Efeito in vitro da terapia fotodinâmica sobre bactérias orais em suspensão e na formação de biofilme : ensaio com azul de metileno e toluidina ativados por luz halógena

Photodynamic therapy (PDT) has been proposed as an alternative method for the treatment of biofilm-dependent oral diseases like dental caries. This therapy consists of simultaneous action of a visible light (L) and a photosensitizer (FS) in the presence of oxygen, which leads to production of different reactive oxygen species that can interact with the bacterial cell components, and promote cell death. This study aims to evaluate the antimicrobial action of PDT on oral bacteria in suspension, as well as the formation of mono and multi-species biofilms, in vitro, from a standard strain of Streptococcus mutans (ATCC 25175) and saliva samples, respectively. The dye methylene blue (MB) and toluidine blue (TB) were used at a concentration of 100 mg/ L and activated by halogen light (600 to 750 nm) from a modified hand held photopolymerizer (Ultralux ®, Dabi Atlante, Ribeirão Preto , São Paulo, Brazil.). Planktonic cultures were prepared and submitted to different experimental conditions: 1. PDT using TB 2. PDT using MB, 3. L+ FS- , 4. TB + L - ; 5. MB+ L-; 6. L- FS- (negative control) and 7. administration of 0.12% chlorhexidine digluconate (positive control) (Periogard ®, Colgate-Palmolive Company, New York, USA). The immediate and mediated action of PDT on bacterial suspensions, as well as its effect on biofilm formation were observed from the number of colony-forming units per milliliter (CFU/mL) and measures optical density (OD). The data were statistically analyzed using the Kruskal-Wallis and Mann-Whitney test for the significance level of 5%. According to the results, the PDT showed no antibacterial action on suspensions of S. mutans, regardless of the dye used. PDT with MB activated by halogen light was able to reduce 86.6% CFU/mL multi-species planktonic cultures, however, this reduction was not significant (p > 0.05). PDT showed antibacterial effect, mediate on multi-species planktonic cultures with TB (p < 0.001) and MB (p < 0.001), activated by halogen light. PDT was able to prevent the formation of multispecies biofilm, through the activation of TB by halogen light (p = 0.01). We conclude that activation of the dye toluidine blue and methylene blue, by halogen light (PDT) showed antimicrobial activity, compared to multi-species planktonic cultures prepared from saliva samples

Análise imunoistoquímica após terapia fotodinâmica com cloro-alumínio ftalocianina em tecidos gengivais de humanos

Although photodynamic therapy have been used as a useful tool over the past 30 years in oncology, few clinical trials have been conducted in dentistry. Photodynamic therapy (PDT) uses non - toxic photosensitizers and selective which are administered in target cells followed by local application of visible light, producing reactive oxygen species capable of causing cell death by apoptosis or necrosis, injured the local vasculature, and exert important effects on the im mune system. New generations of photosensitizing agents, such as nanoparticulate phthalocyanines, has shown excellent results in antitumor and antibacterial activity . In this context, the present work constitutes the first clinical protocol of local appli cation of nanoemulsion chloro - aluminum phthalocyanine (AlClFc) followed by irradiation in human gingiva, and analyzed descriptively and comparatively , by means of immunohistochemistry , the expression of RANK , RANKL , OPG and VEGF in a split - mouth model . Eight healthy volunteers with clinical indication for extraction were included in the study . Seven days before the extraction, was injected in the gingiva of participants, 5 μ M of nanoemulsion AlClFc followed by irra diation with diode laser (660nm , 7 J/cm2 ), the contralateral side was used as control. Tissue specimens were removed seven days after the TFD is performed. Tissues sample were divided into two groups (test and con trol groups) for histological and immunohistochemical analysis. Patients were monitored at days, 0, 7, 14 and 30 to assess adverse effects of the therapy. Vascular alterations were seen in gingival samples that received PDT. Areas of edema and vascular con gestion, and intense vascularization were viewed . Additionally, dystrophic calcification in subepithelial region were observed in the test group. The results showed a similar pattern of immunostaining scores of RANK, RANKL and VEGF between the test and co ntrol groups, with no statistically significant difference (p = 0.317, p = 0.777, p = 0 .814, respectively). RANK and RANKL exhibited weak or absent immunostaining in most specimens analyzed. There was n o immunostaining for OPG. VEGF showed moderate to stro ng immunostaining in specimens from the test group. In addition, the clinical study showed that therapy was well tolerated by all patients. Adverse effects were short - time and completely reversible. Taken together, the results presented in this study showe d that PDT mediated by nanoemulsion containing AlClPc is safe for clinical application in gingival tissue and suggests that a strong immunostaining for VEGF after therapy .

