2 (IL-33Rα) are highly expressed in spinal cord tissue, and ST2 expression is markedly increased in the spinal cords of mice with EAE. Furthermore, ST2-deficient (ST2(-/-) ) mice developed exacerbated EAE compared with wild-type (WT) mice while WT, but not ST2(-/-) EAE mice treated with IL-33 developed significantly attenuated disease. IL-33-treated mice had reduced levels of IL-17 and IFN-γ but produced increased amounts of IL-5 and IL-13. Lymph node and splenic macrophages of IL-33-treated mice showed polarization toward an alternatively activated macrophage (M2) phenotype with significantly increased frequency of MR(+) PD-L2(+) cells. Importantly, adoptive transfer of these IL-33-treated macrophages attenuated EAE development. Our data therefore demonstrate that IL-33 plays a therapeutic role in autoimmune CNS disease by switching a predominantly pathogenic Th17/Th1 response to Th2 activity, and by polarization of anti-inflammatory M2 macrophages.

To investigate the role of regulatory T cells in the modulation of long-term immune dysfunction during experimental sepsis. It is well established that sepsis predisposes to development of a pronounced immunosuppression. Nevertheless, the mechanisms underlying the immune dysfunction after sepsis are still not well understood.Prospective experimental study.University research laboratory.Wild-type mice underwent cecal ligation and puncture and were treated with antibiotic during 3 days after surgery. On days 1, 7, or 15 after cecal ligation and puncture, the frequency of regulatory T cells, proliferation of CD4 T cells and bacterial counts were evaluated. Fifteen days after cecal ligation and puncture, surviving mice underwent secondary pulmonary infection by intranasal inoculation of nonlethal dose of Legionella pneumophila. Some mice received agonistic glucocorticoid-induced tumor necrosis factor receptor antibody (DTA-1) before induction of secondary infection.Mice surviving cecal ligation and puncture showed a markedly increased frequency of regulatory T cells in thymus and spleen, which was associated with reduced proliferation of CD4 T cells. Fifteen days after cecal ligation and puncture, all sepsis-surviving mice succumbed to nonlethal injection of L. pneumophila. Treatment of mice with DTA-1 antibody reduced frequency of regulatory T cells, restored CD4 T cell proliferation, reduced the levels of bacteria in spleen, and markedly improved survival of L. pneumophila infection.These findings suggest that regulatory T cells play an important role in the progression and establishment of immune dysfunction observed in experimental sepsis.

Objectives Interleukin 33 (IL-33) is a new member of the IL-1 family of cytokines which signals via its receptor, ST2 (IL-33R), and has an important role in Th2 and mast cell responses. This study shows that IL-33 orchestrates neutrophil migration in arthritis. Methods and results Methylated bovine serum albumin (mBSA) challenge in the knee joint of mBSA-immunised mice induced local neutrophil migration accompanied by increased IL-33R and IL-33 mRNA expression. Cell migration was inhibited by systemic and local treatments with soluble (s)IL-33R, an IL-33 decoy receptor, and was not evident in IL-33R-deficient mice. IL-33 injection also induced IL-33R-dependent neutrophil migration. Antigen- and IL-33-induced neutrophil migration in the joint was dependent on CXCL1, CCL3, tumour necrosis factor α (TNFα) and IL-1β synthesis. Synovial tissue, macrophages and activated neutrophils expressed IL-33R. IL-33 induces neutrophil migration by activating macrophages to produce chemokines and cytokines and by directly acting on neutrophils. Importantly, neutrophils from patients with rheumatoid arthritis successfully treated with anti-TNFα antibody (infliximab) expressed significantly lower levels of IL-33R than patients treated with methotrexate alone. Only neutrophils from patients treated with methotrexate alone or from normal donors stimulated with TNFα responded to IL-33 in chemotaxis. Conclusions These results suggest that suppression of IL-33R expression in neutrophils, preventing IL-33-induced neutrophil migration, may be an important mechanism of anti-TNFα therapy of inflammation.

