SARS-CoV-2 diagnosis in saliva samples: Usefulness and limitations.

Saliva samples are important for diagnosis, because they are noninvasive and easy to acquire. The objective of this cross-sectional study was to investigate the value saliva samples have in detecting SARS-CoV-2 in comparison to nasal swabs and a new system named CovidCheck. A standard methodology identified the virus in 185 nasopharyngeal swabs and saliva samples revealing a sensitivity, specificity and positive and negative predictive values of 82,100,100 and 94.67%, respectively for saliva samples. Viral presence in saliva samples with the standard methodology in comparison to the CovidCheck system was evaluated in 67 samples with sensitivity, specificity and positive and negative predictive values of 68, 81, 68 and 81%, respectively. In conclusion, our results highlight the usefulness saliva samples have in detecting respiratory viral infections. However, presence of viral inhibitors and viral load in saliva, and the patient's clinical status should be considered as they might affect amplifying systems results.

Systemic inflammatory response to non-surgical treatment in hypertensive patients with periodontal infection.

ABSTRACT: Hypertension is associated with chronic inflammation in the tissues and organs that are involved in the regulation of arterial pressure, such as kidneys and blood vessels. Periodontal disease affects systemic inflammatory markers, leading to endothelial dysfunction, atherosclerotic plaque instability, dyslipidaemia, and insulin resistance. These conditions can also cause an increase in the blood pressure. Nonsurgical periodontal therapies, such as scaling and root planning, can affect systemic markers of inflammation. We evaluated the effect of scaling and root planning on serum levels of inflammation biomarkers in hypertensive patients. The sample consisted of 19 hypertensive patients with Periodontitis. The patients underwent laboratory tests that included glycaemia, cholesterol, triglycerides and blood count. Blood pressure was measured before periodontal therapy, and the second blood pressure recording was obtained at the re-evaluation appointment. Quantification of peripheral blood cytokines was performed using the Milliplex Inflammation Human Cytokine kit (Interleukin 1-ß, Interleukin-4, Interleukin-6, Interleukin-8, Interleukin-10, Interleukin-12 P70, Interleukin-17A, vascular endothelial growth factor and tumor necrosis factor-alpha). All cytokine levels decreased from the initial examination to reassessment. Cytokines that reflected a statistically significant difference included Interleukin-1ß and endothelial vascular growth factor (P = .04 and P = .004). Hypertensive patients with periodontitis undergoing non-surgical periodontal treatment exhibited a decrease in proinflammatory cytokine levels. Non-surgical periodontal treatment decreases the levels of systemic proinflammatory cytokines in controlled hypertensive patients.

Systemic Th17 response in the presence of periodontal inflammation.

BACKGROUND: The relationship between periodontitis and the pathogenesis of other inflammatory diseases, such as diabetes, rheumatoid arthritis and obesity has been an important topic of study in recent decades. The Th17 pathway plays a significant role in how local inflammation can influence systemic inflammation in the absence of systemic pathology. OBJECTIVE: To determine Th17 biased-cells in systemically healthy patients in the presence of generalized chronic periodontitis. METHODOLOGY: A total of 28 patients were recruited without systemic inflammatory pathology, which was determined by clinical history, the Health Assessment Questionnaire (HAQ) and rheumatoid factor detection. Of these patients, 13 were diagnosed as healthy/gingivitis (H/G) and 15 as generalized chronic periodontitis (GCP). Th17 (CD4+CD161+) cells and Th17IL23R+ (CD4+CD161+IL-23R+) cells were quantified by flow cytometry, based on the total cells and on the lymphocyte region, termed the "enriched population" (50,000 events for each). RESULTS: The percentages of Th17 cells of the H/G and periodontitis groups were similar on total cells and enriched population (19 vs 21.8; p=4.134 and 19.6 vs 21.8; p=0.55). However, Th17IL23R+ cells differ significantly between periodontally healthy patients and generalized chronic periodontitis patients in both total cell (0.22% vs 0.65%; p=0.0004) and enriched populations (0.2% vs 0.75%; p=0.0266). CONCLUSIONS: GCP patients (otherwise systemically healthy) were characterized by increased Th17-proinflammatory cell phenotype positive for the IL-23 receptor in peripheral blood. The proportion of Th17 cells that are negative for the IL-23 receptor in the peripheral blood of systemically healthy patients seemed to be unaffected by the presence or absence of chronic periodontitis.

