MicroRNA signatures differentiate Crohn's disease from ulcerative colitis.

BACKGROUND: Excessive and inappropriate immune responses are the hallmark of several autoimmune disorders, including the inflammatory bowel diseases (IBD): Crohn's disease (CD) and ulcerative colitis (UC). A complex etiology involving both environmental and genetic factors influences IBD pathogenesis. The role of microRNAs (miRNAs), noncoding RNAs involved in regulating numerous biological processes, to IBD pathology, in terms of initiation and progression, remains ill-defined. In the present study, we evaluated the relationship between colon, peripheral blood, and saliva whole miRNome expression in IBD patients and non-inflammatory bowel disease (non-IBD) controls to identify miRNAs that could discriminate CD from UC. Quantitative real-time PCR (qRT-PCR) was used to validate and assess miRNA expression. RESULTS: Microarray analysis demonstrated that upwards of twenty six miRNAs were changed in CD and UC colon biopsies relative to the non-IBD controls. CD was associated with the differential expression of 10 miRNAs while UC was associated with 6 miRNAs in matched colon tissues. CD was associated with altered expression of 6 miRNAs while UC was associated with 9 miRNAs in whole blood. Expression of miR-101 in CD patients and miR-21, miR-31, miR-142-3p, and miR-142-5p in UC patients were altered in saliva. CONCLUSIONS: Our results suggest that there is specific miRNA expression patterns associated with UC versus CD in three separate tissue/body fluids (colon, blood, and saliva). Further, the aberrant miRNA expression profiles indicate that miRNAs may be contributory to IBD pathogenesis, or at least reflect the underlying inflammation. Scrutinizing miRNA expression in saliva and blood samples may be beneficial in monitoring or diagnosing disease in IBD patients. A panel of miRNAs (miR-19a, miR-21, miR-31, miR-101, miR-146a, and miR-375) may be used as markers to identify and discriminate between CD and UC.

The Expression of Lung Resistance Protein in Saliva: A Novel Prognostic Indicator Protein for Carcinoma of the Breast.

Considering that saliva is a fluid inundated with proteins, it is possible that solubilized oncogenic proteins may be present in saliva and may be useful in differentiating between healthy and diseased individuals. As a consequence, the purpose of this study was to determine if the solubilized form of LRP was present in stimulated whole saliva and could differentiate between 16 healthy women and 16 women with confirmed Stage I breast cancer. LRP levels were determined using gel electrophoresis and Western blot technology. The results showed LRP at significantly higher concentrations among breast cancer subjects as compared to healthy women.

Assessment of partial-mouth periodontal examination protocols for periodontitis surveillance.

OBJECTIVE: To evaluate bias associated with nine identified partial-mouth periodontal examination (PMPE) protocols in estimating periodontitis prevalence using the periodontitis case definition given by the Centers of Disease Control and Prevention and American Academy of Periodontology (CDC/AAP). MATERIAL AND METHODS: Prevalence from full-mouth examination was determined in a sample of 3667 adults ≥30 years old from the National Health and Nutrition Examination Survey (NHANES) 2009-2010. Prevalence, absolute bias, relative bias, sensitivity and inflation factor were derived for these protocols according to the CDC/AAP definition and half-reduced CDC/AAP definition as ≤50% of sites were measured. RESULTS: Bias in moderate and severe periodontitis prevalence ranged between 11.1-52.5% and 27.1-76.3% for full-mouth mesiobuccal-distolingual protocol and half-mouth mesiobuccal protocol respectively; according to the CDC/AAP definition. With half-reduced CDC/AAP definition, half-mouth four sites protocol provided small absolute bias (3.2%) and relative bias (9.3%) for the estimates of moderate periodontitis prevalence; corresponding biases for severe periodontitis were -1.2% and -10.2%. CONCLUSION: Periodontitis prevalence can be estimated with limited bias when a half-mouth four sites protocol and a half-reduced CDC/AAP case definition are used in combination.

The use of salivary protein secretions as an in vivo model to study Mantel cell lymphoma progression and treatment.

PURPOSE: The objective of this study was to compare the salivary protein profiles from an individual diagnosed with MCL before, during, and after chemotherapy. METHODS: Saliva specimens acquire prior, during, and after chemotherapy. Specimens were analyzed using a LC-MS/MS mass spectrometer. RESULTS: The results of the study suggest salivary protein alterations secondary to MCL and that a number of the proteins were changed as a result of chemotherapeutic intervention suggesting that the treatment, from a proteomic perspective, was efficacious. The study suggests that salivary secretions may be used as in vivo model for studying MCL progression and treatment efficacy.