Homocysteine is an independent risk factor for coronary heart disease, as well as for cerebrovascular and peripheral vascular diseases. The purpose of this study was to investigate the effects of hyperhomocysteinemia (HHcy) on vascular reactivity within carotid artery segments isolated from ovariectomized female rats. Treatment with DL-Hcy thiolactone (1 g/kg body weight per day) reduced the phenylephrine-induced contraction of denuded rings. However, the treatment did not alter KCl-induced contractions, or relaxations induced by sodium nitroprusside or acetylcholine. We report elevated expressions of iNOS, eNOS, and nitrotyrosine in homocysteine-treated rat artery sections. Moreover, the inhibition of NOS by L-NAME, 1,400 W, or L-NNA restored phenylephrine-induced vasoconstriction in carotid artery segments from Hcy-treated rats. In conclusion, our findings show that severe HHCy can promote an acute decrease in the endothelium-independent contractile responses of carotid arteries to adrenergic agonists. This effect was restored by nitric oxide synthase inhibitors, which further supports the involvement of nitric oxide in HHcy-derived vascular dysfunction.

Cytokines mediate the balance between protective and destructive immunity in periodontitis. We sought to investigate the role of IL-33 in periodontitis. The expression of IL-33 in gingival tissue from healthy controls ( n = 10) and patients with chronic periodontitis ( n = 17) was investigated. Based on a murine model of periodontal disease, the function of IL-33 was determined first by administration of exogenous IL-33 and second by inhibition of IL-33 signaling using mice deficient in the IL-33 receptor ST2. Alveolar bone level, serum antibody, and lymphocyte responses were assessed in the murine model. Expression of IL-33 and ST2 was elevated in gingival tissues from patients with chronic periodontitis as compared with healthy tissues ( P < 0.05). Similarly, Il33 expression was higher in periodontal tissues of Porphyromonas gingivalis–infected mice as compared with sham-infected controls ( P < 0.05). IL-33 treatment of P. gingivalis–infected mice significantly exacerbated alveolar bone loss when compared with infection or IL-33 treatment alone ( P < 0.001). Conversely, P. gingivalis infection–induced alveolar bone loss was attenuated in mice lacking ST2. The percentages of T and B lymphocytes expressing nuclear factor κB ligand (RANKL) in the gingival tissues and T lymphocytes expressing RANKL in the cervical draining lymph nodes were higher in IL-33-treated P. gingivalis–infected mice versus phosphate buffered saline–treated P. gingivalis–infected controls (all P < 0.001). Targeting the RANKL pathway by osteoprotegerin administration abrogated periodontal bone destruction in P. gingivalis–infected, IL-33-treated mice. These data demonstrate a previously unrecognized role for IL-33 in exacerbating bone loss in a RANKL-dependent manner in the context of bacterial infection and suggest that this pathway may be amenable to manipulation as a novel therapeutic target in periodontitis.

Background and Objective The angiotensin type 1 (AT1) receptor has been implicated in the pathogenesis of inflammatory bone disorders. This study aimed to investigate the effect of an AT1 receptor antagonist in infection-induced and arthritis-associated alveolar bone loss in mice. Material and Methods Mice were subjected to Aggregatibacter actinomycetemcomitans oral infection or antigen-induced arthritis and treated daily with 10 mg/kg of the prototype AT1 antagonist, losartan. Treatment was conducted for 30 d in the infectious condition and for 17 d and 11 d in the preventive or therapeutic regimens in the arthritic model, respectively. The mice were then killed, and the maxillae, serum and knee joints were collected for histomorphometric and immunoenzymatic assays. In vitro osteoclast assays were performed using RAW 264.7 cells stimulated with A. actinomycetemcomitans lipopolysacharide (LPS). Results Arthritis and A. actinomycetemcomitans infection triggered significant alveolar bone loss in mice and increased the levels of myeloperoxidase and of TRAP+ osteoclasts in periodontal tissues. Losartan abolished such a phenotype, as well as the arthritis joint inflammation. Both arthritis and A. actinomycetemcomitans conditions were associated with the release of tumor necrosis factor alpha (TNF-α), interferon-gamma, interleukin-17 and chemokine (C-X-C motif) ligand 1 and an increased RANKL/osteoprotegerin ratio in periodontal tissues, but such expression decreased after losartan treatment, except for TNF-α. The therapeutic approach was as beneficial as the preventive one. In vitro, losartan prevented LPS-induced osteoclast differentiation and activity. Conclusion The blockade of AT1 receptor exerts anti-inflammatory and anti-osteoclastic effects, thus protecting periodontal tissues in distinct pathophysiological conditions of alveolar bone loss.

