Salivary protein candidates for biomarkers of oral disorders in people with a crack cocaine use disorder

Abstract The use of cocaine and its main derivative, crack, can cause some systemic effects that may lead to the development of some oral disorders. Objective To assess the oral health of people with a crack cocaine use disorder and identify salivary protein candidates for biomarkers of oral disorders. Methodology A total of 40 volunteers hospitalized for rehabilitation for crack cocaine addiction were enrolled; nine were randomly selected for proteomic analysis. Intraoral examination, report of DMFT, gingival and plaque index, xerostomia, and non-stimulated saliva collection were performed. A list of proteins identified was generated from the UniProt database and manually revised. Results The mean age (n=40) was 32 (±8.88; 18-51) years; the mean DMFT index was 16±7.70; the mean plaque and gingival index were 2.07±0.65 and 2.12±0.64, respectively; and 20 (50%) volunteers reported xerostomia. We identified 305 salivary proteins (n=9), of which 23 were classified as candidate for biomarkers associated with 14 oral disorders. The highest number of candidates for biomarkers was associated with carcinoma of head and neck (n=7) and nasopharyngeal carcinoma (n=7), followed by periodontitis (n=6). Conclusions People with a crack cocaine use disorder had an increased risk of dental caries and gingival inflammation; less than half had oral mucosal alterations, and half experienced xerostomia. As possible biomarkers for 14 oral disorders, 23 salivary proteins were identified. Oral cancer and periodontal disease were the most often associated disorders with biomarkers.

Association of salivary alpha-2-macroglobulin with glycemia and glycated hemoglobin in type 2 diabetes mellitus: a systematic review and meta-analysis study

Abstract BACKGROUND: Chronically elevated alpha-2-macroglobulin (A2MG) in the blood has been correlated with diabetes and the HbA1c profile; however, no systematic review has been conducted to evaluate the association of A2MG salivary levels and glycemia or HbA1c levels in diabetes mellitus type 2 (DM2) patients. OBJECTIVE: To evaluate whether A2MG salivary levels are related to the glycemia or HbA1c levels in DM2 patients. DESIGN AND SETTING: Systematic review developed at Universidade Federal de Uberlândia (UFU), Brazil. METHODS: Eight databases were used as research sources. The eligibility criteria included studies that reported data regarding mean salivary A2MG and the correlation between glycemia and/or HbA1c levels of DM2 subjects (uncontrolled and well-controlled) and non-diabetic subjects. The risk of bias of the studies selected was assessed using the Joanna Briggs Institute (JBI) critical appraisal tools for use in JBI systematic reviews. Pooled correlation coefficients were estimated using the Hunter-Schmidt method. Study estimates were weighted according to their sample size, and heterogeneity was calculated using the chi-square statistic. RESULTS: Four studies on DM2 patients were included in this systematic review after careful analysis of 1482 studies. Three studies compared A2MG with HbA1c and glycemia. Overall, the correlation between A2MG and HbA1c was strong (r = 0.838). In contrast, the correlation between A2MG and glycemia was low (r = 0.354). CONCLUSION: The strong association between HbA1C and salivary A2MG suggests that this salivary protein has the potential to be a surrogate for HbA1C, if corroboratory further evidence is obtained through large-scale studies.

Salivary flow and salivary proteins in male and female children with autism spectrum disorder: pilot study / Fluxo salivar e proteínas salivares em crianças dos gêneros masculino e feminino com transtorno do espectro do autismo: estudo piloto

Introduction: autism spectrum disorder is a neurodevelopmental condition that affects the establishment of bonds and communication. Dental care is more difficult for people with this disorder, because in addition to communication difficulties, non-cooperation with respect to oral hygiene and continuous use of medication are common. Greater predisposition to caries, as well as alterations in the flow and concentration of salivary proteins were reported in these individuals. Objective: considering that sex can affect salivary flow and protein concentration, our objective was to analyze these parameters in the saliva of children with autism. Material and method: total unstimulated saliva was obtained from 12 boys and 12 girls aged between 5 and 15 years, with the aid of a catheter, after 2 hours of fasting and oral hygiene. Salivary flow was determined by estimating the mass of saliva. Total protein was determined in the supernatant obtained after centrifugation at 10,000 x g, for 10 minutes, by the Lowry method, with bovine albumin as standard. The results are expressed as mean and standard deviation. The data were submitted to the Shapiro-Wilk and Mann Whitney tests, with a significance level of 5%. Result: salivary flow values for boys (0.3555 ± 0.24 ml/min) and girls (0.2522 ± 0.1727 ml/min), and protein values for boys (1.430 ± 0.7480 mg/mL) and girls (1.075 ± 0.3702 mg/mL) were not significantly different between sexes. Conclusion: in children with autism spectrum disorder, sex does not influence unstimulated flow and salivary protein values.

Introdução: o transtorno do espectro autista é uma condição do neurodesenvolvimento que afeta o estabelecimento de vínculos e a comunicação. Cuidados odontológicos são mais difíceis em portadores desse transtorno, pois além da dificuldade de comunicação são comuns a não cooperação na higiene bucal e uso contínuo de medicamentos. Maior predisposição à cárie, alterações no fluxo e na concentração de proteínas salivares foram relatadas nesses indivíduos. Objetivo: considerando que o gênero pode afetar fluxo salivar e concentração proteica, nosso objetivo foi analisar esses parâmetros na saliva de crianças com autismo. Material e método: saliva total não estimulada foi obtida de 12 meninos e 12 meninas com idades entre 5 e 15 anos, com auxílio de um cateter, após 2 horas de jejum e higienização da cavidade bucal. O fluxo salivar foi determinado estimando-se a massa de saliva e o total de proteínas foi determinado no sobrenadante obtido após centrifugação a 10.000 x g, por 10 minutos, pelo método de Lowry, com albumina bovina como padrão. Os resultados foram expressos como média e desvio padrão, sendo submetidos aos testes de Shapiro-Wilk e Mann Whitney, com nível de significância em 5%. Resultado: os valores de fluxo salivar não foram significativamente diferentes quando comparados meninos (0.3555 ± 0.24 ml/min) e meninas (0.2522 ± 0.1727 mL/min), bem como os valores de proteínas (meninos: 1.430 ± 0.7480 mg/mL; meninas 1,075 ± 0,3702 mg/mL). Conclusão: em crianças com transtorno do espectro autista o gênero não influencia os valores de fluxo não estimulado e proteínas salivares.

