Avaliação de herpesvírus em sangue e saliva de pacientes cirróticos / Evaluation of herpesvirus in blood and saliva of cirrhotic patients

Pacientes cirróticos em fila de transplante hepático possuem inúmeros prejuízos em vias fisiológicas, como a evidente deficiência imune. O dano ao sistema reticuloendotelial, o comprometimento das vias e células de defesa tornam o paciente hepatopata mais propenso ao desenvolvimento de infecções que influem em sua qualidade de vida e podem piorar o seu prognóstico. Ainda neste grupo, a existência de compostos tóxicos não excretados pelo fígado cirrótico pode favorecer quadros inflamatórios e a latência viral. A família dos Herpesvírus humanos (HHV) possui o atributo de desenvolver latência vitalícia nos indivíduos infectados e seus episódios de reativação podem ser precipitados por quadros de imunossupressão. O uso da saliva para identificação de HHV é viável e pode demonstrar os 8 subtipos virais em diferentes níveis. O objetivo deste estudo foi identificar a presença de HHV-1, HHV-2, VZV, EBV, CMV, HHV-6A, HHV-6B, HHV-7 e HHV-8 em amostras de plasma sanguíneo e saliva de pacientes com cirrose hepática em fila de espera pelo transplante e verificar se a presença desses vírus tem relação com dados da cirrose, como doença descompensada, complicações e etiologia, além de dados advindos de exames sanguíneos, como a população de leucócitos circulantes. Foi desenvolvido um estudo transversal observacional com dados demográficos e médicos de prontuários e amostras de sangue e saliva de 72 indivíduos cirróticos provindas do Biobanco da Faculdade de Odontologia da Universidade de São Paulo, onde permaneceram congeladas em ultrafreezer até o momento da técnica da detecção da cadeia de polimerase. Foram utilizados dois protocolos distintos, o primeiro denominado HSV-Pan, que amplifica o DNA de HHV-1, HHV-2, EBV, CMV e HHV-8, e o segundo denominado VZV-Pan responsável pela amplificação de VZV, HHV-6 tipo A e B e HHV-7. Para as amostras positivas realizou-se subsequente digestão enzimática para identificação do subtipo de HHV. A amostra foi composta majoritariamente por indivíduos do sexo masculino (n=51, 70%), com média de idade 54 anos (DP=10). A mediana do índice MELD foi 15 (DP=6) e 59% foram classificados com cirrose descompensada. 47,2% (n=34) apresentavam leucopenia. Não foram identificados HHV em amostras sanguíneas. Em saliva, os principais HHV identificados foram HHV-7 (n=42, 62%) e EBV (n=30, 41%). Houve correlação estatística entre a presença de HSV-Pan e a identificação de alterações em série branca sanguínea (p=0,019), especificamente com a população total de leucócitos (p=0,025), mas não houve correlação com subpopulações como neutrófilos e linfócitos. Essa correlação aconteceu, provavelmente, às expensas do EBV presente na saliva, já que apenas 5 pacientes apresentaram positividade para outros vírus. De fato, a correlação entre o EBV com leucócitos totais e com linfócitos apresentou-se estatisticamente significante (p=0,038 e p=0,047, respectivamente). Não houve correlação entre a presença viral e outras variáveis independentes, como complicações da cirrose ou doença descompensada. Conclui-se que, na população estudada, a identificação de EBV na saliva está vinculada ao estado de imunidade circulante do paciente cirrótico. É possível que a deficiência imune apresentada pelos pacientes com cirrose possa ter um papel no shedding do EBV em saliva.

Salivary Metabolites in Patients with Mucopolysaccharidosis

ABSTRACT Objective: To identify the salivary metabolites profile of Mucopolysaccharidosis (MPS) types I, II, IV, and VI patients. Material and Methods: The participants were asked to refrain from eating and drinking for one hour before sampling, performed between 7:30 and 9:00 a.m. Samples were centrifuged at 10.000 × g for 60 min at 4°C, and the supernatants (500µl) were stored at −80°C until NMR analysis. The salivary proton nuclear magnetic resonance (1H-NMR) spectra were acquired in a 500 MHz spectrometer, and TOCSY experiments were used to confirm and assign metabolites. Data were analyzed descriptively. Results: Differences in salivary metabolites were found among MPS types and the control, such as lactate, propionate, alanine, and N-acetyl sugar. Understanding these metabolite changes may contribute to precision medicine and early detection of mucopolysaccharidosis and its monitoring. Conclusion: The composition of low molecular weight salivary metabolites of mucopolysaccharidosis subjects may present specific features compared to healthy controls.

Índice de Dieta Mediterránea y pH Salival en Mujeres Adultas. Estudio Observacional / Mediterranean Diet Index and Salivary pH in Adult Women. Observational Study

El ser humano en el transcurso de la evolución ha cambiado la percepción de la alimentación dejando de lado el consumo de carne y sus derivados por factores éticos y morales, donde se sabe la dieta tiene gran relevancia en la salud general, siendo un factor de riesgo para la aparición de diversas enfermedades motivo por cual la saliva se ha utilizado como instrumento de diagnóstico Describir y comparar el índice de Dieta mediterránea según pH salival en mujeres estudiantes de Odontología de entre 20 a 30 años. Estudio descriptivo transversal comparativo. Participaron 17 mujeres adultas seleccionadas por criterio no probabilístico en la carrera de Odontología, quienes fueron evaluadas en su índice de Dieta mediterránea y pH salival. Se aplicó la prueba T de Student para muestras independientes y U Mann-Whitney para comparar características entre grupos en función del pH salival. Existen diferencias significativas entre grupos para la frecuencia de consumo de cereales integrales (p = 0,026), carnes magras (p = 0,021) y pH salival (p < 0,001). Existe un predominio hacia la baja adherencia a una dieta mediterránea, existiendo diferencias significativas en la frecuencia de consumo de cereales integrales y carnes magras, siendo el consumo de estos un indicador del riesgo de la cavidad oral determinado por pH.

