CD3+CD4+LAP+Foxp3-regulatory cells of the colonic lamina propria limit disease extension in ulcerative colitis

Background and Aims: In ulcerative colitis (UC), inflammation begins in the rectum and can extend proximally throughout the entire colon. The extension of inflammation is an important determinant of disease course, and may be limited by the action of regulatory T cells (Tregs). In this cross-sectional study, we evaluated the relationship between UC extension and the proportions of CD3+CD4+Foxp3+ and CD3+CD4+LAP+Foxp3- Tregs in the colonic lamina propria (LP) of 79 UC patients and 29 controls. The role of these cells in UC extension was also investigated in the murine oxazolone-induced colitis model. Methods: Patients: Disease extension was classified according to the Montreal classification. Where possible, endoscopic biopsies of involved and uninvolved tissue were obtained from UC patients. Mouse model: Colitis was induced by intrarectal oxazolone administration. Lamina propria mononuclear cells were isolated from patient biopsies and mouse colon tissue using enzymatic method and the percentage of CD3+CD4+Foxp3+ and CD3+CD4+LAP+Foxp3-cells evaluated by immunofluorescence. Confocal microscopy was applied for the visualization and quantification of CD4+LAP+ cells on tissue histological sections. Results: In UC patients with distal colitis the proportion of LP CD3+CD4+Foxp3+ Tregs was significantly higher in inflamed tissue than uninvolved tissue. As opposite, the proportion of LP CD3+CD4+LAP+ Tregs was significantly higher in uninvolved tissue than involved tissue. Both LP CD3+CD4+Foxp3+ and LP CD3+CD4+LAP+ Tregs proportion in involved tissue was significantly higher than in controls irrespective of the extension of inflammation. In mice with oxazolone-induced distal colitis, treatment with LAP-depleting antibody was associated with the development of extensive colitis. Conclusions: Our findings suggest that CD3+CD4+LAP+Foxp3-Tregs limit the extension of inflammatory lesions in UC patients

3D Evaluation of Upper Airway Morphological Changes in Growing Patients with Class II Malocclusion Using Sander Bite Jumping Appliance

Class II malocclusion due to mandibular retrognathia is associated with a posterior positioning of the tongue and the hyoid bone, reducing the oropharyngeal volume. This could be a contributing factor to the development of respiratory and cardiovascular problems. This study evaluates the oropharyngeal volume variation in 13 patients with class II malocclusion undergoing functional orthopedic treatment with Sander Bite Jumping Appliance (SBJ). CBCT scans were performed before treatment (T0) and approximately after 12.5 months (T1): the retropalatal volume and retroglossal volume were quantified in mm(3) using a segmentation software. At T1, the retropalatal volume increased in 2523 +/- 2088 mm(3), and the retroglossal volume increased in 2258 +/- 1717 mm(3). Both values were statistically significant (p < 0.05). This widening of the airways may allow prevention and treatment of sleep-disordered breathing, including obstructive sleep apnea syndrome

Evaluation of the Efficacy of Low-Particle-Size Toothpastes against Extrinsic Pigmentations: A Randomized Controlled Clinical Trial

Stain-removing domiciliary protocols are focused on the elimination of dental extrinsic pigmentations by the application of abrasive toothpastes, extensively available in commerce. The goal of the present study is to evaluate the efficacy of two different stain removal molecule-formulated toothpastes by the reduction of clinical parameters: the micro-cleaning crystals and activated charcoal. A total of 40 participants with extrinsic dental pigmentations were enrolled and divided into two groups: a Control group, assigned to a toothpaste with micro-cleaning crystals (Colgate Sensation White); and a Trial group, with microparticle-activated charcoal toothpaste (Coswell Blanx Black). At T0 (baseline), T1 (10 days), T2 (1 month), and T3 (3 months), clinical parameters, including Lobene stain index calculated for intensity and extension, plaque control record, and bleeding on probing, were measured. Statistically significant differences were found in both groups (p < 0.05): a reduction of extrinsic pigmentation, both in intensity and extension, was obtained in the Control group, but their total elimination could be achieved only in the Trial group with the activated charcoal molecule, though without significant difference between the groups (p > 0.05). No intergroup differences were found for each timeframe for PCR, BoP, LSI-I, and LSI-E. Both tested toothpastes can be recommended for domiciliary oral hygiene of patients with extrinsic pigmentations

