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.

Tempo de viabilidade da saliva humana em meio externo para fins de extração e purificação do DNA: uma revisão de escopo / Viability time of human saliva in the external environment for DNA extraction and purification: a scoping review

Introdução: A análise do DNA salivar é uma dos métodos de identificação humana relacionados à Odontologia Legal. A coleta do material genético salivar consiste num processo simples e pouco invasivo, por possuir grande potencial discriminatório tem sido amplamente aplicado em investigações criminais. Objetivo: Por meio de uma revisão de escopo, objetivou-se mapear o tempo de viabilidade da saliva humana em meio externo para fins de extração e purificação de DNA. Material e Métodos: A revisão foi conduzida seguindo o protocolo JBI e registrada (doi: 10.17605/OSF.IO/PN9ET). A estratégia de busca foi adaptada para as bases: Pubmed, Web of Science, Scopus, LILACS, Cochrane e Google Scholar, sem restrições sobre período de publicação ou idioma. Foram excluídos estudos que exploraram apenas as metodologias e materiais relacionados a extração de DNA , também aqueles que estudaram DNA de outros componentes que não a saliva. Resultados: Identificou-se 283 estudos. Após triagem inicial, 15 referências foram lidas na íntegra, sendo 6 incluídas na revisão, em função da confirmação da elegibilidade. Bitucas de cigarro, próteses dentárias, pastilhas de compressão dentária, cavidade oral e cartões de FTA foram os substratos descritos como fonte de saliva coletada. A viabilidade do DNA foi verificada em tempos que variaram de 1 dia à 11 anos. Conclusão: O protocolo de coleta e armazenamento das amostras é um fator que pode influenciar a quantidade e qualidade do material examinado, todavia, observou-se DNA viável em análise realizada uma década após a coleta da saliva e esse foi o tempo máximo de acompanhamento relatado nos estudos

Introduction: Salivary DNA analysis is one of the human identification methods related to forensic dentistry. Collection of salivary genetic material consists of a simple and poorly invasive process because it has great discriminatory potential has been widely applied in criminal investigations. Objective: Through a scope review, the feasibility time of human saliva was mapped in the external environment for DNA extraction and purification purposes. Material and Methods: The review was conducted following the JBI protocol and registered (doi: 10.17605/OSF.IO/PN9ET). The search strategy was adapted for the databases: Pubmed, Web of Science, Scopus, LILACS, Cochrane and Google Scholar, with no restrictions on period of publication or language. Studies that explored only methodologies and materials related to DNA extraction were excluded, as well as those that studied DNA from components other than saliva. Results: 283 studies were identified. After initial screening, 15 references were read in full, 6 of which were included in the review. Cigarette butts, dentures, dental compression tablets, oral cavity and FTA cards were the substrates described as a source of collected saliva. DNA viability was verified in times ranging from 1 day to 11 years. Conclusion: The sample collection and storage protocol is a factor that can influence the quantity and quality of the material examined, however, viable DNA was observed in an analysis performed a decade after saliva collection and this was the maximum reported follow-up time in the studies

Study on Biochemical Perspectives of Antioxidant and Oxidant Indices in Oral Squamous Cell Carcinomas (OSCCs) and Other Oral Potentially Malignant Disorders (OPMDs)

Background-Oxidative stress in biological systems is a complex process that is characterized by an inequity between the production of free radicals (FR) and the ability of the body to eliminate these reactive species through the use of endogenous and exogenous antioxidants. The pathogenesis of oral cancer has been linked to alterations in the antioxidant defense mechanism. Materials & Methods- Saliva from twenty patients with OSCC, forty patients with OPMDs and twenty healthy subjects in the age group of thirty five to seventy five years was analyzed for levels of nitric oxide, vitamin C, total sialic acid and GSH using spectrophotometry. Results - The levels of salivary vitamin C and glutathione were significantly reduced and those of nitric oxide and sialic acid were raised in patients having OPMD's and oral squamous cell carcinoma. The co- relation between the AOI and calculated ratios indicated that antioxidant potential of the saliva was decreased and was statistically related to (p <0.001) development of OPMD, which further may progress to oral cancer, notably OSCC. Conclusion- The current study demonstrated that the estimation of vitamin C, nitric oxide, sialic acid and GSH in saliva could be used as an early potential diagnostic biomarker in the screening of oral cancer. The antioxidant-oxidant indices (AOI's) can be used as a reliable tool for predicting the oral microenvironment and its predicted change towards development of oral cancer. This optimized developed protocol was also found to be simple and cost effective.

