Teeth with acute apical abscess vs. teeth with chronic apical periodontitis: a quantitative and qualitative proteomic analysis.

OBJECTIVE: To quantitatively and qualitatively analyze the proteomic profile of teeth with acute apical abscesses (AAA) compared with teeth with chronic apical periodontitis (CAP) and to correlate the expression of detected human proteins with their main biological functions. MATERIALS AND METHODS: Samples were obtained from root canals of 9 patients diagnosed with AAA and 9 with CAP. Samples were analyzed by reversed-phase liquid chromatography coupled to mass spectrometry. Label-free quantitative proteomic analysis was performed by Protein Lynx Global Service software. Differences in protein expression were calculated using the t-test (p < 0.05). RESULTS: In total, 246 human proteins were identified from all samples. Proteins exclusively found in the AAA group were mainly associated with the immunoinflammatory response and oxidative stress response. In the quantitative analysis, 17 proteins were upregulated (p < 0.05) in the AAA group, including alpha-1-acid glycoprotein, hemopexin, fibrinogen gamma chain, and immunoglobulin. Additionally, 61 proteins were downregulated (p < 0.05), comprising cathepsin G, moesin, gelsolin, and transketolase. Most of the proteins were from the extracellular matrix, cytoplasm, and nucleus. CONCLUSIONS: The common proteins between the groups were mainly associated with the immune response at both expression levels. Upregulated proteins mostly belonged to the acute-phase proteins, while the downregulated proteins were associated with DNA/RNA regulation and repair, and structural function. CLINICAL RELEVANCE: The host response is directly related to the development of apical abscesses. Thus, understanding the behavior of human proteins against the endodontic pathogens involved in this condition might contribute to the study of new approaches related to the treatment of this disease.

Mismatch Negativity is associated with affective social behavior in microcephaly.

Some children with severe microcephaly related to Zika virus infection show affective social-like behavior, such as smiling and rejection to a stranger's lap. Our objective was to check the association between this behavior and the occurrence of Mismatch Response (MMR) in event-related potentials. Twenty eight microcephalic children, aged 1-3 years, were divided in Affect(+) and Affect(-) groups, according to either the presence or absence of affective social-like behavior, respectively, and underwent the OddBall paradigm with vowels as auditory stimuli. MMR was statistically estimated comparing MMR sample means between both groups. The Affect(+) group significantly differed from the Affect(-) group and, as opposed to the latter, showed MMR as Mismatch Negativity (MMN) in the left occipital, left and right posterior temporal, and (especially) the right and median parietal leads. The relationship observed between MMN and affective social-like behavior suggests that these children may have cognitive mechanisms capable of providing some social interaction, despite their profound neurological dysfunction. MMN diagnostic techniques seem to be promising for the triage of microcephalic subjects regarding cognitive functions and for choosing a strategy for some social adaptation.

Proteomic analysis of infected root canals with apical periodontitis in patients with type 2 diabetes mellitus: A cross-sectional study.

AIM: This study aimed to quantitatively and qualitatively determine the proteomic profile of apical periodontitis (AP) in type 2 diabetes mellitus (T2DM) patients in comparison with systemically noncompromised patients and to correlate the protein expression of both groups with their biological functions. METHODOLOGY: The sample consisted of 18 patients with asymptomatic AP divided into two groups according to the presence of T2DM: diabetic group-patients with T2DM (n = 9) and control group-systemically healthy patients (n = 9). After sample collection, the root canal samples were prepared for proteomic analysis using reverse-phase liquid chromatography-mass spectrometry. Label-free quantitative proteomic analysis was performed by Protein Lynx Global Service software. Differences in protein expression between groups were calculated using t-test (p < .05). Biological functions were analysed using the Homo sapiens UniProt database. RESULTS: A total of 727 human proteins were identified in all samples. Among them, 124 proteins common to both groups were quantified, out of which 65 proteins from the diabetic group showed significant differences compared with the control: 43 upregulated (p < .05) and 22 downregulated (p < .05) proteins. No significant differences in protein expression were seen for the remaining 59 proteins (p > .05). Most proteins with differences in expression were related to immune/inflammatory response. Neutrophil gelatinase-associated lipocalin, Plastin-2, Lactotransferrin and 13 isoforms of immunoglobulins were upregulated. In contrast, Protein S100-A8, Protein S100-A9, Histone H2B, Neutrophil defensin 1, Neutrophil defensin 3 and Prolactin-inducible protein were downregulated. CONCLUSIONS: Quantitative differences were demonstrated in the expression of proteins common to diabetic and control groups, mainly related to immune response, oxidative stress, apoptosis and proteolysis. These findings revealed biological pathways that provide the basis to support clinical findings on the relationship between AP and T2DM.

