ABSTRACT Background: Depressive symptomatology in health professionals associated with viral crises is highly prevalent globally, being its detection a priority. Therefore, the objective of this research was to analyze the convergent validity and internal consistency of the Patient Health Questionnaire (PHQ-2) in healthcare professionals. Method: E-Health study where cross-sectional data was collected online (n = 725), from 281 general practitioners, 237 specialist doctors and 207 nurses during the Colombian quarantine, between April 20 and August 10, 2020. Average age 41.3 years (± 8.76). 38.4% were men (278) and 61.6% women (447). 66.1% of health personnel treated patients infected with coronavirus and 33.9% did not provide these services. The 9-item version of the PHQ was administered, validated in the Colombian population together with the 2-item version of the PHQ. Results: A high correlation was found between the scales (r = .860, P <0.001), demonstrating the convergent validity of the PHQ-2 to measure depressive symptomatology. The internal consistency of the PHQ-2 was adequate, with a Cronbach’s Alpha of 0.80 (I.C. = 0.76 - 0.83). Conclusions: The PHQ-2 has adequate psychometric standards of reliability and validity, so its rapid administration, easy qualification and interpretation, makes it a reliable and valid instrument for the rapid detection, without work overload, of depressive symptoms in doctors and nurses whether or not they care for patients with viral outbreaks.

RESUMEN: Introducción: La sintomatologia depresiva en profesionales de la salud asociado a crisis virales es de alta prevalencia a nivel global, siendo su detección una prioridad por lo cual, el objetivo de esta investigación fue analizar la validez convergente y consistencia interna del cuestionario de salud del paciente-2 (PHQ-2) en profesionales sanitarios. Método: Estudio eSalud donde se recopilaron datos transversales en línea (n=725), de 281 médicos generales, 237 médicos especialistas y 207 enfermeras durante la cuarentena colombiana, entre el 20 de abril y el 10 de agosto de 2020. Edad promedio 41,3 años (± 8,76). El 38,4% eran hombres (278) y el 61.6% mujeres (447). El 66.1% del personal sanitario atendió pacientes contagiados por coronavirus y el 33,9% no prestó estos servicios. Se administró la versión de 9 ítems del PHQ, validada en población colombiana junto a la versión de 2 ítems del PHQ. Resultados: Se encontró una alta correlación entre las escalas (r=.860, P<0.001), demostrando la validez convergente del PHQ-2 para medir la sintomatología depresiva. La consistencia interna del PHQ-2 fue adecuada, con un Alpha de Cronbach de 0.80 (I.C.= 0.76 - 0.83). Conclusiones: El PHQ-2 presenta adecuados estándares psicométricos de confiabilidad y validez, por lo que su rápida administración, fácil calificación e interpretación, lo convierte en un instrumento confiable y valido para la detección rápida, sin sobrecargas laborales, de los síntomas depresivos en médicos y enfermeras que atiendan o no pacientes en condiciones de brotes virales.

The correlation between iron distribution in octahedral or tetrahedral sites in synthetic magnetite and zinc spinel ferrites and their catalytic properties were investigated in this work. The zinc-doped nanomagnetites were prepared by co-precipitation method and annealed at 420 °C. X-ray diffraction, UV-Vis, X-ray absorption and 57Fe Mössbauer spectroscopies were used to identify iron sites. The obtained results showed that ZnII replaces the relatively smaller FeIII cation in the tetrahedral sites, increasing the cubic lattice dimension, and this replacement remarkably enhances the catalytic effectiveness of the Zn-spinel ferrite towards the indigo carmine degradation compared to undoped magnetite. The indigo carmine degradation experiments were carried out using photo-Fenton method in which 50 mL of indigo carmine dye aqueous solutions (20 mg L−1, pH 3.0), 20 mg ZnxFe(3-x)O4, and 2.0 mL H2O2 (aqueous solution at 0.028 mol L−1) were placed under UV light at 25 °C. Under these experimental conditions the degradation rate of indigo carmine was superior to 98%.