Role of Saliva in Point of Care Technology

Introduction: Saliva has been used as sample for point of care monitoring. In addition to its feasibility of use, the advantages of saliva when compared to other body fluids, the new technically advanced methods to detect the biomarkers of interest in saliva rendered the point of care testing (POCT) for wider applications. Significance of POCT: An accurate, quick and portable POCT is ideally required for hospital, laboratory and personal ap- plications. Together with point of care diagnostics, the scientifically validated numerous upcoming biomarkers will extend their applicability for real time health tracking and personalized precision medicine in different areas of health care systems. The utilization of salivary POC analytical procedures is perhaps in its emerging stage. Conclusion: This paper summarizes the overall perception regarding the salivary point of care testing and its applications.

Diagnostic utility of salivary pepsin in laryngopharyngeal reflux: a systematic review and meta-analysis

Abstract Objective: Salivary pepsin has emerged as a biomarker for Laryngopharyngeal Reflux (LPR), which, however, has been questioned for its efficacy due to a lack of supporting medical data. Therefore, this study analyzed the diagnostic value of salivary pepsin for LPR and assessed a better cutoff value. Methods: Studies were searched in PubMed, Embase, and Cochrane Library from their receptions to October 1, 2021. Then, RevMan 5.3 and Stata 14.0 were utilized to summarize the diagnostic indexes for further meta-analysis. Data were separately extracted by two reviewers according to the trial data extraction form of the Cochrane Handbook. The risk of bias in Randomized Control Trials (RCTs) was evaluated with the Cochrane Risk of Bias Tool. Results: A total of 16 studies matched the criteria and were subjected to meta-analysis. The results revealed a pooled sensitivity of 61% (95% CI 50%-71%), a pooled specificity of 67% (95% CI 48%-81%), a positive likelihood ratio of 2 (95% CI 1.2-2.8), a negative likelihood ratio of 0.58 (95% CI 0.47-0.72), and the area under the receiver operating characteristic curve of 0.67 (95% CI 0.63-0.71). Subgroup analyses indicated that the cutoff value of pepsin at 50 ng/mL had a higher degree of diagnostic accuracy than that of pepsin at 16 ng/mL in cohort studies. Conclusion: The review demonstrated low diagnostic performance of salivary pepsin for LPR and that the cutoff value of 50 ng/mL pepsin had superior diagnostic accuracy. Nevertheless, the diagnostic value may vary dependent on the utilized diagnostic criteria. Therefore, additional research is needed on the improved way of identifying salivary pepsin in the diagnosis of LPR, and also longer-term and more rigorous RCTs are warranted to further assess the effectiveness of salivary pepsin.

The potential of salivary albumin to degrade composite resin / Albumina salivar: investigando seu potencial de degradar resinas compostas

