Oral toxicity to high level sodium fluoride causes impairment of autophagy.

Autophagy is a highly conserved intracellular digestion process that degrades damaged proteins and organelles but the biological roles of autophagy in pathological aspects of oral tissues remain largely unknown. We sought to elucidate the function of autophagy, especially its interplay with apoptosis and oxidative stress, in the oral toxicity induced by exposure to 5 mM sodium fluoride (NaF). Human cementoblasts (HCEM-2) in culture were exposed to 5 mM NaF for 5 min, after which cell viability and cell apoptosis were assessed using the MTS assay and an Annexin V-FITC/PI apoptosis detection kit, respectively. Quantitative RT-PCR and Western blotting were performed to characterize the expression levels of markers for autophagy, apoptosis, and oxidative stress. Senescence-resistant (SAMR1) mice were exposed to 5 mM NaF in their drinking water from 12 to 58 weeks. Micro-computed tomography was used to measure changes in their alveolar bone while immunohistochemistry and immunofluorescence staining was used to evaluate protein expression levels. HCEM-2 cells exposed to 5 mM NaF had decreased levels of autophagy, as shown by reduced expression levels of ATG5, Beclin-1 and LC3-II, elicited apoptosis, which in turn induced oxidative stress and inflammation, as manifested by elevated levels of Bax, cleaved caspase-3, SOD1 and phospho NF-κB. Treatment of mice with 5 mM NaF resulted in histological abnormalities in periodontal tissues, induced excessive oxidative stress and apoptosis, and reduced autophagy. Micro-computed tomography analysis demonstrated that 5 mM NaF caused a decrease in bone areas of mice compared with controls. Exposure to 5 mM NaF induced RANKL (receptor activator of nuclear factor κB ligand) and cathepsin K expression in periodontal tissues, while ATG5 and Beclin-1 expression was abrogated by 5 mM NaF. Taken together, our findings suggest that 5 mM NaF elicits oral toxicity that contributes to excessive apoptosis, oxidative stress, and defective autophagy, which aggravates periodontal tissue damage.

Phenytoin-induced gingival overgrowth caused by death receptor pathway malfunction.

OBJECTIVE: In this study, we investigated the role of phenytoin (PHT) in death receptor-induced apoptosis of gingival fibroblasts to clarify the mechanism of PHT-induced gingival overgrowth. METHODS: Human gingival fibroblasts were cultured to semiconfluence and treated with PHT (0.025, 0.1, 0.25, and 1.0 µM) for 48 h, and then, the apoptotic cell numbers were relatively determined by absorptiometry. After 24 h of 0.25 µM PHT treatment, caspase activity was measured by absorptiometry, apoptotic and cell cycle phase distribution was analyzed by flow cytometry, expression levels of apoptotic genes were quantified by real-time qPCR, and expression of apoptotic proteins was detected by Western blot analysis. After 48 h of 0.25 µM PHT treatment, appearance of apoptotic cells was detected by TUNEL assay. RESULTS: PHT treatment decreased the proportion of apoptotic cells in gingival fibroblasts compared to a serum-free control culture in response to the protein changes as follows: PHT upregulated c-FLIP and, in turn, downregulated FADD, caspase-8, and caspase-3; PHT upregulated c-IAP2 and downregulated TRAF2; PHT downregulated caspase-9 and caspase-3 via decreased RIPK1 activity and increased Bcl-2 activity. CONCLUSION: PHT-induced gingival overgrowth may result from the above-mentioned mechanisms involving apoptosis inhibition in gingival fibroblasts.

Tooth fragment reattachment: a case report.

The aim of this article is to present a case report for the multidisciplinary treatment of anterior tooth fractures with invasion of the biologic width and pulpectomy. Successful esthetic and functional results were achieved by bonding the crown fragment, without any form of preparation or the utilization of intracanal posts.

Nosocomial outbreak of epidemic keratoconjunctivitis accompanying environmental contamination with adenoviruses.

An outbreak of acute keratoconjunctivitis involving 27 patients occurred in the Department of Ophthalmology, Kurume University Hospital. Adenoviral DNA was detected in four inpatients, one outpatient and one healthcare worker. Sequence-based typing of adenoviral DNA indicated serotype 3 from one inpatient, the rest being serotype 37. At a later stage of the outbreak adenoviral DNA types 37 and/or 3 were also detected from almost all environmental instruments and commonly used eye drops, despite thorough disinfection of the environment and enforcement of various infection control measures. The detection rate of adenoviral DNA in environmental swabs was 81%. A further second disinfection of the environment reduced the detection rate of adenoviral DNA to 38%. The outbreak ceased after closing the ophthalmology ward and outpatient consulting room, accompanied by enhanced cleaning of environmental instruments and the introduction of disposable eye drops for individual patients.