The human being in the course of evolution has changed the perception of food, leaving aside the consumption of meat and its derivatives due to ethical and moral factors, where it is known that diet has great relevance in general health, being a factor of risk for the appearance of various diseases, which is why saliva has been used as a diagnostic tool. To describe and compare the Mediterranean Diet index according to salivary pH in female dentistry students between 20 and 30 years old. Comparative cross-sectional descriptive study. Seventeen adult women selected by non-probabilistic criteria participated in the Dentistry career, who were evaluated in their Mediterranean Diet index and salivary pH. Student's T test for independent samples and Mann-Whitney U test were applied to compare characteristics between groups based on salivary pH. There are significant differences between groups for the frequency of consumption of whole grains (p = 0.026), lean meats (p = 0.021) and salivary pH (p < 0.001). There is a predominance towards low adherence to a Mediterranean diet, with significant differences in the frequency of consumption of whole grains and lean meats, the consumption of these being an indicator of oral risk determined by pH.

A multilevel analysis model for dental caries determinants in independently-living elderly

To evaluate the prevalence of untreated caries and its association with biological, individual, and environmental variables in independently-living elderly people. Methods: This cross-sectional study included 72 elderly (≥60 years) patients of a university dental clinic in Belo Horizonte, Brazil. Sociodemographic data, systemic diseases, medications, and free sugar intake were collected. Visible plaque, Decayed, Missing, and Filled Teeth (DMFT), and Decayed and Filled Root (DFR) indexes were assessed through clinical examination. Unstimulated saliva was collected to determine salivary flow, pH, and buffering capacity. Descriptive analysis and multilevel logistic regression analysis were performed following a dental caries theoretical model (p <0.05, 95% CI). Results: The mean DMFT and DFR were 24.44 (SD=4.59) and 3.21 (SD=2.93), respectively. The prevalence of untreated caries was 61.11%. In the adjusted multilevel regression model involving 1639 teeth, untreated dental caries was significantly associated with the presence of biofilm (OR = 1.84; 95% CI: 1.24­2.74), salivary buffering capacity (OR = 0.87; 95% CI: 0.77­0.99) and per capita income (OR = 0.06; 95% CI: 0.004­0.74). Conclusion: The experience of dental caries was widespread among independently-living elderly patients, and its variability was best explained by the presence of biofilm, reduced salivary buffering capacity, and low per capita income. A comprehensive assessment is needed of the biological, individual, and environmental factors related to the presence of dental caries in independently-living elderly people

Cárie dentária e lesões cervicais não cariosas em idosos independentes: características individuais, ambientais e salivares / Dental caries and noncarious cervical lesions in independent elderly: individual, environmental, and salivary characteristics

O envelhecimento populacional característico das mudanças demográficas atuais vem sendo acompanhado por maior preservação dentária. Pessoas idosas possuem particularidades que as tornam mais susceptíveis ao desenvolvimento de alterações bucais como cárie dentária e lesões cervicais não cariosas (LCNC). Com a idade avançada, a prevalência da cárie dentária aumenta, e as LCNC tornam-se mais comuns e graves. O objetivo desse estudo foi avaliar a frequência de cárie dentária e de LCNC em idosos independentes, além de investigar associações entre proteínas salivares e a presença de cárie, e entre LCNC e fatores oclusais em dois artigos distintos. Neste estudo transversal, parte dos dados foi coletada usando entrevista estruturada. A experiência de cárie e de LCNC foi determinada pelos índices de Dentes Cariados, Perdidos e Obturados (CPO-D), Raízes Cariadas e Obturadas (CO- R) e Índice de Desgaste Dentário (TWI), coletados em exame clínico. Ao total, foram avaliados 127 indivíduos, com idade entre 60 e 84 anos. A média do CPO-D foi 23,86 e frequência de cárie dentária foi 65,4%. Para LCNC, a frequência de lesões com profundidade superior a 1mm foi 65,4%. Modelos 3D foram obtidos a partir de escaneamento intraoral para análise oclusal, morfologia e morfometria das LCNC, em 53 idosos, totalizando 1245 dentes com frequência absoluta de 352 (28,27%) dentes com LCNC. Lesões arredondadas possuíam maior altura, enquanto lesões anguladas apresentaram maior profundidade. Dentes com LCNC apresentaram maior proporção de facetas de desgaste, contatos oclusais intensos e contatos de interferência oclusal. Saliva em repouso foi coletada durante 5 minutos para quantificação do fluxo, pH, capacidade de tamponamento e proteínas salivares por meio de análise proteômica realizada em 80 participantes. Os indivíduos foram subdivididos em um grupo sem cárie ativa (47,5%) e outros dois grupos com lesões de cárie divididos pela mediana de dentes acometidos: £2 dentes (31,3%) e >2 dentes com cárie ativa (21,3%). Na análise proteômica, foram quantificadas 665 proteínas, envolvidas em processos de imunidade inata, modulação enzimática, atividade dos íons cálcio e adesão celular. Dessas, 68 proteínas demonstraram diferenças significativas de abundância entre os grupos. O conjunto de dados proteômicos foi correlacionado com índice de placa dentária visível, experiência de cárie radicular, fluxo salivar, capacidade tampão, pH e porcentagem de ingestão de açúcar. Com as correlações, 87 candidatos a biomarcadores foram identificados e 19 repetiram a significância da análise anterior. Dessa forma, os resultados deste estudo indicam potenciais biomarcadores e características individualizadas do paciente idoso que podem contribuir para o direcionamento do manejo clínico desse estrato populacional.

