Correlation of quantitative light-induced fluorescence and qualitative visual rating in infiltrated post-orthodontic white spot lesions.

OBJECTIVES: The aim of this prospective, clinical single-centre study was to evaluate the masking efficacy of post-orthodontic resin infiltration after 12-month follow-up and correlate quantitative and qualitative outcome measures. METHODS: Patients with completed fixed orthodontic treatment and the presence of one or more vestibular active non-cavitated white spot lesion/s (WSL) [ICDAS 1 or 2 (International Caries Detection and Assessment System)] were provided with resin infiltration 3-12 months after bracket removal. All patients (n = 31) participating before (t0) intervention were invited again and examined after 12 months (t2). Enamel demineralization was scored using quantitative light-induced fluorescence [QLF (DeltaF[flourescence], DeltaQ[lesion volume], White Spot Area)] and qualitative visual rating [11-point Likert-scale from 0 (no lesions visible on any tooth) to 10 (all teeth affected on the entire vestibular surface)]. RESULTS: In 17 patients (7 female and 10 male) 112 WSL (ICDAS 1: n = 1; ICDAS 2: n = 111) in 112 teeth were (re)examined. Before treatment (t0) a significant, weak (DeltaF), and moderate (DeltaQ, White Spot Area) correlation was observed between the quantitative and the qualitative rating (P < 0.002) [median DeltaF: -7.31 (-10.4/-6.58)%; DeltaQ:-2.25 (-10.8/-0.41)% mm2; White Spot Area: 0.34 (0.05/1.16) mm2; visual rating:3.7 ± 1.2]. Resin infiltration led to significantly increased fluorescence and decreased visual scores (P < 0.001) 7 days (t1) and 12 months (t2) after treatment. No significant changes based on DeltaF [-6.55 (-7.29/-6.08)%] and on visual ratings [1.0 ± 1.0] were observed between t1 and t2 (P = 1.000). After 7 days (t1) the correlation between the quantitative and the qualitative ratings remained significant, weak to moderate (P < 0.002). After 12 months (t2) the correlation was (non-)significant and weak for DeltaF, DeltaQ, and White Spot Area (P ≤ 0.097). LIMITATIONS: Since the overall masking efficacy of resin infiltration has been shown previously, an untreated control group was omitted. CONCLUSIONS: When assessing the masking efficacy of infiltrated post-orthodontic WSL only a weak to moderate correlation was found between QLF values and visual ratings. Furthermore, over time this correlation decreased. Thus, it remains unclear if QLF is a viable method to assess and quantify infiltrated post-orthodontic WSL over time. TRIAL REGISTRATION: German Clinical Trials Register (DRKS-ID:DRKS00005067).

Spatial agreement of demineralized areas in quantitative light-induced fluorescence images and digital photographs.

OBJECTIVES: Previous work has shown qualitatively that detection of demineralized tooth areas (white spot lesions, WSLs) is more reliable in digital photographs (DP) as in quantitative light-induced fluorescence (QLF) images. Based on non-rigid, multimodal image registration, we now quantitatively compare manual and automatic markings in both modalities. METHODS: After braces removal, pairs of DP and QLF were acquired from 124 teeth of 31 patients. Three experienced raters marked the WSL on both DP and QLF images, each of which was presented twice in randomized order. For each tooth and each modality, a ground truth (GT) was established using the simultaneous truth and performance level estimation algorithm on the total of six manual markings per image. DP and QLF image pairs were spatially registered, by aligning the outline of the tooth area in DPs to that of the corresponding tooth area in QLF. Between all pairs of markings for all teeth, position and size were compared quantitatively by the Dice coefficient and the novel coefficient of inclusion. RESULTS: Our hypotheses: (i) the clinical inspection supported by DP is more sensitive to WSL as that by QLF, disregarding whether the automatic analysis or the experts' manual assessment of QLF is applied, and (ii) detected lesions in QLF are included in those of DP, were confirmed and not confirmed, respectively. CONCLUSION: DP and QLF are valuable methods to detect WSL in demineralized teeth. Combining both modalities can provide additional information on early lesion assessment.

Quantitative light-induced fluorescence images and digital photographs - Reproducibility of manually marked demineralisations.

OBJECTIVE: Hard tooth tissue demineralisation is an undesirable side effect of orthodontic treatment with fixed appliances. Whereas both clinically and in digital photographs (DP), demineralisations appear as white spot lesions, WSLs appear as dark areas when quantitative light-induced fluorescence (QLF) imaging is used. This study aims at comparing the reproducibility of the detection of decalcified tooth areas in DP and QLF. MATERIALS AND METHODS: DP and QLF pairs were acquired from 139 teeth of 32 patients after braces removal. Three raters manually marked the decalcified area on both DP and QLF images. The markings were repeated after 2 weeks. A ground truth was estimated for each tooth and modality using the simultaneous truth and performance level estimation (STAPLE) algorithm. The Dice coefficients (DC) of each rater marking to the ground truth were calculated for all teeth and modalities to quantify the spatial agreement. A three-way repeated measures analysis of variance (ANOVA) was used to compare the means of the DCs for both modalities ([Formula: see text]). Intra-observer and intercycle variabilities were assessed comparing the means across the raters and the cycles for both modalities. RESULTS: ANOVA revealed a statistical significant difference between the modalities [[Formula: see text], [Formula: see text]]. The standard deviation of the DC for the photographs are lower than those for the QLF images. Intra-observer and intercycle differences are rather small as compared to the intermodality differences. CONCLUSIONS: The results indicate a higher spatial reproducibility in identifying a decalcified area on a tooth surface using visual inspection of DP rather than QLF images.