Systemic Th17 response in the presence of periodontal inflammation

Abstract The relationship between periodontitis and the pathogenesis of other inflammatory diseases, such as diabetes, rheumatoid arthritis and obesity has been an important topic of study in recent decades. The Th17 pathway plays a significant role in how local inflammation can influence systemic inflammation in the absence of systemic pathology. Objective: To determine Th17 biased-cells in systemically healthy patients in the presence of generalized chronic periodontitis. Methodology: A total of 28 patients were recruited without systemic inflammatory pathology, which was determined by clinical history, the Health Assessment Questionnaire (HAQ) and rheumatoid factor detection. Of these patients, 13 were diagnosed as healthy/gingivitis (H/G) and 15 as generalized chronic periodontitis (GCP). Th17 (CD4+CD161+) cells and Th17IL23R+ (CD4+CD161+IL-23R+) cells were quantified by flow cytometry, based on the total cells and on the lymphocyte region, termed the "enriched population" (50,000 events for each). Results: The percentages of Th17 cells of the H/G and periodontitis groups were similar on total cells and enriched population (19 vs 21.8; p=4.134 and 19.6 vs 21.8; p=0.55). However, Th17IL23R+ cells differ significantly between periodontally healthy patients and generalized chronic periodontitis patients in both total cell (0.22% vs 0.65%; p=0.0004) and enriched populations (0.2% vs 0.75%; p=0.0266). Conclusions: GCP patients (otherwise systemically healthy) were characterized by increased Th17-proinflammatory cell phenotype positive for the IL-23 receptor in peripheral blood. The proportion of Th17 cells that are negative for the IL-23 receptor in the peripheral blood of systemically healthy patients seemed to be unaffected by the presence or absence of chronic periodontitis.

Periodontal Disease, Inflammatory Cytokines, and PGE2 in Pregnant Patients at Risk of Preterm Delivery: A Pilot Study.

Periodontal disease is an infection that, in pregnant women, can act as a risk factor for preterm delivery by increasing local and systemic inflammatory responses. Objective. To analyze the presence of periodontal disease, proinflammatory cytokines, and prostaglandin E 2 (PGE2) in pregnant patients at high risk for preterm delivery. Materials and Methods. Pilot study for a case-control study. We included 46 pregnant patients (23 patients at risk of preterm delivery as cases and 23 patients without risk of preterm delivery as controls). We excluded patients who received periodontal treatment, antibiotics, or antimicrobials over the last 3 months as well as those with infections or diseases such as diabetes or hypercholesterolemia. The patients underwent a periodontal assessment, and their levels of cytokines (interleukin- [IL-] 2, IL-6, IL-10, and tumor necrosis factor- [TNF-] α) and prostaglandin E2 (PGE2) were quantified. Results. Patients with periodontal disease showed higher levels of cytokines (IL-2, IL-6, IL-10, and TNF-α) and PGE 2 . Patients at high risk for preterm birth showed higher IL levels compared with patients at low risk for preterm delivery. PGE 2 increased with the severity of periodontal disease. PGE 2 was higher in patients at low risk for preterm delivery, although this difference was not significant. Conclusion. Periodontal disease can increase the systemic inflammatory response as well as the levels of PGE 2 and inflammatory cytokines in pregnant patients.

Differential reactivity of salivary igA and igG against streptococcus mutans proteins in humans with different caries experience.

