Synthesis and Biological Characterization of a New Norbormide Derived Bodipy FL-Conjugated Fluorescent Probe for Cell Imaging

Background: Norbormide (NRB) is a selective rat toxicant endowed with vasoconstrictor activity confined to the rat peripheral arteries. In a recent work we used a fluorescent derivative of NRB (NRB-AF12), obtained by coupling the NBD fluorophore to the parent molecule via a linker, in order to gain information about the possible site of action of the unlabeled compound. We found that NRB-AF12 labeled intracellular organelles in both NRB-sensitive and -insensitive cells and we accordingly proposed its use as a scaffold for the development of a new class of fluorescent probes. In this study, we examined the fluorescent properties of a BODIPY FL-conjugated NRB probe (MC009) developed: (A) to verify if NRB distribution could be influenced by the attached fluorophore; (B) to improve the fluorescent performance of NRB-AF12. Methods: MC009 characteristics were investigated by confocal fluorescence microscopy, in freshly isolated rat caudal artery myocytes (FIRCAM) and in LX2 cells, representative of NRB-sensitive and insensitive cells, respectively. Main results: In both FIRCAM and LX2 cells MC009 stained endoplasmic reticulum, mitochondria, Golgi apparatus and lipid droplets, revealing the same intracellular distribution as NRB-AF12, and, at the same time, had both improved photostability and gave a more intense fluorescent signal at lower concentrations than was possible with NRB-AF12, which resulted in a better and finer visualization of intracellular structures. Furthermore, MC009 was effective in cellular labeling in both living and fixed cells. At the concentration used to stain the cells, MC009 did not show any cytotoxic effect and did not affect the regular progression of cell cycle and division. Conclusions: This study demonstrates that the distribution of fluorescently labeled NRB is not affected by the type of fluorophore attached to the parent compound, supporting the idea that the localization of the fluorescent derivatives may reasonably reflect that of the parent compound. In addition, we observed a marked improvement in the fluorescent properties of BODIPY FL-conjugated NRB (MC009) over its NBD-derived counterpart (NRB-AF12), confirming NRB as a scaffold for the development of new, high performance, non-toxic fluorescent probes for the labeling of intracellular structures in both living and fixed cells

The relationship between the bruxism and the severity of symptoms in the temporomandibular disorder

PURPOSE: to check the occurrence of bruxism in female gender adult subject, with temporomandibular jaw dysfunction and its association with the level of temporomandibular symptoms. METHODS: 28 women, with ages between 19 and 56 years, who had temporomandibular jaw dysfunction and had not been submitted to a preview treatment, made part of this study. All subjects answered a questionnaire written by Fonseca et al (1994) entitled Anamnesic Index, that enabled us to classify temporomandibular jaw symptoms level and also to check the symptoms of this oral parafunction behavioral bruxism; the subjects were submitted an odontological evaluation, that was composed by an exam of masticatory muscles, though the digital touch in the oral and extra-oral muscles, examination in the temporomandibular joint and a teeth examination. The results were analyzed in a descriptive form, and in order to check the relationship between the severity level of temporomandibular dysfunction with the bruxism occurrence Independent Square-Qui test and the Fisher test were used with a statistic significance of 5%. RESULTS: it was observed the parafuntional habit was presented in 50% of the cases with temporomandibular dysfunction. It wasn't possible a relationship between bruxism and the level of symptoms in temporomandibular jaw dysfunction, by Anamnesic Index. CONCLUSION: the results of this study suggest that subjects with symptoms of temporomandibular jaw dysfunction must be asked and evaluated about bruxism, regardless of the level of temporomandibular dysfunction. Besides this, it is important to investigate the diagnosis and the treatment of bruxism also in patients without temporomandibular jaw dysfunction in order to prevent development of lesions in the temporomandibular joints and the other structures of the stomatognatic system.OBJETIVO: verificar a ocorrência de bruxismo em sujeitos adultos do sexo feminino com queixa de disfunção temporomandibular e sua relação com o grau de sintomatologia da disfunção. MÉTODOS: participaram deste estudo 28 mulheres na faixa etária de 19 a 56 anos, que apresentavam sintomas de disfunção temporomandibular e que não haviam realizado tratamento anterior. Todas responderam o questionário de Índice Anamnésico proposto por Fonseca et al (1994), o qual possibilita a classificação do grau de sintomatologia de disfunção temporomandibular e verificação da queixa do hábito parafuncional bruxismo; e realizaram uma avaliação odontológica, constituída de exame da musculatura mastigatória, por meio de palpação digital intra e extra-oral, inspeção das articulações temporomandibulares e exame dental. Os resultados foram analisados descritivamente e, para verificar a relação entre o grau de severidade da disfunção temporomandibular com a ocorrência de bruxismo, foram realizados o Teste de Independência do Qui-quadrado e o Teste Exato de Fisher, ambos ao nível de significância de 5%. RESULTADOS: verificou-se que o bruxismo esteve presente em 50% dos casos de disfunção temporomandibular. Não houve relação entre o bruxismo e o grau de sintomatologia de disfunção temporomandibular, estabelecido pelo Índice Anamnésico. CONCLUSÃO: os resultados deste estudo sugerem que sujeitos com sintomas de disfunção temporomandibular devem ser questionados e avaliados quanto à presença de bruxismo, independentemente do grau de sintomatologia da disfunção. Assim como deve ser realizado diagnóstico e tratamento do bruxismo em sujeitos assintomáticos de disfunção temporomandibular como forma de prevenir o desenvolvimento de lesões nas articulações temporomandibulares e demais estruturas do sistema estomatognático.Secretaria Municipal de Saúde Centro de Referência em Saúde do TrabalhadorUniversidade Federal de Santa Maria Associação dos Pais e Amigos do ExcepcionalUniversidade Federal de Santa MariaUniversidade Federal de São Paulo (UNIFESP) Universidade Federal de Santa Maria Departamento de FonoaudiologiaUNIFESP, Universidade Federal de Santa Maria Depto. de FonoaudiologiaSciEL