Abstract This study aims to evaluate the in silico genomic characteristics of Streptococcus infantarius subsp. infantarius, isolated from Coalho cheese from Paraíba, Brazil, with a view to application in lactic fermentations. rRNA sequences from the 16S ribosomal region were used as input to GenBank, in the search for patterns that could reveal a non-pathogenic behavior of S. infantarius subsp. infantarius, comparing mobile genetic elements, antibiotic resistance genes, pan-genome analysis and multi-genome alignment among related species. S. infantarius subsp. infantarius CJ18 was the only complete genome reported by BLAST/NCBI with high similarity and after comparative genetics with complete genomes of Streptococcus agalactiae (SAG153, NJ1606) and Streptococcus thermophilus (ST106, CS18, IDCC2201, APC151) revealed that CJ18 showed a low number of transposases and integrases, infection by phage bacteria of the Streptococcus genus, absence of antibiotic resistance genes and presence of bacteriocin, folate and riboflavin producing genes. The genome alignment revealed that the collinear blocks of S. thermophilus ST106 and S. agalactiae SAG153 have inverted blocks when compared to the CJ18 genome due to gene positioning, insertions and deletions. Therefore, the strains of S. infantarius subsp. infantarius isolated from Coalho cheese from Paraíba showed genomic similarity with CJ18 and the mobility of genes analyzed in silico showed absence of pathogenicity throughout the genome of CJ18, indicating the potential of these strains for the dairy industry.

Abstract Fibrinolytic enzymes are considered promising alternative in the treatment of cardiovascular diseases by preventing fibrin clots. A protease from Mucor subtilissimus UCP 1262 was obtained by solid state fermentation and purified by ion exchange chromatography. The purified extract was administered at an acute dose of 2000 mg/mL to evaluate its toxic effects to the lungs of mice. After 14 days of treatment, a histomorphometric study was performed by the type 1 and 2 pneumocyte count and the evaluation of the lung area. As result, the experimental group showed a significant decrease of type 2 pneumocyte and although a decrease in the alveolar area was observed in relation to the control group, no significant pulmonary toxicity, emphysema, and fibrosis characteristics were detected. The in vitro tests suggest possible clinical applications for the enzyme.

Abstract Objective Allergic sensitization is one of the key components for the development of allergies. Polysensitization seems to be related to the persistence and severity of allergic diseases. Furthermore, allergic sensitization has a predictive role in the development of allergies. The aim of this study was to characterize the pattern of sensitization of atopic patients treated at different pediatric allergy referral centers in Brazil. Methods A nation-wide transversal multicenter study collected data on patients attended in Brazil. Peripheral blood samples were collected to determine the serum levels of allergen-specific IgE. If allergen-specific IgE was higher than 0.1 kUA/L, the following specific components were quantified. Results A total of 470 individuals were enrolled in the study. Mite sensitization was the most frequent kind in all participants. A high frequency of sensitization to furry animals and grasses featured in the respiratory allergies. Regarding components, there was a predominance of sensitization to Der p 1 and Der p 2. It has been verified that having a food allergy, atopic dermatitis, or multimorbidity are risk factors for the development of more severe allergic disease. Conclusion Studies on the pattern of allergic sensitization to a specific population offer tools for the more effectual prevention, diagnosis, and treatment of allergic diseases. Sensitization to dust mites house was the most prevalent in the evaluated sample. High rates of sensitization to furry animals also stand out. Patients with food allergy, atopic dermatitis, or multimorbidity appear to be at greater risk for developing more severe allergic diseases.