Capacity of ocular infiltrating T helper type 1 cells of patients with non-infectious uveitis to produce chemokines.

BACKGROUND/AIMS: Chemokines are key molecules that initiate leucocyte infiltration to the inflammatory site. The involvement of chemokines in uveitis is well studied, yet the source of this molecule in the inflamed eye is not clearly identified. The possible sources of chemokines are ocular resident cells or the inflammatory cells infiltrated to the eye. Here the authors examined whether ocular infiltrating T cells of uveitis patients do produce chemokines. METHODS: T cell clones (TCCs) were established from ocular infiltrating cells of patients with non-infectious uveitis. TCCs were characterised using flow cytometry. Spontaneous production of chemokines by TCCs was evaluated by ELISA. RESULTS: TCCs from ocular infiltrating cells were revealed to be memory activated Th1 type CD4 positive cells. Those TCCs produced larger amounts of chemokines than TCCs from peripheral blood mononuclear cells of uveitis or healthy donors. CONCLUSIONS: The present data indicate that ocular infiltrating T cells of patients with non-infectious uveitis produce chemokines and recruit further infiltrating lymphoid cells. Such T cells may have roles in the prolonged/chronic state of non-infectious uveitis.

Estudo do benefício da reabilitalção vestibular nas síndromes vestibulares periféricas / Study about the benefits of vestibular rehabilitation in peripheral vestibular disorders

BACKGROUND: Vestibular rehabilitation in vestibular disorders. AIM: To study the benefits of Vestibular Rehabilitation through the Brazilian Dizziness Handicap Inventory-DHI. METHOD: Application of the Vestibular Rehabilitation proposed by Zee (1985) and analysis using the Sign; T-Student and Kendall's T tests. RESULTS: A change in the diagnosis of vestibular peripheral disorder to normal vestibular patterns was obtained in 75% of the cases. CONCLUSION: Our results indicated a change of the diagnosis and in the quality of life of the tested individuals. The use of the DHI demonstrated to be an excellent tool for qualitative studies.

Tema: reabilitação vestibular nas lesões vestibulares. Objetivo: estudar o benefício da Reabilitação Vestibular por meio do Dizziness Handicap Inventory (DHI) - Brasileiro. Método: aplicação da Reabilitação Vestibular proposta por Zee (1985) e análise pelo teste de sinais; teste t-student e teste t de Kendall. Resultados: obtivemos a mudança dodiagnóstico de Síndrome Irritativa para Exame Vestibular Normal em 75% dos nossos casos. Conclusão: nossos resultados apontaram a mudança do diagnóstico e da qualidade de vida dos indivíduos avaliados, e a aplicação do DHI se mostrou uma excelente ferramenta para estudo qualitativo.

Macrophage migration inhibitory factor in ocular fluids of patients with uveitis.

AIMS: To investigate the levels of macrophage migration inhibitory factor (MIF) in intraocular fluids of uveitis patients, the capacity of intraocular infiltrating lymphocytes to produce MIF, and the correlation between MIF levels in the eye and intraocular inflammatory activity. METHODS: MIF levels were measured by enzyme linked immunosorbent assay (ELISA) using (1) aqueous humour (AH) of 12 uveitis patients and eight control patients with cataract, (2) vitreous fluid of 15 uveitis patients and eight control patients with idiopathic macular hole, and (3) culture supernatants of T cell clones (TCCs) established from intraocular fluids of uveitis patients. MIF expression on infiltrating cells was determined by a double staining immunofluorescence technique using a flow cytometry. RESULTS: Significant levels of MIF were detected from intraocular fluids of uveitis patients (4.0 (SD 3.0) ng/ml in AH and 16.5 (24.7) ng/ml in vitreous), whereas MIF levels in control groups were below the detectable levels. There was a significant correlation between MIF levels and vitreous inflammation (29.7 (30.0) ng/ml in active uveitis v 3.3 (2.6) ng/ml in inactive uveitis, p< 0.05). Significant levels of MIF were detected in culture supernatants of TCCs from ocular fluids of uveitis patients. MIF was expressed on infiltrating CD4+ lymphocytes from vitreous of uveitis patients. CONCLUSION: Significant levels of MIF are present in intraocular fluids of patients with uveitis. Lymphocytes infiltrating in the eye are capable of producing MIF. MIF levels in vitreous fluid are correlated with vitreous inflammation activity. These data thus indicate that MIF in the eye has a significant role in the pathophysiology of ocular inflammation.