ABSTRACT Albumin is a salivary enzyme capable of cleaving ester linkages and catalyzing degradation of resin-based dental materials. However, the effect of concentration-dependent esterolytic action on composite resins as yet remains unexplored. Aim: The purpose of this study was to evaluate whether artificial saliva formulations with different concentrations of albumin affected the surface roughness, flexural strength and microhardness of a composite resin. Materials and Method: Specimens (25x2x2mm) of a nanofilled composite (Filtek Z350XT, 3M/ESPE) were prepared and analyzed for average surface roughness (Ra/pm). The specimens were then allocated to 6 groups (n=30), to be treated with different salivary albumin concentrations: 0, 10, 50, 100, 200, 400 pg/mL. The specimens were stored in their respective artificial saliva groups, half of them for 24 h and the remainder for 180 days (artificial saliva renewed weekly), after which they were submitted to a new Ra reading, and tested for three-point flexural strength (FS, MPa). The specimens stored for 180 days were analyzed for Knoop microhardness (KH, Kg/mm2). Data were submitted to two-way ANOVA (Ra and FS) and one-way ANOVA (KH). Results: Although Ra increased (p < 0.001) and FS decreased (p < 0.001) from 24 hours to 180 days of storage, the albumin concentration did not significantly affectRa (p = 0.168), FS (p = 0.477) or KH (p = 0.378). Conclusion: The esterolytic action of albumin did not increase the artificial-saliva-induced hydrolytic degradation of the composite resin.

RESUMO Albumina, uma enzima encontrada na saliva, é capaz de clivar ligagoes éster e catalisar a degradando de materiais dentários á base resina. Apesar da agao esterolítica ser potencialmente concentragao-dependente, a investigando desse efeito sobre resinas compostas ainda permanence inexplorado. Objetivo: O objetivo deste estudo foi avaliar se formulagoes de saliva artificial contendo diferentes concentragoes de albumina afetariam a rugosidade superficial, a resistencia flexural e a microdureza de uma resina composta. Materials e Método: Corpos de pro-va em barra (25x2x2mm) foram confeccionados a partir de uma resina composta nanoparticulada (Filtek Z350XT, 3M/ESPE) e foram submetidos á leitura de rugosidade superficial média inicial (Ra, pm), em rugosímetro. Entao, as amostras foram divididas em 6 grupos (n=30) de acordo com a concentrando de albumina na saliva: 0, 10, 50, 100, 200, 400 pg/mL. As amostras foram armazenadas nas formulagoes de saliva artificial correspondente ao seu grupo, metade por 24 h e as demais por 180 dias (com trocas de saliva semanais). As amostras foram submetidas a no-vas leituras de rugosidade (Rafinal) e avaliadas quanto á resistencia flexural de tres pontos (RF, MPa). As amostras armazenadas por 180 dias foram avaliadas quanto á microdureza Knoop (KH, Kg/mm2). Os dados foram submetidos a análises de variáncia a dois critérios (Ra e RF) e a um critério (KH). Resultados: Apesar de haver aumento na Ra (p < 0,001) e uma diminuigao da RF (p < 0,001) de 24 h para 180 dias, a concentragao de albumina nao afetou significativamente a Ra (p = 0,168), a RF (p = 0,477) ou a KH (p = 0,378). Conclusoes: A agdo esterolítica da albumina nao aumentou a degradagao hidrolítica da resina composta induzida pela saliva artificial.

Association of Hyper-Low-Density Lipoprotein and Hypo-High-Density Lipoprotein Cholesterolemia with Low Saliva Flow Rates in Japanese Community-Dwelling Elders

Abstract Introduction The associations of low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein (HDL-C) with reduced saliva flow rates have not been previously reported. Objective The present study aimed to assess the association of cholesterolemia with reduced saliva flow rates in community-dwelling elderly subjects. Methods The present study analyzed 342 participants (170 males and 172 females aged between 78 and 79 years old). Unstimulated salivary flow rate (USFR) was assessed using a cotton roll method. Low-USFR was defined as 0.10 g/30 seconds. Stimulated salivary flow rate (SSFR) was assessed by having the participants chew tasteless gum for 3minutes. Low-SSFR was defined as 1.0 mL/minute. Blood samples were collected for the measurement of LDL-C, HDL-C, rheumatoid factor, hemoglobin A1c, and creatinine. To assess depression, the General Health Questionnaire 30 was used. A standardized questionnaire was completed, covering the current and previous medications of the participants and smoking status. We stratified the serum LDL-C levels of the participants as normal, moderate or severe hyper-LDL cholesterolemia and serum HDL-C levels as normal or hypo-HDL cholesterolemia. Multivariate logistic regression models were established and low-USFR or low-SSFR were set as dependent variables in the aforementioned models. Results After controlling for the effects of the other variables, the odds ratios (ORs) (95% confidence intervals [CIs]) for low-USFR were 2.25 (1.10-4.61) for moderate and 5.69 (1.55-20.8) for severe hyper-LDL cholesterolemia, while that of hypo-HDL cholesterolemia was 3.40 (1.33-8.69). Severe hyper-LDL cholesterolemia was also associated with low-SSFR with an OR of 3.89 (1.39-10.88). Conclusion Elderly patients with cholesterolemia have a risk of reduced salivary flow rate.