Citrus biomass is a source of biobased products and presents extensive chemical diversity. Among the structural macromolecules, lignins have been used in resins, bioplastics, and nanoencapsulation. Brazil has a large source of lignins, which are orange trees removed from groves that need to be renewed. However, the knowledge about these lignins is limited and little explored. Thus, this work aimed to evaluate the lignins from Citrus sinensis and to provide information on the interesting use of orange trees as a source of macromolecules. The lignins were isolated from the trunk, central and secondary roots from the orange tree by Technical Association of the Pulp and Paper Industry (TAPPI)-methods. The lignin contents for the extractive-free samples were 19.8 ± 0.19, 21.9 ± 0.36, and 19.5 ± 0.41% (m/m) in the trunk, central and secondary roots, respectively. Infrared (IR) and 13C nuclear magnetic resonance (NMR) analyses confirmed typical bands and chemical shifts to syringyl and guaiacyl units. These results are consistent with the features of lignins previously reported for orange tree branches and hardwood roots. The greatest amount of lignin in the orange tree was found in the central root. The lignins extractive-free by TAPPI-T264 cm-97 presented low solubility in organic solvents, due to their compaction and folding, which was confirmed by 13C NMR in the solid-state, differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA).

Abstract Fibrinolytic proteases are a promising alternative in the pharmaceutical industry, they are used in the treatment of cardiovascular diseases, especially thrombosis. Microorganisms are the most interesting source of fibrinolytic proteases. The aim of this study was the production of fibrinolytic protease from Streptomyces parvulus DPUA 1573, the recovery of the protease by aqueous two-phase system and partial biochemical characterization of the enzyme. The aqueous two-phase system was performed according to a 24-full factorial design using polyethylene glycol molar mass, polyethylene glycol concentration, citrate concentration and pH as independent variables. It was analyzed the effect of different ions, surfactants, inhibitors, pH and temperature on enzyme activity. The best conditions for purifying the enzyme were 17.5% polyethylene glycol 8,000, 15% Phosphate and pH 8.0, it was obtained a partition coefficient of 7.33, a yield of 57.49% and a purification factor of 2.10-fold. There was an increase in enzyme activity in the presence of Fe2+ and a decrease in the presence of $\beta$-Mercaptoethanol, phenylmethylsulfonyl fluoride and Iodoacetic acid. The optimum pH was 7.0 and the optimum temperature was 40 ºC. The purified protease exhibited a molecular mass of 41 kDa. The fibrinolytic protease from Streptomyces parvulus proved to be a viable option for the development of a possible drug with fibrinolytic action.

Abstract The industrial demand for proteolytic enzymes is stimulating the search for new enzyme sources. Fungal enzymes are preferred over bacterial enzymes, and more effective and easier to extract. The aim of this work was to evaluate the potential of protease production by solid state fermentation (SSF) of Mucor subtilissimus UCP 1262, evaluate different specific activities, purify and partially characterize the enzyme in terms of biochemical as to the optimal pH and temperature. Initially, the enzyme crude extract was screened for 3 different proteolytic activities, collagenolytic (161.4 U/mL), keratinolytic (39.6 U/mL) and fibrinolytic (26.1 U/mL) in addition to conventional proteinase activity. After ammonium sulfate precipitation, the active fractions with fibrinolytic activity were dialyzed in 15 mM Tris-HCl buffer, pH 8, loaded onto DEAE-Sephadex A50 ion-exchange column and gel filtrated through Superdex 75 HR10/300. The enzyme showed a fibrinolytic maximum activity at 40 C and pH 9,0. The purified enzyme showed activity against a chromogenic chymotrypsin substrate, SDS-PAGE showing a molecular mass of approximately 70 kDa and, the specific activity of 25.93 U/mg. These characteristics suggest that the enzyme could be and efficiently produced in a simple and low-cost way using Mucor subtilissimus UCP 1262 in SSF.