Efeitos da terapia fotodinâmica com aluminio – cloro ftalocianina sobre a peroxidação lipídica e produtos antioxidantes em tecidos inflamados animais

Photodynamic therapy (PDT) consists of a non-toxic photosensitizing agent (FS) administration followed by a laser source resulting in a sequence of photochemical and photobiological processes that generate reactive oxygen species (ROS) that damaging cells. The present work evaluated the effects of PDT nanoemulsion-aluminum chloride phthalocyanine (AlClFc) mediated on malondialdehyde (MDA), glutathione (GSH), superoxide dismutase (SOD) and glutathione peroxidase (GPx) levels, which represent indicators involved in oxidative stress and antioxidant defenses. For this purpose, this study used 120 female rats of the Rattus norvegicus species, Wistar race, divided into 5 groups: Healthy (H), with periodontal disease (PD), with periodontal disease and treatment with FS (F), with periodontal disease and treatment with the laser (L); and periodontal disease and treatment with PDT (FL). An experimental model for represent periodontal disease (PD) was induced by ligature (split-mouth). Seven days later the induction of PD, the treatments were instituted according to the groups. In the group treated with PDT was applied 40μl FS (5μM) followed by laser irradiation diode InGaAlP (660nm, 100J / cm2). The rats were sacrificed on the 7th and 28th day after treatment and tissue specimens were removed and subjected to histological, immunohistochemical methods and enzymatic colorimetric measurements with detection by UV / VIS spectroscopy. Inflammatory changes, connective tissue disorganization and alveolar bone loss were displaying in groups with PD induced. The enzyme dosages showed that MDA levels were higher in PD induced groups, with no statistically significant differences (p> 0.05). High levels of GSH were found in groups L (p = 0.028) and FL (p = 0.028) compared with PD group, with statistically significant differences. Immunohistochemistry for SOD showed higher immunostaining in L and FL groups, compared to the PD group without statistically significant differences (p> 0.05). GPx showed lower immunoreactivity in the DP group when compared to the other groups and statistically significant differences were observed between the DPxL groups (p <0.05). TFD administered in this experiment did not induce elevation of MDA levels significantly increased the GSH levels and showed intense immunostaining pada SOD and GPx, showing that this therapy does not accentuated lipid peroxidation, however, it was able to induce effects on the antioxidant defenses processes. The LBI therapy appeared to show photomodulatory promoting effects reduction of the MDA levels, increasing GSH levels and with intense immunostaining for SOD and GPx, demonstrating that laser therapy induced antioxidant effects.

Modulação da expressão da proteína XPA em resposta ao tratamento com azul de metileno fotossensibilizado

Reactive oxygen species (ROS) are produced by aerobic metabolism and react with biomolecules, such as lipids, proteins and DNA. In high concentration, they lead to oxidative stress. Among ROS, singlet oxygen (1O2) is one of the main ROS involved in oxidative stress and is one of the most reactive forms of molecular oxygen. The exposure of some dyes, such as methylene blue (MB) to light (MB+VL), is able to generate 1O2 and it is the principle involved in photodynamic therapy (PDT). 1O2 e other ROS have caused toxic and carcinogenic effects and have been associated with ageing, neurodegenerative diseases and cancer. Oxidative DNA damage is mainly repaired by base excision repair (BER) pathway. However, recent studies have observed the involvement of nucleotide excision repair (NER) factors in the repair of this type of injury. One of these factors is the Xeroderma Pigmentosum Complementation Group A (XPA) protein, which acts with other proteins in DNA damage recognition and in the recruitment of other repair factors. Moreover, oxidative agents such as 1O2 can induce gene expression. In this context, this study aimed at evaluating the response of XPA-deficient cells after treatment with photosensitized MB. For this purpose, we analyzed the cell viability and occurrence of oxidative DNA damage in cells lines proficient and deficient in XPA after treatment with MB+VL, and evaluated the expression of this enzyme in proficient and complemented cells. Our results indicate an increased resistance to treatment of complemented cells and a higher level of oxidative damage in the deficient cell lines. Furthermore, the treatment was able to modulate the XPA expression up to 24 hours later. These results indicate a direct evidence for the involvement of NER enzymes in the repair of oxidative damage. Besides, a better understanding of the effects of PDT on the induction of gene expression could be provided