Evaluation of Salivary Flow Rate in Patients undergoing Chemotherapy for Solid Tumors / Avaliação de Fluxo Salivar em Pacientes Submetidos à Quimioterapia para Tumores Sólidos

Chemotherapy for cancer treatment offers the possibility of eradicating neoplastic cells, however, its use can result in systemic and oral adverse effects, with emphasis on the qualitative-quantitative modification of saliva. To evaluate the stimulated salivary flow rate (SFR) in patients undergoing chemotherapy for treatment of solid malignancies. This was a cross-sectional study with twenty patients diagnosed with solid tumors undergoing chemotherapy with cytotoxic drugs. Subjects were submitted to stimulated sialometry in the 1st cycle of chemotherapy (baseline, 7th day and 14th day). Additionally, sociodemographic data regarding these patients were collected in a standardized questionnaire. Descriptive and inferential statistical analysis were performed, with a significance level of p<0.05. There was a predominance of women (75%) aged over 40 years old (90%). The most prevalent tumor location was breast (55%), followed by ovaries (15%), and the most used drug was cyclophosphamide (50%). In the sample, no statistically significant result was observed with regard to SFR (p>0.05) in the studied periods. The data suggest that the 1st cycle of chemotherapy was not able to induce significant changes in the SFR. New studies need to be performed for the prospective identification of potential changes in salivary parameters.(AU)

A quimioterapia para tratamento do câncer oferece possibilidade de erradicação das células neoplásicas, todavia, seu emprego pode resultar em efeitos adversos sistêmicos e em cavidade oral, com destaque para modificação quali-quantitativa da saliva. Avaliar a velocidade de fluxo salivar (VFS) estimulado em pacientes sob quimioterapia para tratamento de neoplasias malignas sólidas. Tratou-se de um estudo transversal, no qual vinte pacientes com tumores sólidos sob regime quimioterápico com drogas sabidamente citotóxicas, foram submetidos à sialometria estimulada no 1º ciclo de quimioterapia (baseline, 7º dia e 14º dia). Adicionalmente, foram coletados dados referentes às características sociodemográficas desses pacientes. Foi realizada análise estatística descritiva e inferencial, com nível de significância p<0,05. Observou-se predominância de mulheres (75%) em idade acima dos 40 anos (90%). A localização do tumor mais prevalente foi a mama (55%), seguida de ovários (15%) e a droga mais utilizada foi a ciclofosfamida (50%). Na amostra não foi observado resultado estatisticamente significativo no que diz respeito à variável VFS (p>0,05) nos períodos estudados. Os dados sugerem que o 1° ciclo de quimioterapia não foi capaz de induzir alterações significativas na VFS. Novos estudos precisam ser realizados para a identificação prospectiva de potenciais alterações nos parâmetros salivares.(AU)

A Mouthwash with Cetylpyridinium Chlorideis reducing salivary SARS-CoV-2 viral load in +COVID-19 / Un enjuague con Cloruro de Cetilpiridinio redujo la carga salival del virus SARSCoV-2 en pacientes con Covid-19

