The preparation of supported lipid bilayers and its functional study / 生物医学工程学杂志

Supported lipid bilayers (SLBs) have been widely used in biomedical and bioengineering research because its structure and function are similar to natural cell membrane. A fluorescence recovery after photobleaching (FRAP) technique was used to measure the lateral diffusion of the SLBs composed of 1, 2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and 1, 2-dioleoyl-sn-glycero-3-［(N-(5-amino-1-carboxyp-entyl) iminodiacetic acid)］ (DGS-NTA) on the glass slide, and the effects of the DOPC-to-DGS-NTA ratio, small unilamellar vesicles (SUV) producing method, sizes of bleaching areas and concentrations of loading proteins on the SLBs fluidity and diffusion coefficient were studied systematically in this paper. The results demonstrated that: (1) SUV made by probe sonication exhibited more uniform and smaller size compared with that made by film extrusion, but the whole process of SLBs formation must not be exposed to air. (2) The fluorescence recovery rate and diffusion coefficient of the SLBs decreased with the increasing bleaching area size. With the mole ratio of DOPC to DGS-NTA decreasing from 98∶2 to 84∶16, the fluidity and fluorescence recovery degree decreased gradually, and the SLBs would lose its fluidity if the ratio reached to 82∶18. (3) The average fluorescence intensity of SLBs increased linearly with the loading protein concentration (10-40 nmol·L ), and the protein showed good mobility on the SLBs. The study would provide a good platform of bio-membrane for further research on interactions among cell membrane molecules and subsequent signals response.

Alterações morfológicas e químicas do esmalte humano após o clareamento dental: avaliação in situ / Morphological and chemical changes of human enamel after tooth bleaching: in situ evaluation

Introdução: A alteração de cor dos dentes apresenta-se como um dos fatores que mais concorrem para o desequilíbrio do sorriso, sendo o clareamento dental amplamente difundido e solicitado pelos pacientes. Objetivo: Este estudo in situ teve como objetivo avaliar as mudanças morfológicas e químicas do esmalte quando submetido a três agentes clareadores à base de peróxido de hidrogênio ativados com fonte de luz híbrida e um agente placebo (gel sem peróxido de hidrogênio), por meio do uso da espectrometria de energia dispersiva de raios-X (EDS). Metodologia: Fragmentos de terceiros molares humanos foram divididos em quatro grupos (n=12), para a realização de uma sessão de clareamento com cinco aplicações de oito minutos dos géis clareadores: Placebo (Plac); Lase Peroxide Flex 35% e 15% (DMC) (LPF35LH e LPF15LH); Gel experimental a 10% (DMC) (EXP10LHV), e foram fotocatalizados com luz híbrida: LED azul/laser de diodo (LH) (Whitening Lase II, DMC) ou LED violeta/laser de diodo (LHV) (luz experimental, DMC). Após o clareamento, os espécimes foram fixados a dispositivos intraorais usados pelos participantes durante 15 dias. A composição inorgânica e topografia da superfície de esmalte foram analisadas antes e após o clareamento, e depois de 3, 7 e 15 dias de exposição à saliva. Os valores elementares da composição foram analisados por ANOVA a um critério de medidas repetidas e teste de Tukey. Para a topografia os escores foram determinados por três examinadores previamente calibrados pelo teste Kappa e foi aplicado o teste estatístico de Friedman e Kruskal-Wallis, e as comparações individuais foram realizadas pelo teste de Dunn ( = 0,05). Resultados: De maneira geral, não houve alterações significativas na porcentagem elementar do esmalte nos diferentes períodos estudados. Ao analisar os dois principais elementos, o grupo LPF35HL obteve o menor valor de cálcio (Ca), possuindo diferença estatisticamente significante quando comparado com o grupo EXP10LHV, enquanto os valores de fosfato (P) permaneceram constantes. Morfologicamente somente o grupo EXP10LHV demostrou maior planificação da superfície quando comparado o período de 7 dias com 15 dias. Conclusão: Os diferentes protocolos clareadores empregados, demonstraram alterações pontuais na variação dos elementos químicos e na morfologia do esmalte dental ao longo do período de avaliação.(AU)