Comparative Analysis of the Proteomic Profile of the Dental Pulp in Different Conditions. A Pilot Study.

This study aimed to quantitatively compare the difference in protein expression in the progression of pulp pathogenesis, as well as to describe the biological functions of proteins identified in pulp tissue. Samples were obtained from six patients treated at the Araçatuba School of Dentistry and were divided into three groups: normal pulp - from teeth extracted for orthodontic indication; inflamed pulp and necrotic pulp - from patients diagnosed with irreversible pulpitis and chronic apical periodontitis, respectively. After previous proteomic preparation, dental pulp samples were processed for label-free quantitative proteomic analysis in a nanoACQUITY UPLC-Xevo QTof MS system. The difference in expression between the groups was calculated using the Protein Lynx Global Service software using the Monte Carlo algorithm. A total of 465 human proteins were identified in all groups. The most expressed proteins in the inflamed pulp group in relation to the normal pulp group were hemoglobin, peroxiredoxins and immunoglobulins, whereas the less expressed were the tubulins. Expression levels of albumins, immunoglobulins and alpha-2-macroglobulin were higher in the necrotic pulp group than in the inflamed pulp group. As for the qualitative analysis, the most prevalent protein functions in the normal pulp group were metabolic and energetic pathways; in the inflamed pulp group: cellular communication and signal transduction; and regulation and repair of DNA/RNA, while in the necrotic pulp group proteins were associated with the immune response. Thus, proteomic analysis showed quantitative and qualitative differences in protein expression in different types of pulp conditions.

Comparison of two rotary systems in bacteria/lps removal from endodontic infections: randomized clinical trial.

This clinical study compared the effectiveness of two rotary systems: HyFlex CM (Coltene-Whaledent, Altstetten, Switzerland) and ProTaper Next (Dentsply Sirona, Ballaigues, Switzerland) on the removal of cultivable bacteria and endotoxins from primarily infected root canals. This study was designed as a randomized single-blinded, 2-arm, clinical trial. Twenty-four primarily infected root canals were selected and randomly divided into 2 groups: HyFlex CM (n = 12); and ProTaper Next (n = 12). Samples were collected before and after the biomechanical preparation and inoculated in specific flasks. Irrigation was performed using 2.5% sodium hypochlorite. A kinetic turbidimetric lysate assay of limulus amoebocytes was used to quantify endotoxins. Microbiological culture technique was used to determine the count of bacterial colony forming units (CFU/mL). Data collected were statistically analyzed using SigmaPlot 12.0 for Windows. The Two-Way ANOVA statistical test was performed and the level of significance was 5%. In the samples before the biomechanical preparation, cultivable bacteria and endotoxins were evidenced in 100% of the cases. The culture analysis revealed that there was no statistically significant difference in the bacterial reduction between the two instrumentation systems. Endotoxins were present in 100% of the canals after instrumentation and there was no statistical difference between the two systems in endotoxin reduction. Thus, it was concluded that both instrumentation systems were effective in reducing root canal bacteria and endotoxins with primary endodontic infection and that there was no statistical difference between them. However, no system was able to eliminate 100% of the bacteria and their by-products.

Análise comparativa do perfil proteômico da polpa dentária em condição normal, inflamada e necrótica / Comparative analysis of the proteomic profile of dental pulp under normal, inflamed and necrotic conditions