Assessing periodontitis in populations: a systematic review of the validity of partial-mouth examination protocols.

OBJECTIVE: To estimate bias associated with partial-mouth periodontal examination (PMPE) protocols regarding estimates of prevalence, severity and extent of clinical attachment loss (CAL), pocket depth (PD) and gingival recession (REC). MATERIAL AND METHODS: A search was made for articles published in English, from 1946 to 2012, which compared PMPE versus full-mouth periodontal examination protocols for CAL or PD ≥ 4 mm or REC ≥3 mm thresholds. PMPE protocols were evaluated for sensitivity of estimates of periodontitis prevalence, relative biases for severity and extent estimates. RESULTS: A review of the literature identified 12 studies which reported 32 PMPE protocols. Three PMPE protocols which had sensitivities ≥85% and relative biases ≤0.05 in absolute values for severity and extent estimates were as follows: (1) half-mouth six-sites, (2) diagonal quadrants six-sites and (3) full-mouth mesiobuccal-midbuccal-distobuccal (MB-B-DB). Two other PMPE protocols (full-mouth and half-mouth mesiobuccal-midbuccal-distolingual) performed well for prevalence and severity of periodontitis; however, their performance in estimates of extent was unknown. CONCLUSIONS: Among the 32 PMPE protocols listed, the half-mouth six-sites and full-mouth MB-B-DB protocols had the highest sensitivities for prevalence estimates and lowest relative biases for severity and extent estimates.

Thermal damage and excision time of micro and super pulsed diode lasers: A comparative ex vivo analysis.

OBJECTIVES: The primary aim of this ex vivo study was to evaluate thermal damage and cutting efficiency of micro and super pulsed diode lasers. The secondary aim was to suggest a guideline to perform simple surgical excisions adequate for histopathological evaluation. MATERIAL AND METHODS: Ten groups of 10 specimens of pig tongues were excised using a blade (G1), a micro pulsed (G2-G9), and a super pulsed diode (G10) lasers. Different output power, pulse duration, pulse interval, and duty cycle were tested. Quantitative measures of thermal damage and excision times were recorded. Statistical analysis was performed at a significance level of 5%. RESULTS: The control group (G1) presented no thermal damage. Within the laser groups (G2-G10), no statistically significant differences in depth of thermal damage (µm) were noted. G3 showed significantly less area of thermal damage (mm2 ) when compared with G7 and G9 (p < .05). The median excision time of the control group and super pulsed diode laser group were significantly lower (p < .001) than the micro pulsed diode laser groups. CONCLUSIONS: The cutting efficiency of the super pulsed diode laser is comparable to traditional blade, and with appropriate parameters, these lasers can produce predictable surgical outcomes with less collateral damage.

Proteomics, morphoproteomics, saliva and breast cancer: an emerging approach to guide the delivery of individualised thermal therapy, thermochemotherapy and monitor therapy response.

The field of proteomics is in its infancy; however the discipline, its technology, and our abilities to translate the proteomic data are rapidly evolving. In the near future proteomics should significantly improve our ability to make early cancer diagnoses, direct appropriate personalised therapy, and monitor response to therapy, including thermal therapy. The potential role of proteomics in breast cancer early diagnosis, prediction of aggressiveness is clear. Its potential importance in guiding treatment choice and prediction of treatment response is especially intriguing. This paper reviews the varied methodologies used in the field of proteomics, including gel-free, label-free proteomics, quantitative proteomics, phosphoproteomics, protein extraction from formalin-fixed, paraffin-embedded tissue sections (FFPE) proteomics, laser capture microdissection proteomics, and targeted proteomics. It also discusses two new areas, morphoproteomics and salivary proteomics cancer diagnostics, as well as selected pre-clinical and clinical analyses using the described methodologies. Morphoproteomics defines which signal transduction pathways exist within the tumour cells and the surrounding tissue comprising a patient's cancer biopsy specimen. Morphoproteomics, and the other histology-based proteomic techniques are actually beginning to clinically make possible individualised treatment of breast cancer. Salivary proteomics, in part because it is non-invasive, is a new area of breast cancer diagnostics that can be used to non-invasively monitor an individual patient's response to treatment with every treatment cycle. The current literature demonstrates that a diagnosis of breast cancer can be readily made using proteomic methodologies, and that proteomics can also define cancers with a poor prognosis at the time of diagnosis. With such early prognostic information we expect proteomics will soon be a science that on the basis of prognosis, guides individualised therapy and as well, have the ability to monitor the results of thermal therapy, radiation, and chemotherapy treatment during therapy.

Methods for analyzing saliva proteins for systemic disease detection.