Avaliação ecológico-econômica do manguezal de Macau/RN e a importância da aplicação de práticas preservacionistas pela indústria petrolífera local

This work presents the results of a survey in oil-producing region of the Macau City, northern coast of Rio Grande do Norte. All work was performed under the Project for Monitoring Environmental Change and the Influence of Hydrodynamic forcing on Morphology Beach Grass Fields, Serra Potiguar in Macau, with the support of the Laboratory of Geoprocessing, linked to PRH22 - Training Program in Geology Geophysics and Information Technology Oil and Gas - Department of Geology/CCET/UFRN and the Post-Graduation in Science and Engineering Oil/PPGCEP/UFRN. Within the economic-ecological context, this paper assesses the importance of mangrove ecosystem in the region of Macau and its surroundings as well as in the following investigative exploration of potential areas for projects involving reforestation and / or Environmental Restoration. At first it was confirmed the ecological potential of mangrove forests, with primary functions: (i) protection and stabilization of the shoreline, (ii) nursery of marine life, and (iii) source of organic matter to aquatic ecosystems, (iv) refuge of species, among others. In the second phase, using Landsat imagery and techniques of Digital Image Processing (DIP), I came across about 18,000 acres of land that can be worked on environmental projects, being inserted in the rules signed the Kyoto Protocol to the market carbon. The results also revealed a total area of 14,723.75 hectares of activity of shrimp production and salting that can be harnessed for the social, economic and environmental potential of the region, considering that over 60% of this area, ie, 8,800 acres, may be used in the planting of the genus Avicennia considered by the literature that the species best sequesters atmospheric carbon, reaching a mean value of 59.79 tons / ha of mangrove

Efetividade do uso tópico de fluoreto e da escovação no controle de cáries produzidas "in vivo"

The objective of this clinical study was to evaluate the effectiveness of the toothbrushing with and without fluoride and the daily fluoride rinse (NaF 0.05%) on produced white spot, in vivo. This was a clinical study, controlled, randomized and triple blind. Thirty patients were selected for orthodontics reasons from Orthodontics Specialization Course at the Brazilian Dental Association - Section of Rio Grande do Norte. In this study it was used 4 bicuspid upper and lower. They had orthodontic reason for extractions, in 35 days, at least. The sample had one hundred and twenty teeth that received orthodontic bands. The bands were fixed with polycarboxylate cement, and there was a space standardized between bands and one surface of teeth. The four bicuspid of each patients were randomized and nominated as A, B, C and D. These nominations determinated the sequence of the extractions and what was done in each tooth. All the patients had been submitted to the toothbrushing with or without fluoride for 35 days. After this period, the A tooth of each patient was extracted to serve as control. The others teeth (B, C and D) were extracted one by each week. The entire sample was analyzed through the clinical examination and by laser fluorescence (DIAGNOdent®) in three different times: before orthodontic bands, 28 days after fixed and then removed the bands and, the last one, 07 days after one of the three treatments (toothbrushing with or without fluoride, tooth paste with fluoride and mouth rinse with fluoride). At the beginning all groups (A, B, C and D) had the same conditions, no significant difference was found. The same situation was found in a clinical examination. The results of the DIAGNOdent® for the groups that used tooth paste without fluoride, with fluoride and mouth rinse with fluoride, after 28 days, there was no significant difference. Clinically, the white spot was formed in all teeth after 28 days. When it was compared the three treated groups, the group without fluoride in tooth paste had worst result than the others groups. But there was no significant association between the number of active and inactive white spots and the type of treatment that the teeth had received. The demineralization of the enamel surface, under the orthodontic bands, it happened in a few weeks. The exposition of the white spots in oral environmental resulted in an improvement, but it was not enough to return to the values from the base line, either for the toothbrushing and/or the use of fluorite mouth rinse. Mouth rinse and toothpaste with fluoride have showed to reduce the incidence of demineralization in the enamel, but none seems to be superior to another one in an in vivo study