The rabbit carotid artery, injured by silicone collar, presents a perivascular inflammatory response and alterations in vascular responsiveness. Considering that angiotensin II (Ang II) plays an important role in cardiovascular physiology and pathology and that cardiovascular disease increases in postmenopausal women, the aim of this study was to investigate whether the Ang II contractile response in ovariectomized rat carotid artery is modified after a vascular injury by silicone collar. The positioning of the silicone collar around the common carotid artery for 14 days leads to an increased cross-sectional area of adventitial layer with inflammatory cells and an extensive angiogenesis. The Ang II-induced contraction was significantly decreased in collared arteries when compared with contralateral arteries. The reduction in the constrictor effect of Ang II in collared arteries was not modified by the presence of indomethacin (a non-selective inhibitor of cyclooxygenase) or PD 123,319 (a selective antagonist of the Ang II AT 2 receptor). Moreover, while endothelium removal induced an increase in the Ang II responsiveness of both arteries (collared and contralateral), the Emax induced by Ang II was still lower in collared arteries. However, the "in vitro" pretreatment of the arteries with an inhibitor of nitric oxide synthase enzyme (L-NAME) significantly enhanced the maximal contractions response to Ang II only in injured arteries. Furthermore, the expression of iNOS (inducible nitric oxide synthase) was observed in the adventitial layer of collared arteries, indicating that the NO formed in the adventitial layer has an important role in injured arteries. Moreover, our data show impairment of extracellular calcium mobilization, mediated by Ang II, in the collared artery, although the intracellular calcium mobilization was not modified by the injury. In conclusion, the increased production of NO and a decrease in the calcium influx displayed by Ang II in the collared artery appears to counteract and reduce the biologic effect of Ang II.

Background The first response to perivascular injury is observed in the adventitial layer. The purpose of this study was to determine the time course of vascular reactivity alterations after collar injury in rats. We also analyzed the relation between adventitial layer injury and vascular responsiveness to vasoconstrictor agents. Methods Wistar rats had a silicone collar positioned around the carotid artery. The ipsilateral and contralateral arteries were morphologically analyzed 4, 7, 14 and 28 days after injury, and cumulative concentration–response curves to phenylephrine (Phe), angiotensin II (Ang II) and KCl were obtained for arteries isolated from collared and sham-operated groups. Results Inflammatory cells and fibroblasts were observed in the adventitial layer of collared arteries 4, 7, 14 and 28 days after injury. Intimal thickening was observed in collared arteries only 14 and 28 days after perivascular injury. A decrease in maximum effect values (Emax) for Phe, Ang II and KCl was observed in the collared artery when compared with the contralateral artery at all times after injury, whereas an increase in vascular responsiveness was observed in the contralateral artery 4 days after injury. Conclusions The impairment of the contractile response preceded the intimal thickening. The compromise of vascular reactivity coincided with the presence of inflammatory cells and angiogenesis in the adventitial layer. The enhancement of the efficacy and potency of Ang II and Phe in collared-contralateral arteries 4 days after collar placement may be related to a receptor-mediated compensatory mechanism stimulated by the collar injury.

This paper presents a probability density function formula for predicting the polarization dependent loss (PDL) in an optical transmission system composed of passive devices and connecting fibers. A new calculation technique, which enables the probability density function formula to be obtained theoretically, is used instead of the most complicated part of the Muller-matrix or Jones-matrix calculations, which has been thought to be necessary for analyzing PDL. This technique involves calculation of the transmission coefficients of the transmission system and its devices from their PDLs. In the theoretical development, the central limit theorem is used as the sole approximation. A Monte Carlo numerical simulation was done to verify the validity of the analytical theory. Very good agreement between simulation and analytical theory is obtained when the number of devices having PDL is four or more. An experiment also demonstrated the validity of the analytical theory. The theory can also explain some phenomena that occur in systems composed of optical amplifiers, even though it had originally been developed to explain PDL-related phenomena in systems composed of passive devices only.