Aim: The aim of this randomised, double-blind, placebo-controlled pilot clinical trial is to evaluate the capacity of a mouthwash to reduce SARS-CoV-2 viral load in the saliva of patients with COVID-19. Methods: Twenty-three symptomatic SARS-CoV-2-positive outpatients were selected andrandomised into two groups and registered at NTC 04563689. Both groups rinsed and gargled for one minute with either distilled water (Placebo) or with 0.05% Cetylpyridinium chloride (CPC) plus 0.12% Chlorhexidine (CHX) mouthwash (PERIOAID Intensive Care). Saliva samples were collected before the use of placebo or mouthwash and after 15 minutes and 1 and 2 hours of either of the above treatment. A saliva sample was also taken five days after regular use of placebo or mouthwash twice daily. The virus was detected by qRT-PCR. Results: A great heterogeneity in the viral load values was observed at baseline in both groups for nasopharyngeal and saliva samples. Most of the patients who used the mouthwash (8/12) had a significant decrease in baseline viral load after 15 min (greater than 99% reduction). This inhibitory effect was maintained for up to two hours in 10 of the 12 patients. At five days, SARS-CoV-2 RNA was detectedin only 1 patient from the mouthwash group and in 5 from the placebo group. Conclusions: This study points out that a CPC mouthwash can reduce the viral load in saliva of COVID-positive patients. This finding may be important in transmission control of SARS-CoV-2. Nevertheless, the clinical relevance of CPC mouthwash-reduction on SARS-CoV-2 shedding in saliva requires further study.

Objetivo: El objetivo de este ensayo clínico piloto aleatorizado, doble ciego y controlado con placebo es evaluar la capacidad de un enjuague bucal para reducir la carga viral del SARS-CoV-2 en la saliva de pacientes con COVID-19. Materiales y métodos:Veintitrés pacientes ambulatorios positivos para SARS-CoV-2 sintomáticos fueron seleccionados y aleatorizados en dos grupos y registrados en el NTC 04563689. Ambos grupos se enjuagaron y hicieron gárgaras durante un minuto con agua destilada (placebo) o con cloruro decetilpiridinio al 0 ,05 % (CPC). ) más enjuague bucal con Clorhexidina (CHX) al 0,12% (PERIOAID Intensive Care). Se recolectaron muestras de saliva antes del uso de placebo o enjuague bucal y después de 15 minutos y 1 y 2 horas de cualquiera de los tratamientos anteriores. También se tomó una muestra de saliva cinco días después del uso regular de placebo o enjuague bucal dos veces al día. El virus fue detectado por qRT-PCR. Resultados:Se demostró una gran heterogeneidad en los valores de carga viral al inicio del estudio en grupos ambos para muestras de nasofaringe y saliva. La mayoría de los pacientes que usaron el enjuague bucal (8/12) tuvieron una disminución significativa en la carga viral inicial después de 15 minutos (reducción superior al 99 %). Este efecto inhibidor se mantuvo hasta dos horas en 10 de los 12 pacientes. A los cinco días, se detectó ARN del SARS-CoV-2 en solo 1 paciente del grupo de enjuague bucal y en 5 del grupo de placebo. Conclusiones:Este señala que un enjuague bucal CPC puedereducir la carga viral en saliva de pacientes COVID positivos. Este hallazgo puede ser importante en el control de la transmisión del SARS-CoV-2. Sin embargo, la relevancia clínica de la reducción del enjuague bucal con CPC en la excreción de SARS-CoV-2 en la saliva requiere más estudios.

Correlación entre la glucosa salival con la glucosa de ayuno, la Hemoglobina glicada y el Péptido-C en personas con diabetes mellitus tipo 2