Resumo: Estudos realizados nos países em desenvolvimento descrevem um aumento progressivo dos transtornos mentais, com grandes repercussões sociais e econômicas. O presente estudo se propõe a analisar a prevalência dos transtornos mentais comuns (TMC) e seus fatores associados nos moradores da área urbana da cidade de São Paulo, Brasil. A partir dos dados levantados pelo Inquérito de Saúde do Município de São Paulo (ISA-Capital) de 2015, foi identificada a presença de TMC com a utilização do Self-Reporting Questionnaire (SRQ-20). A associação de TMC com as variáveis sociodemográficas e de condições de saúde foi analisada pela frequência relativa, corrigida pelos respectivos pesos decorrentes de uma amostragem de conglomerados, estimando-se a prevalência e intervalos de 95% de confiança (IC95%) em cada situação e avaliando-se a significância da associação pelo qui-quadrado corrigido pela distribuição F. A prevalência de TMC encontrada foi de 19,7% (IC95%: 18,2-21,4), sendo maior nas seguintes categorias: mulheres (24,3%); pessoas com 60 anos ou mais (25,3%); praticantes da umbanda/candomblé (37,8%); viúvos (30,4%); quem nunca frequentou a escola (31,4%); inativos/desempregados (28,3%); e quem possuía renda familiar de até um salário mínimo (28,8%). Em relação às variáveis de condição de saúde, a prevalência de TMC foi maior nos indivíduos que referiram: morbidade nos últimos 15 dias (36,9%); deficiência física (21,6%); deficiência mental ou intelectual (44,4%); problema emocional ou mental (48,9%); cefaleia (33,63%); e nos portadores de uma ou mais doenças crônicas (24,1%). As informações desta pesquisa reafirmam a relevância da prevalência de TMC assim como sua associação com os grupos sociais mais vulneráveis, o que reforça a necessidade da implementação de medidas públicas de saúde mental.

Abstract: Studies in developing countries report a steady increase in mental disorders, with major social and economic repercussions. The current study proposes to analyze the prevalence of common mental disorders (CMDs) and associated factors in urban residents of São Paulo, Brazil. Based on data collected in the Health Survey in São Paulo City (ISA-Capital) in 2015, the study identified the presence of CMDs using the Self-Reporting Questionnaire (SRQ-20). The association of CMDs with sociodemographic variables and health conditions was analyzed as relative frequency, corrected by the respective weights resulting from cluster sampling, estimating the prevalence and 95% confidence intervals (95%CI) and assessing the association’s significance by the chi-square test, corrected by the F distribution. Prevalence of CMDs was 19.7% (95%CI: 18.2-21.4), higher in women (24.3%); persons 60 years or older (25.3%); followers of the African-Brazilian umbanda or candomblé religions (37.8%); widows/widowers (30.4%); individuals that had never attended school (31.4%); unemployed (28.3%); those with family income up to one minimum wage (28.8%); individuals that reported illness in the previous 15 days (36.9%); those with physical disabilities (21.6%); mental or intellectual disabilities (44.4%); emotional or mental problems (48.9%); headache (33.63%); and individuals with one or more chronic diseases (24.1%). The information in this study reaffirms the relevance of the prevalence of CMDs and their association with the most vulnerable social groups, corroborating the need to implement public measures in mental health.

Resumen: Estudios realizados en los países en desarrollo describen un aumento progresivo de los trastornos mentales, con grandes repercusiones sociales y económicas. El presente estudio se propone analizar la prevalencia de los trastornos mentales comunes (TMC) y sus factores asociados en los habitantes del área urbana de la ciudad de São Paulo, Brasil. A partir de los datos recabados por la Encuesta de Salud en el Municipio de São Paulo (ISA-Capital) de 2015, se identificó la presencia de TMC con la utilización del Self-Reporting Questionnaire (SRQ-20). La asociación de TMC con las variables sociodemográficas y de condiciones de salud se analizó a través de la frecuencia relativa, corregida por los respectivos pesos derivados de una muestra de conglomerados, estimándose la prevalencia y intervalos de confianza (IC95%) y evaluándose la significancia de la asociación por el chi-cuadrado, corregido por la distribución F. La prevalencia de TMC encontrada fue de 19,7% (IC95%: 18,2-21,4), siendo mayor en mujeres (24,3%); personas con 60 años o más (25,3%); practicantes de umbanda/candomblé (37,8%); viudos (30,4%); que nunca asistieron a la escuela (31,4%); inactivos/desempleados (28,3%); quienes tenía una renta familiar de hasta un salario mínimo (28,8%); individuos que informaron morbilidad durante los últimos 15 días (36,9%); deficiencia física (21,6%); deficiencia mental o intelectual (44,4%); problema emocional o mental (48,9%); cefalea (33,63%); y en los portadores de una o más enfermedades crónicas (24,1%). La información de esta investigación reafirma la relevancia de la prevalencia de TMC, así como su asociación con los grupos sociales más vulnerables, lo que refuerza la necesidad de la implementación de medidas públicas de salud mental.