This paper proposes a new optical access network system; the multi-bit rate passive double star (multi-bit rate-PDS) system. The multi-bit rate-PDS system passes multi-bit rate data streams between the optical service unit (OSU) and the optical network unit (ONU) using the time-unit packet (TP). This simplifies the provision of user flexibility and provides very economical network services to users. We describe the architecture and frame structure of the multi-bit rate-PDS system, and elaborate the advantage of its user accommodation. Furthermore, we show how to decide the multiplexed bit rates in the multi-bit rate-PDS system and how to realize the receiver circuit for multi-bit rate data packet streams.

Objective: To report the results of prenatal triple marker screening on a population of Japanese pregnant women. Methods: From April 1994 through March 1999, a total of 32,925 native Japanese women with singleton pregnancies requested a triple marker-screening test. Multiples of the median values for 3 markers and individual risks for each patient were calculated following adjustment for the Japanese weight correction factor. The risk cut-off values used for Down syndrome (T21), open spina bifida (OSB) and trisomy 18 (T18) were 1:295, 1:290, and 1:100, respectively. Follow-up information was collected postpartum and statistically analyzed. Results: Detection rates (DR) of T21 for women less than 35 years, over 35 years and overall were 58, 94, and 83%, respectively. DR of T18 for women less than 35 years, over 35 years and overall were 75, 79, and 79%, respectively. DR of open neural tube defects (ONTD) was 100%. Conclusions: The first cumulative data of an intervention program and prospective follow-up studies in Japan have proven to be similar to other published reports. Individual risk values were calculated for each pregnancy for T21, T18 and ONTD. This screening program is more effective than age-dependent screening for detecting T21, T18 and ONTD pregnancies.

We monitored 30 laboratory hemostatic parameters in an attempt to better comprehend alterations in coagulation and fibrinolysis in 10 patients with hematological malignancies subjected to autologous peripheral blood stem cell transplantation (APBSCT). These parameters were assessed before and just after high-dose conditioning chemotherapy, on days 1, 7, 14 and 28. Although, clinical manifestations associated with fibrino-coagulation disorders never occurred, including veno-occlusive disease, a statistically significant increase was seen in 7 of 30 parameters, compared to values seen before conditioning chemotherapy. These were subdivided into early and late phase parameters. The early phase parameters, which increased during the first day after the conditioning chemotherapy was given, then returned to baseline values, included protein C, plasma tissue factor and tissue-plasminogen activator. The late phase parameters, which increased over baseline values during days 7 to 28, included free-protein S, fibrinogen, plasmin-alpha2-plasmin inhibitor complex and soluble-thrombomodulin. The increase of early phase parameters, as produced by the liver and by endothelial cells, may reflect tissue damage by conditioning chemotherapy. Late phase parameters increased in parallel with C-reactive protein, which suggests a correlation with the degree of inflammation, such as the presence of infective disease during neutropenia. These subclinical alterations in coagulation and fibrinolysis which take on a biphasic pattern during the course of APBSCT should be kept in mind by the attending physicians during therapy.

The oral cavity is a complex environment that may harbor more than 750 bacterial species. Proper oral hygiene is essential to maintain the equilibrium of microbial community and oral health. The ecological balance can be compromised in inadequate microbial control situations and an oral infection can be evoked. The bacteria can aid in the formation of dental plaque and caries, leading to periodontal disease (PD) and periapical lesion (PL). PD is the most common chronic inflammatory disorder of microbial origin that affects toothsupporting tissues including the periodontal ligament and the alveolar bone. Dental caries is characterized by demineralization of enamel and dentine produced by microorganisms’ acids. This process can cause pulp necrosis and root canal infection and the progression through the root apex can induce PL. PD and PLs constitute inflammatory and immune response against oral pathogens. Both processes encompass pathogenic mechanisms of inflammation-mediated soft tissue destruction and bone resorption. The etiopathogenesis of these diseases have been extensively investigated over the last decades and the role of several cell types, cytokines and pathways has been described (Graves, 2008, Graves et al., 2011a, Nair, 1997). Last decades research have documented the importance and commitment of immune system to protect the host from pathogen and also the paradoxical effect accounting for the bone resorption observed in these diseases. More recently, the pattern of immune cell response involved in the lesions progression (i.e. Th1, Th2, Th17, Th9 or T regulatory) has received particular attention (Cardoso et al., 2009, Colic et al., 2009a, Gaffen & Hajishengallis, 2008, Ohlrich et al., 2009, Queiroz-Junior et al, 2011). Although chemokines and cytokines are pivotal to determine these Th patterns, not much is known regarding the expression of these markers in the regulation of bone resorption in sites of PD and PL. This