Soluble Fas ligand and soluble Fas in ocular fluid of patients with uveitis.

AIMS: To investigate the presence of soluble Fas ligand (sFasL) and soluble Fas (sFas) in ocular fluid of patients with uveitis. METHODS: Samples of aqueous humour (AH, n=17), vitreous fluid (n=9), and serum (n=60) were collected from patients with uveitis which included Behçet's disease, Vogt-Koyanagi-Harada disease, sarcoidosis, human T lymphotropic virus type 1 (HTLV-I) uveitis, sympathetic ophthalmia, HLA-B27 associated acute anterior uveitis, and ocular toxoplasmosis. The AH of patients with age related cataract without uveitis obtained during cataract surgery was used as controls (n=20). The amounts of sFasL and sFas were measured by enzyme linked immunosorbent assay. RESULTS: Significant amounts of sFasL were detected in AH of patients with age related cataract (non-uveitis group). sFasL was also detected in AH of patients with uveitis, though the amounts were slightly lower than those in the non-uveitis group. On the other hand, the levels of sFas in AH of patients with uveitis were significantly higher than those in controls. As for the disease activity, the levels of sFasL and sFas in the vitreous fluid of patients with active uveitis were significantly higher than those in inactive uveitis. sFasL in the serum of healthy donors and patients with uveitis was below detectable levels, except for patients with HTLV-I uveitis who had significant amounts of sFasL in the serum. The levels of sFas in the serum of patients with Behçet's disease, sarcoidosis, and HTLV-I uveitis were significantly higher than those of healthy donors. CONCLUSIONS: sFasL is present in the AH of non-uveitic eyes with age related cataract. Intraocular levels of sFasL and sFas are significantly increased in uveitis, particularly in active uveitis. These data suggest that intraocular sFasL and sFas may have a regulatory role in uveitis.

Polymorphism of the 5'-flanking region of the tumor necrosis factor (TNF)-alpha gene and susceptibility to human T-cell lymphotropic virus type I (HTLV-I) uveitis.

The genetic background of human T-cell lymphotropic virus type I (HTLV-I) uveitis (HU) was investigated by studying the distribution of 5 polymorphisms of the 5'-flanking promoter/enhancer region of the tumor necrosis factor (TNF)-alpha gene in patients with HU, together with patients with adult T-cell leukemia (ATL), asymptomatic HTLV-I carriers, and healthy controls. The frequencies of the -1,031C allele (T-->C transition at position -1,031) and -863A allele (C-->A transition at position -863) in the HU patients, but neither in the ATL patients nor in the carriers, were significantly higher than those in the controls. The -1,031C and -863A alleles, in the absence of the HLA B61 or the DRB1*0901 allele which is in linkage disequilibrium with these alleles, were associated with increased susceptibility to HU. These results suggest that the -1,031C and -863A alleles might be genetic risk factors for HU.

[A report of two cases suggesting positive influence of pregnancy on uveitis activity].

BACKGROUND: Little is known about the influence of pregnancy on uveitis activity. We report two cases suggesting a favorable influence of pregnancy on the clinical course of uveitis. CASE: A 30-year-old woman who was three months pregnant was suspected Vogt-Koyanagi-Harada disease based on the systemic symptoms and ocular findings of iritis and multi-focal serous retinal detachment. She was positive to human leukocyte antigen (HLA)-DR 4. She was treated only with topical corticosteroids. One month later, the retinal detachment disappeared. Six month later, a healthy baby was born. The fundus of both eyes took on a sunset glow appearance and there has been no recurrence of uveitis. The other case was a 23-year-old woman with Behçet's disease who had several episodes of uveitis in a year even on tacrolimus. Because of pregnancy, all systemic drugs including tacrolimus were discontinued since then. Interestingly, the frequency of uveitis was remarkably decreased during the pregnancy. A normal healthy baby was born. The uveitis has almost completely disappeared since parturition until now. CONCLUSION: It is considered that the increase of intrinsic hormone, especially corticosteroid, and some other factors with pregnancy may give the suppressive influence on uveitis in our cases.