Estimation of salivary proteins in early childhood caries before and after treatment using gel electrophoresis

Background: Saliva being an important biological fluid of our body contains both specific and nonspecific protective factors which form an integral part of our immune system. Salivary proteins play a substantial role in protecting humans against infection. Their level in oral cavity is subject to constant variations which is dependent on various factors. Purpose: The purpose of the study was to compare the levels of salivary proline-rich proteins (PRPs), glycoproteins, amylase bands, and salivary pH in children with early childhood caries before and after treatment using gel electrophoresis. Materials and Methods: The whole salivary pH, mean protein concentrations, and electrophoretic profiles of the salivary proteins were measured using sodium dodecylsulfate–polyacrylamide gel electrophoresis in both pre- and posttreatment groups. Statistical analysis was done using Statistical Package for Social Sciences (SPSS) version 15.0 software. Chi-square test and independent t-test were used to further compare the results. Results: The results were statistically significant in all the groups. There was variation in pre- and posttreatment values seen. Conclusion: Salivary proteins (glycoproteins, PRPs, and amylase) establish an imperative defense mechanism as antimicrobial agents.

Biomarcadores salivares do estresse oxidativo em crianças com cárie dentária: revisão sistemática e meta-análises / Salivary biomarkers of oxidative stress in children with dental caries: systematic review and meta-analysis

Objetivo. Avaliar a relação entre biomarcadores salivares de estresse oxidativo e cárie dentária em crianças. Métodos. Estudos realizados em crianças de até 12 anos comparando biomarcadores salivares de estresse oxidativo como malondialdeído (MDA), superóxido dismutase (SOD), ácido úrico, capacidade antioxidante total (TAC) e proteína total, considerando crianças com lesões de cárie dentária e sem cárie foram selecionados. Além disso, parâmetros salivares como fluxo salivar, pH, capacidade tampão e níveis de cálcio foram avaliados. Uma revisão sistemática da literatura foi realizada em 8 bases de dados. A diferença média padronizada (SMD) foi medida usando variância inversa como método estatístico e efeitos aleatórios como modelo de análise, correspondendo a um intervalo de confiança (IC) de 95%. Resultados. Os níveis de TAC foram maiores em crianças afetadas por cárie dentária em comparação com as sem cárie (grupo controle), independentemente da idade (SMD 2,66; IC 1,33; 3,98) ou sexo (SMD 0,98; IC 0,56; 1,39). Quando ajustados para proteína normalizada, os níveis de MDA foram menores no grupo de cárie dentária do que no grupo controle (SMD -16,5; IC - 29,02; -4,00), e os níveis de SOD foram maiores no grupo de cárie dentária (SMD 5,09; IC 0,01; 10,18). A concentração de proteína total na saliva de crianças com cárie dentária foi maior do que no grupo controle, independentemente da idade (SMD 0,98; IC 0,27; 1,69) ou sexo (SMD 0,77; IC 0,45; 1,10). Os parâmetros salivares avaliados apresentaram níveis mais baixos em crianças com cárie dentária (p< 0,05). Conclusões. Os níveis de biomarcadores de estresse oxidativo e parâmetros salivares estão alterados na saliva de crianças com cárie dentária(AU)

Objective. To assess the relationship between salivary biomarkers of oxidative stress and dental caries in children. Methods. Studies conducted in children up to 12 years old comparing salivary biomarkers of oxidative stress such as malondialdehyde (MDA), superoxide dismutase (SOD), uric acid, total antioxidant capacity (TAC), and total protein, considering children with dental caries lesions and caries-free ones were selected. In addition, salivary parameters such as salivary flow, pH, buffering capacity, and calcium levels were evaluated. A systematic literature review was carried out in 8 databases. The standardized mean difference (SMD) was measured using inverse variance as a statistical method and random effects as an analysis model, corresponding to a 95% confidence interval (CI). Results. The TAC levels were higher in children affected by dental caries compared to caries-free ones (control group), regardless of age (SMD 2.66; CI 1.33; 3.98), or gender (SMD 0.98; CI 0.56; 1.39). When adjusted for normalized protein, MDA levels were lower in the dental caries group than in the control group (SMD -16.51; CI -29.02; -4.00), and SOD levels were higher in the dental caries group (SMD 5.09; CI 0.01; 10.18). The total protein concentration in saliva of children with dental caries was higher than in the control group, regardless of age (SMD 0.98; CI 0.27; 1.69), or gender (SMD 0.77; CI 0.45; 1.10). The salivary parameters assessed had lower levels in children affected by dental caries (p<0.05). Conclusions. The levels of oxidative stress biomarkers and salivary parameters are altered in saliva of children with dental caries(AU)

Estimation of salivary ferritin in children with childhood caries ­ a cross sectional study / Estimativa da ferritina salivar em crianças com cárie na infância - um estudo transversal