Objetivo : Determinar la correlación entre la glucosa salival con la glucosa en ayunas, HbA1c y el péptido C en personas con Diabetes Mellitus tipo 2 (DM2). Materiales y métodos : Estudio transversal llevado a cabo en el Centro de Investigación en Diabetes, Obesidad y Nutrición (CIDON) en Lima, Perú durante el año 2021. Se categorizó en buen control metabólico (HbA1c<7 %) y mal control metabólico (HbA1c≥7 %). Se midió la glucosa basal, HbA1c y el péptido C en sangre. La glucosa salival se midió con el método glucosa oxidasa. La correlación de Spearman fue usada para determinar la asociación entre la glucosa salival con la glucosa en ayunas, HbA1c y el péptido- C. Resultados : Participaron un total de 142 personas con DM2. La concentración de glucosa salival fue significativamente más elevada en DM2 con mal control metabólico (p<0.01). Se observó una correlación positiva débil significativa entre la glucosa salival y la glucosa basal (r=0.23, p=0.04) y HbA1c (r=0.26, p=0.02) en DM2 con mal control metabólico y una correlación negativa insignificante (r=-0.08; p=0.47) con el péptido C. Conclusiones : La glucosa salival presenta una asociación significativa y positiva con la glucosa en sangre y la HbA1c, pero no con el péptido C en personas con DM2 con mal control metabólico. Sin embargo, hay muchos factores que deben ser considerados y analizados más a fondo para determinar su posible uso.

Objetivo : To determine the correlation between salivary glucose levels with fasting blood glucose, HbA1c, and C-peptide in patients with type 2 diabetes mellitus (T2DM). Materials and methods : This is a cross-sectional study performed at the Centro de Investigación en Diabetes, Obesidad y Nutrición (CIDON) in Lima, Peru, during 2021. Patients were categorized as those with good metabolic control (HbA1c<7 %), and poor metabolic control (HbA1c≥7 %). Baseline fasting blood glucose, as well as blood HbA1c and C-peptide values were measured. Salivary glucose was measured using the glucose oxidase method. Spearman's correlation was used for determining an association between salivary glucose levels and fasting blood glucose, HbA1c, and C-peptide. Results : One-hundred and forty-two subjects with T2DM participated in the study. Salivary glucose was significantly higher in T2DM subjects with poor metabolic control (p<0.01). A weak positive correlation between salivary glucose and fasting blood glucose (r= 0.23, p= 0.04) and HbA1c (r= 0.26, p= 0.02) was observed in subjects with T2DM and poor metabolic control, and also a non-significant negative correlation (r=-0.08; p= 0.47) with C-peptide. Conclusions : Salivary glucose levels show significant and positive association with fasting blood glucose and HbA1c, but not with C-peptide in persons with T2DM and poor metabolic control. However, there are many factors that should be considered and analyzed in detail aiming to determine its potential use.

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

Accuracy of a raw saliva-based COVID-19 RT-LAMP diagnostic assay

ABSTRACT The Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) pandemic demanded rapid diagnosis to isolate new COVID-19 cases and prevent disease transmission. Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) rapidly became the gold standard for diagnosis. However, due to the high cost and delay of the results, other types of diagnosis were implemented, such as COVID-19 Ag Rapid Tests and Reverse Transcription Technique followed by Loop-Mediated isothermal Amplification (RT-LAMP). In this work, we validated the use of RT-LAMP in saliva samples rather than nasopharyngeal swabs, as the collection is more comfortable. First, we selected 5 primer sets based on the limit of detection for SARS-CoV-2 RNA, then validated their sensitivity and specificity in patient samples. A total of 117 samples were analyzed by fluorometric RT-LAMP and compared with qRT-PCR results. Our results show that the use of a high-sensitive primer ORF1-a, together with a low-sensitive primer set Gene E (time to threshold of 22.9 and 36.4 minutes, respectively, using 200 copies of viral RNA), achieved sensitivity in purified RNA from saliva samples of 95.2% (95% CI 76.1-99.8) with 90.5% specificity (95% CI 69.6-98.8) (n = 42).As RNA purification increases the turnaround time, we tested the outcome of RT-LAMP utilizing raw saliva samples without purification. The test achieved a sensitivity of 81.8% (95% CI 59.7-94.8) and a specificity of 90.9% (95% CI 70.8-98.8). As a result, the accuracy of 92.9% (95% CI 80.5-98.5) in purified RNA-saliva samples was lowered to an acceptable level of 86.4% (95% CI 72.6-94.8) in raw saliva. Although mass vaccination has been implemented, new strains and low vaccination progress helped to spread COVID-19. This study shows that it is feasible to track new COVID-19 cases in a large population with the use of raw saliva as sample in RT-LAMP assay which yields accurate results and offers a less invasive test.