Introduction: The tooth color change is one of the factors that contributes most to the smile imbalance, and dental bleaching is widely diffused and requested by the patients. Objective: The aimed of this in situ study is to evaluate the morphological and chemical changes of the enamel when submitted to three activated hydrogen peroxide bleaching agents with hybrid light source and a placebo agent (gel without hydrogen peroxide), using energy-dispersive X-ray spectroscopy (EDS). Methodology: Fragments of human third molars were divided into four groups (n = 12) to perform a bleaching session with five eight minute applications of bleaching gels: Placebo (Plac); Lase Peroxide Flex 35% and 15% (DMC) (LPF35LH and LPF15LH); 10% experimental gel (DMC) (EXP10LHV), and photocatalyzed with hybrid light: blue LED / diode laser (LH) (Whitening Lase II, DMC) or violet LED / diode laser (LHV). After bleaching, the specimens were fixed to intraoral devices used by participants for 15 days. The inorganic composition and topography of the enamel surface were analyzed before and after bleaching, and after 3, 7 and 15 days of exposure to saliva. The elementary values of the composition were analyzed by one-way ANOVA at a repeated measures and Tukey's test. For the topography the scores were determined by three examiners previously calibrated by the Kappa test and the Friedman and Kruskal-Wallis statistical test were applied, and the individual comparisons were performed by the Dunn test ( = 0.05). Results: In general, there were no significant changes in the elemental percentage of enamel in the different periods studied. When analyzing the two main elements, the LPF35HL group had the lowest calcium (Ca) value, which had a statistically significant difference when compared to the EXP10LHV group, while the phosphate (P) values remained constant. Morphologically, only the EXP10LHV group showed greater surface planning when compared to the period of 7 days with 15 days. Conclusion: The different bleaching protocols employed showed specific alterations in the variation of the chemical elements and the morphology of the dental enamel during the evaluation period.(AU)

Rapid and simple method of photobleaching to reduce background autofluorescence in lung tissue sections.

Autofluorescence exhibited by tissues often interferes with immunofluorescence. Using imaging and spectral analysis, we observed remarkable reduction of autofluorescence of formalin fixed paraffin embedded tissues irradiated with light prior to incubation with immunofluorescent dyes. The technique of photobleaching offers significant improvement in the quality and specificity of immunofluorescence. This has the potential for better techniques for disease diagnosis.

Avaliação in vivo da efetividade e do pH de géis clareadores no clareamento em consultório em 12 meses de acompanhamento / In vivo evaluation of the effectiveness and pH of bleaching gels for in office whitening- 12 months follow-up

O objetivo deste estudo in vivo, internacional, randomizado e duplo cedo foiavaliar comparativamente a efetividade e o pH de diferentes géis clareadores natécnica de clareamento em consultório, com e sem o emprego de fonte de luzhíbrida em função do grau de alteração de cor, sensibilidade e manutenção dotratamento ao longo de 12 meses de acompanhamento. Foram selecionados 48voluntários de acordo com os critérios de inclusão e exclusão. Os pacientes foramdivididos, de forma randomizada, em 4 grupos de 12 participantes cada, onde:Grupo EXP10 –5 aplicações do gel de peróxido de hidrogênio a 10% (GelExperimental – DMC Equipamentos) e ativação de luz híbrida de LED (violeta)/Laser(Experimental – DMC Equipamentos) com 7’ e 30” por aplicação, com tempo total de37’30; Grupo LP15 – 5 aplicações do gel de peróxido de hidrogênio 15% (LasePeroxide Lite – DMC Equipamentos) seguindo mesmo protocolo do grupo EXP10;Grupo TB35LH – 3 aplicações do gel de peróxido de hidrogênio a 35% (Total BlancOffice - DFL) e ativação de luz híbrida de LED (azul)/Laser (Whitening Lase II – DMCEquipamentos) de 7’ e 30” por aplicação, com tempo total de 22’30”; Grupo TB35 – 3aplicações do gel de peróxido de hidrogênio a 35% (Total Blanc Office - DFL) semativação com fonte de luz, totalizando 45”. A determinação dos valores de pH foirealizada com o peagômetro digital (Sentron Model 1001, Sentron) nos temposinicial e após o término do protocolo clareador. A aferição da cor foi feita comespectofotômetro VITA Easyshade antes do clareamento, após 24 horas, 1 semana,1, 6 e 12 meses. A sensibilidade dentária e grau de satisfação dos pacientes foramavaliados por meio do questionário VAS e IPS antes, imediatamente após oclareamento, 24 horas e uma semana após. Os resultados da alteração do pHreceberam tratamento estatístico pela ANOVA e teste de Bonferroni a 0,05%...