New technological developments, coupled with the limitations of existing methodologies for the detection of disease, are propelling the field of salivary diagnostics forward at unprecedented rates. Advancements in proteomics and nanotechnology are paving the way for diagnostic tests that will be capable of rapid multi-analyte detection in both laboratory and nonlaboratory settings. Technological advancements have also benefited biomarker research to the point where saliva is now recognized as an excellent diagnostic medium that can be collected simply and noninvasively. This article reviews the varying nanotechnological platforms and how they will utilize saliva as the diagnostic medium.

Impact of simulated patients on students' self-assessment of competency in practice of geriatric dentistry.

PURPOSE: The evaluation of the effectiveness of simulation training in dentistry has previously been focused primarily on psychomotor hand skills. This study explored the impact of simulated patient (SP) encounters in simulation on students' self-assessment of competency in their clinical and communication abilities with geriatric patients. METHODS: Students from 2 cohorts were recruited for this study. Cohort 1 (n = 30) participated in the standard curriculum with no simulation training and served as the control group. Cohort 2 (n = 34) participated in a SP experience, simulating the initial stages of a care visit for 2 nursing home patients. Students' perceptions of competency to perform these clinical and communication tasks were assessed. A group debriefing session was held 5 weeks post-simulation where Cohort 2 completed a student feedback form. RESULTS: A statistically significant change (P < 0.00001) was noted for both cohorts in their self-reported competence to perform clinical tasks following exposure to an independent clinical experience. In addition to this gain, individuals in Cohort 2 demonstrated improvements following simulation and expressed different responses of impact to questions related to treatment, pharmacology, and managing a complex medical history. CONCLUSIONS: This study suggests that simulation of patient interactions using SPs can strengthen students' self-assessment of competency in their abilities, leading to more genuine interactions with actual patients. These findings will help inform the design of future SP encounters as a component of an evolving humanistic curriculum.

Breast cancer related proteins are present in saliva and are modulated secondary to ductal carcinoma in situ of the breast.

OBJECTIVE: The objective of this study was to determine if protein-by-products secondary to cancer related oncogenes appear in the saliva of breast cancer patients. METHODS: Three pooled (n = 10 subjects/pool) stimulated whole saliva specimens from women were analyzed. One pooled specimen was from healthy women, another pooled specimen from women diagnosed with a benign breast tumor and the other one pooled specimen was from women diagnosed with ductal carcinoma in situ (DCIS). Differential expression of proteins was measured by isotopically tagging proteins in the tumor groups and comparing them to the healthy control group. Experimentally, saliva from each of the pooled samples was trypsinized and the peptide digests labeled with the appropriate iTRAQ reagent. Labeled peptides from each of the digests were combined and analyzed by reverse phase (C18) capillary chromatography on an Applied Biosystems QStar LC-MS/MS mass spectrometer equipped with an LC-Packings HPLC. RESULTS: The results of the salivary analyses in this population of patients yielded approximately 130 proteins in the saliva specimens. Forty-nine proteins were differentially expressed between the healthy control pool and the benign and cancer patient groups. CONCLUSIONS: The study suggests that saliva is a fluid suffused with solubilized by-products of oncogenic expression and that these proteins may be modulated secondary to DCIS. Additionally, there may be salivary protein profiles that are unique to both DCIS and fibroadenoma tumors.

Maxillary frenectomy using a carbon dioxide laser in a pediatric patient: a case report.

Dental lasers contribute significantly to the field of cosmetic dentistry, providing an invaluable resource for clinicians who perform different types of esthetic procedures. An increasing number of general dentists are using the carbon dioxide (CO2) laser for surgical procedures. However, the literature about using a CO2 laser to perform oral soft tissue ablation in the pediatric patient is limited. This case report presents a successful case of a maxillary labial frenectomy in a pediatric patient using a CO2 laser.

Pre-analytic saliva processing affect proteomic results and biomarker screening of head and neck squamous carcinoma.

The objective of this study was to assess the effect of pre-analytical processing on proteomic analysis of saliva and to identify salivary biomarkers for potential clinical applications. Saliva samples from five healthy individuals and three head and neck squamous carcinoma (HNSC) patients were initially depleted of major protein constituents. Saliva from healthy subjects was divided and processed by three different methods prior to liquid chromatography and tandem mass spectrometry technique (LC-MS/MS) analysis. The results showed marked differences amongst the methods. The SDS-PAGE separation and in-gel digestion method yielded the highest number of proteins that included the majority of those identified by the other two methods. The in gel-digestion method was used in the LC-MS/MS analysis of saliva from three HNSC patients and the results were compared with those from healthy subjects. Our analysis identified two proteins, alpha-1-B-glycoprotein and complement factor B proteins, to be present in patients but not in normal specimens. Paradoxically, cystatin S, parotid secretory factor, and poly-4-hydrolase beta-subunit proteins were detected in most normal salivas but not in patient specimens. Subsequent analysis of complement factor B by Western blotting showed strong immunoreactive bands of complement factor B in HNSC patients' and negative or weakly positive in normal saliva samples. We conclude that: 1) initial saliva processing affects protein analysis, 2) in-gel digestion followed by LC-MS/MS detects the most saliva proteins, 3) certain proteins are differentially found in patient and normal salivas and 4) a small set of proteins can be targeted for future validation for clinical investigation.