Objective: Early childhood caries is the presence of dental caries in a child upto seventy one months of age. Saliva plays a major role in maintaining good oral health. The composition of saliva acts as a marker for oral health and the salivary proteins help in modulating the oral microflora in the oral cavity. Some salivary biomarkers help in detecting caries risk and can also predict their prognosis. Ferritin is one of the major biomarkers present in the saliva which acts as an iron binding protein and also as a monitoring tool in children suffering from iron deficiency. The ferritin levels are in increased in serum as well as in saliva to balance the deficiency of iron in the body. Material and methods: Sixty children were selected for the study aged between three to six years. The saliva sample was collected using standard spit method in a sterile container and Ferritin was tested in the samples by Chemiluminescence microparticle immunoassay(CMIA). Results: Salivary ferritin was found to be higher in the saliva of children with early childhood caries(mean value= 5.867) than in children without early childhood caries(mean value= 3.412). Conclusion: A direct association is present between salivary ferritin levels and dental caries. Increased level of ferritin is observed in children with Early childhood caries. Clinical relevance: The level of salivary ferritin is found to be raised in the present study in children with Early childhood caries. The exact mechanism is although not known it can be assumed that the children with early childhood caries might have deficiency of iron which has led to the increased amount of salivary ferritin in the saliva. (AU)

Objetivo: A cárie precoce é definida como a presença de cárie dentária em uma criança de até setenta e um meses de idade. A saliva desempenha um papel importante na manutenção de uma boa saúde bucal. A composição da saliva atua como um marcador para a saúde bucal e as proteínas salivares auxiliam na modulação da microflora oral na cavidade oral. Alguns biomarcadores salivares ajudam a detectar o risco de cárie e também podem prever seu prognóstico. A ferritina é um dos principais biomarcadores presentes na saliva, que atua como uma proteína ligadora de ferro e também como uma ferramenta de monitoramento em crianças com deficiência de ferro. Os níveis de ferritina aumentam tanto no soro quanto na saliva para equilibrar a deficiência de ferro no corpo. Material e Métodos: foram selecionadas para o estudo 60 crianças com idades entre três e seis anos. A amostra de saliva foi coletada pelo método padrão de cuspir em um recipiente estéril e a ferritina foi testada nas amostras através de um imunoensaio de micropartículas por quimioluminescência (CMIA). Resultados: A ferritina salivar foi maior na saliva de crianças com cárie na primeira infância (valor médio = 5,867) do que em crianças sem cárie na primeira infância (valor médio = 3,412). Conclusão: Existe uma associação direta entre os níveis de ferritina salivar e a cárie dentária. Nível elevado de ferritina é observado em crianças com cárie na primeira infância. Relevância clínica: O nível de ferritina salivar está elevado no presente estudo em crianças com cárie na primeira infância. Embora o mecanismo exato seja desconhecido, pode-se presumir que crianças com cárie na primeira infância podem ter deficiência de ferro, o que levou a um aumento na quantidade de ferritina salivar na saliva. (AU)

Los patrones electroforéticos de proteínas salivales permiten diferenciar los grupos transandino y cisandino de las especies de Rhodnius de Colombia / Salivary proteins electrophoretic patterns enabled differentiating Colombian Rhodnius Trans-Andean and Cis-Andean groups

Introducción. Las especies Rhodnius (Hemiptera: Reduviidae: Triatominae) están conformadas por insectos hematófagos vectores de Trypanosoma cruzi, agente etiológico de la enfermedad de Chagas, y T. rangeli, parásito infectivo pero no patógeno para el vertebrado. El estudio de la diversidad proteica de la saliva de estos insectos permite la obtención de perfiles electroforéticos unidimensionales característicos de algunas especies de triatominos. Sin embargo, el reporte de los patrones electroforéticos de proteínas salivales de las especies de Rhodnius ha sido escaso. Objetivo. Hacer un análisis comparativo de los perfiles electroforéticos unidimensionales de las proteínas salivales de R. colombiensis, R. pallescens, R. pictipes, R. prolixus y R. robustus. Materiales y métodos. Se obtuvieron los perfiles electroforéticos de la saliva de las especies en estudio mediante electroforesis en gel de poliacrilamida con dodecilsulfato sódico (Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis, SDS-PAGE) y se construyó un fenograma mediante el método UPGMA (Unweighted Pair Group Method Using Arithmetic Averages). Resultados. Los perfiles electroforéticos de las proteínas solubles de saliva presentaron bandas en un rango de masa aproximado de 15 a 45 kDa, los cuales permitieron diferenciar las cinco especies estudiadas. El fenograma reveló la existencia de dos grupos principales: uno conformado por los grupos cisandinos Pictipes y Prolixus y otro constituido por el grupo transandino Pallescens. Conclusiones. Existen diferencias en los perfiles electroforéticos de las proteínas salivales entre R. colombiensis, R. pallescens, R. pictipes, R. prolixus y R. robustus, cuya variabilidad permitió construir un fenograma congruente con los grupos del género Rhodnius.

Introduction: Rhodnius (Hemiptera: Reduviidae: Triatominae) species are made up of haematophagous insect vectors for Trypanosoma cruzi (Chagas' disease aetiological agent) and T. rangeli, an infective parasite that is not pathogenic for vertebrate hosts. The study of their salivary protein diversity enables the obtention of characteristic one-dimensional electrophoretic profiles of some triatomine species; however, few reports have dealt with Rhodnius species salivary proteins electrophoretic patterns. Objective: To compare R. colombiensis, R. pallescens, R. pictipes, R. prolixus, and R. robustus' salivary proteins one-dimensional electrophoretic profiles. Materials and methods: SDS-PAGE was used for obtaining electrophoretic profiles of saliva from the species under study. The unweighted pair group method with arithmetic mean (UPGMA) was used for constructing a phenogram. Results: Electrophoretic profiles of soluble saliva had protein bands ranging from 15 to 45 kDa, thereby enabling the five species studied to be differentiated. The phenogram revealed two main groups, one formed by the Pictipes and Prolixus cis-Andean groups and another consisting of the Pallescens trans-Andean group. Conclusion: Differences were revealed regarding R. colombiensis, R. pallescens, R. pictipes, R. prolixus, and R. robustus electrophoretic profiles of salivary proteins; their variability facilitated constructing a phenogram which was taxonomically congruent with the groups from the genus Rhodnius.