The aim of the present in vivo study, international, randomized and doubleearly was to comparatively evaluate the effectiveness and pH of different bleachingagents for in office bleaching techniques, with and without the use of a hybrid lightsource depending on the degree of color change, sensitivity and maintenancetreatment over a 12-month follow-up. Selected were 48 volunteers according to theinclusion and exclusion criteria. The patients were randomly divided into 4 groups of12 participants each, where: EXP Group10- 5 applications of 10% hydrogen peroxidegel (Experimental Gel - Equipment DMC) and activation of hybrid LED light (violet) /Laser (Experimental - Equipment DMC) with 7 "and 30" per application, with a totaltime of 37'30; LP15 Group- 5 applications of 15% hydrogen peroxide gel (LasePeroxide Lite - Equipment DMC) following the same protocol of the EXP10 Group;TB35LH Group- 3 applications of 35% hydrogen peroxide gel (of Blanc Office - DFL)and activation of hybrid LED light (blue) / Laser (Whitening Lase II - DMC Equipment)7 "and 30" per application, with a total time of 22'30; TB35 Group - 3 applications of35% hydrogen peroxide gel (of Blanc Office - DFL) without light activation, totaling45 ". The determination of pH was carried out with a digital pH meter (Sentron Model1001, Sentron) in the initial times and after the bleaching protocol. The colormeasurement was made with a VITA Easyshade spectrophotometer before thetreatment, after 24 hours, 1 week, 1, 6 and 12 months. Tooth sensitivity and degreeof patient satisfaction were assessed by the VAS IPS questionnaire before,immediately after bleaching, 24 hours and one week after. The pH change resultswere statistically processed by the ANOVA and Bonferroni tests at 0.05%...

Avaliação in vivo da efetividade e do pH de géis clareadores no clareamento em consultório em 12 meses de acompanhamento / In vivo evaluation of the effectiveness and pH of bleaching gels for in office whitening- 12 months follow-up

O objetivo deste estudo in vivo, internacional, randomizado e duplo cedo foiavaliar comparativamente a efetividade e o pH de diferentes géis clareadores natécnica de clareamento em consultório, com e sem o emprego de fonte de luzhíbrida em função do grau de alteração de cor, sensibilidade e manutenção dotratamento ao longo de 12 meses de acompanhamento. Foram selecionados 48voluntários de acordo com os critérios de inclusão e exclusão. Os pacientes foramdivididos, de forma randomizada, em 4 grupos de 12 participantes cada, onde:Grupo EXP10 –5 aplicações do gel de peróxido de hidrogênio a 10% (GelExperimental – DMC Equipamentos) e ativação de luz híbrida de LED (violeta)/Laser(Experimental – DMC Equipamentos) com 7’ e 30” por aplicação, com tempo total de37’30; Grupo LP15 – 5 aplicações do gel de peróxido de hidrogênio 15% (LasePeroxide Lite – DMC Equipamentos) seguindo mesmo protocolo do grupo EXP10;Grupo TB35LH – 3 aplicações do gel de peróxido de hidrogênio a 35% (Total BlancOffice - DFL) e ativação de luz híbrida de LED (azul)/Laser (Whitening Lase II – DMCEquipamentos) de 7’ e 30” por aplicação, com tempo total de 22’30”; Grupo TB35 – 3aplicações do gel de peróxido de hidrogênio a 35% (Total Blanc Office - DFL) semativação com fonte de luz, totalizando 45”. A determinação dos valores de pH foirealizada com o peagômetro digital (Sentron Model 1001, Sentron) nos temposinicial e após o término do protocolo clareador. A aferição da cor foi feita comespectofotômetro VITA Easyshade antes do clareamento, após 24 horas, 1 semana,1, 6 e 12 meses. A sensibilidade dentária e grau de satisfação dos pacientes foramavaliados por meio do questionário VAS e IPS antes, imediatamente após oclareamento, 24 horas e uma semana após. Os resultados da alteração do pHreceberam tratamento estatístico pela ANOVA e teste de Bonferroni a 0,05%...