Population aging, characteristic of current demographic changes, has been accompanied by greater dental preservation. Older individuals possess particularities that make them more susceptible to the development of oral conditions such as dental caries and noncarious cervical lesions (NCCL). With advanced age, the prevalence of dental caries increases, and NCCL becomes more common and severe. The aim of this study was to assess the frequency of dental caries and NCCL in independent older individuals, as well as to investigate associations between salivary proteins and the presence of caries, and between NCCL and occlusal factors in two distinct articles. In this cross-sectional study, part of the data was collected using a structured interview. Dental caries and NCCL experience were determined by the Decayed, Missing, and Filled Teeth (DMFT) index, Decayed and Filled Roots (DFR) index, and Tooth Wear Index (TWI), collected through a clinical examination. In total, 127 individuals aged between 60 and 84 years were evaluated. The mean DMFT was 23.86, and the frequency of dental caries was 65.4%. For NCCL, the frequency of lesions with a depth greater than 1mm was 65.4%. 3D models were obtained from intraoral scanning for occlusal analysis, morphology, and morphometry of NCCL, in 53 elderly individuals, totaling 1245 teeth with a frequency of 352 (28.27%) teeth with NCCL. Saucer lesions had greater height, while wedge-shaped lesions showed greater depth. Teeth with NCCL presented a higher proportion of wear facets, intense occlusal contacts, and occlusal interference contacts. Unstimulated saliva was collected for 5 minutes to quantify flow, pH, buffering capacity, and salivary proteins through proteomic analysis performed on 80 participants. Individuals were subdivided into a group without active caries (47.5%) and two other groups with caries lesions divided by the median of affected teeth: ≤2 teeth (31.3%) and >2 teeth with active caries (21.3%). In the proteomic analysis, 665 proteins involved in innate immunity, enzymatic modulation, calcium ion activity, and cell adhesion processes were quantified. Of these, 68 proteins showed significant differences in abundance between the groups. The proteomic dataset was correlated with visible dental plaque index, radicular caries experience, salivary flow, buffering capacity, pH, and percentage of sugar intake. Through these correlations, 87 potential biomarker candidates were identified, and 19 replicated the significance of the previous analysis. Thus, the results of this study indicate potential biomarkers and individualized characteristics of elderly patients that may contribute to directing the clinical management of this population stratum.

Sensitivity and specificity of salivary pipecolic acid in head and neck squamous cell carcinoma

Aim: The aim of the present preliminary case-control study was to test the sensitivity and specificity of salivary pipecolic acid in predicting head and neck squamous cell carcinoma (HNSCC). Methods: High-performance liquid chromatography was used for the analysis of non-stimulated saliva samples from 40 individuals: 20 in the case group (recently diagnosed with untreated HNSCC) and 20 in the control group (individuals without cancer). Both groups included patients taking daily oral hypoglycemic drugs (comorbidity). The case and control groups were matched at a proportion of 1:1 for sex and comorbidity. Results: Mean salivary levels of pipecolic acid were 169.38 ng/ mL in the case group and 114.66 ng/mL in the control group (p<0.001). Individuals who took oral hypoglycemic drugs had higher levels of pipecolic acid in both the case and control groups (p<0.001). The receiver operating characteristic curve analysis revealed 90% sensitivity and 65% specificity for head and neck cancer, with an area under the curve of 0.838 between the case and control groups. Conclusions: Pipecolic acid had high sensitivity for the diagnosis of HNSCC but low specificity in the sample analyzed. Our findings suggest that salivary pipecolic acid levels are associated with glucose homeostasis. Studies with larger samples are required to evaluate the specificity of this metabolite

Establishment of a saliva donor selection for in vitro biofilm growth / Seleção de doadores de saliva para crescimento de biofilme in vitro

Introdução: O emprego de biofilmes polimicrobianos, utilizando a saliva como inóculo, é um modelo promissor para o estudo de biofilmes cariogênicos in vitro. Entretanto, ainda não existe uma padronização para seleção de doadores de saliva. Objetivo: O objetivo deste estudo foi estabelecer uma metodologia para seleção de doadores de saliva utilizando fatores salivares microbianos e características in vitro do biofilme. Material e método: Para doação de saliva foram selecionados vinte voluntários. Os voluntários permaneceram 24 horas sem escovar os dentes e ficaram em jejum por 2 horas antes da coleta da saliva. Foram avaliados os seguintes parâmetros: viabilidade das bactérias anaeróbias totais e mutans streptococci; concentração inibitória mínima (CIM) e concentração bactericida mínima (CBM) da clorexidina; capacidade de formação de biofilme por meio da biomassa; e a suscetibilidade dos biofilmes à clorexidina. Resultado: A viabilidade bacteriana da saliva, a capacidade de formação de biofilme e a suscetibilidade do biofilme à clorexidina foram apresentadas como média e intervalo de confiança (95%). A diferença entre a viabilidade do biofilme (mutans streptococci e bactérias totais) após tratamento com NaCl 0,9% e diacetato de clorexidina 0,2% foi comparada pelo teste t de Student com nível de significância estabelecido em 5%. A viabilidade total de bactérias anaeróbias (mediana) foi de 7,28 log 1+UFC/mL (unidades formadoras de colônia/mL). A viabilidade dos mutans streptococci na saliva apresentou mediana de 5,47 log 1+UFC/mL. Para capacidade de formação de biofilme a mediana da biomassa foi de 0,1172 A570. Conclusão: O tratamento com clorexidina reduziu significativamente os mutans streptococci e a viabilidade total das bactérias. A metodologia para seleção do doador de saliva foi estabelecida com sucesso.