Chicken eggs as a surveillance tool for malaria and leishmaniasis vector presence

Abstract INTRODUCTION: Malaria and leishmaniases are transmitted by vectors during blood-feeding. Vector-infected animals develop antibodies against the vector's saliva. This study evaluated IgY antibody detection in the chicken eggs exposed to bites from Migonemyia migonei, Lutzomyia longipalpis and Anopheles aquasalis. METHODS: We used ELISA to quantify the antibody levels in the sera and exposed chicken eggs. RESULTS: High IgY levels were observed following immunization; furthermore, higher reactivity was observed in the eggs and species-specific immune response was observed post final immunization. CONCLUSIONS: Chicken eggs can be used as sentinels to surveil vector saliva antibodies.

Acción Inmediata de los Enjuagues Orales en la Saliva / Immediate Action of Mouthwashes on Whole Saliva

RESUMEN: Numerosos estudios confirman la efectividad de los enjuagatorios orales sobre la viabilidad de los microorganismos que producen gingivitis y halitosis, pero poco se conoce sobre la influencia de los mismos en el medio ambiente oral. El objetivo del siguiente trabajo fue analizar In vivo e In vitro el efecto de enjuagatorios orales sobre la saliva total no estimulada. Se trabajó con saliva de individuos sanos. Para el estudio in vivo se recogieron las muestras antes y después del enjuague oral a diferentes tiempos (1, 5, 10, 15, 30, 45 y 60 minutos). Para el ensayo in vitro, se incubó la saliva con igual volumen de la solución enjuagatoria a 37 ºC con agitación a diferentes tiempos (1, 5, 10 y 15 minutos). Se determinó pH inmediatamente recogidas las muestras. Posteriormente fueron centrifugadas y determinados flujo salival y proteínas totales. La separación de proteínas por electroforesis en SDS-PAGE se realizó solo en el ensayo in vivo. Los resultados mostraron que los enjuagatorios fluorurados poco alteran la fisiología oral a través de flujo salival, pH y proteínas totales. La combinación fluoruro de sodio/xilitol produjo mayor estimulación del flujo salival. La mezcla de aceites esenciales provocó un incremento del flujo salival y de pH, redujo el contenido de proteínas totales, evidenciando por SDS-PAGE que las comprometidas fueron particularmente las de mediano y bajo peso molecular. Clorhexidina debido a su elevada sustantividad, incrementó significativamente flujo salival y pH in vivo. In vitro, fuera del medioambiente oral, los enjuagatorios estudiados ejercieron un efecto similar sobre proteínas totales. Los enjuagatorios de uso frecuente alteraron parámetros salivales, por lo que podría estudiarse la acción que ejercen sobre otros componentes de la saliva con actividad biológica importante en cavidad oral.

ABSTRACT: Numerous studies confirm the effectiveness of mouthwashes on the viability of microorganisms that produce gingivitis and halitosis, but little is known about their influence on the oral environment. The objective of the following work was to analyze In vivo and In vitro the effect of mouthwashes on total non-stimulated saliva. We worked with saliva from healthy individuals. For In vivo study, samples were collected before and after oral rinsing at different times (1, 5, 10, 15, 30, 45 and 60 minutes). For the In vitro assay, the saliva was incubated with equal volume of the rinse solution at 37 ° C with shaking at different times (1, 5, 10 and 15 minutes). PH was determined immediately collected samples. Subsequently they were centrifuged and determined salivary flow and total proteins. Separation of proteins by SDS-PAGE electrophoresis was performed only in the In vivo assay. The results showed that fluoridated rinses hardly alter oral physiology through salivary flow, pH and total proteins. The combination of sodium fluoride / xylitol produced greater stimulation of salivary flow. The mixture of essential oils caused an increase in salivary flow and pH, reduced the total protein content, evidencing by SDS-PAGE that those involved were particularly those of medium and low molecular weight. Chlorhexidine due to its high substantivity, significantly increased salivary flow and pH In vivo. In vitro, outside the oral environment, the rinses studied had a similar effect on total proteins. Rinses used frequently altered salivary parameters, so that the action they exert on other components of saliva with important biological activity in the oral cavity could be studied.

Peptídeos peptidomiméticos da película adquirida do esmalte: efeitos no crescimento de cristal de hidroxiapatita / Peptidomimetics of acquired enamel pellicle peptides: effects on hydroxyapatite crystal growth