The aim of the present in vivo study, international, randomized and doubleearly was to comparatively evaluate the effectiveness and pH of different bleachingagents for in office bleaching techniques, with and without the use of a hybrid lightsource depending on the degree of color change, sensitivity and maintenancetreatment over a 12-month follow-up. Selected were 48 volunteers according to theinclusion and exclusion criteria. The patients were randomly divided into 4 groups of12 participants each, where: EXP Group10- 5 applications of 10% hydrogen peroxidegel (Experimental Gel - Equipment DMC) and activation of hybrid LED light (violet) /Laser (Experimental - Equipment DMC) with 7 "and 30" per application, with a totaltime of 37'30; LP15 Group- 5 applications of 15% hydrogen peroxide gel (LasePeroxide Lite - Equipment DMC) following the same protocol of the EXP10 Group;TB35LH Group- 3 applications of 35% hydrogen peroxide gel (of Blanc Office - DFL)and activation of hybrid LED light (blue) / Laser (Whitening Lase II - DMC Equipment)7 "and 30" per application, with a total time of 22'30; TB35 Group - 3 applications of35% hydrogen peroxide gel (of Blanc Office - DFL) without light activation, totaling45 ". The determination of pH was carried out with a digital pH meter (Sentron Model1001, Sentron) in the initial times and after the bleaching protocol. The colormeasurement was made with a VITA Easyshade spectrophotometer before thetreatment, after 24 hours, 1 week, 1, 6 and 12 months. Tooth sensitivity and degreeof patient satisfaction were assessed by the VAS IPS questionnaire before,immediately after bleaching, 24 hours and one week after. The pH change resultswere statistically processed by the ANOVA and Bonferroni tests at 0.05%...

Neuronal RNA granule contains ApCPEB1, a novel cytoplasmic polyadenylation element binding protein, in Aplysia sensory neuron

The cytoplasmic polyadenylation element (CPE)-binding protein (CPEB) binds to CPE containing mRNAs on their 3' untranslated regions (3'UTRs). This RNA binding protein comes out many important tasks, especially in learning and memory, by modifying the translational efficiency of target mRNAs via poly (A) tailing. Overexpressed CPEB has been reported to induce the formation of stress granules (SGs), a sort of RNA granule in mammalian cell lines. RNA granule is considered to be a potentially important factor in learning and memory. However, there is no study about RNA granule in Aplysia. To examine whether an Aplysia CPEB, ApCPEB1, forms RNA granules, we overexpressed ApCPEB1-EGFP in Aplysia sensory neurons. Consistent with the localization of mammalian CPEB, overexpressed ApCPEB1 formed granular structures, and was colocalized with RNAs and another RNA binding protein, ApCPEB, showing that ApCPEB1 positive granules are RNA-protein complexes. In addition, ApCPEB1 has a high turnover rate in RNA granules which were mobile structures. Thus, our results indicate that overexpressed ApCPEB1 is incorporated into RNA granule which is a dynamic structure in Aplysia sensory neuron. We propose that ApCPEB1 granule might modulate translation, as other RNA granules do, and furthermore, influence memory.

