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The rotavirus infection mechanism seems to be a multi-step process which is still not fully understood. The heat shock cognate protein hsc70 has been proposed as being a co-receptor molecule for rotavirus entry into susceptible cells. In this work, an attempt was made to determine the existence of possible domains for VP4 and VP6 binding to hsc70. We selected amino acid sequences 531-554 from VP4 and 280-297 from VP6 on the basis of already recognized sequences for binding to hsc70. This study determined that DLPs and synthetic peptides from VP6 (aa 280-297) and VP4 (aa 531-554), individually or in combination, inhibited rotavirus RRV, YM and WA entry into MA104 and Caco-2 cells in an additive and dose-dependent manner. Hyperimmune sera against these synthetic peptides blocked infection by infectious TLPs. Capture ELISA results showed that DLPs interact with hsc70, probably through VP6 as the specific interaction between hcs70 and DLPs was disrupted by a VP6 peptide. These results suggest that VP6 takes part during rotavirus cell entry by binding to hsc70. This, as well as previous work, provides insight concerning the function of hsc70 within a multi-step model of rotavirus entry.

These results suggest Aa-induced proinflammatory endothelial responses are regulated by rosuvastatin in a mechanism that appears to involve KLF2 activation. Use of rosuvastatin to prevent cardiovascular disease may reduce risk of endothelial activation by bacterial antigens.

Eikenella corrodens is a gram‐negative bacterium, and although primarily associated with periodontal infections or infective endocarditis, it has been identified in coronary atheromatous plaques. The effect of its lipopolysaccharide (LPS) on human coronary artery endothelial cells (HCAECs) is unknown. Our aim was to examine the mechanism underlying the inflammatory response in HCAECs stimulated with E. corrodens‐LPS and to evaluate monocyte adhesion. Endothelial responses were determined by measuring the levels of chemokines and cytokines using flow cytometry. The surface expression of intercellular adhesion molecule 1 (ICAM‐1) was determined using a cell‐based ELISA, and the adhesion of THP‐1 monocytes to HCAECs was also monitored. The involvement of toll‐like receptors (TLRs) 2 and 4 was examined using TLR‐neutralizing antibodies, and activation of extracellular signal‐regulated kinase (ERK)1/2 and nuclear factor‐kappa B (NF‐κB) p65 were measured by western blotting and ELISA, respectively. Eikenella corrodens‐LPS increased secretion of interleukin‐8 (IL‐8), monocyte chemotactic protein 1 (MCP‐1), and granulocyte–macrophage colony‐stimulating factor (GM‐CSF), and expression of ICAM‐1 on the surface of HCAECs, consistent with the increased adhesion of THP‐1 cells. Moreover, E. corrodens‐LPS interacted with TLR4, a key receptor able to maintain the levels of IL‐8, MCP‐1, and GM‐CSF in HCAECs. Phosphorylation of ERK1/2 and activation of NF‐κB p65 were also increased. The results indicate that E. corrodens‐LPS activates HCAECs through TLR4, ERK, and NF‐κB p65, triggering a pro‐atherosclerotic endothelial response and enhancing monocyte adhesion.

Studies have been conducted on the pathogenicity of periodontopathogens in cultures of endothelial cells on two‐dimensional (2D) polystyrene surfaces, where the monolayer formed is not exposed to proteins of the subendothelial matrix. In this work, we developed a culture system by seeding human coronary artery endothelial cells (HCAECs) onto three‐dimensional (3D) scaffolds of collagen type I, a subendothelial protein. The inflammatory responses of the HCAEC monolayers, formed either on 3D scaffolds or directly on a 2D polystyrene plate, to lipopolysaccharide (LPS) from Aggregatibacter actinomycetemcomitans (Aa) and Porphyromonas gingivalis (Pg) were evaluated. The transcription of 3 genes, the secretion of 40 cytokines and 2 prostanoids, and the adhesion of monocytes to 2D and 3D cultures with or without exposure to lipopolysaccharides (control) were assessed. HCAECs exhibited differences in transcriptional and secretory profiles between the 3D and 2D models. In addition, the inflammatory responses of HCAEC to Aa‐LPS and Pg‐LPS differed between the two models. In 3D cultures treated with Aa‐LPS, the levels of IL‐8, RANTES, G‐CSF, ICAM‐1, IL‐6, and TXA2 were significantly higher than those in the controls. In 2D cultures treated with Aa‐LPS, IL‐8, RANTES, G‐CSF, ICAM‐1, TNF‐RI, PGI2, and TXA2 levels were significantly higher than those in their controls. In the presence of Aa‐LPS, monocyte adhesion did not differ between treated and control 3D cultures but was significantly higher in treated 2D cultures than in the controls. In response to Pg‐LPS, cytokine‐prostaglandin secretion and monocyte adhesion did not differ between 3D and 2D cultures. These data indicate that HCAECs respond differently to these two types of LPS.