Introduction Periapical lesions are chronic inflammatory disorders of periradicular tissues caused by etiologic agents of endodontic origin. The inflammatory chemokines are thought to be involved in the latter observed osteolysis. With a murine model of experimental periapical lesion, the objective of this study was to evaluate the role of the chemokine receptor CCR2 in the lesion progression, osteoclast differentiation and activation, and expression of inflammatory osteolysis-related mediators. Methods For lesion induction, right mandibular first molars were opened surgically with a ¼ carbine bur, and 4 bacterial strains were inoculated in the exposed dental pulp; left mandibular first molars were used as controls. Animals were killed at 3, 7, 14, and 21 days after surgeries to evaluate the kinetics of lesion development. Results CCR2 KO mice showed wider lesions than WT mice. CCR2 KO mice also expressed higher levels of the osteoclastogenic and osteolytic factors, receptor activator of nuclear factor kappa B ligand (RANKL) and cathepsin K, of the proinflammatory cytokine tumor necrosis factor–alpha, and of the neutrophil migration related chemokine, KC. Conclusions These results suggest that CCR2 is important in host protection to periapical osteolysis.

An inline amplifier system was constructed with erbium-doped fiber amplifiers spaced at 100 km and 80 km intervals. The system transmits 2.5 Gb/s signals over 2500 km with continuous-phase frequency-shift-keying heterodyne detection and over 4500 km with intensity-modulation direct detection. With respect to amplifier output signal power levels, it is experimentally shown that there exists a dynamic range within which long-distance signal transmission can be achieved with only small receiver sensitivity degradation. The range's upper and lower limits are determined by fiber nonlinearities and amplifier noise characteristics, respectively. >

Objective. To investigate the mechanism underlying neutrophil migration into the articular cavity in experimental arthritis and, by extension, human inflammatory synovitis. Methods. Antigen-induced arthritis (AIA) was generated in mice with methylated bovine serum albumin (mBSA). Migration assays and histologic analysis were used to evaluate neutrophil recruitment to knee joints. Levels of inflammatory mediators were measured by enzyme-linked immunosorbent assay. Antibodies and pharmacologic inhibitors were used in vivo to determine the role of specific disease mediators. Samples of synovial tissue and synovial fluid from rheumatoid arthritis (RA) or osteoarthritis patients were evaluated for CXCL1 and CXCL5 expression. Results. High levels of CXCL1, CXCL5, and leukotriene B4 (LTB4) were expressed in the joints of arthritic mice. Confirming their respective functional roles, repertaxin (a CXCR1/CXCR2 receptor antagonist), anti-CXCL1 antibody, anti-CXCL5 antibody, and MK886 (a leukotriene synthesis inhibitor) reduced mBSA-induced neutrophil migration to knee joints. Repertaxin reduced LTB4 production in joint tissue, and neutrophil recruitment induced by CXCL1 or CXCL5 was inhibited by MK886, suggesting a sequential mechanism. Levels of both CXCL1 and CXCL5 were elevated in synovial fluid and were released in vitro by RA synovial tissues. Moreover, RA synovial fluid neutrophils stimulated with CXCL1 or CXCL5 released significant amounts of LTB4. Conclusion. Our data implicate CXCL1, CXCL5, and LTB4, acting sequentially, in neutrophil migration in AIA. Elevated levels of CXCL1 and CXCL5 in the synovial compartment of RA patients provide robust comparative data indicating that this mechanism plays a role in inflammatory joint disease. Together, these results suggest that inhibition of CXCL1, CXCL5, or LTB4 may represent a potential therapeutic strategy in RA.

Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-27T11:22:42Z No. of bitstreams: 0Bitstream added on 2014-05-27T14:33:52Z : No. of bitstreams: 1 2-s2.0-42449127014.pdf: 609396 bytes, checksum: 1d7f5f941cb20a900121085eb2c496f7 (MD5) Made available in DSpace on 2014-05-27T11:22:42Z (GMT). No. of bitstreams: 0 Previous issue date: 2007-12-01 The aim of this article is to show the clinical and histochemical alterations of the first periodontal ligament, on the right side, after upper molars teeth extraction on the left side in gerbils. After two months, the periodontal ligaments were removed and processed for histochemical analysis. The data showed that TRAP reaction was able to evidence the osteoclastic activity in the hyperfunction hemimandible, right side, explaining the functional changes in the periodontal ligament after teeth extraction, and a little gingival recession and radicular exposure of teeth without function was observed at inferior molars of the left side. Faculty of Dentistry of Ribeirão Preto University of São Paulo, São Paulo Biosciences Institute- Molecular and Cellular Biology (UNESP), Rio Claro, São Paulo Faculty of Medicine of Ribeirão Preto University of São Paulo, São Paulo Faculdade de Odontologia de Ribeirão Preto USP Departamento de Morfologia, Estomatologia e Fisiologia, Av. Café S/N, CEP: 14040-904, Ribeirão Preto SP Biosciences Institute- Molecular and Cellular Biology (UNESP), Rio Claro, São Paulo

AC ionizer with high frequency became more common in semiconductor industries. Neutralization mechanism of such equipment is, however, not well understood because the measurement of extremely small ion current is interfered by AC induction. Our attempt is to use the Faraday cage that is directly connected to an AC ionizer. At frequencies of 1 Hz and 10 Hz, clear pulse shape ion currents were measured. The amounts of transported charges increase with both applied voltage and pressure. The effect of pressure is more significant than voltage. At higher frequency, the measured current changes from pulse to sinusoidal shape. From the delay time of 1 kHz, the average air velocity between the needle and Faraday cage was calculated. From DC corona measurement inside the nozzle, it became clear that the portion of transported charge to the Faraday cage is quite small.

Objective The aim of this study was to compare the expression of nuclear factor κB (RANK), RANK ligand (RANKL), and osteoprotegerin (OPG) in odontogenic epithelial tumors and dentigerous cysts. Study design The expression of these molecules was evaluated in solid/multicystic ameloblastomas (n = 12) and unicystic ameloblastomas (n = 8), keratocystic odontogenic tumors (n = 19), dentigerous cysts (n = 9), and dental follicles (n = 9) by immunohistochemistry. Results In odontogenic epithelium, a similar expression of RANK, RANKL, and OPG was observed in all samples. With regard to stroma, the number of RANK-positive and RANKL-positive cells was higher in solid/multicystic ameloblastomas compared with dentigerous cysts. Dental follicles had lower numbers of RANK-positive, RANKL-positive, and OPG-positive cells than solid/multicystic ameloblastomas and keratocystic odontogenic tumors. The majority of solid/multicystic ameloblastomas (75%) and unicystic ameloblastomas (62.5%) had higher numbers of RANKL-positive than OPG-positive cells. In contrast, 62.4% of keratocystic odontogenic tumors and 100% of dentigerous cysts exhibited a higher content of OPG-positive than RANKL-positive cells. Conclusion Our results indicate differences in the RANK, RANKL, and OPG expression in odontogenic epithelial tumors. The imbalance of these factors could contribute to the differential bone/tooth resorption activity in these lesions.

In this paper, we illustrate a decision support system of modeling the room designs suited for personal preference. The preference to colors is differed to individual personality. Nevertheless, modeling the room design is depended on the designer's sensitivity until now. Therefore, the technique of modeling the various room designs people satisfy is needed. In this paper, we propose automatic modeling system of the room design using interactive genetic algorithm (IGA). IGA has the advantage of learning the user's interactive evaluation, which is applied to the production of new room designs suited for the personal preference. The proposed system can help a decision support of modeling the various room designs, and reduce the burdens of designers significantly. In proposed system, the design evolution of IGA can reflect the user preference, and we present the effectiveness of the system by simulations.