Os peptídeos da estaterina (DR9) e da histatina 3 (RR14), que ocorrem naturalmente na película in vivo, amplificam o efeito inibitório do crescimento de cristais de hidroxiapatita, função relacionada à remineralizarão do esmalte e formação de cálculos dentários. A hipótese da duplicação/hibridação de domínios funcionais dos peptídeos DR9 da estaterina e RR14 da histatina 3 foi testada. Para isto, os peptídeos peptidomiméticos (DR9-DR9, DR9-RR14), além deles individualmente e suas proteínas intactas (DR9, RR14, estaterina e histatina 3) foram estudados em sete concentrações diferentes para avaliar o efeito da inibição do crescimento de cristais de hidroxiapatita. Foi utilizado um ensaio colorimétrico de microplaca para quantificar o crescimento de cristais de hidroxiapatita. As experiências foram feitas em triplicata e a concentração inibitória (IC50) foi estabelecida para cada grupo. A IC50 foi calculada para todos os peptídeos e proteínas testados. A histatina 3 e o RR14 não atingiram o valor de IC50. O DR9- RR14 atingiu o valor de IC50 a 3,80 M. Como esperado, DR9 e DR9-DR9 demonstraram um efeito inibitório significativo na atividade de crescimento de cristais, atingindo o valor de IC50 a 2,82 M e 1,07 M, respectivamente. A estaterina atingiu o valor de IC50 a 2,50 M. Na análise estatística, foram aplicados os testes ANOVA e Student-Newman-Keuls para comparações por pares, para comparar os valores entre os grupos. O DR9-DR9 amplificou o efeito inibitório do crescimento de cristais de hidroxiapatita quando comparado com DR9 único (p <0,05), demonstrando que a multiplicação do domínio funcional é uma forte tendência evolutiva da proteína. De forma interessante, o peptídeo híbrido DR9-RR14 demonstrou um efeito inibitório intermediário quando comparado com outros dois grupos: DR9 único e DR9-DR9. Este estudo utilizou a abordagem peptidomimética para investigar uma via potencial de evolução da proteína relacionada com a duplicação/hibridação dos constituintes peptídicos naturais da película adquirida de esmalte. O conhecimento obtido por meio dos resultados deste trabalho pode fornecer uma base para o desenvolvimento de peptídeos sintéticos para uso terapêutico, tanto contra cárie dentária, como para a doença periodontal.(AU)

The statherin and histatin 3 peptides (DR9 and RR14 respectively), which occur naturally in the film in vivo, amplify the inhibitory effect for the growth of hydroxyapatite crystals, a function related to remineralization of the enamel and formation of dental calculi. The hypothesis of duplication/hybridization of functional domains of the DR9 peptides of the statherin and RR14 of histatin 3 was tested. For this, the peptidomimetic peptides (DR9-DR9, DR9-RR14), in addition to them individually and their intact proteins (DR9, RR14, statherin and histatin 3) were studied at seven different concentrations to evaluate the effect of growth inhibition of hydroxyapatite crystals. A colorimetric assay of microplate was used to quantify the growth of hydroxyapatite crystals. The experiments were done in triplicate and the inhibitory concentration (IC50) was established for each group. The IC50 was calculated for all peptides and proteins tested. Histatin 3 and RR14 did not reach the IC50 value. DR9-RR14 reached the IC50 value at 3.80 M. As expected, DR9 and DR9-DR9 demonstrated a significant inhibitory effect on crystal growth activity, reaching the IC50 value at 2.82 M and 1.07 M, respectively. Statherin reached the IC50 value at 2.50 M. ANOVA and Student-Newman-Keuls tests for paired comparisons were applied to compare the values between the groups. DR9-DR9 amplified the inhibitory effect of hydroxyapatite crystal growth when compared to single DR9 (p <0.05), demonstrating that the multiplication of the functional domain is a strong protein evolution pathway. Interestingly, the hybrid peptide DR9-RR14 demonstrated an intermediate inhibitory effect when compared to other two groups: single DR9 and DR9-DR9. This study utilized the peptidomimetic approach to investigate a potential pathway of protein evolution related to duplication/hybridization of the natural peptidic constituents of the acquired enamel film. The knowledge obtained through the results of this work can provide a basis for the development of synthetic peptides for therapeutic use, both against dental caries and for periodontal disease.(AU)

Salivary protein characteristics from saliva of carious lesion-free and high caries adults

It has been argued that specific salivary proteins could have a protective effect against caries, but data from the many available studies are rather contradictory. The purpose of this study was to analyze whether there is a relationship between protein concentration, electrophoretic profile and concentration of salivary IgA and the presence or absence of caries in adults. Adults with high caries activity (HC) and without caries lesions (CF), assessed by ICDAS criteria, were asked to provide unstimulated saliva samples. Protein concentration (μg/mL) was determined using the Bradford method. Western blotting was used to detect IgA. Data were compared using Student’s t test at p<0.05. Total protein concentration in CF was higher (50.65±7.5 μg/mL) than in HC individuals (26.80±2.5 μg/mL) (p=0.001). More protein bands were visualized in the gels from CF than the HC group (p=0.001). CF subjects showed higher salivary IgA concentration (11.27±0.5 μg) than HC individuals (1.71±0.2μg) (p=0.001).Salivary composition in high caries experience and cariesfree young adults seems to differ in terms of the type and amount of proteins. Further research is needed to expand these findings.

Se ha descrito que proteínas salivales específicas podrían tener un efecto protector sobre la caries, sin embargo, los datos de los numerosos estudios disponibles son contradictorios. El propósito de este trabajo fue analizar si existe una relación entre la concentración total de proteínas, perfil electroforético y la concentración de IgA salival y la presencia o ausencia de lesiones de caries en adultos. Se obtuvieron muestras de flujo salival no estimulado de adultos con alta actividad de caries (HC) y sin lesiones de caries (CF), evaluados según criterios ICDAS. La concentración total de proteínas (mg / ml) se determinó utilizando el método de Bradford. Para detección de IgA se empleó Western Blot. Los datos se compararon mediante la prueba t student, estableciendo diferencias significativas si p<0,05. La concentración total de proteínas en CF fue mayor (50,65 ± 7,5 mg / ml) que en individuos HC (26,80 ± 2,5 mg / ml) (p=0,001). En los geles, se visualizó un mayor número de bandas de proteínas en CF que en el grupo HC (p=0,001). Los Sujetos CF mostraron mayor concentración de IgA salival (11,27 ± 0,5 μg) que los individuos HC (1,71 ± 0,2 μg) (p=0,001). La composición salival de sujetos adultos jóvenes con alta experiencia y libres de caries, parece ser diferente en función del tipo y la cantidad de proteínas. Se requiere de más investigación para profundizar estos resultados.