Introduction: The utilization of polymicrobial biofilms, with saliva as an inoculum, represents a promising model for in vitro studies on cariogenic biofilms. However, there is still no standardization for selecting saliva donors. Objective: The aim of this study is to establish a methodology for the selection of saliva donors using microbial salivary factors and in vitro biofilm characteristics. Material and method: For saliva donation, twenty volunteers were selected. Volunteers remained 24 h without brushing their teeth and fasted for 2 h before saliva collection. The following parameters were evaluated: total anaerobic bacteria and mutans streptococci viability; minimum inhibitory concentration (MIC) and minimum bactericide concentration (MBC) of chlorhexidine; biofilm forming capacity by biomass assessment; and the susceptibility of biofilms to chlorhexidine. Result: Saliva bacterial viability, biofilm forming capacity and biofilm susceptibility to chlorhexidine were presented as mean and confidence interval (95%). The difference between biofilm (mutans streptococci and Total bacteria) viability after treatment with NaCl 0.9% and 0.2% chlorhexidine diacetate was compared using the Student t-test with a significance level established at 5%. Total anaerobic bacteria viability (median) was 7.28 log 1+CFU/mL (colony forming units/ mL). Mutans streptococci viability in the saliva showed a median of 5.47 log 1+CFU/mL. Biofilm forming capacity showed that biomass had a median of 0.1172 A570. Conclusion: Treatment with chlorhexidine significantly reduced mutans streptococci and total bacteria viability. The methodology for the selection of the saliva donor was successfully established.

Promising applications of human-derived saliva biomarker testing in clinical diagnostics / 国际口腔科学杂志·英文版

Saliva testing is a vital method for clinical applications, for its noninvasive features, richness in substances, and the huge amount. Due to its direct anatomical connection with oral, digestive, and endocrine systems, clinical usage of saliva testing for these diseases is promising. Furthermore, for other diseases that seeming to have no correlations with saliva, such as neurodegenerative diseases and psychological diseases, researchers also reckon saliva informative. Tremendous papers are being produced in this field. Updated summaries of recent literature give newcomers a shortcut to have a grasp of this topic. Here, we focused on recent research about saliva biomarkers that are derived from humans, not from other organisms. The review mostly addresses the proceedings from 2016 to 2022, to shed light on the promising usage of saliva testing in clinical diagnostics. We recap the recent advances following the category of different types of biomarkers, such as intracellular DNA, RNA, proteins and intercellular exosomes, cell-free DNA, to give a comprehensive impression of saliva biomarker testing.

A preliminary study on salivary microbiota of patients with laryngopharyngeal reflux / 中华耳鼻咽喉头颈外科杂志

Objective: To investigate the characteristics of salivary microbiota in patients with laryngopharyngeal reflux (LPR). Methods: A case-control study was applied to enroll 60 patients and healthy subjects who were outpatients of the Department of Otorhinolaryngology Head and Neck Surgery of the Eighth Medical Center of the PLA General Hospital from December 2020 to March 2021, including 35 males and 25 females, aged from 21 to 80 (33.75±11.10) years. Thirty patients with suspected laryngopharyngeal reflux were selected as study group and thirty healthy volunteers without pharyngeal symptoms were selected as control group. Their salivary samples were collected, and the salivary microbiota was detected and analyzed by 16S rDNA sequencing. SPSS 18.0 software was used for statistical analysis. Results: There was no significant difference in the diversity of salivary microbiota between the two groups. At the phylum classification level, the relative abundance of Bacteroidetes in the study group was higher than that in the control group[37.86(31.15, 41.54)% vs 30.24(25.51, 34.18)%,Z=-3.46,P<0.01]. And the relative abundance of Proteobacteria in the study group was lower than that in the control group [15.76(11.81, 20.17)% vs 20.63(13.98, 28.82)%, Z=-1.98,P<0.05]. At the genus level, the relative abundance of Prevotella, Lactobacillus, Parascardovia and Sphingobium in the study group was higher than that in the control group(Z values were-2.92, -2.69, -2.05, -2.31, respectively, P<0.05).And the relative abundance of Streptococcus, Cardiobacterium, Klebsiella and Uruburuella of study group was lower than that of control group(Z values were -2.43, -2.32, -2.17, -2.32, respectively, P<0.05). LEfSe difference analysis showed that there were 39 bacteria with significant differences between the two groups, including Bacteroidetes, Prevotellaceae and Prevotella, which were enriched in the study group, and Streptococcaceae, Streptococcus and other taxa, which were enriched in the control group(P<0.05). Conclusion: The changes of the microflora in the saliva between LPR patients and healthy people suggest that the dysbacteriosis might exist in LPR patients, which may play an important role in the pathogenesis and development of LPR.

Saliva microbiota and metabolite in individuals with caries or periodontitis / 中华口腔医学杂志

Objective: To detect and analyze the characteristics of oral microbiota in species composition, function and metabolism among caries, periodontitis and oral healthy individuals, hunting for the microbiome-derived biomarkers with specificity and sensitivity to estimate the occurrence of these two diseases. Methods: Saliva samples were collected from 10 patients with high caries risk [decayed-missing-filled teeth (DMFT)≥6, HC group] in Department of Endodontics, 10 patients with periodontitis of grade Ⅱ A-Ⅲ C (PG group) in Department of Periodontology and 10 oral healthy individuals (HH group) from School of Stomatology, The Fourth Military Medical University during from March 2022 to June 2022. A baseline examination was conducted on all participants, including their oral conditions of caries and periodontal health. Metagenomic sequencing (Illumina PE150 platform) and liquid chromatography-mass spectrometry were used to detect microorganisms and their metabolites in the samples respectively. The sequencing data were analyzed to obtain the information of microbial taxonomic composition, functional genes and metabolites in each group of samples. The basic oral conditions and saliva samples of subjects in each group were evaluated and collected by the same professional endodontist. Results: There were no significant difference in baseline characteristics such as age and sex among the subjects in each group (P>0.05). DMFT in HC group (9.0±1.7) was significantly higher than that in HH group (0) and PG group (0) (F=243.00, P<0.001). Sequencing data analysis showed that the taxonomic compositions of salivary microbiota in each group were mainly Firmicutes, Bacteroidetes, Proteobacteria, Actinobacteria and Fusobacteria at the phylum level, and Streptococcus, Neisseria, Rothia, Prevotella at the genus level. Differential analysis showed that, compared with the HH group, HC group and PG group had significant differences in taxonomic composition (P<0.05), and the most significant among them was Prevotella. At the species level, Prevotella pallens was the most significant change in HC group, and Porphyromonas gingivalis in PG group. Metabolite analysis showed that there were significant differences in metabolites between HC group and PG group. The results showed that, compared with the HH group, the most significant metabolite change was 3-hydroxy-1, 5-diphenylpentan-1-one in HC group (P=0.001) and N1 acetylspermine in PG group (P=0.002) respectively. Compared with the PG group, the metabolite of HC group with the most significant difference is D-glucosamine 6-phosphate (P=0.006). The metabolism gene function analysis showed that, the enrichment of carbohydrate metabolism related genes was highest in HC group, followed with HH group, and it was lowest in PG group. In addition, compared with the HH group, the abundance of functional genes related to glucose metabolism, such as ABC transporter and phosphotransferase system, were significantly decreased in PG group (P<0.05), but significantly increased in HC group (P<0.05). Conclusions: There is a significant correlation between the alternation of carbohydrate metabolism of salivary microbiota with the occurrence of caries and periodontitis. In the future, Prevotella pallens and 3-hydroxy-1, 5-diphenylpentan-1-one may be the potential biomarkers of caries; while Porphyromonas gingivalis and N1 acetylspermine work in the predictions of periodontitis.