Hyperhomocysteinemia is a known risk factor for cardiovascular diseases, but the underlying mechanisms of this pathology are complex. We aimed to evaluate the effect of hyperhomocysteinemia in vasorelaxations induced by alpha(1D)-adrenoceptor agonists. Vascular reactivity of rat carotid artery to the alpha-adrenoceptor agonist, phenylephrine, was enhanced in hyperhomocysteinemia. Mechanical removal of endothelium did not modify the carotid responsiveness to phenylephrine, compared to control. Phenylephrine induces endothelium-dependent relaxation, in the presence of 5-methyl urapidil (alpha(1A)-adrenoceptor antagonist). We hypothesised that endothelial-relaxant alpha(1)-adrenoceptors are impaired by hyperhomocysteinemia. Incubation with prazosin (selective alpha(1)-adrenoceptor antagonist) or BMY7378 (8-[2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl]-8-azaspiro[4,5]decane-7, 9-dione dihydrochloride) (selective alpha(1D)-adrenoceptor antagonist), similarly inhibited phenylephrine-induced relaxations in both control and hyperhomocysteinemic carotids. Immunohistochemistry showed enhanced immunoreactivity for eNOS and iNOS in hyperhomocysteinemic rats. In carotid arteries from hyperhomocysteinemic rats there was a decrease in superoxide dismutase activity and enhanced superoxide anion production. We conclude that alpha(1D)-adrenoceptors mediate endothelium-dependent relaxation triggered by phenylephrine in rat carotid artery and affect the final tone. Furthermore, the enhanced phenylephrine-induced contraction in carotid artery due to hyperhomocysteinemia is endothelium-dependent and involves a loss of the inhibitory effect of relaxant alpha(1D)-adrenoceptors by reducing NO biodisponibility.

Summary form only given. This paper reports the results of WDM transmission experiments done using the FSA commercial submarine amplifier system: which manages the fiber cable dispersion to transmit a single-NRZ l0-Gbit/s signal.

The authors report 2.5 and 10 Gbit/s transoceanic distance transmission in a commercial FSA optical amplifier system which has 90 km repeater spacing. The unique fibre dispersion management technique and amplifier passband management scheme used in the system are successfully demonstrated. The SNR improvement achieved with polarisation scrambling is also reported.

For multi-media communications, high capacity and flexible data transmission systems, such as the Passive Optical Network (PON), are in strong demand. In such point-multipoint communication systems, however, it is necessary to handle data packets at the same bit-rate. This precondition makes the PON system inflexible. If the PON system could handle multi-bit-rate packets, the user could choose the type of Optical Network Unit (ONU) needed making it easy to accommodate multiple users. Accordingly, we developed a 3R regeneration circuit for multi-bit-rate packets for use at the Optical Line Termination (OLT) point. It will greatly improve the flexibility of the PON system. We intend to extend the circuit to handle wider ranges of bit rates.

A 20-year-old Japanese man developed generalized, subcutaneous, painless nodules, fever, abnormal liver function, serosal effusions, hepatosplenomegaly, lymphadenopathy and anemia. Skin biopsies revealed lobular panniculitis with a morphologically benign histiocytic infiltration and prominent phagocytosis. Atypical T lymphocytes were also present in the skin and liver. The diagnosis given was aggressive cytophagic histiocytic panniculitis (CHP) or aggressive subcutaneous panniculitic T cell lymphoma (SPTCL). He received cyclophosphamide, doxorubicin, and vincristine on day 1, prednisolone on days 1-5, and etoposide on days 1, 3 and 5 (CHOP-E), with the support of granulocyte colony-stimulating factor. This regimen was repeated every 2 weeks and complete clinical remission (CCR) was attained after three cycles of CHOP-E. As the clinical course of aggressive CHP is recurrent and often fatal, he was given high-dose chemotherapy followed by autologous peripheral blood stem cell transplantation (APBSCT), after five cycles of CHOP-E. He has remained in CCR for 12 months after APBSCT. High-dose chemotherapy followed by APBSCT is considered to be one of the most beneficial therapies for patients with aggressive CHP and aggressive phase SPTCL.