Imunização de cães com produtos oriundos de Lutzomyia Longipalpis em duas diferentes abordagens: Canarypoxvirus sp. Expressando o gene que codifica para a proteína salivar LJM17 e/ou LJL143, e a proteína do intestino médio luloper1 como vacina bloqueadora de transmissão / dog immunization with products from Lutzomyia longipalpis in two different approaches: Canarypoxvirus sp. Expressing the gene that encodes the protein salivary LJM17 and / or LJL143 and the midgut protein luloper1 as a transmission blocking vaccine

As interações entre flebótomo, parasita e hospedeiro desempenham um papel importante na transmissão da leishmaniose. As moléculas provenientes do vetor são relevantes para estas interações e incluem as proteínas da saliva e do intestino médio. Nos flebótomos, as Leishmanias passam por um ciclo de desenvolvimento complexo dentro do intestino médio sob a proteção da matriz peritrófica, necessário para a geração de formas metacíclicas infectantes. As Leishmanias são transmitidas pelos flebótomos que co-injetam parasitas juntamente com a saliva, na derme do hospedeiro. Estudos anteriores demonstraram que a imunização de cães com duas proteínas salivares (LJM17 ou LJL143) de L. longipalpis, resultaram em uma imunidade mediada por células Th1 sistêmica e local afetando a sobrevivência do parasita in vitro. Assim, o objetivo deste trabalho foi avaliar a imunidade conferida pela imunização de cães com DNA e rCanarypoxvirus expressando o gene que codifica para as proteínas salivares de L. longipalpis (LJM17 e/ou LJL143)...

Sand fly, parasite, host interactions play an important role in the transmission of leishmaniasis. Vector molecules are relevant for such interactions and include midgut and salivary proteins. In vector sand fly species, Leishmania parasites undergo a complex developmental cycle within the midgut, protected by the peritrophic matrix that is necessary for generation of infectious metacyclics. Leishmania parasites are transmitted by sand flies that co-inject parasites and saliva, in the host’s skin. Previous studies showed immunization of dogs with two proteins (LJM17 or LJL143) from Lutzomyia longipalpis, resulted in a systemic and local Th1 cell-mediated immunity affecting parasite survival in vitro. In this work we evaluated the immunity conferredbyimmunization of dogs with DNA and recombinant Canarypoxvírusexpressing the gene encoding the salivary ofL.longipalpis (LJM17and/orLJL143). Immunization with both LJL143 and LJM17...

Análise microbiológica e imunologica do fluido gengival e proteoma salivar de individuos com Sindrome de Down com doença periodontal / Microbiological and immunological analysis in the gingival crevicular fluid and salivary proteome analysis in individuals with Down syndrome with periodontal disease

O objetivo deste estudo foi avaliar a quantidade de periodontopatógenos e de citocinas inflamatórias no fluido gengival, bem como as proteínas salivares em indivíduos com Síndrome de Down (SD) com Doença Periodontal (DP), comparando-os com indivíduos cromossomicamente normais, antes e 45 dias após o tratamento periodontal não-cirúrgico. Para detectar e quantificar as bactérias (Porphyromonas gingivalis, Tannerella forsythia e Treponema denticola), o fluido gengival foi coletado de 35 indivíduos com DP, sendo 23 indivíduos com SD e 12 não-sindrômicos (controle). Para quantificar as citocinas inflamatórias e as proteínas salivares foram coletados fluido gengival e saliva de 30 indivíduos com DP, sendo 20 indivíduos com SD e 10 não-sindrômicos (controle). Os efeitos do tratamento nos parâmetros clínicos foram positivos para o índice de placa, sangramento à sondagem, profundidade de sondagem e nível de inserção, em ambos os grupos. Porém, a contagem dos periodontopatógenos foi maior nos indivíduos com SD comparados com o grupo controle, antes e 45 dias após o tratamento periodontal. As citocinas Th1, Th2 e Th17 também foram encontradas em maiores quantidades nos indivíduos com SD do que nos controle, mesmo depois do tratamento periodontal. Adicionalmente, maiores quantidades de proteínas salivares com propriedades antimicrobianas, lubrificação, metabolismo, organização celular, resposta imune e transporte foram encontradas em indivíduos com SD depois do tratamento periodontal. Conclui-se que os resultados desta pesquisa podem contribuir para uma compreensão mais aprofundada do comportamento microbiológico, imunológico e do proteoma salivar de indivíduos com SD, e, consequentemente, explicar a alta prevalência e severidade da doença periodontal nesses indivíduos.

The aim of this study was to quantify the periodontopathogens, inflammatory cytokines and salivary proteins in subjects with Down syndrome (DS) and normal subjects, both with periodontal disease (PD), before and 45 days after non-surgical periodontal therapy. To detect and quantify bacteria (Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola), crevicular gingival fluid (CGF) was collected from 35 individuals with PD, 23 with DS and 12 non-syndromic (control). To quantify the inflammatory cytokines and salivary proteins, CGF and saliva of 30 individuals with PD, 20 with SD and 10 non-syndromic (control) were collected. The effects of the non-surgical periodontal therapy on clinical parameters were positive in both groups. However, the count of periodontopathogens was higher in individuals with DS compared with the control group, before and after periodontal therapy.Th1, Th2 and Th17 cytokines were also found in higher amounts in individuals with DS even after periodontal therapy compared with control patients. Furthermore, higher amounts of salivary proteins with antimicrobial properties, lubrication, metabolism, cellular organization, immune response and transport were quantified in individuals with DS after periodontal therapy. Despite of clinical parameters improvement after non-surgical periodontal therapy in subjects with DS, it is concluded that the results of this study may contribute to a more profound understanding of microbiological and immunological behavior, as well as knowledge of the salivary proteome in individuals with Down syndrome, and also might explain the high prevalence and severity of periodontal disease in these individuals

Principales proteínas salivales: estructura, función y mecanismos de acción / Salivary proteins: structure, function and mechanisms of action