Resumo: Estudos realizados nos países em desenvolvimento descrevem um aumento progressivo dos transtornos mentais, com grandes repercussões sociais e econômicas. O presente estudo se propõe a analisar a prevalência dos transtornos mentais comuns (TMC) e seus fatores associados nos moradores da área urbana da cidade de São Paulo, Brasil. A partir dos dados levantados pelo Inquérito de Saúde do Município de São Paulo (ISA-Capital) de 2015, foi identificada a presença de TMC com a utilização do Self-Reporting Questionnaire (SRQ-20). A associação de TMC com as variáveis sociodemográficas e de condições de saúde foi analisada pela frequência relativa, corrigida pelos respectivos pesos decorrentes de uma amostragem de conglomerados, estimando-se a prevalência e intervalos de 95% de confiança (IC95%) em cada situação e avaliando-se a significância da associação pelo qui-quadrado corrigido pela distribuição F. A prevalência de TMC encontrada foi de 19,7% (IC95%: 18,2-21,4), sendo maior nas seguintes categorias: mulheres (24,3%); pessoas com 60 anos ou mais (25,3%); praticantes da umbanda/candomblé (37,8%); viúvos (30,4%); quem nunca frequentou a escola (31,4%); inativos/desempregados (28,3%); e quem possuía renda familiar de até um salário mínimo (28,8%). Em relação às variáveis de condição de saúde, a prevalência de TMC foi maior nos indivíduos que referiram: morbidade nos últimos 15 dias (36,9%); deficiência física (21,6%); deficiência mental ou intelectual (44,4%); problema emocional ou mental (48,9%); cefaleia (33,63%); e nos portadores de uma ou mais doenças crônicas (24,1%). As informações desta pesquisa reafirmam a relevância da prevalência de TMC assim como sua associação com os grupos sociais mais vulneráveis, o que reforça a necessidade da implementação de medidas públicas de saúde mental.

Abstract: Studies in developing countries report a steady increase in mental disorders, with major social and economic repercussions. The current study proposes to analyze the prevalence of common mental disorders (CMDs) and associated factors in urban residents of São Paulo, Brazil. Based on data collected in the Health Survey in São Paulo City (ISA-Capital) in 2015, the study identified the presence of CMDs using the Self-Reporting Questionnaire (SRQ-20). The association of CMDs with sociodemographic variables and health conditions was analyzed as relative frequency, corrected by the respective weights resulting from cluster sampling, estimating the prevalence and 95% confidence intervals (95%CI) and assessing the association’s significance by the chi-square test, corrected by the F distribution. Prevalence of CMDs was 19.7% (95%CI: 18.2-21.4), higher in women (24.3%); persons 60 years or older (25.3%); followers of the African-Brazilian umbanda or candomblé religions (37.8%); widows/widowers (30.4%); individuals that had never attended school (31.4%); unemployed (28.3%); those with family income up to one minimum wage (28.8%); individuals that reported illness in the previous 15 days (36.9%); those with physical disabilities (21.6%); mental or intellectual disabilities (44.4%); emotional or mental problems (48.9%); headache (33.63%); and individuals with one or more chronic diseases (24.1%). The information in this study reaffirms the relevance of the prevalence of CMDs and their association with the most vulnerable social groups, corroborating the need to implement public measures in mental health.