Sistema auxiliar na análise de mamogramas digitais

This work proposes the development of a Computer System for Analysis of Mammograms SCAM, that aids the doctor specialist in the identification and analysis of existent lesions in digital mammograms. The computer system for digital mammograms processing will make use of a group of techniques of Digital Image Processing (DIP), with the purpose of aiding the medical professional to extract the information contained in the mammogram. This system possesses an interface of easy use for the user, allowing, starting from the supplied mammogram, a group of processing operations, such as, the enrich of the images through filtering techniques, the segmentation of areas of the mammogram, the calculation the area of the lesions, thresholding the lesion, and other important tools for the medical professional's diagnosis. The Wavelet Transform will used and integrated into the computer system, with the objective of allowing a multiresolution analysis, thus supplying a method for identifying and analyzing microcalcifications

Análise molecular dos éxons 8 a 11 do gene da p53 em amostras de câncer de colo de útero no Rio Grande do Norte

Mutations on TP53 gene are common in human cancer but not in cervical cancer where they are rarely found and the inactivation and degradation of p53 protein are attributed to the action of E6 viral oncogene from high risk human papillomavirus (HPV). Analysis of cervical cancer cell lines suggests that HPV negative samples shows mutation on TP53, but clinical approaches didn t confirmed this hypothesis. However, in most TP53 mutations studies on cervical cancer, only the exons 5 to 8 were analyzed. Approximately 90% of mutations described are on this region. Recent studies on several cancer suggests that mutation frequency in the other exons must be considered. The aim of this work was to verify whether mutations on coding and non-coding regions occur in cancer tissue from cervical cancer in patients from Rio Grande do Norte using Denaturing Gradient Gel Electrophoresis (DGGE) as screening tool. Exons 8 to 11 were analyzed including some introns from 80 tumor samples and 8 peripheral blood samples from healthy women. DNA were submitted to PCR using primers with GC clamp on the end of one of them. The results were observed for each region after DGGE and silver staining. It was observed no amplified fragment with different migration profile from those obtained from DNA of peripheral blood. These results agree with those from literature where TP53 mutations in cervical cancer have been described in a very low frequency

Uso de técnicas de Geoprocessamento e Sensoriamento Remoto no levantamento e integração de dados necessários a gestão ambiental dos campos de extração de óleo e gás do Canto do Amaro e Alto da Pedra no município de Mossoró - RN

The objective of this work is to identify, to chart and to explain the evolution of the soil occupation and the envirionment vulnerability of the areas of Canto do Amaro and Alto da Pedra, in the city of Mossoró-RN, having as base analyzes it multiweather of images of orbital remote sensors, the accomplishment of extensive integrated works of field to a Geographic Information System (GIS). With the use of inserted techniques of it analyzes space inserted in a (GIS), and related with the interpretation and analyzes of products that comes from the Remote Sensoriamento (RS.), make possible resulted significant to reach the objectives of this works. Having as support for the management of the information, the data set gotten of the most varied sources and stored in digital environment, it comes to constitute the geographic data base of this research. The previous knowledge of the spectral behavior of the natural or artificial targets, and the use of algorithms of Processing of Digital images (DIP), it facilitates the interpretation task sufficiently and searchs of new information on the spectral level. Use as background these data, was generated a varied thematic cartography was: Maps of Geology, Geomorfológicals Units soils, Vegetation and Use and Occupation of the soil. The crossing in environment SIG, of the above-mentioned maps, generated the maps of Natural and Vulnerability envirionmental of the petroliferous fields of I Canto do Amaro and Alto da Pedra-RN, working in an ambient centered in the management of waters and solid residuos, as well as the analysis of the spatial data, making possible then a more complex analysis of the studied area