Visualization of serotonin 1A receptor trafficking in neuron-like PC12 cells / 基础医学与临床

Objective To explore the mechanisms of trafficking and signaling of serotonin 1A receptor(5-HT_(1A))and its spatiotemporal distribution in living cells.Methods The mouse 5-HT_(1A) gene amplified by RT-PCR was recombined into pEGFP-N1 vector and the EGFP coding sequence was located in-frame at the C-terminal end of the 5-HT_(1A) receptor.The 5-HT_(1A)-EGFP was transfected into neuron-like PC12 cells as well as HEK293.The transfected cells were visualized using confocal microscopy,the mobility of 5-HT_(1A)-EGFP was monitored by live measurements and fluorescence recovery after photobleaching.Results The 5-HT_(1A) gene was identitical with the published gene sequence NM_008308.4 and a 5-HT_(1A)-EGFP fusion construct was created.After stable transfection of the plasimd into a PC12 cell line and analysis with a confocal laser scanning microscopy,the EGFP-tagged 5-HT_(1A) was predominantly associated with the plasma membrane,but some intracellular vesicles in the perinuclear region also contained the fusion protein.The predominant localization of 5-HT_(1A)-EGFP at the plasma membrane was confirmed in transiently transfected HEK293 cells.Bleached fluorescence was partialy recovered in 100 seconds,indicating that the 5-HT_(1A)-EGFP was mobiled on the membrane.Conclusion Spatiotemporal distribution and mobility of 5-HT_(1A) tagged with EGFP can be monitored in the 5-HT_(1A)-EGFP stable PC12 cell line,which could be an excellent neuron-like experimental cell model for research of 5-HT_(1A) trafficking and signaling.

Alterações biomecânicas da córnea porcina com riboflavina fotossensibilizada por luz não-ultravioleta / Biomechanical changes on porcine corneas after riboflavin induced crosslink technique with non-UV light

OBJETIVO: Demonstrar a possibilidade de alterar a plasticidade do estroma corneano através da utilização do agente fotossensível riboflavina associado ao uso de iluminação não-ultravioleta. MÉTODOS: Experimento prospectivo duplo cego. Vinte e cinco olhos de porcos enucleados até 24 horas antes do experimento, foram divididos nos seguintes grupos: Grupo RB01+L-Riboflavina 0,1 por cento com irradiação de luz azul; Grupo RB01-Riboflavina 0,1 por cento sem irradiação de luz azul; Grupo RB05+L-Riboflavina 0,5 por cento com irradiação de luz azul; Grupo RB05-Riboflavina 0,5 por cento sem irradiação; Grupo L-Solução salina balanceada e irradiação de luz azul. RESULTADOS: Após o tratamento das informações dos grupos estudados, obtivemos diferenças estatisticamente significantes no grupos RB01+L e RB05+L(p<0,05). Os grupos sem irradiação de luz azul e o grupo somente com irradiação da luz azul sem riboflavina, não apresentaram diferenças estatisticamente significantes. CONCLUSÃO: A utilização de riboflavina para efetuar o processo de aumento de ligações covalentes entre as fibrilas colágenas pode ser uma das chaves para o controle de doenças da córnea como o ceratocone. Nas concentrações estudadas e doses de irradiância concebidas, o processo de aumento das características biomecânicas da córnea foram obtidas com sucesso.

PURPOSE: Demonstrate the possibility of changing the biomechanical behaviour of cornea stroma by usage of riboflavin associated with non-ultraviolet light. METHODS: Double blind prospective study. Twenty five porcine eyes enucleated 24 hours before the experiment, have been divided on the following groups : Riboflavin 0,1 percent with irradiation of blue light, Riboflavin 0,1 percent without irradiation of light , Riboflavin 0,5 percent with irradiation of blue light, Riboflavin 0,5 percent without irradiation of light and BSS with irradiation of light. RESULTS: Differences where noticed in groups 1 and 3 (p<0,05). The groups without irradiation and the group with only irradiation, had no significant difference on their results. CONCLUSION: The usage of riboflavin appears to increase the crosslink connections among collagen fibrills. On the studied concentrations studied and radiant parameters the increase of biomechanical characteristics of cornea have been obtained successfully.