Resumen: Estudios realizados en los países en desarrollo describen un aumento progresivo de los trastornos mentales, con grandes repercusiones sociales y económicas. El presente estudio se propone analizar la prevalencia de los trastornos mentales comunes (TMC) y sus factores asociados en los habitantes del área urbana de la ciudad de São Paulo, Brasil. A partir de los datos recabados por la Encuesta de Salud en el Municipio de São Paulo (ISA-Capital) de 2015, se identificó la presencia de TMC con la utilización del Self-Reporting Questionnaire (SRQ-20). La asociación de TMC con las variables sociodemográficas y de condiciones de salud se analizó a través de la frecuencia relativa, corregida por los respectivos pesos derivados de una muestra de conglomerados, estimándose la prevalencia y intervalos de confianza (IC95%) y evaluándose la significancia de la asociación por el chi-cuadrado, corregido por la distribución F. La prevalencia de TMC encontrada fue de 19,7% (IC95%: 18,2-21,4), siendo mayor en mujeres (24,3%); personas con 60 años o más (25,3%); practicantes de umbanda/candomblé (37,8%); viudos (30,4%); que nunca asistieron a la escuela (31,4%); inactivos/desempleados (28,3%); quienes tenía una renta familiar de hasta un salario mínimo (28,8%); individuos que informaron morbilidad durante los últimos 15 días (36,9%); deficiencia física (21,6%); deficiencia mental o intelectual (44,4%); problema emocional o mental (48,9%); cefalea (33,63%); y en los portadores de una o más enfermedades crónicas (24,1%). La información de esta investigación reafirma la relevancia de la prevalencia de TMC, así como su asociación con los grupos sociales más vulnerables, lo que refuerza la necesidad de la implementación de medidas públicas de salud mental.

Currently, most food waste is used as animal feed and this process does not take advantage of the available chemical composition. A Brazilian example is the citrus fruit processing wastes (CFPW), which have begun to draw attention due to their biological importance. In order to access the main compounds of this matrix, an efficient and environmentally friendly procedure was tested. From this extract, flavonoids as naringenin, hesperitin, chrysoeriol, sinensetin, 3,5,6,7,3',4'-hexamethoxyflavone, nobiletin, 5-methoxysalvigenin, 3,5,6,7,8,3',4'-heptamethoxyflavone, 3,5,6,7,4'-pentamethoxyflavone and isosakuranetin were identified by high performance liquid chromatography (HPLC-UV), liquid chromatography-mass spectrometry (LC-MS) and liquid chromatography-nuclear magnetic resonance (LC-NMR). This study describes the potential use of citrus waste as a source of biologically active compounds. The extraction method proposed for this work was not expensive and the flavonoids were obtained in large amounts, thus, this extraction method is being developed using pilot plant scale-up techniques and will soon be available to the industry at a low cost.

RESUMO A ocorrência de doenças radiculares representa uma das principais causas de perda de rendimento na cultura do melão, com destaque para patógenos causadores das podridões de raízes e colos, como os fungos Fusarium solani, Macrophomina phaseolina e Rhizoctonia solani, observados no meloeiro tanto de forma isolada quanto associada. A utilização de material vegetal e restos de culturas, além de melhorar as propriedades físicas do solo e favorecer a atividade microbiana neste ambiente, pode interferir negativamente sobre a população de patógenos. Portanto, o objetivo deste trabalho foi verificar o efeito da utilização do Feijão-de-porco [Canavalia ensiformis (L.) DC] no manejo da podridão radicular do meloeiro (Cucumis melo L.) causada pelos patógenos F. solani, M. phaseolina e R. solani, quando sozinhos ou associados. O experimento foi realizado em casa de vegetação no delineamento de blocos casualizados, com esquema fatorial 8 x 4, sendo 8 tipos de fungos/associações e 4 formas de manejo, com 4 repetições. As características avaliadas foram: incidência de podridão radicular das plantas de melão no final do ciclo, prevalência dos patógenos e a massa da matéria seca do meloeiro. O feijão-de-porco em cobertura proporciona menor incidência de podridão radicular no meloeiro quando Fusarium solani está sozinho. Macrophomina phaseolina é o fungo que prevalece na maioria das associações nas quais está presente. Aumento na massa da matéria seca do meloeiro foi observado quando aplicado o mulch (filme de polietileno preto) nos tratamentos com associação de F. solani + R. solani e no solo não infestado.