IL-13 mRNA tissue content identifies two subsets of adult ulcerative colitis patients with different clinical and mucosa-associated microbiota profiles

BACKGROUND AND AIM: A personalized approach to therapy has great promise to improve disease outcomes. To this end, the identification of different subsets of patients according with the prevalent pathogenic process might guide in the choice of therapeutic strategy. We hypothesize that UC patients might be stratified according to distinctive cytokine profiles and/or to a specific mucosa-associated microbiota. METHODS: In a cohort of clinically and endoscopic active UC patients and controls, we analyzed by qPCR the mucosal cytokine mRNA content and the mucosa-associated microbiota composition assessed by the 16SrRNA gene sequencing. RESULTS: We demonstrate, by means of data-driven approach, the existence of a specific UC patient subgroup characterized by elevated IL-13mRNA tissue content separated by patients with low IL-13 mRNA tissue content. The two subsets differ in clinical-pathological characteristics. High IL-13mRNA patients are younger at diagnosis and show higher prevalence of extensive colitis than low IL-13mRNA ones. They also show a more frequent use of steroid/immunosuppressant/anti-TNFα therapy during a one-year follow-up. The two subgroups show a differential enrichment of mucosa associated microbiota genera with prevalence of Prevotella in patients with high IL-13mRNA tissue content and Sutterella and Acidaminococcus in patients with low IL-13mRNA tissue content. CONCLUSION: Assessment of mucosal IL-13mRNA might help in the identification of the patients' subgroup that might benefit from a therapeutic approach modulating IL-13

The Mutyh Base Excision Repair Gene Influences the Inflammatory Response in a Mouse Model of Ulcerative Colitis

BACKGROUND: The Mutyh DNA glycosylase is involved in the repair of oxidized DNA bases. Mutations in the human MUTYH gene are responsible for colorectal cancer in familial adenomatous polyposis. Since defective DNA repair genes might contribute to the increased cancer risk associated with inflammatory bowel diseases, we compared the inflammatory response of wild-type and Mutyh(-/-) mice to oxidative stress. METHODOLOGY/PRINCIPAL FINDINGS: The severity of colitis, changes in expression of genes involved in DNA repair and inflammation, DNA 8-oxoguanine levels and microsatellite instability were analysed in colon of mice treated with dextran sulfate sodium (DSS). The Mutyh(-/-) phenotype was associated with a significant accumulation of 8-oxoguanine in colon DNA of treated mice. A single DSS cycle induced severe acute ulcerative colitis in wild-type mice, whereas lesions were modest in Mutyh(-/-) mice, and this was associated with moderate variations in the expression of several cytokines. Eight DSS cycles caused chronic colitis in both wild-type and Mutyh(-/-) mice. Lymphoid hyperplasia and a significant reduction in Foxp3(+) regulatory T cells were observed only in Mutyh(-/-) mice. CONCLUSIONS: The findings indicate that, in this model of ulcerative colitis, Mutyh plays a major role in maintaining intestinal integrity by affecting the inflammatory response

Role of regulatory cells and tissue cytokines in patients with ulcerative colitis: implications for patient-oriented therapy