Dental caries is an infectious disease which still constitutes a public health concern. It begins at an early age and is caused mainly Streptococcus mutans (S. mutans). The aim of this study was to characterize the salivary humor immune response to S. mutans proteins in patients with caries, with history of caries and without caries, in order to determine which S. mutans proteins participate in the immunological response in subjects with different caries experience. Saliva was collected by spontaneous salivation for 5 minutes from 60 subjects aged 18 to 30 years, classified according to their caries experience as: without caries (Group I), with active caries (Group II) and with history of caries (Group III). The antigens derived from S. mutans by sonication were recognized by salivary IgA and IgG by Western Blot. The results showed that all the individuals studied recognized S. mutans proteins with molecular weights in the range of 8 to 191 kDa, with similar recognition profiles for salivary IgA and IgG. Subjects without caries recognized the 29 kDa protein, also known as S. mutans Antigen A, via salivary IgA, differing from patients with caries and history of caries, who recognized it via IgG. The protective response against S. mutans is mediated by IgA. To conclude, a differential response to the 29 kDa protein between study individuals may be indicative of resistance to dental caries and may have a protective role in the induction of IgA antibodies against dental caries, as found in the group without caries, in contrast to subjects with active caries and history of caries.

Differential reactivity of salivary igA and igG against streptococcus mutans proteins in humans with different caries experience

Dental caries is an infectious disease which still constitutes a public health concern. It begins at an early age and is caused mainly Streptococcus mutans (S. mutans). The aim of this study was to characterize the salivary humor immune response to S. mutans proteins in patients with caries, with history of caries and without caries, in order to determine which S. mutans proteins participate in the immunological response in subjects with different caries experience. Saliva was collected by spontaneous salivation for 5 minutes from 60 subjects aged 18 to 30 years, classified according to their caries experience as: without caries (Group I), with active caries (Group II) and with history of caries (Group III). The antigens derived from S. mutans by sonication were recognized by salivary IgA and IgG by Western Blot. The results showed that all the individuals studied recognized S. mutans proteins with molecular weights in the range of 8 to 191 kDa, with similar recognition profiles for salivary IgA and IgG. Subjects without caries recognized the 29 kDa protein, also known as S. mutans Antigen A, via salivary IgA, differing from patients with caries and history of caries, who recognized it via IgG. The protective response against S. mutans is mediated by IgA. To conclude, a differential response to the 29 kDa protein between study individuals may be indicative of resistance to dental caries and may have a protective role in the induction of IgA antibodies against dental caries, as found in the group without caries, in contrast to subjects with active caries and history of caries.

La caries dental es una enfermedad infecciosa que continua siendo un problema de salud publica, inicia a temprana edad y es causada principalmente por Streptococcus mutans (S. mutans). El objetivo de este estudio fue caracterizar la respuesta inmune humoral salival, ante las proteinas de S. mutans, en pacientes con caries, historia de caries e individuos libres de caries, para asi establecer que proteinas de S. mutans participan en la respuesta inmunologica en los diferentes estadios de caries. La saliva de 60 individuos entre 18 y 30 anos de edad, clasificados de acuerdo al estado de caries: libres de caries (grupo I), caries activa (grupo II) e historia de caries (grupo III), se colecto por salivacion espontanea durante 5 minutos. Los antigenos derivados de S. mutans por sonicacion, fueron reconocidos por IgA e IgG salivales por Western Blot. Los resultados mostraron que todos los individuos estudiados reconocen las proteinas de S. mutans en el rango de 8 a 191 kDa de peso molecular con perfiles de reconocimiento similares para IgA e IgG salival. Se encontro que los sujetos libres de caries reconocen por IgA salival la proteina de 29 kDa, tambien llamada Antigeno A de S. mutans, de manera diferente que los pacientes con caries e historia de caries quienes reconocieron la proteina via IgG. La respuesta protectora frente a S. mutans es mediada por IgA. En conclusion, una respuesta diferencial a la proteina de 29 kDa entre los individuos estudiados, puede ser indicativo de resistencia a la caries dental y tener un papel protector en la induccion de anticuerpos IgA frente a la caries dental, como se encontro en el grupo libre de caries, a diferencia de los sujetos con historia de caries y caries activa.

The carboxy-terminal region of CD5 is required for c-CBL mediated TCR signaling downmodulation in thymocytes.