ABSTRACT The occurrence of root diseases represent one of the major causes of yield loss in melon crop, especially considering pathogens that cause root and stem rot, such as the fungi Fusarium solani, Macrophomina phaseolina and Rhizoctonia solani, observed in muskmelon either alone or in association. The use of plant materials and crop residues, besides improving the soil physical properties and favoring the microbial activity in this environment, negatively interferes in the population of pathogens. Therefore, the aim of this study was to verify the effect of using jack beans [Canavalia ensiformis (L.) DC] in the management of root rot of muskmelon (Cucumis melo L.) caused by the pathogens F. solani, M. phaseolina and R. solani either alone or in association. The experiment was carried out in a greenhouse in randomized block design, with 8 x 4 factorial arrangement, 8 types of fungi/associations and 4 management systems, and 4 replicates. The evaluated characteristics were: incidence of root rot in melon plants at the end of the cycle, prevalence of pathogens and dry weight of muskmelon. Jack beans in coverage provides lower incidence of root rot in muskmelon when Fusarium solani is alone. Macrophomina phaseolina is the fungus that prevails in most associations in which it is present. Increased dry weight of muskmelon was observed when mulch (the black polyethylene film) was applied in the treatments with F. solani + R. solani association and in non-infested soil.

Resumo Exposição a altos níveis de flúor durante a amelogênese causa fluorose no esmalte. Este estudo tem como objetivo determinar e comparar as sequências de aminoácidos presentes no esmalte de dentes controles e fluoróticos. A investigação incluiu amostras de esmalte obtidas de terceiros molares erupcionados e não erupcionados, ambas ou com grau de fluorose TF 4-6 (n=7) ou sem sinais de fluorose (controles, n=7), congelados a -20 oC até a extração das proteínas. As amostras sofreram ataque ácido e foram processadas utilizando um coquetel de inibidores de proteinases, sendo imediatamente analisadas. MALDI-TOF/TOF seguido pela pesquisa com MASCOT foram utilizados para a análise dos peptídeos. Os peptídeos mais abundantes foram das amelogeninas com sequências N-terminal WYQSIRPPYP (que é codificada especificamente pela amelogenina X) e MPLPPHPGHPGYINF (que não apresenta dimorfismo sexual algum), não havendo diferenças entre dentes fluoróticos e controles. Nenhuma alteração na proteólise ocorreu nas amostras fluoróticas, o que sugere que o aumento na quantidade de proteínas existentes nas amostras fluoróticas não está correlacionada a habilidade das proteinases em clivar as proteínas em humanos. Este estudo mostrou como extrair com sucesso peptídeos do esmalte superficial. Um número relativamente baixo de dentes foram suficientes para se obter ótimos dados a respeito de peptídeos encontrados no esmalte maduro.

Abstract Exposure to high fluoride levels during amelogenesis causes enamel fluorosis. This study aimed to determine and compare the amino acid sequences in the enamel of fluorotic and control teeth. This investigation included enamel samples obtained from erupted and non-erupted third molars with either TF grade 4-6 (n=7) fluorosis or no sign of fluorosis (controls, n=7). The samples were kept frozen at -20 °C until protein extraction. Samples were etched and processed with a cocktail of proteinase inhibitors and immediately analyzed. Matrix Assisted Laser Desorption/Ionization-Time-Of-Flight/Time-of-Flight Mass Spectrometry (MALDI-TOF/TOF) followed by MASCOT search aided the peptides analysis. The more abundant peptides bore the N-terminal amelogenin sequences WYQSIRPPYP (which is specific for the X-encoded amelogenin) and MPLPPHPGHPGYINF (which does not show sexual dimorphism) were not different in control or fluorotic enamel. There was no missing proteolytic cleavage in the fluorotic samples, which suggested that the increased amount of protein described in fluorotic enamel did not stem from the decreased ability of proteinases to cleave the proteins in humans. This study showed how to successfully obtain peptide from superficial enamel. A relatively low number of teeth was sufficient to provide good data on the actual peptides found in mature enamel.