Assessment of salivary opiorphin in oral lichen planus

Opiorphin is a pentapeptide, which could be isolated from human fluids and has a decreasing effect on pain. Aim: Since lichen planus is a chronic mucocutaneous disease, which causes pain or burning feeling in the oral mucosa, this study aimed to compare salivary opiorphin levels of oral lichen planus (OLP) patients with healthy subjects. Methods: This case-control study, was performed on 24 patients with OLP lesions and 21 healthy subjects. After collecting unstimulated saliva, opiorphin levels were compared between two groups through statistical analyses. Results: There was not any significant difference between OLP patients and healthy subjects according to salivary opiorphin concentration (p=0.378). Also, in the OLP group, opiorphin concentration was not significantly different between males and females (p=0.601). Analytical analysis could not show any remarkable difference between various severity of OLP lesions regarding to salivary opiorphin levels (p=0.653). Conclusion: In this study, salivary opiorphin levels was not significantly different between patients with OLP and healthy subjects; however, more studies are suggested for better assessment of salivary opiorphin levels in various types of OLP lesions and its correlation with pain severity

Lineage and phylogenetic analysis of HPV-16, -18 in saliva of HNSCC patients

Aim: Head and Neck Squamous Cell Carcinoma (HNSCC) is a global health problem whose incidence varies by geographic region and race according to risk factors. Human papillomavirus (HPV) infection is a significant risk factor for HNSCC. HPV-16 and HPV-18 are two forms of HPV that are carcinogenic. HNSCCs that are HPV positive have a better prognosis rather than HPV negative. The purpose of this research was to characterize HPV-16, -18 variations in the saliva of HNSCC patients by examining the genetic diversity of HPV-16, -18 utilizing the full E6, E7, and L1 genes. Methods:The case-control research included 15 patients with HNSCC and 15 healthy volunteers. Unstimulated entire saliva samples were obtained from the case and control groups by spitting method. Genomic DNA was isolated from all saliva samples. A PCR reaction was used to determine the presence of HPV in saliva. HPV-positive samples were genotyped and data were analyzed. We conducted a variant study on the HPV-16, -18 E6, and E7 genes. Results: Three patients with HNSCC were HPV-positive for two HPV genotypes out of 30 people diagnosed with HPV-DNA. HPV-16 and -18 were the most common genotypes. The HPV-16, -18 E6, and E7 genes were sequenced and compared to the HPV-16, -18 (E6, E7) prototype sequence. In all, HPV-16 lineages A1 and HPV-18 lineages A3 were discovered. Conclusion: Regarding the variation of HPV found in Iranian HNSCC patients, the need for further studies in HPV genotyping was seen. Sequencing HPV genes in HNSCC may help answer questions about HPV genotyping in the Iranian population. HPV genotype analysis aids in the development of vaccinations against HNSCC, halting disease progression and preventing HPV-associated HNSCC

O papel da saliva na pandemia por COVID-19: alternativa diagnóstica para um problema de saúde pública global / The role of saliva in the COVID-19 pandemic: diagnostic alternative for a global public health problem

O objetivo desse estudo foi relatar as evidências disponíveis sobre o papel da saliva no diagnóstico da Covid-19. Trata-se de uma revisão narrativa da literatura de cunho qualitativo com buscas realizadas a partir das bases de dados eletrônicas PubMed, Google Acadêmico e Scielo, nos idiomas português e inglês utilizando os termos "Covid-19", "Diagnóstico" e "Saliva". Foi observado a utilização da saliva com a finalidade de se monitorar o estado de saúde e da doença de uma pessoa cujo objetivo tem se tornado extremamente desejável quanto à promoção da saúde e à pesquisa de cuidados em saúde. Nesse sentido, amostras de saliva podem ser usadas no diagnóstico da Covid-19. Um obstáculo crítico para o diagnóstico salivar pode ser a validação de amplo espectro em pacientes com Covid-19 durante o período de incubação, a fase de resposta viral e a fase inflamatória do hospedeiro de indivíduos assintomáticos e sintomáticos. Conclui-se que a saliva pode ter aplicações potenciais no contexto da Covid-19, mas, são necessários mais estudos para investigar o potencial diagnóstico da Covid-19 na saliva e seu impacto na transmissão desse vírus.

The aim of this study was to review the available evidence on the role of saliva in the diagnosis of Covid-19. This is a narrative review of the literature of a qualitative nature whose search was carried out from the digital databases PubMed, Academic Google and Scielo, in Portuguese and English, using the terms "Covid-19", "Diagnosis" and "Saliva". It was observed that the use of saliva in order to monitor a person's state of health and illness it became an objective extremely desirable in terms of health promotion and health care research. In this sense, saliva samples can be used in the diagnosis of Covid-19. A critical obstacle for salivary diagnosis may be broad-spectrum validation in patients with Covid-19 during the incubation period, the viral response phase, and the host inflammatory phase in asymptomatic and symptomatic patients. It is concluded that saliva may have potential applications in the context of Covid-19, but further studies are needed to investigate the diagnostic potential of Covid-19 in saliva and its impact on the transmission of this virus.