Avaliação de lesões brancas de cáries: um estudo de prognóstico

Objective: The aim of the present study is to assess the current situation of white enamel lesions on vestibular surfaces of permanent upper incisors, diagnosed 6 years ago, without clinical intervention. Methods: A prospective study reassessed 53 students of both sexes, aged between 13 and 18 years old, all attending the public school system in Natal, Brazil. Data collection was performed by duly calibrated examiners, and a clinical chart consisting of demographic data on dental caries, oral hygiene, and gingival condition was prepared. A tactile-visual examination was conducted using a clinical mirror and periodontal probe. Data compilation and analysis were carried out using a SPSS software. In this analysis the chi-squared test was used for qualitative independent variables. To identify the net effect of treatment, multiple logistic analysis with forward stepwise model selection was performed. Results: The final sample was composed of 106 lesions in the 53 individuals, with mean age of 15.02 years, visible plaque index (VPI) of 23.34%, and gingival blood indices (GBI) of 25.92%. A statistically significant relationship (p = 0.003) was found between initial DMFS and prognosis of white enamel lesion. Conclusions: We observed that past caries experience and dental plaque were the main predictive factors for negative lesion outcomes, demonstrating the need for oral hygiene control through continuing preventive measures

Diferenças funcionais das regiões hipocampais na formação da memória do tipo o quê", "quando" e "onde" em ratos

Episodic memory refers to the recollection of what, where and when a specific event occurred. Hippocampus is a key structure in this type of memory. Computational models suggest that the dentate gyrus (DG) and the CA3 hippocampal subregions are involved in pattern separation and the rapid acquisition of episodic memories, while CA1 is involved in memory consolidation. However there are few studies with animal models that access simultaneously the aspects ‗what-where-when . Recently, an object recognition episodic-like memory task in rodents was proposed. This task consists of two sample trials and a test phase. In sample trial one, the rat is exposed to four copies of an object. In sample trial two, one hour later, the rat is exposed to four copies of a different object. In the test phase, 1 h later, two copies of each of the objects previously used are presented. One copy of the object used in sample trial one is located in a different place, and therefore it is expected to be the most explored object.However, the short retention delay of the task narrows its applications. This study verifies if this task can be evoked after 24h and whether the pharmacological inactivation of the DG/CA3 and CA1 subregions could differentially impair the acquisition of the task described. Validation of the task with a longer interval (24h) was accomplished (animals showed spatiotemporal object discrimination and scopolamine (1 mg/kg, ip) injected pos-training impaired performance). Afterwards, the GABA agonist muscimol, (0,250 μg/μl; volume = 0,5 μl) or saline were injected in the hippocampal subregions fifteen minutes before training. Pre-training inactivation of the DG/CA3 subregions impaired the spatial discrimination of the objects (‗where ), while the temporal discrimination (‗when ) was preserved. Rats treated with muscimol in the CA1 subregion explored all the objects equally well, irrespective of place or presentation time. Our results corroborate the computational models that postulate a role for DG/CA3 in spatial pattern separation, and a role for CA1 in the consolidation process of different mnemonic episodes

Índice de aromaticidade baseado na deslocalização, degenerescência e densidade (D3BIA) para acenos e moléculas homoaromáticas

The aromaticity index is an important tool for the investigation of aromatic molecules. This work consists on new applications of the aromaticity index developed by teacher Caio Lima Firme, so-called D3BIA (density, delocalization, degeneracy-based index of aromaticity). It was investigated its correlation with other well-known aromaticity indexes, such as HOMA (harmonic oscillator model of aromaticity), NICS (nucleus independent chemical shielding), PDI (para-delocalization index), magnetic susceptibility (), and energetic factor in the study of aromaticity of acenes and homoaromatic species based on bisnoradamantanyl cage. The density functional theory (DFT) was used for optimization calculations and for obtaining energetic factors associated with aromaticity and indexes HOMA and NICS. From quantum theory of atoms in molecules (QTAIM) it was obtained the indexes D3BIA, PDI and . For acenes, when the over-mentioned indexes were applied it was observed no correlation except for D3BIA and HOMA (R2=0.752). For bisnoradamantenyl dication and its derivatives, it was obtained a good correlation between D3BIA and NICS. Moreover, it was evaluated solely one of the factors used on D3BIA calculation, the delocalization index uniformity (DIU), so as to investigate its possible influence on stability of chemical species. Then, the DIU was compared with the formation Gibbs free energy of some pairs of carbocations, isomers or not, which each pair had small difference in point group symmetry and no difference among other well-known stability factors. The obtained results indicate that DIU is a new stability factor related to carbocations, that is, the more uniform the electron density delocalization, the more stable the is carbocation. The results of this work validate D3BIA and show its importance on the concept of aromaticity, indicating that it can be understood from degeneracy of atoms belonging the aromatic site, the electronic density in the aromatic site and the degree of uniformity of electron delocalization