Este estudo teve como objetivo comparar quantitativamente a diferença de expressão proteica na progressão da patogênese pulpar, bem como correlacionar as funções biológicas das proteínas identificadas no tecido pulpar normal, inflamado ou necrótico. As amostras foram obtidas de pacientes atendidos na Clínica Endodôntica da Faculdade de Odontologia de Araçatuba para tratamento endodôntico, sendo divididos em três grupos: grupo de polpa normal, com amostras do tecido pulpar obtidas a partir de dentes extraídos por indicação ortodôntica (n = 2); grupo de polpa inflamada, com amostras obtidas de pacientes com diagnóstico de pulpite irreversível (n = 2) e grupo de polpa necrótica, cujas amostras foram obtidas de pacientes com diagnóstico de periodontite apical crônica (n = 2). Após o preparo proteômico prévio, as amostras de polpa dentária foram processadas para análise proteômica quantitativa livre de marcadores em um sistema nanoACQUITY UPLC-Xevo QTof MS. A diferença na expressão entre os grupos de polpa normal e inflamada e grupos de polpa inflamada e necrótica foi calculada com o software Protein Lynx Global Service, usando o algoritmo Monte-Carlo, e expressa como p <0,05 para proteínas presentes em menor abundância e 1-p> 0,95 para proteínas presentes em maior abundância. Um total de 465 proteínas humanas foram identificadas em todos os grupos. Nos grupos normal, inflamado e necrótico, foram encontradas 241, 240 e 124 proteínas, respectivamente. Na análise quantitativa, as proteínas mais expressas foram hemoglobinas, peroxirredoxinas e imunoglobulinas, enquanto as menos expressas foram as tubulinas no grupo de polpa inflamada em relação ao grupo de polpa normal. Já, no grupo de polpa necrótica em relação ao de polpa normal, foram encontradas em expressão aumentada as albuminas, imunoglobulinas e alpha-2- macroglobulina, enquanto as menos expressas foram hemoglobinas e actinas. Quanto a análise qualitativa, as proteínas identificadas no grupo pulpar normal estavam envolvidas nas vias metabólicas e energéticas. No grupo de polpa inflamado, as funções proteicas mais prevalentes foram: comunicação celular e transdução de sinal; e regulação e reparo de DNA/RNA, enquanto no grupo da polpa necrótica prevaleceram proteínas associadas à resposta imune. Sendo assim, a análise proteômica mostrou diferenças quantitativas na expressão proteica em diferentes tipos de condições pulpares e revelou que a inflamação pulpar induz à maior expressão de proteínas relacionadas a comunicação e transdução de sinal. No entanto, com o avanço para a necrose pulpar as proteínas estavam associadas à resposta imunológica(AU)

This study aimed to quantitatively compare the difference in protein expression in the progression of pulp pathogenesis, as well as to correlate the biological functions of proteins identified in normal, inflamed or necrotic pulp tissue. The samples were obtained from patients treated at the Endodontic Clinic of the Araçatuba Dental School for endodontic treatment, and were divided into three groups: normal pulp group with pulp tissue samples obtained from orthodontic teeth (n = 2) ; inflamed pulp group, whose samples were obtained from patients diagnosed with irreversible pulpitis (n = 2) and necrotic pulp group, whose samples were obtained from patients diagnosed with chronic apical periodontitis (n = 2). After previous proteomic preparation, dental pulp samples were processed for label-free quantitative proteomic analysis in a nanoACQUITY UPLC-Xevo QTof MS system. The difference in expression between the normal and inflamed pulp groups and groups of inflamed and necrotic pulp was calculated using the Protein Lynx Global Service software using the Monte Carlo algorithm and expressed as p <0.05 for proteins present in lower abundance and 1-p> 0.95 for proteins present in greater abundance. A total of 465 human proteins were identified in all groups. In the normal, inflamed and necrotic groups, 241, 240 and 124 proteins were found, respectively. In the quantitative analysis, the most expressed proteins were hemoglobin, peroxiredoxins and immunoglobulins, whereas the less expressed were the tubulins in the inflamed pulp group in relation to the normal pulp group. Expression of albumins, immunoglobulins and alpha-2-macroglobulin were increased in the necrotic pulp group when compared to normal pulp, whereas hemoglobin and actin were less expressed. As for the qualitative analysis, the proteins identified in the normal pulp group were involved in the metabolic and energetic pathways. In the inflamed group the most prevalent protein functions were: cellular communication and signal transduction; and regulation and repair of DNA / RNA, while in the necrotic pulp group proteins associated with the immune response prevailed. Thus, proteomic analysis showed quantitative differences in protein expression in different types of pulp conditions, and revealed that pulp inflammation induced increased expression of proteins related to cellular communication and signal transduction. Nevertheless, with the progression to pulp necrosis, the proteins were associated with immune response(AU)