Influence of NaHCO3 powder on color and roughness of the composite submitted to bleaching and euterpe oleracea / Influencia del polvo de NaHCO 3 en el color y rugosidad del composite sometido a blanqueo y euterpe oleracea

Aim: This study aims to evaluate the surface roughness and susceptibility to staining of bleached composite resin with 22% carbamide peroxide, as well as the effect of subsequent prophylaxis with NaHCO3 powder. Material and Methods: Forty disk-shaped (2 × 6 mm) specimens of composite resin (Z250 XT) were prepared. Half of the specimens were subjected to bleaching with 22% carbamide peroxide, and the other half were stored in artificial saliva. In sequence, all specimens were immersed in acai juice (Euterpe oleracea) for 4 h for 14 days, and subdivided into two groups. Group 1 samples were subjected to prophylaxis treatment, while group 2 samples were subjected to treatment with artificial saliva. Surface roughness (Ra) and color (ΔE*) were measured after polishing (T0), bleaching (T1), immersion in acai juice (T2), and application of NaHCO3 powder (T3) using a profilometer and a spectrophotometer. Results: Statistical analyses (analysis of variance and Tukey's test, p≤0.05) revealed that regarding color there was statistically significance for the factors in isolation, except for the factor bleaching. For both color and surface roughness there was statistically significant difference for the interaction, except for the interaction between NaHCO3 power and bleaching. Conclusion: The NaHCO3 air-powder polishing decreases the staining of the composite resin; however, it increases the surface roughness. With respect to the color variable, the whitening factor had no significant effect on the tested material; however, it increases surface roughness.

Objetivo: Este estudio tiene como objetivo evaluar la rugosidad de la superficie y la susceptibilidad a la tinción de la resina compuesta blanqueada con peróxido de carbamida al 22%, así como el efecto de la profilaxis posterior con polvo de NaHCO3. Material y Métodos: Se prepararon cuarenta especímenes en forma de disco (2 × 6 mm) de resina compuesta (Z250 XT). La mitad de los especímenes se sometieron a blanqueo con peróxido de carbamida al 22% y la otra mitad se almacenó en saliva artificial. En secuencia, todos los especímenes se sumergieron en jugo de acai (Euterpe oleracea) durante 4h durante 14 días, y se subdividieron en dos grupos. Las muestras del grupo 1 se sometieron a tratamiento profiláctico, mientras que las muestras del grupo 2 se sometieron a tratamiento con saliva artificial. La rugosidad de la superficie (Ra) y el color (ΔE*) se midieron después del pulido (T0), el blanqueo (T1), la inmersión en jugo de acai (T2) y la aplicación de polvo de NaHCO3 (T3) utilizando un perfilómetro y un espectrofotómetro. Resultados: Los análisis estadísticos (análisis de varianza y prueba de Tukey, p≤0.05) revelaron que en relación al color hubo significancia estadística para los factores en forma aislada, excepto para el factor blanqueamiento. Tanto para el color como para la rugosidad de la superficie hubo una diferencia estadísticamente significativa para la interacción, excepto para la interacción entre el poder de NaHCO3 y el blanqueo. Conclusión: El pulido al aire con polvo de NaHCO3 disminuye el manchado de la resina compuesta; sin embargo, aumenta la rugosidad de la superficie. Con respecto a la variable color, el factor de blanqueamiento no tuvo efecto significativo sobre el material ensayado; sin embargo, aumenta la rugosidad de la superficie.

Correlation between salivary biomarkers and plaque-induced gingivitis exacerbated by pregnancy

Objetive: To evaluate the correlation between salivary biomarkers (the salivary antioxidant ability, salivary level of polyphenols, and other antioxidants) with plaque-induced gingivitis exacerbated by pregnancy in pregnant and nonpregnant women. Material and Methods: For this observational study, medical records, dental examinations, and analyses of saliva samples were carried out in pregnant and nonpregnant women. A p-value of <0.05 was considered significant. Results: The pregnant women (n =17) exhibited a lower antioxidant capacity (p-value=0.0041), higher levels of polyphenols, gingival index, bleeding on probing, and subjects consuming mineral-enriched products (p-value from <0.0001 to 0.0466), and unchanged levels of phosphotungstic acid reactive substances, proteins, oral hygienic habits, plaque index and probing depth (p-value from 0.0683 to 0.8358), in comparison with the nonpregnant women (n=9). Also, a positive correlation between the gingival index and salivary polyphenol content was observed (r-value = 0.4087, p-value = 0.0202). Conclusion: The salivary polyphenols correlate with plaque-induced gingivitis exacerbated by pregnancy, suggesting a deficiency of salivary antioxidant protection.

Objetivo: Evaluar la correlación entre los biomarcadores salivales (la capacidad antioxidante salival, el nivel salival de polifenoles y otros antioxidantes) con la gingivitis inducida por placa exacerbada por el embarazo en mujeres embarazadas y no embarazadas. Material y Métodos: Para este estudio observacional, se realizaron registros médicos, exámenes dentales y análisis de muestras de saliva en mujeres embarazadas y no embarazadas. Se consideró significativo un valor de p<0,05. Resultados: Las gestantes (n=17) presentaron menor capacidad antioxidante (p=0,0041), mayores niveles de polifenoles, índice gingival, sangrado al sondaje y los sujetos que consumían productos enriquecidos con minerales (p<0,0001 a p<0,0466), y no hubo diferencias en los niveles de sustancias reactivas al ácido fosfotúngstico, proteínas, hábitos de higiene bucal, índice de placa y profundidad de sondaje (p=0,0683 a 0,8358), en comparación con las mujeres no embarazadas (n=19). Además, se observó una correlación positiva entre el índice gingival y elcontenido de polifenoles salivales (r = 0,4087, p= 0,0202). Conclusión: Los polifenoles salivales se correlacionan con la gingivitis inducida por placa y exacerbada por el embarazo, lo que sugiere una deficiencia de protección antioxidante salival.