A temática ambiental na educação superior : políticas, gestão acadêmica e projetos de formação nos cursos da Universidade do Estado do Rio Grande do Norte

Conselho Nacional de Desenvolvimento Científico e Tecnológico

Caracterização morfológica e sedimentologica da plataforma, continental brasileira adjacente aos municípios de Fortim, Aracati e Icapuí- CE

The study area is located on the Brazilian Continental Shelf adjacent to Ceará State, inserted in the submerged Potiguar Basin. This area was submitted to extensional efforts during Upper Cretaceous, associated to the begining of the rifting that resulted in African and South American Continent separation. The main goal of this research was to better understand the sedimentary and geomorphological characteristics of the continental shelf adjacent to Fortim, Aracati and Icapuí (Ceará State). The used data base included geophysical (sides scan sonar and bathymetry studies) and sedimentological survey, associated to satellite image processing and interpretation. Inferences about suspended material and longshore drift was possible using satellite images, and differente bedforms were characterized such as: different kinds of dunes (longitudinal, cross and oblique), bioclastic banks, paleochannels, flat and rock bottom. The researched area comprehended about 2509,13 km2, where 6 different sedimentary facies, based on sediment composition and texture, could be recognized, such as: Bioclastic Sand, Siliciclastic Sand, Biosiliciclastic Sand, Bioclastic gravel, Biosiliciclastic sand with granule and gravel, and Silicibioclastic sand with granule and gravel. The integration of bathymetric, satellite image, side scan sonar and sedimentological data allow us a better characterization of this continental shelf area, as to advance in the knowledge of the continental shelf of the state of Ceara, a very important area to the oil industry because of its potential exploration and e exploitation, and to environmental survey as well

Análise geoambiental da plataforma continental Rasa Equatorial, por meio de sensoriamento remoto e dados sedimentológicos: parrachos de Rio do Fogo-RN, Brasil

In the shallow continental shelf in Northeastern Rio Grande do Norte - Brazil, important underwater geomorphological features can be found 6km from the coastline. They are coral reefs, locally known as “parrachos”. The present study aims to characterize and analyze the geomorphological feature as well as the ones of the benthic surface, and the distribution of biogenic sediments found in parrachos at Rio do Fogo and associated shallow platforms, by using remote sensing products and in situ data collections. This was made possible due to sedimentological, bathymetric and geomorphological maps elaborated from composite bands of images from the satellite sensors ETM+/Landsat-7, OLI/Landsat-8, MS/GeoEye and PAN/WordView-1, and analysis of bottom sediments samples. These maps were analyzed, integrally interpreted and validated in fieldwork, thus permitting the generation of a new geomorphological zoning of the shallow shelf in study and a geoenvironmental map of the Parrachos in Rio do Fogo. The images used were subject to Digital Image Processing techniques. All obtained data and information were stored in a Geographic Information System (GIS) and can become available to the scientific community. This shallow platform has a carbonate bottom composed mostly by algae. Collected and analyzed sediment samples can be classified as biogenic carbonatic sands, as they are composed 75% by calcareous algae, according to the found samples. The most abundant classes are green algae, red algae, nonbiogenic sediments (mineral grains), ancient algae and molluscs. At the parrachos the following was mapped: Barreta Channel, intertidal reefs, submerged reefs, the spur and grooves, the pools, the sandy bank, the bank of algae, sea grass, submerged roads and Rio do Fogo Channel. This work presents new information about geomorphology and evolution in the study area, and will be guiding future decision making in the handling and environmental management of the region