Study on Cx43 expression and function of GJIC in ectopic and eutopic endometrial stromal cells in vitro / 重庆医科大学学报

Objective:To investigate Cx43 expression and gap junctional intercellular communication(GJIC) in ectopic and eutopic endometrial stromal cells in endometriosis(EMs),and to explore the influence of aberrant GJIC in stromal cells on pathogenesis of EMs. Methods:The stromal cells were isolated from samples including 24 ectopic endometriotic tissues located in ovaries,41 eutopic endometria with endometriosis and 30 normal endometria. The endometrial stromal cells models were established in vitro by being cultured in manual conditions mimicked with estrogen and progesterone. Laser scanning confocal microscopy(LSCM) was used to determine the expression of Cx43 protein and the function of GJIC in three groups stromal cells. Results:The success rate of isolation and culture of endometriotic stromal cells was 45.8%(11/24);of eutopic endometrial stromal cells with EMs was 92.7(38/41);of normal endometrial stromal cells was 93.3%(28/30). The purities of ectopic and eutopic endometrial stromal cells were 95% and 98% respectively. The level of Cx43 protein and the function of GJIC in stromal cells from ectopic endometrial tissues were much lower than those from the other two groups,the highest level of Cx43 protein and the function of GJIC were observed in normal endometrial stromal cells group,and the differences among these groups were significant (P﹤0.01). Conclusions:(1) It will be helpful to establish models of normal,ectopic and eutopic endometrial stromal cells in vitro simultaneously when investigating the pathgenesis of EMs. (2)Downregulation of Cx43 expression and aberrant function of GJIC are related to pathogenesis of EMs. Regulation of Cx43 or GJIC in endometrial stromal cells is implied to be a potential strategy to treat EMs.

Application of fluorescence recovery after photobleaching in the detection of gap junctional intercellular communication in bladder smooth muscle / 第三军医大学学报

Objective To explore the functional changes of gap junctional intercellular communication(GJIC) in bladder smooth muscle. Methods The functions of GJIC in bladder smooth muscle were detected by fluorescence recovery after photobleaching(FRAP). The mean fluorescence recovery rates of the bladder smooth muscle cells in the experimental group and the control group were compared. Results The mean fluorescence recovery rate of the experimental group was significantly higher than that in the control group( P

Interstitial cells of Cajal regulates excitability of detrusor myocytes: structure evidence / 第三军医大学学报

Objective To investigate the role of bladder interstitial cells of Cajal(ICCs) in regulating the excitability of detrusor myocytes.Methods 1)Ultrastructural connection between ICCs and detrusor myocytes was detected by transmission electron microscope(TEM).2) Fluorescence redistribution after photobleaching(FRAP) was carried out to detect the intercellular communication between detrusor myocytes and its neighboring ICCs.Results 1)Gap junction between the 2 kinds of cells was confirmed by TEM.2)After target detrusor myocyte was photobleached,fluorescence intensity in detrusor myocytes was recovered gradually.In the meantime,fluorescence intensity in the ICCs neighboring to the target detrusor myocyte was decreased.This indicates a signal transmission from the neighboring ICCs to the target detrusor myocyte.Conclusion Structural connection is seen between ICCs and detrusor myocytes.Excitability signals might be transferred from ICCs to detrusor myocyte.

Study on gap junctional mediated intercellular communication in detrusor instability / 中华泌尿外科杂志

Objective To study the functional changes of gap junctional mediated intercellular communication in detrusor instability so as to demonstrate the feasibility of blocking excitatory communication as the target of therapy for DI. Methods The function of GJIC in the cultured bladder detrusor cells were detected by FRAP. Results At the fourth minutes after bleaching,the mean fluorescences recovery rates of the DI groups bladder detrusor cells was (35.791?0.836)%,the control groups (8.645?0.673)%.The mean fluorescences recovery rates of the DI groups were significant higher than those in control groups (P