Objective To determine the capacity of salivary biomarkers in the early diagnosis of oral squamous cell carcinoma. Study design A systematic review of the literature was performed based on the English titles listed in the PubMed, EBSCO, Cochrane, Science Direct, ISI web Science and SciELO databases using the following search descriptors: Oral cancer, diagnosis, biomarkers, saliva and oral squamous cell carcinoma. Abstracts and full-text articles were assessed independently by two reviewers. International checklists for assessment of methodological quality were used. Levels of evidence and grades of recommendation through the Scottish Intercollegiate Guidelines Network (SIGN) template were recognized. The units of analysis were identified through a reference matrix. Results Through the research strategy and after application of different filters and considering choosing criteria, six studies were obtained for analysis. Salivary biomarkers for oral cancer most frequently found were mRNA and proteins for IL-8, CD44, MMP-1 and MMP-3. New peptide-biomarkers such as Cyfra 21-1 and ZNF510 were found. ZNF 510 was the only biomarker which increased in the population with tumour stage T1 + T2 and T3 + T4. Only one study showed a sensitivity and specificity of 96% when the biomarker ZNF 510 is employed to discriminate early and late tumour stages. Conclusions There is no sufficient scientific evidence to support the capacity of the identified salivary biomarkers for the early diagnosis of oral cancer (sub-clinical stages of the pathogenic period before cancer phenotypes are manifested). Salivary biomarkers, however, may be employed to discriminate between healthy and cancer patients.

Epidemiological studies have established a clinical association between periodontal disease and atherosclerosis. Bacteremia and endotoxemia episodes in patients with periodontitis appear to link these two diseases by inducing a body-wide production of cardiovascular markers. The presence of oral bacteria in atherosclerotic lesions in patients with periodontitis suggests that bacteria, or their antigenic components, induce alterations in the endothelium associated with atherosclerosis. Therefore, a causal mechanism explaining the association between both diseases can be constructed using in vitro models. This review presents current experimental approaches based on in vitro cell models used to shed light on the mechanism by which periodontal pathogenic microorganisms, and their antigenic components, induce proatherosclerotic endothelial activity. Monolayer cultures of endothelial vascular or arterial cells have been used to assess periodontal pathogenic bacteria and their antigenic compounds and endothelial activation. However, these models are not capable of reflecting the physiological characteristics of the endothelium inside vascularized tissue. Therefore, the shift from two-dimensional (2D) cellular models toward three-dimensional (3D) models of endothelial cells resembling an environment close to the physiological environment of the endothelial cell within the endothelium is useful for evaluating the physiological relevance of results regarding the endothelial dysfunction induced by periodontopathogens that are currently obtained from 2D models. The use of in vitro 3D cellular models can also be relevant to the search for therapeutic agents for chronic inflammatory diseases such as atherosclerosis. Here, we present some strategies for the assembly of 3D cultures with endothelial cells, which is useful for the study of periodontopathogen-mediated disease.

BackgroundThe validity of the surveys on self-reported smoking status is often questioned because smokers underestimate cigarette use and deny the habit. It has been suggested that self-report should be accompanied by cotinine test. This report evaluates the usefulness of serum cotinine test to assess the association between smoking and periodontal status in a study with a large sample population to be used in studies with other serum markers in epidemiologic and periodontal medicine researches.Material and Methods578 patients who were part of a multicenter study on blood biomarkers were evaluated about smoking and its relation to periodontal disease. Severity of periodontal disease was determinate using clinical attachment loss (CAL). Smoking was assessed by a questionnaire and a blood sample drawn for serum cotinine determination.ResultsThe optimal cut-off point for serum cotinine was 10 ng/ml. Serum cotinine showed greater association with severity of CAL than self-report for mild-moderate CAL [OR 2.03 (CI95% 1.16-3.53) vs. OR 1.08 (CI95% 0.62-1.87) ] advanced periodontitis [OR 2.36 (CI95% 1.30- 4.31) vs. OR 2.06 (CI95% 0.97-4.38) ] and extension of CAL > 3 mm [ OR 1.78 (CI95% 1.16-1.71) vs. 1.37 (CI95% 0.89-2.11)]. When the two tests were evaluated together were not shown to be better than serum cotinine test.ConclusionsSelf-reported smoking and serum cotinine test ≥ 10ng/ml are accurate, complementary and more reliable methods to assess the patient’s smoking status and could be used in studies evaluating serum samples in large population and multicenter studies. Clinical Relevance: The serum cotinine level is more reliable to make associations with the patient’s periodontal status than self-report questionnaire and could be used in multicenter and periodontal medicine studies. Key words:Biological markers, serum, cotinine, periodontitis, smoking.