Salivary analysis in the diagnosis and treatment of breast cancer: a role for the general dentist.

Saliva testing is an attractive area of research for the general dentist, as it offers a great opportunity to utilize an easily accessible fluid for the diagnosis of disease. Diseases that may be difficult to detect, such as breast cancer, are an area of particular interest. Breast cancer is the second leading cause of death among women in the U.S. and early detection is critical to patient survival. Frequent and inexpensive testing is the key to early detection. The general dentist is in the perfect position to take salivary samples from patients during routine checkups or procedures and to refer patients depending on the results.

Oral mucositis: review of pathogenesis, diagnosis, prevention, and management.

Oral mucositis is a debilitating and morbid condition among cancer patients that affects their quality of life and their overall ability to respond to treatment. The dentist plays an important role in the multidisciplinary health care team for the overall management of this condition. This article summarizes preventive and therapeutic treatment modalities available to dentists, based on the latest literature.

Grading Class I Preparations in Preclinical Dental Education: E4D Compare Software vs. the Traditional Standard.

The aim of this study was to compare the effectiveness of a novel assessment software system with the traditional grading protocol used in the University of Texas School of Dentistry at Houston operative dentistry preclinical curriculum. In the study, conducted in 2016, 98 Class I preparations were evaluated both traditionally and digitally by two teams of calibrated preclinical faculty members (two evaluators for each team). Scores from each faculty pair were averaged for the traditional and the digital grading systems, and the scores for the two grading systems were compared. The analysis found no significant difference between the two grading systems with respect to isthmus width (p=0.073) and remaining marginal ridge (p=0.5841), but there was a significant difference with respect to pulpal floor depth assessment (p<0.0001). The data suggested that both grading techniques can be used with repeatable confidence for two out of three grading criteria: isthmus width and remaining marginal ridge. In addition, the software offers a self-assessment tool for students to perfect their psychomotor skills while promoting independence and immediate feedback.

A Catalogue of Altered Salivary Proteins Secondary to Invasive Ductal Carcinoma: A Novel In Vivo Paradigm to Assess Breast Cancer Progression.

The objective of this manuscript is to introduce a catalogue of salivary proteins that are altered secondary to carcinoma of the breast. The catalogue of salivary proteins is a compilation of twenty years of research by the authors and consists of 233 high and low abundant proteins which have been identified by LC-MS/MS mass spectrometry, 2D-gel analysis and by enzyme-linked immunosorbent assay. The body of research suggests that saliva is a fluid suffused with solubilized by-products of oncogenic expression and that these proteins may be useful in the study of breast cancer progress, treatment efficacy and the tailoring of individualized patient care.

Partial-mouth periodontal examination protocol for estimating periodontitis extent and severity in a US population.

We have previously demonstrated that half-mouth four-site periodontal examination protocol performed well in estimating periodontitis prevalence. This study aimed to assess biases associated with this same protocol in estimating periodontitis extent and severity in a United States population. Periodontitis extent as determined by percentage of sites with clinical attachment loss (CAL) ≥3, and ≥5 mm and severity as determined by mean CAL were calculated for full-mouth examination and half-mouth four-site protocol based on 3734 adults sampled from the National Health and Nutrition Examination Survey 2009-2010. Probing depth was excluded because of low data reliability. The comparison between full-mouth and half-mouth assessments was based on bias, relative bias, Wilcoxon signed-rank test, and intra-class correlation coefficient (ICC). For full-mouth examination, periodontitis extent was 21.2% for CAL ≥3 mm and 6.9% for CAL ≥5 mm; periodontitis severity (mean CAL) was 1.73 mm. Half-mouth four-site protocol provided bias -1.2% and relative bias -5.7% for extent (CAL ≥3 mm). Corresponding numbers were -0.3% and 4.3% for extent (CAL ≥5 mm), -0.05 mm and -2.9% for severity. Although the difference between full-mouth and half-mouth assessments was statistically significant, ICCs between them were ≥0.96 for extent (CAL ≥3, 5 mm), and severity (mean CAL). Half-mouth four-site protocol performed well in estimating periodontitis extent and severity based on CAL. Therefore, this protocol should be considered for periodontitis surveillance.