Introducción: Las proteínas salivales son elementos fundamentales en las importantes funciones que desempeña la saliva en el mantenimiento de la salud bucal. Objetivo: Fundamentar las funciones de las principales proteínas salivales teniendo en cuenta la relación estructura-función de las macromoléculas. Métodos: Se realizó una revisión bibliográfica sobre el tema a través de la consulta de bases de datos (PubMed, Hinari, SeCimed, Scielo y EBSCO) y se seleccionaron 30 referencias relevantes. Desarrollo: Se describen las estructuras moleculares, funciones y mecanismos de acción de las principales proteínas salivales: mucinas, aglutinina, proteínas ricas en prolina, inmunoglobulinas, lisozima, peroxidasa humana salival, lactoferrina, estaterina, cistatinas e histatinas. Conclusiones: Los mecanismos de acción de las proteínas salivales evidencian la relación estructura-función que caracteriza a las macromoléculas y fundamentan su papel dentro de las funciones de la saliva relacionadas con el mantenimiento de la salud bucal.

Introduction: Salivary proteins are key elements in the important functions saliva plays in the maintenance of oral health. Objective: To lay the foundations of the functions for the principal salivary proteins according to the structure-function relationship of macromolecules. Methods: A review was performed about the subject through the consultation of data bases PubMed, Hinari, Secimed, Scielo and EBSCO. Results: The molecular structures, functions and mechanisms were described for the salivary proteins: mucins, agglutinin, proline rich proteins, immunoglobulins, lysozyme, salival peroxidase, lactoferrin, statherins, cystatins and histatins. Conclusions: The mechanisms of salivary proteins make evident the structure-function relationship of macromolecules and fundament their role in saliva functions related to the maintenance of oral health.

Comparación de la concentración total de proteínas salivales de adultos y adultos mayores / Comparison of total salivary protein concentration in adults and older adults

Objetivo: Entre las moléculas orgánicas que componen la saliva se han descrito un gran número de proteínas. No existe evidencia suficiente que permita especular sobre los cambios en el contenido proteico salival asociados al envejecimiento normal. El objetivo fue determinar si existen diferencias en la concentración total de proteínas en saliva de adultos y adultos mayores. Método: Se obtuvieron muestras de saliva estimulada y no estimulada de individuos pertenecientes a dos grupos etarios, adultos (A) (n=30) (edad promedio: 25 años) y adultos mayores (AM) (n=30) (edad promedio: 68 años). Se excluyeron aquellos sujetos que presentaron alguna patología oral/sistémica o consumo de fármacos/sustancias relacionados con alteraciones del flujo o composición salival. Cada muestra fue centrifugada. A 10 ul del sobrenadante se agregaron 190 ul de solución de Bradford. Se realizó el recuento mediante lectura con espectrofotómetro (595 nm). La concentración de proteínas fue comparada entre los grupos en estudio utilizando la prueba t de Student (p<0.05). Resultados: La concentración de proteínas totales de saliva no estimulada fue mayor en AM (5.5+/-2.6 ug/ul) que en A (3.2+/-1.7 ug/ul) (p<0.05). La concentración de proteínas totales de saliva estimulada fue mayor en AM (4.9+/-0.4 ug/ul) que en A (4.0+/-0.3 ug/ul) (p<0.05). Conclusión: La concentración de proteínas en saliva aumenta en adultos mayores. Se requieren nuevos estudios para caracterizar estos cambios salivales con la edad.

Objective: A high number of proteins has been described in saliva. Scarce evidence allows to speculate about the variations on protein content associated with aging. The aim was to determine if there is a difference in total protein concentration between adults and older adults. Methods: Samples were collected from unstimulated and stimulated saliva of individuals from two age groups, adults (A) (n=30) (mean age 25 years) and older adults (OA) (n=30)(mean age: 68 years). Subjects with oral/systemic diseases or consuming medications that alter salivary flow were excluded from the study. Each sample was centrifuged and 10 ul of the supernatant was added to 190 ul of Bradford solution. Protein concentration was obtained by spectrophotometry (595 nm) Protein concentration was compared between the groups using student’s t test (p<0.05). Results: Total protein concentration from unstimulated saliva was higher in OA (5.5+/-2.6 ug/ul) than in A (3.2 +/- 1.7 ug/ul) (p<0.05). Total protein concentration of stimulated saliva was higher in OA (4.9+/-0.4 ug/ul) than in A (4.0+/-0.3 ug/ul) (p<0.05). Conclusion: Salivary protein concentration increases in older adults. Further studies are suggested to characterize these changes.

Sandfly saliva of Lutzomyia ovallesi (Diptera: Psychodidae) as a possible marker for the transmission of Leishmania in Venezuela Andes region.

Background & objectives: The saliva of the Phlebotominae is highly immunogenic to the vertebrate host and is a determining factor in the Leishmania infection. The aim of this work was to study the saliva of Lutzomyia ovallesi as a possible risk marker for the transmission of Leishmania. Methods: Two populations of L. ovallesi from different geographical areas and subjected to different environmental conditions were compared by geometric morphometry of the wings, by protein profile analysis of salivary glands and by assessing the presence of anti-saliva protein in human sera confronted with laboratory L. ovallesi saliva. Results: The results showed differences in the isometric size and structure of the wings but no allometric effects. Protein profiles of salivary glands of both the L. ovallesi populations studied were found to be similar, based on 11 protein bands with molecular weights ranging from 16 to 99 kDa. Anti-saliva antibodies were present in human sera, but human sera infected and uninfected with leishmaniasis could not be differentiated. Interpretation & conclusion: We conclude that the saliva of laboratory-reared L. ovallesi is representative of that of the wild population. It is suggested to study the presence of anti-saliva antibodies in other species of sandflies and mosquitoes.