Association of salivary parameters and erosive tooth wear in preschool children

Abstract The aims of this study were to 1) assess the association between erosive tooth wear (ETW) according to the BEWE (Basic Erosive Wear Examination) scoring system and salivary parameters and 2) compare salivary parameters according to ETW severity in the subgroup of children with ETW. This cross-sectional study included 52 preschool children aged 5 years paired by sex. A calibrated examiner assessed ETW using BEWE criteria, and stimulated saliva was collected to determine salivary flow, osmolality, pH, and buffering capacity. The children were divided into two groups: without ETW (n=26; BEWE score 0) and with ETW (n=26; BEWE scores 1 and 2). Logistic regression analysis was used to determine the magnitude of the association between ETW and salivary parameters and estimates of odds ratios (OR). In the unadjusted analysis, the ETW group was more likely to have lower salivary flow (OR = 0.079; 95%CI = 0.013-0.469; p = 0.005) and lower osmolality (OR = 0.993; 95%CI = 0.985-1.000; p = 0.049). In the adjusted analysis, salivary flow remained significantly associated with ETW (OR = 0.087; 95%CI = 0.014-0.533; p = 0.008). Lower salivary osmolality values were observed in ETW, especially in preschool children with more severe ETW (BEWE score 2). Receiver operating characteristic (ROC) determined a salivary osmolality cutoff point of < 201 for the presence of ETW. In conclusion, salivary flow rate was significantly associated with ETW prevalence. Lower values of salivary osmolality were observed in preschool children with distinct erosive lesions with significant loss of tooth structure (BEWE 2).

Dose-response relationship between toothpaste soluble fluoride absorbed in the gastrointestinal tract and saliva fluoride secretion

Aim: This study aimed to evaluate if there is a dose-response relationship between toothpaste chemically soluble fluoride absorbed in the gastrointestinal tract and fluoride secreted by saliva, giving support to the use of saliva as surrogate for plasma fluoride. Methods: A 4-phase single blind study was conducted, in which 10 participants were subjected in each phase to one of the assigned treatment groups: group I: fresh sample of a Na2FPO3/CaCO3-based toothpaste with 1,334 µg F/g of total soluble fluoride (TSF) and groups II­IV: aged samples of this toothpaste presenting TSF concentrations of 1,128, 808, and 687 µg F/g, respectively. In all phases, the participants ingested an amount of toothpaste equivalent to 70.0 µg F/Kg body weight, as total fluoride (TF). Saliva and blood samples were collected before (baseline) and up to 180 min after toothpaste ingestion as indicator of fluoride bioavailability. F concentration in saliva and blood plasma was determined with a fluoride ion-specific electrode. The areas under the curve (AUC) of F concentration versus time (AUC = ng F/mL × min) and the peaks of fluoride concentration (Cmax) in saliva and plasma were calculated. Results: A significant correlation between mg of TSF ingested and the AUC (r=0.47; p<0.01), and Cmax (r=0.59; p<0.01) in saliva was found; for TF, the correlation was not significant (p>0.05). In addition, the correlations between plasma and saliva fluoride concentrations were statistically significant for AUC (r=0.55; p<0.01) as for Cmax (r=0.68; p<0.01). Conclusion: The findings support that saliva can be used as a systemic biomarker of bioavailable fluoride present in Na2FPO3/CaCO3-based toothpaste

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

Effect of maternal cortisol levels on fetal heart rate patterns in primiparous pregnant women in the third trimester