Altered Mucosal Microbiome Diversity and Disease Severity in Sjögren Syndrome.

There is mounting evidence that the microbiome has potent immunoregulatory functions. We assessed the effects of intestinal dysbiosis in a model of Sjögren syndrome (SS) by subjecting mice to desiccating stress (DS) and antibiotics (ABX). We characterized the conjunctival, tongue and fecal microbiome profiles of patients with SS. Severity of ocular surface and systemic disease was graded. 16S ribosomal RNA gene sequencing characterized the microbiota. ABX + DS mice had a significantly worse dry eye phenotype compared to controls, a decrease in Clostridium and an increase in Enterobacter, Escherichia/Shigella, and Pseudomonas in stool after ABX + DS for 10 days. Goblet cell density was significantly lower in ABX treated groups compared to controls. Stool from SS subjects had greater relative abundances of Pseudobutyrivibrio, Escherichia/Shigella, Blautia, and Streptococcus, while relative abundance of Bacteroides, Parabacteroides, Faecalibacterium, and Prevotella was reduced compared to controls. The severity of SS ocular and systemic disease was inversely correlated with microbial diversity. These findings suggest that SS is marked by a dysbiotic intestinal microbiome driven by low relative abundance of commensal bacteria and high relative abundance of potentially pathogenic genera that is associated with worse ocular mucosal disease in a mouse model of SS and in SS patients.

Impact of Technique-Specific Operative Videos on First-Year Dental Students' Performance of Restorative Procedures.

The aim of this study was to examine the impact of psychomotor operative video demonstrations on first-year dental students who are performing specific procedures for the first time in a preclinical setting. The class was randomly divided into two groups, and three restorative procedures were selected. On the date on which each procedure was to be performed in the preclinical laboratory for the first time, one group (experimental, n=50) was shown a technique video for that specific procedure immediately before commencing the exercise; the control cohort (n=50) did not view the video. Technical performance on procedures was evaluated by students and two calibrated and blinded examiners. The students' perceptions of the experience were also collected in a survey. All first-year students participated in the study, for a 100% response rate. A Mann-Whitney U test did not show any group differences in technical performance (mean values on preparation: 77.1 vs. 77.8; amalgam: 82.7 vs. 82.8; composite: 79.7 vs. 78.0). A Spearman rho test revealed a significantly higher correlation in 13 out of 25 evaluation categories between student self-assessment and blinded examiner assessment for the experimental group. A chi-square test of questionnaire responses revealed a positive student perception of administering these videos for the preparation (X(2)=4.8, p<0.03), the amalgam restoration (X(2)=12.4, p<0.001), and the composite restoration (X(2)=11.3, p<0.001). The psychomotor video demonstrations did not immediately improve student performance on preclinical operative procedures, but they were well received by students and augmented self-assessment ability. These findings suggest that videos can be a useful teaching aid in a preclinical environment, especially regarding comprehension of concepts.

Predictors of tooth loss in two US adult populations.

OBJECTIVES: This study determines tooth loss rate over a 10-year period and identifies predictors of tooth loss in two separate US adult longitudinal study populations. METHODS: Subjects from the Baltimore Longitudinal Study of Aging (BLSA), consisting of 47 men and 47 women, ages ranging from 30 to 69 years, were compared to subjects from the VA Dental Longitudinal Study (VADLS) in Boston, MA, consisting of 481 men in the same age range. Baseline and follow-up examinations were performed on each cohort over a 10-year period. Using multivariate regression models, significant predictors of tooth loss were identified. RESULTS: A mean rate of tooth loss of 1.5 teeth lost per 10 years was noted in the VADLS cohort compared to 0.6 teeth lost per 10 years in the BLSA (P < .001). Combining subjects from both populations, significant predictors of tooth loss were baseline values of: percent of teeth with restorations, mean probing pocket depth score, age, tobacco use, alcohol consumption, number of teeth present, and male sex. However, the set of significant predictor variables differed between the two populations and sexes. In BLSA men, number of teeth present, percent of teeth with restorations, mean probing pocket depth score, and alcohol consumption, but not age, were significant, while in BLSA women, only age was a significant predictor. CONCLUSIONS: Over a 10-year period, the incidence of tooth loss, the rates of tooth loss, and the predictors of tooth loss were found to vary by population and by sex. These results illustrate the limits of generalizing tooth loss findings across different study cohorts and indicate that there may exist important differences in risk factors for tooth loss among US adult populations.