The effectiveness of the polarisation scrambling technique for suppressing BER fluctuation is confirmed experimentally with a 2900 km optical in-line amplifier transmission system. The technique suppresses the BER fluctuation by three orders of magnitude, from 10/sup -5/-10/sup -9/ to 10/sup -8/10/sup -9/.

A 2.5Gbit/s, 50km span straight line transmission over 10 000 km using 199 Er-doped fibre amplifiers was successfully demonstrated. The simultaneous transmission or supervisory control and response signals was also confirmed

Thirty three cases of posterior dislocations and posterior fracture-dislocations of the hip were followed-up one to five years. The satiefactory results were obtained in seventeen cases.The clinical results of seventeen cases out of hirty three were analysed as regard the intervals between dislocation and reduction by closed or open method, roentgenographic findings, age of patients, and complications such as a sciatic nerve palsy.It was concluded that in simple dislocations, early closed reduction was acceptable, but in fracture-dislocations early primary open reduction with removal of lose fragment or internal fixation of the fragment was better.

Strain-measuring tests were performed with strain gages on rectangular coupons taken from perforated plate with triangular pitch under both uniaxial tensile loads and pure bending loads. The effective Young’s modulus obtained from the tests have strong correlation with the values recommended by the ASME Code. Next, to evaluate the stiffening effects of tubes, similar strain-gage tests were performed for different types of perforated coupons. One type had the tubes strength welded into the penetration holes of the test coupons. Another type had the tubes both expanded and strength welded into the test coupons. Stiffening effects of tubes are clearly obtained from these tests. Judging from the effective Young’s modulus of triangular pitch obtained by the testing, the recommended minimum credit given to the tube wall is 50 percent under elastic load condition. In addition, for experimental tests on an actual large-sized shell-and-tube heat exchanger under hydraulic test condition, good correlation was obtained between calculated and measured stress when full credit was taken for the tube wall in the calculation.

Parmi les 18 raies de Stokes attendues par l'analyse theorique du groupe d'espace de D 2h 18 , une est absente. Chacune est successivement attribuee a la representation irreductive du groupe ponctuel d 2h , ce qui confirme la symetrie cristalline orthorhombique du cristal de K 2 CuF 4 a un seul domaine. Les spectres d'une structure cristalline a plusieurs domaines presentent des traces de cette symetrie orthorhombique

The energy spectrum of diffuse hard X-rays measured in the range 10–40 keV shows a rather sharp change of slope. The logarithmic derivative of the spectrum changes around 20–30 keV by the increment significantly greater than 0.5 within an interval smaller than 50 keV.

This paper describes a bin picking system being installed two characteristic algorithms; one automatically constructs two dimensional models of structural industrial parts and the other recognizes parts having cylindrical components as main ones, using their models. Models are constructed automatically as much as possible and are modified interactively, if necessary. Parts may be stacked randomly. The recognition algorithm first extracts the main components and next searches for other components, utilizing their models. When models are too simple to detect orientations of parts, each part is looked by the other camera after being picked out from a bin. Our system uses only one image for a scene of stacked parts. The three dimensional positions of parts are obtained using both the results of recognition and a touch detector equipped in the gripper of a robot. Experimental results are presented finally.

An important and difficult problem in image processing is to segment an image into several different kinds of regions. Unless we isolate regions, we cannot perform further processing for pattern recognition, scene analysis, image interpretation, etc. Besides, if the segmentation phase can be done at high speed, this reduces the total processing time greatly.

The inflammatory oral diseases are characterized by the persistent migration of polymorphonuclear leukocytes, monocytes, lymphocytes, plasma and mast cells, and osteoblasts and osteoclasts. In the last decade, there has been a great interest in the mediators responsible for the selective recruitment and activation of these cell types at inflammatory sites. Of these mediators, the chemokines have received particular attention in recent years. Chemokine messages are decoded by specific receptors that initiate signal transduction events, leading to a multitude of cellular responses, including chemotaxis and activation of inflammatory and bone cells. However, little is known about their role in the pathogenesis of inflammatory oral diseases. The purpose of this review is to summarize the findings regarding the role of chemokines in periapical and periodontal tissue inflammation, and the integration, into experimental models, of the information about the role of chemokines in human diseases.