CD5 functions as a negative regulator of TCR signaling during thymocyte development, however, the molecular mechanisms involved in this process remain elusive. A key molecule involved in the down modulation of TCR signaling is c-Cbl, an ubiquitin ligase that physically associates with CD5. Crosslinking of TCR in thymocytes leads to ubiquitylation and lysosomal/proteasomal degradation of TCR downstream signaling effectors and CD5 itself. The present report shows that co-engagement of CD3 with CD5 enhanced c-Cbl phosphorylation, which was not affected by the deletion of the pseudo-ITAM domain of CD5, the putative binding site for c-Cbl. However, amino acids present in the carboxy-terminal region of CD5, were necessary for this effect, indicating that ITAM-independent sites were involved in the interaction of c-Cbl with CD5. The carboxy-terminal region of CD5 was also required for Vav degradation, a well-known target for c-Cbl-dependent ubiquitylation. These results support the notion that the distal cytoplasmic domain of CD5, including Y463, plays a relevant role in the downmodulation of TCR signals in thymocytes via c-Cbl.

Immunoglobulin A, G and M levels in saliva in children between 3 - 12 years of age, healthy and with gingivitis.

UNLABELLED: The aim of this study was to measure the level of immunoglobulin A, G and M in saliva of 3- to 12-year-old children, both healthy and diagnosed with gingivitis. METHODS: A sample of 177 children was selected, of whom 24 were healthy and 153 were diagnosed with gingivitis according to Loe 's index. Samples of saliva were taken and the ELISA test was applied to obtain the immunoglobulin concentrations expressed in microg/ml. A relationship was established between the immunoglobulin levels, the disease (gingival index) and Loe 's bacterial plaque index. IgG levels were higher in healthy children. In the group with gingivitis, 95.8% of the children had incipient gingivitis with a low average index of bacterial plaque (1.33). A direct correlation was found between age and gingival index, while an inverse correlation was found between age and bacterial plaque index. The analysis of the behavior of immunoglobulin according to age showed that age was only significantly correlated to IgA levels. The analysis comparing immunoglobulin levels and clinical parameters for gingivitis showed a direct correlation between gingival index and lgM The gingival index was found that to increase with the age of the subject, even though bacterial plaque decreases. It was also found that age is a better predictor of lgA level than gingival index and bacterial plaque index are; and that gingival index is a better predictor of lgM level than age and bacterial plaque index are.

Immunoglobulin A, G and M levels in saliva in children between 3-12 years of age, healthy and with gingivitis

El objetivo de este estudio cuantificar los niveles de inmunoglobulinas A, G y M en saliva de niños entre 3 – 12 añossanos y con gingivitis. Métodos: la muestra fue de 177 niños distribuidos en dos grupos: 24 sanos y 153 con diagnóstico de gingivitis según el índice de Loe a quienes se les tomaron muestras de saliva y por medio de la prueba de ELISA se obtuvieronlas concentraciones de las inmunoglobulinas expresadas enµg/ml. Resultados: Se encontró que en la saliva de los niños sanos los niveles de IgG son significativamente mayores que en los niños con gingivitis. El grupo gingivitis estuvo conformado por un 95.8 por ciento de niños con diagnóstico de gingivitis incipiente que presentó un promedio bajo de índice de placabacteriana. Al hacer análisis de correlación entre las variables estudiadas, se encontró una correlación directa entre la edad y el índice gingival, una correlación inversa entre la edad y el índice de placa bacteriana, correlación directa entre los niveles de IgA y la edad y correlación directa entre el índice gingival y la IgM. Conclusiones: Se encontró que en la medida en que el individuo crece aumenta el índice gingival, aunque se presenta menor cantidad de placa bacteriana. También seconcluyó que la edad es mejor predictor de los niveles de IgA que el índice gingival y el índice de placa bacteriana y que el índice gingival es mejor predictor de los niveles de IgM que la edad y el índice de placa bacteriana.

Association of salivary proteins with dental caries in a Colombian population.