Biomarcadores salivares do carcinoma de células escamosas / Salivary biomarkers of squamous cell carcinoma

O carcinoma de células escamosas (OSCC) é uma neoplasia maligna que atinge a cavidade oral, lábios e orofaringe com uma das maiores taxa de mortalidade em todo o mundo, quando em comparação com outros carcinomas, o que o torna um problema de saúde pública (MORO, 2018). Devido à sua grande prevalência, os pesquisadores buscam por mecanismos para aprimorar o diagnóstico da doença em sua fase inicial, no intuito de possibilitar melhor qualidade de vida e sobrevida a esses pacientes (CHENG et al., 2014). Tem-se desenvolvido estudos através da expressão de proteínas, envolvendo o mecanismo biomolecular da carcinogênese oral na busca de identificação de biomarcadores que tenham potencial preditivo e um bom prognóstico para OSCC (CARVALHO; OLIVEIRA, 2015; LOUSADA-FERNANDEZ et at., 2018). Em uma revisão sobre o genoma salivar do câncer oral, este destacou-se justamente pelo fato de suas proteínas estarem localmente expressas (SHAH et at., 2011), tornando seus biomarcadores salivares de fácil e rápida coleta, já que a proliferação desordenada de células malignas deixa derivados de DNA, RNA, vesiculas (exossomos) e marcadores proteicos nos fluidos creviculares, podendo demostrar de forma preventiva o câncer. Com isso, esses biomarcadores podem ser identificados na fase inicial da doença, ao contrário do exame clínico, já que as manifestações clínicas são tardias, o que gera um prognóstico ruim (LOUSADA-FERNANDEZ et al., 2018). Diante desse quadro, o objetivo deste trabalho foi fazer uma revisão de literatura acerca do papel dos biomarcadores tumorais e discutir sobre a atual situação do câncer bucal, justificando o uso de biomarcadores salivares e entendendo seu papel no diagnóstico precoce. Para isso foi feita uma busca detalhada nas bases de dados Pubmed-Medline, Biblioteca Virtual em Saúde, Lilacs e Scielo, para identificar os mais recentes biomarcadores citados nas literaturas ultilizando as palavras-chaves: carcinoma, biomarcadores, saliva. Também foram incluídos dados coletados da Organização Mundial da Saúde e do Instituto Nacional do Câncer.

Effects of acid stimulation on saliva flow rate and compositions of human parotid and submandibular glands / 北京大学学报(医学版)

OBJECTIVE@#To investigate the effect of acid stimulation on salivary flow rate and compositions of human parotid and submandibular glands, so as to provide basis for comprehensive evaluation of salivary gland function in both health and disease status.@*METHODS@#In the study, 210 healthy participants' whole saliva samples were collected under passive drooling, and their parotid gland and submandibular gland secretions were collected by negative pressure suction. 2% citric acid was dropped on the tip of tongue every 1 min for acid stimulation for a total of 5 times to collect stimulated whole saliva, parotid and submandibular gland saliva. The collected saliva was weighed and saliva flow rate was calculated. The K+, Na+, Cl-, Ca2+, total protein, total phosphorus and α-amylase in saliva samples were detected by biochemical analyzer, and the changing features of flow rate and compositions of different kinds of saliva were compared and analyzed.@*RESULTS@#After acid stimulation, saliva flow rate significantly increased. The increase proportion of parotid gland saliva (10.7 folds) was much higher than that of submandibular gland saliva (2.9 folds). The concentrations of Na+, Cl-, Ca2+, total protein and α-amylase in parotid gland saliva increased significantly (P < 0.05), but there was no significant difference in total phosphorus and K+ (P=0.89, P=0.34). The concentration of Na+ and Ca2+ in saliva of submandibular gland increased significantly(P < 0.05), the concentration of total phosphorus decreased significantly(P < 0.05), and the concentration of Cl- increased, but the difference was not significant(P=0.068). There was no significant difference in total protein, K+ and α-amylase (P=0.85, P=0.07, P=0.95). The compound secretion rate of total phosphorus in saliva of submandibular gland remained unchanged(P=0.066), while the secretion rate of K+, Na+, Cl-, Ca2+, total protein and α-amylase significantly increased(P < 0.01). The compound secretion rate of K+, Na+, Cl-, Ca2+, total protein and total phosphorus and α-amylase in parotid gland saliva increased(P < 0.01). The concentrations of Na+, Cl-, K+, total phosphorus, total protein and α-amylase in parotid were higher than those in submandibular gland (P < 0.01), and the concentration of Ca2+ in submandibular gland saliva was significantly higher than that in parotid (P < 0.001).@*CONCLUSION@#The response of parotid to acid stimulation is stronger, and the secretion of submandibular gland is more stable. Acid stimulation significantly influences the concentrations of electrolytes in saliva, and the composited secretion rate is an evaluation index to reflect both flow rate and composition concentration of saliva. The parotid gland plays an important role in the secretion of total protein, total phosphorus and α-amylase in saliva, and the submandibular gland is the main source of Ca2+ in saliva.

Application and Prospect of RNA Profiling Analysis in Forensic Body Fluid Identification / 法医学杂志

In forensic physical evidence identification, the accurate identification of the individual origin and their body fluid composition of the biological samples obtained from the crime scene play a critical role in determining the nature of a crime. In recent years, RNA profiling has become one of the fastest developing methods for body fluids identification. Due to the characteristics of tissue or body fluid specific expression, various types of RNA markers have been proven to be promising candidate markers for body fluids identification in previous studies. This review summarizes the research progress of RNA markers in body fluids identification, including the RNA markers that have been effectively verified in current research and their advantages and disadvantages. Meanwhile, this review prospects the application of RNA markers in forensic medicine.