SUMMARY OBJECTIVE: This study aimed to determine whether maternal cortisol levels affect fetal heart rate patterns in primiparous pregnant women in the third trimester. METHODS: This cross-sectional descriptive study included 400 primiparous pregnant women with uncomplicated pregnancies between November and December 2022. The study included primiparous pregnant women over 18 years old in the third trimester who had not exercised for at least 2 h before the fetal heart rate monitoring and had a healthy pregnancy without consuming any food or drink. Fetuses with decelerating heartbeats and pregnant women who showed uterine contraction and cervical dilation during the fetal heart rate monitoring were excluded from the study. Research data were collected with the data collection form. The fetal heart rate data were collected using a cardiotocograph. At least two accelerations during the 20-min nonstress test period were the basis for diagnosing a reactive nonstress test. About 5 mL of maternal saliva for cortisol measurements was collected before fetal heart rate monitoring. Research data were analyzed with IBM SPSS Statistics for Macintosh, Version 28.0. A p-value of <0.05 was considered significant. RESULTS: There were no significant differences in the comparison of the groups in terms of education and income status, family type, fetal gender, pregnancy planning status, BMI and age averages, or gestational week averages (p>0.05). The number of at least two accelerations required for the diagnosis of reactive NST was also higher in Group 1 (maternal salivary cortisol level ≤24.20). A moderately positive relationship between fetal heart rate and maternal salivary cortisol was observed (r=0.448, p=0.000). In total, 11.9% of the total change in fetal heart rate level is explained by maternal cortisol (R2=0.119). Maternal cortisol increases fetal heart rate level (ß=0.349). CONCLUSION: These findings suggest that stress in primiparous pregnant women with high cortisol levels may influence fetal heart rate patterns. It was revealed that the increase in cortisol level, considered a stress hormone, may be a harbinger of fetal tachycardia.

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.

Identification of Human Body Fluid Stains and Non-Biological Stains by Three-Dimensional Fluorescence Spectroscopy / 法医学杂志

OBJECTIVES@#To establish a rapid and nondestructive identification method for human body fluid stains and non-biological stains using three-dimensional fluorescence spectroscopy.@*METHODS@#The collected three-dimensional fluorescence spectrum data of human saliva, 3% blood, coffee and Fanta® stains were processed with dimensionality reduction. After wavelet transform, spectral denoising and feature extraction, the classification formula was established. The Fisher discriminant was used for spectrum matching and recognition to establish the analysis method to distinguish stain types.@*RESULTS@#According to the results of data training and comparison, all the recognition accuracies of Fanta®, coffee, saliva and blood were more than 91.39%. Among them, saliva reached 100% recognition accuracy.@*CONCLUSIONS@#Three-dimensional fluorescence spectroscopy is a potential method for rapid and nondestructive identification of biological and non-biological stains.

Research and exploration of salivary biological markers for hepatitis B-related hepatocellular carcinoma / 中华肝脏病杂志

Objective: To study using isotope-labeled relative and absolute quantitative proteomics methodologies to screen for salivary biological markers as a simple, non-invasive tool for identifying hepatitis B-related HCC at an early stage. Methods: Saliva samples were collected to extract salivary proteins. Isotope-labeled relative and absolute quantitative proteomics were used to analyze the differentially expressed proteins between the hepatocellular carcinoma (HCC) and non-HCC groups. Western blotting, immunohistochemistry, and enzyme-linked immunosorbent assays were used to verify differential proteins and identify markers in liver cancer tissues and saliva. Statistical analysis was used to analyze the diagnostic efficiency of salivary biomarkers. Results: 152 differentially expressed salivary proteins were screened out between the HCC and non-HCC groups. Western blot, immunohistochemistry, and enzyme-linked immunosorbent assays validated that the expressions of α-1-acid glycoprotein 1 (ORM1) and alpha-fetoprotein (AFP) were significantly increased in HCC (P < 0.05). There was a significant correlation between salivary AFP and serum AFP (P < 0.05). HCC was diagnosed when salivary α-1-acid glycoprotein 1 combined with AFP. The area under the receiver operating characteristic curve was 0.8726 (95% confidence interval: 0.8104 ~ 0.9347), the sensitivity was 78.3%, and the specificity was 88%. Conclusion: Salivary AFP and α-1-acid glycoprotein 1 can serve as potential biomarkers for hepatitis B-related hepatocellular carcinoma.