UNLABELLED: Saliva has an important role in maintaining normal conditions of the oral tissues. Variability in salivary composition determines its protective characteristics against dental caries. Knowledge of the molecular content of saliva in humans is important to better understand its protective properties. The aim of the present work was to recognize protein composition in whole saliva of subjects with active caries (AC), History of caries (HC), and free of caries (H) in a Colombian population, by electrophoretic pattern, and to correlate these results with clinical diagnoses. Patients over 18 years old were selected after clinical examinations, and classified into three study groups. After patients signed the informed consent form, whole saliva samples were collected. Total protein determinations were made using the Bradford method. Individual saliva samples were analyzed by SDS-PAGE and related to DMFT indexes. The gels were analyzed by Quantity One 1-D software (BIO-RAD). No statistically significant difference was found between the total protein concentration and absence of prior experience of dental decay. Total protein content was higher in female subjects (p = 0.0028) than male, and regarding the disease, it was higher in women with HC and AC. Salivary proteins present in the majority of individuals were 101, 77, 62, 55, 44, 22 and 13 kDa in size. Association was found between 17 kDa salivary protein and AC in men. CONCLUSIONS: whole salivary proteins are very similar in the three groups, except for the 17 kDa salivary protein, which might be risk marker for dental caries.

Cytokines produced by CD4+ T cells against a synthetic GTF-I(1301-1322) peptide of Streptococcus mutans in naturally sensitized humans.

Streptococcus mutans (S. mutans) is the main etiological agent in dental caries. Its virulence factors are the proteins PAc and glucosyltransferase (GTF), which are related to its physiopathogenia and have been used in research for a dental caries vaccine. It was reported that using experimental animal models, GTF-I(1301-1322) synthetic peptide from the GLU region of the GTFs has Tepitopes, induces production of serum antibodies in saliva and reduces the presence of caries, but little is known about the cellular response in naturally sensitized humans. The aim of this study was to observe whether GTF-I(1301-1322) peptide is capable of activating CD4+ T cells in PBMC from naturally sensitized humans, to classify the response and to establish the relationship with dental caries. The study was conducted on 30 individuals classified into the following 3 groups: active caries (AC), History of Caries (HC), and free of caries (H). A blood sample was drawn from each individual. Specific antigen stimulation and flow cytometry analyses were used to determine cells producing the cytokines IFN-gamma (type 1 cytokine) and IL-13 (type 2 cytokine). Cell memory response to GTF-I(1301-1322) peptide was found in naturally sensitized humans. Three different responses were detected: TH0, TH1, and NR. The percentage of CD4+ T cells producing the cytokines IFN-gamma (type 1 cytokine) was greater than the percentage producing IL-13 (p=0.006). No statistically significant differences were found among the three groups for the other variables studied (p < or = 0.05). In conclusion, specific cellular immune responses against the GTF-I(1301-1322) peptide of S. mutans does not differ between individuals who are naturally sensitized, caries- resistant or with caries.

Association of salivary proteins with dental caries in a Colombian population

La saliva tiene un importante papel en el mantenimiento de las condiciones normales de los tejidos orales. La variabilidad en la composición salival determina su característica protectoracontra la caries dental. Conocer el contenido molecular de la saliva en humanos es importante para el entendimiento de sus propiedades biológicas. El objetivo de este trabajo fue reconocer la composición proteica de la saliva completa de sujetoscon caries activa (AC), historia de caries (HC) y libres de caries (H); en una población colombiana por medio del patrón electroforético y su correlación con el diagnóstico clínico. Cientocuarenta y cinco pacientes mayores de 18 años de edad fueron clasificados en los grupos de estudio por examinación clínica, las muestras de saliva fueron recolectadas, las determinacionesde proteína total fueron hechas por el método Bradford; la saliva individual fue separada por SDS-PAGE y relacionadas con el índice COP. Los geles fueron analizados por el software Quantity One® 1-D (BIO-RAD). No se encontraron diferencias estadísticamente significativas entre las concentraciones de proteína total y la experiencia o no de caries dental. La cantidad de proteína total fue mayor en mujeres que en hombres (p= 0.0028); y con respecto a la enfermedad fue mayor en mujeres con HC y AC. Las proteínas salivares comunes entre los individuos fueron de 101, 77, 62, 55, 44, 22 y 13 kDa. Se encontró asociación entre la proteína salival de 17 kDa y la AC enhombres. Conclusiones: Las proteínas presentes en la muestra de saliva completa son muy similares en los tres grupos de estudio, excepto por la proteína de 17 kDa la cual puede ser unmarcador de riesgo de caries dental.