Application of Duplex Droplet Digital PCR Detection of miR-888 and miR-891a in Semen Identification / 法医学杂志

OBJECTIVES@#To establish a system for simultaneous detection of miR-888 and miR-891a by droplet digital PCR (ddPCR), and to evaluate its application value in semen identification.@*METHODS@#The hydrolysis probes with different fluorescence modified reporter groups were designed to realize the detection of miR-888 and miR-891a by duplex ddPCR. A total of 75 samples of 5 body fluids (including peripheral blood, menstrual blood, semen, saliva and vaginal secretion) were detected. The difference analysis was conducted by Mann-Whitney U test. The semen differentiation ability of miR-888 and miR-891a was evaluated by ROC curve analysis and the optimal cut-off value was obtained.@*RESULTS@#There was no significant difference between the dual-plex assay and the single assay in this system. The detection sensitivity was up to 0.1 ng total RNA, and the intra- and inter-batch coefficients of variation were less than 15%. The expression levels of miR-888 and miR-891a detected by duplex ddPCR in semen were both higher than those in other body fluids. ROC curve analysis showed that the AUC of miR-888 was 0.976, the optimal cut-off value was 2.250 copies/μL, and the discrimination accuracy was 97.33%; the AUC of miR-891a was 1.000, the optimal cut-off value was 1.100 copies/μL, and the discrimination accuracy was 100%.@*CONCLUSIONS@#In this study, a method for detection of miR-888 and miR-891a by duplex ddPCR was successfully established. The system has good stability and repeatability and can be used for semen identification. Both miR-888 and miR-891a have high ability to identify semen, and the discrimination accuracy of miR-891a is higher.

Psychological stress factors and salivary cortisol in nursing students throughout their training / Factores de estrés psicológico y cortisol salivar en alumnos de enfermería a lo largo de la formación / Fatores de estresse psicológico e cortisol salivar em alunos de enfermagem ao longo da formação

ABSTRACT Objective: to analyze psychological stress factors and salivary cortisol concentration in nursing undergraduates throughout their training. Method: a cross-sectional, analytical, and comparative study carried out in an evening course using a sociodemographic questionnaire, an Instrument to Assess Stress in Nursing Students, and salivary cortisol analysis. The study included descriptive and comparative analyses and a multiple linear regression model. Results: 187 participants answered the questionnaires, and 129 had their cortisol quantified. The domains Practical Activities Execution, Professional Communication, and Professional Training represented the stress factors with the highest mean values for 3rd, 4th, and 5th-year students compared to 1st and 2nd year. For the 5th year, it was the domains Professional Communication and Professional Training compared to the 3rd year and Environment compared to the 1st and 3rd year. A significant result was obtained between the times of cortisol collections for males (p < 0.0001), females (p < 0.0001), and for 1st (p = 0.0319) 2nd (p = 0.0245), and 5th (p < 0.0001) years. Conclusion: Students in years 3 through 5 had higher exposure to stressors, and there were adjustments in cortisol production rhythmicity for students in years 1, 2, and 5.

RESUMEN Objetivo: analizar los factores de estrés psicológico y la concentración de cortisol salivar de estudiantes de enfermería a lo largo de su formación. Método: es un estudio transversal, analítico y comparativo realizado en un curso nocturno mediante cuestionario sociodemográfico, Instrumento para Evaluación del Estrés en Estudiantes de Enfermería y análisis del cortisol salivar. Se llevaron a cabo diversos análisis descriptivos, comparativos y modelo de regresión linear múltiple. Resultados: 187 respondieron a los cuestionarios y a 129 se les cuantificó el cortisol. Los dominios Realización de Actividades Prácticas, Comunicación Profesional y Formación Profesional representaron los factores de estrés con los valores medios más altos en los estudiantes de 3er, 4º y 5º año en comparación con los de 1er y 2º año. Para el 5º año fueron los dominios Comunicación Profesional y Formación Profesional en comparación con el 3er año y Medio Ambiente en comparación con el 1er y 3er año. Se obtuvo un resultado significativo entre los momentos de las recogidas de cortisol de los hombres (p < 0,0001), de las mujeres (p < 0,0001), y del 1er (p = 0,0319), 2º (p = 0,0245) y 5º (p < 0,0001) año. Conclusión: los alumnos de 3er a 5º año tuvieron una mayor exposición a los estresores y hubo ajustes en la ritmicidad de la producción de cortisol en los alumnos de 1er, 2º y 5º año.

RESUMO Objetivo: analisar os fatores de estresse psicológico e a concentração de cortisol salivar de graduandos de Enfermagem ao longo da formação. Método: estudo transversal, analítico e comparativo realizado em curso noturno por meio de questionário sociodemográfico, Instrumento para Avaliação de Estresse em Estudantes de Enfermagem e análise do cortisol salivar. Foram feitas análises descritivas, comparativas e modelo de regressão linear múltipla. Resultados: um total de 187 responderam aos questionários, e 129 tiveram o cortisol quantificado. Os domínios Realização das Atividades Práticas, Comunicação Profissional e Formação Profissional representaram os fatores de estresse com os maiores valores médios para alunos do 3º, 4º e 5º anos em comparação ao 1º e 2º anos. Para o 5º ano, foram os domínios Comunicação Profissional e Formação Profissional em relação ao 3º ano e Ambiente em comparação ao 1º e 3º anos. Obteve-se resultado significativo entre os horários das coletas de cortisol para homens (p < 0,0001), mulheres (p < 0,0001) e para o 1º (p = 0,0319) 2º (p = 0,0245) e 5º (p < 0,0001) anos. Conclusão alunos do 3º ao 5º ano tiveram maior exposição aos fatores de estresse, e houve ajustes na ritmicidade de produção do cortisol para alunos do 1º, 2º e 5º anos.