Relationship between the severity of temporomandibular disorder and voice

BACKGROUND: temporomandibular dysfunction is one of the most complex disorders of the body, capable of unchaining alterations in the mandibular movements which in turn can cause damage not only to speech articulation but also to the quality of voice. In the literature, the relationship between the symptomatology severity levels of this dysfunction and their influence on vocal production has been scarcely studied. Aim: to verify the relationship between the symptomatology severity levels of temporomandibular dysfunction and vocal production. METHOD: participants of this study were 24 females, with ages between 16 and 56 years, who were submitted to an anamnesis index questionnaire (Fonseca et al., 1994), to odontological and otolaryngological examinations and to a hearing evaluation. Later, the participants were submitted to a voice recording, using a digital recorder, for a perceptive-auditive analysis of voice parameters such as: vocal type, resonance, quality of emission, pitch and loudness and for the analysis of the acoustic parameters of the wide and narrow band spectrography and of the Multi Dimensional Voice Program (MDVP) - Key Elementrics Real Time software. RESULTS: considering the parameters of the perceptive-auditive evaluation, it was observed that only individuals with severe symptomatology presented a statistically significant decrease in loudness (p = 0.013). The hoarse vocal quality was the most frequent type among the participants, with severities classified as mild and severe, followed by the breathy type. In the wide band spectrography, there was a statistically significant difference for the anti-resonance increase (p = 0.013) in individuals with severe temporomandibular dysfunction. CONCLUSION: it was verified that the severity level causes a decrease in loudness, an increase of noise and alterations in voice resonance, interfering in the vocal quality of these individuals.TEMA: a disfunção temporomandibular é uma das desordens mais complexas do organismo capaz de desencadear alterações nos movimentos mandibulares que provocam prejuízos tanto na articulação da fala como na qualidade da voz. Na literatura a relação entre o grau de severidade da sintomatologia desta disfunção e a influência desta na produção vocal tem sido pouco estudada. OBJETIVO: verificar a relação entre o grau de severidade de sintomatologia da disfunção temporomandibular com a produção vocal. MÉTODO: participaram deste estudo 24 sujeitos, do gênero feminino, com idade variando entre 16 e 56 anos que foram submetidos à aplicação do questionário de índice anamnésico Fonseca et al. (1994), a exame odontológico, exame otorrinolaringológico e avaliação audiológica. Posteriormente os 24 sujeitos da pesquisa foram submetidos à gravação da voz, em gravador digital para posterior análise perceptivo-auditiva dos parâmetros da voz como: tipo vocal, ressonância, qualidade da emissão, pitch e loudness, e para análise dos parâmetros acústicos da espectrografia de banda larga, banda estreita e dos parâmetros acústicos por meio do Multi Dimensional Voice Program (MDVP) da Key Elementrics Real Time. RESULTADOS: verificou-se que de todos os parâmetros da avaliação perceptivo-auditiva da voz o grau de sintomatologia severo apresentou significância estatística para diminuição da loudness (p = 0,013). A qualidade vocal rouca foi a que mais apareceu nos sujeitos com grau leve e severo, seguida pela soprosa. Na espectrografia de banda larga houve significância estatística para o aumento da anti-ressonância (p = 0,013) no grau severo de disfunção temporomandibular. CONCLUSÃO: verificou-se que o grau de severidade ocasiona diminuição da loudness, aumento de ruído e alteração na ressonância da voz interferindo na qualidade vocal desses sujeitos.Universidade Federal de São Paulo (UNIFESP)Universidade Federal de Santa Maria Departamento de Fonoaudiologia Programa de Pós-Graduação em Distúrbios da Comunicação HumanaUniversidade Federal de Santa MariaPontifícia Universidade Católica do Rio Grande do SulUNIFESPSciEL