Ulcerative colitis (UC) is an inflammatory bowel disease that results in a chronic inflammation and ulcers of the mucosa of the colon. In UC, two important issues, specifically related to the prognosis, remain to be clarified. A) Extension of disease greatly influences the prognosis and biological factors involved in the limitation of the disease extent are presently unknown B) Due to increased availability of biological drugs targeting different cytokines, biological therapy need to be optimized on the key cytokine(s) relevant for the inflammatory process. Aim: A) We hypothesized that regulatory T cells might be relevant in limiting the extension of inflammation and that B) UC patients might be stratified according to distinctive cytokine profiles. Methods: In a cross-sectional study, we evaluated the relationship between UC extension and the proportions of CD3+CD4+Foxp3+ and CD3+CD4+LAP+Foxp3-Tregs in the colonic lamina propria (LP) in UC patients undergoing colonoscopy for clinical relapse and controls undergoing colonoscopy for non-inflammatory conditions. The role of these cells in UC extension was also investigated in the murine oxazolone-induced colitis model. B) At the same time, we analyzed tissue cytokine by RTqPCR in endoscopic biopsies from uninvolved and involved tissue of UC patients and controls. Mucosal microbiota analysis was also performed. Results: A) Involved and uninvolved tissue show different and opposite frequency of LP CD3+CD4+Foxp3+ and LP CD3+CD4+LAP+ Tregs with the latter significantly increased in uninvolved vs involved tissue. In mice with oxazolone-induced distal colitis, treatment with α-LAP-depleting antibody was associated with the development of extensive colitis; B) We quantified by RTqPCR TNF-α, IFN-γ, IL-10, IL-6, IL-17, and IL-13 and analyzed the results by r square of the k means for cluster solution. IL-17A and IL-13 mRNA tissue content show classificatory power and their distribution points to the existence of discrete clusters of subjects. Six clusters define the optimal partition of data explaining 91% of the total information present in the data set (R-square=0.91). UC patients were distributed in two clusters characterized by high/low IL-13 mRNA tissue content in the context of high IL-17A mRNA tissue content. The two subsets differ in clinical-pathological characteristics. High IL-13mRNA patients are younger at diagnosis and show higher prevalence of extensive colitis than low IL-13mRNA ones. They also show a more frequent use of steroid/immunosuppressant/anti-TNFα therapy during a one-year follow-up. The two subgroups show a differential enrichment of mucosa associated microbiota genera with prevalence of Prevotella in patients with high IL-13mRNA tissue content and Sutterella and Acidaminococcus in patients with low IL-13mRNA tissue content. Conclusion: A) LP CD3+CD4+LAP+ Tregs efficiently limit the extension of inflammatory lesions. Therapeutic strategy aimed at their expansion may help in containing the spreading of inflammation and possibly in preventing inflammatory relapse. B) Evaluation of IL-13 and IL-17 mRNA content allows the identification of different UC patients’ subsets with associated different clinic phenotypes. Therapeutic options might be optimized according to the patients’ tissue cytokine profile

Photodynamic Therapy in Non-Surgical Treatment of Periodontitis: A Systematic Review and Meta-Analysis

Aim: to evaluate the adjunctive effects of photodynamic therapy (aPDT) on nonsurgical mechanical treatment in patients with periodontitis. Materials and methods: The search strategy was conducted according to the PRISMA guidelines to answer research questions regarding the effectiveness of aPDT in association with non-surgical periodontal therapy. The mean values and standard deviations were collected by data extraction. A descriptive comparison between aPDT in association with periodontal treatment and periodontal treatment alone was performed, and meta-analyses of PPD were also performed. Both randomized controlled clinical trials (RCTs) and controlled clinical trials (CCTs) were included. Results: Out of 2059 records, 14 articles on adjunctive photodynamic therapy were included because they met the eligibility criteria. A comparison between the aPDT data and the control group showed improved PPD for photodynamic therapy (SMD −0.76, p = 0.003; I2 = 88%). Statistical analysis was then applied to the three PPD subgroups. The first group included studies that used indocyanine green in association with a wavelength of 810 nm (SMD −1.79, p 2 = 88%). The second group included studies that used phenothiazine chloride at a wavelength of 660 nm (SMD −0.03, p = 0.84, I2 = 0%). The last group included studies that used methylene blue photosensitizers treated with a wavelength 628–670 nm were included (SMD −0.13, p = 0.38; I2 = 0%). Conclusions: despite the limited number of RCTs and the great heterogeneity between them, it can be concluded that aPDT in association with nonsurgical periodontal treatment improved the clinical parameters at 3 months