Evaluación de un método de extracción de glicoproteína de glándulas submaxilares de bovino / Evaluation of an extraction method glycoprotein bovine submaxillary glands

Los métodos utilizados para la obtención de glicoproteínas de glándulas submaxilares de bovino consisten en esquemas que implican la eliminación de compuestos acuoinsolubles, aislamiento y purificación de las glicoproteínas. El propósito de este estudio fue evaluar un método de obtención de glicoproteínas (Escalona y col. 1989) a partir de glándulas de bovino para obtener un extracto mucínico destinado a la elaboración de un sustituto salival, con el fin de conocer el tipo y cantidad de proteínas presentes y su asociación con la viscosidad. Se analizaron 6 fases acuosas posteriores a la primera centrifugación. De cada una de las fases acuosas se tomó una alícuota y se determinó la cantidad de proteínas totales, y la viscosidad. La viscosidad aparente se midió con un viscosímetro Marca Brokfield modelo LVT con un eje Nº4 a una velocidad de desplazamiento a 73.42seg-1, y las proteínas se caracterizaron por electroforesis en geles de SDS-PAGE al 8 por ciento. Cada uno de los geles se escaneo y la densitometría se realizó en forma computarizada (un programa MultiAnalystTM /PC -Bio Rad- Versión 1.1). Se observó que a medida que se avanzaba en el esquema de extracción de glicoproteínas disminuía la cantidad de proteínas totales, el patrón electroforético mostró diferencias cualitativas entre las muestras evaluadas lo cual se reflejó en una disminución estadísticamente significativa (p<0,05) de la viscosidad aparente. Al expresar la viscosidad con relación al contenido proteico ésta aumentaba, lo que nos permitió inferir que a pesar que hubo una perdida de proteínas se preservaron las glicoproteínas viscosas a expensa del rendimiento proteico

For the obtainment of glycoproteins from bovine submaxillary glands methods are used that encompass the elimination of insoluble compounds, and fractionation and purification procedures. This study was designed to evaluate a particular method (Escalona et al. 1989) for the obtainment of glycoprotein to be used in the preparation of a salivary substitute, and aimed to gathering information on the type and amount of present proteins and their association with viscosity. For this purpose, the six aqueous phases following a centrifugation step in the fractionation scheme were analyzed for protein content (Lowry et al 1951), aparent viscosity (Brookfield viscometer, model LVT), and 8 percent SDS-PAGE electrophoresis with electronic processing of the densitograms (Bio-Rad/multi Analyst). The protein content, aparent viscosity (statistically significant P<0.05), and number of electrophoretic bands showed a decrease with the progress of the fractionation procedure. However, when aparent visxosity was ex pressed in terms of protein content it showed an increase, which is consonous with the relative enrichmentain of the fractions glycoprotein along the purification procedure

Componentes das imunidades inata e adaptativa presentes na saliva humana / Components of the innate and adaptative immunity present in human saliva

A cavidade bucal é a porta principal de entrada de patógenos para o corpo humano. No entanto, devido a um complexo mecanismo de defesa, os inúmeros agentes infecciosos que colonizam ou penetram a cavidade bucal não ocasionam patologias. A saliva, além de desempenharas funções de hidratação e lubrificação dos tecidos da cavidade bucal, atua diretamente na regulação da microbiota e na proteção contra microrganismos. A função de proteção é desempenhada por componentes celulares e moleculares pertencentes às imunidades inata e adaptativa que atuam sobre bactérias, fungos e vírus. As proteínas da imunidade inata constituem a primeira linha de defesa do organismo, e as imunoglobulinas, secretadas pelos linfócitos B1 da imunidade inata e pelos linfócitos B da imunidade adaptativa, potencializam esse mecanismo protetor. Nesta revisão foram destacados os principais componentes celulares e moleculares pertencentes ao Sistema Imune Inato e ao Sistema Imune Adaptativo que atuam na proteção e na manutenção da homeostasia da cavidade bucal

The oral cavity is the main pathway through which pathogens enter the human body. However, as a result of a complex defense mechanism, the numerous infectious agents that enter and colonize the oral cavity do not cause pathologies. Besides moisturizing and lubricating the oral cavity tissues, the saliva acts directly in the regulation of the microbiota and in the protection against microorganisms. The protective function is performed by the cellular and molecular components of the innate and adaptive immunity, which act against bacteria, fungi and viruses. The proteins of the innate immunity are the organism first line of defense and the immunoglobulin, which are secreted by the B1 lymphocytes of the innate immunity and B lymphocytes of the adaptive immunity, boost the protective mechanism. In this review we emphasize the main cellular and molecular components that belong to the Innate Immune System and to the Adaptive Immune System, which act in the protection and the maintenance of the oral cavity homeostasis

Salivary proteins in children aged from 3 to 4 years / 实用口腔医学杂志

Objective: To explore levels of salivary IgA, lysozyme, lactate dehydrogenase, alkaline phosphatase and total proteins in unstimulated (UWS) and stimulated (SWS) whole saliva of caries-free children. Methods: 94 caries-free children, aged from 42 to 54 months, were recruited from urban kindergartens in Beijing. 2 ml UWS and 2 ml SWS were collected from each child to measure salivary IgA, lysozyme, lactate dehydrogenase, alkaline phosphatase and total proteins. Results:1) There were no differences in salivary proteins between boys and girls in both UWS and SWS, except total proteins. In UWS, the concentration of total proteins in girls was lower than that in boys (P0.05). 2) The concentration of IgA in SWS was significantly lower than that in UWS (P0.05). Conclusion:In 3-to 4-year old children, the protein composition in stimulated whole saliva is different from that in unstimulated whole saliva, and it is also different between genders. Therefore, it is more reliable to measure salivary proteins in both UWS and SWS when studying saliva of children.