Effects of anti-adhesive therapy on kidney biomarkers of ischemia reperfusion injury in human deceased donor kidney allografts.

INTRODUCTION: Molecular biomarkers validated previously in animal models are increasingly being studied in conjunction with traditional clinical endpoints in therapeutic trials. PATIENT AND METHODS: We hypothesized that human kidneys would exhibit a brisk, gene-specific inflammatory response during ischemia reperfusion injury (IRI), which would be modified by anti-adhesive therapy. Forty deceased-donor kidneys were biopsied prior to implantation and ∼1 h after reperfusion during an intervention trial with the selectin antagonist YSPSL (recombinant P-selectin glycoprotein ligand Ig). Ten inflammatory genes were measured by RT-PCR and normalized to three housekeeping genes. RESULTS: Pre-implantation kidney biopsies were already significantly inflamed relative to healthy tissue, with transcripts encoding IL-6, IL-8, and CD25 > 10-fold elevated. After reperfusion, IL-6 and IL-8 increased additional 60- and 120-fold (p < 0.05), while already elevated CD25-levels remained stable. Furthermore, transcripts encoding MCP-1, E-selectin, and TNFα were also induced significantly upon reperfusion (p < 0.0005). Systemic treatment of the recipient with YSPSL pre-reperfusion, with or without pre-implantation YSPSL flush of the donor organ, attenuated the post-reperfusion increase in MCP-1 and TGFß (p < 0.05), E-selectin and hemoxygenase 1 transcripts (p < 0.1). CONCLUSIONS: Our data in humans demonstrate a robust increase in inflammatory gene transcript levels during kidney transplantation IRI and reduction thereof by inhibition of leukocyte adhesion.

Transcription of proteinase 3 and related myelopoiesis genes in peripheral blood mononuclear cells of patients with active Wegener's granulomatosis.

OBJECTIVE: Wegener's granulomatosis (WG) is a systemic inflammatory disease that is associated with substantial morbidity. The aim of this study was to understand the biology underlying WG and to discover markers of disease activity that would be useful for prognosis and treatment guidance. METHODS: Gene expression profiling was performed using total RNA from peripheral blood mononuclear cells (PBMCs) and granulocyte fractions from 41 patients with WG and 23 healthy control subjects. Gene set enrichment analysis (GSEA) was performed to search for candidate WG-associated molecular pathways and disease activity biomarkers. Principal components analysis was used to visualize relationships between subgroups of WG patients and controls. Longitudinal changes in proteinase 3 (PR3) gene expression were evaluated using reverse transcription-polymerase chain reaction, and clinical outcomes, including remission status and disease activity, were determined using the Birmingham Vasculitis Activity Score for WG (BVAS-WG). RESULTS: Eighty-six genes in WG PBMCs and 40 in WG polymorphonuclear neutrophils (PMNs) were significantly up-regulated relative to controls. Genes up-regulated in WG PBMCs were involved in myeloid differentiation, and these included the WG autoantigen PR3. The coordinated regulation of myeloid differentiation genes was confirmed by GSEA. The median expression values of the 86 up-regulated genes in WG PBMCs were associated with disease activity (P = 1.3 x 10(-4)), and WG patients with low-level expression of the WG signature genes showed expression profiles that were only modestly different from that in healthy controls (P = 0.07). PR3 transcription was significantly up-regulated in WG PBMCs (P = 1.3 x 10(-5), false discovery rate [FDR] 0.002), but not in WG PMNs (P = 0.03, FDR 0.28), and a preliminary longitudinal analysis showed that the fold change in PR3 RNA levels in WG PBMCs corresponded to changes in the BVAS-WG score over time. CONCLUSION: Transcription of PR3 and related myeloid differentiation genes in PBMCs may represent novel markers of disease activity in WG.

Polymorphisms in the novel gene acyloxyacyl hydroxylase (AOAH) are associated with asthma and associated phenotypes.

BACKGROUND: The gene encoding acyloxyacyl hydroxylase (AOAH), an enzyme that hydrolyzes secondary fatty acyl chains of LPS, is localized on chromosome 7p14-p12, where evidence for linkage to total IgE (tIgE) concentrations and asthma has been previously reported. OBJECTIVE: We hypothesized that variants in AOAH are associated with asthma and related phenotypes. Because both AOAH and soluble CD14 respond to LPS, we tested for gene-gene interaction. METHODS: We investigated the association between 28 single nucleotide polymorphisms throughout the AOAH gene and asthma, concentrations of tIgE, the ratio of IL-13/IFN-gamma, and soluble CD14 levels among 125 African Caribbean, multiplex asthmatic pedigrees (n = 834). Real-time PCR was used to assess whether AOAH cDNA expression differed with AOAH genotype. RESULTS: Significant effects were observed for all 4 phenotypes and AOAH markers in 3 distinct regions (promoter, introns 1-6, and the intron 12/exon 13 boundary/intron 13 region) by means of single-marker and haplotype analyses, with the strongest evidence for a 2-single-nucleotide-polymorphism haplotype and log[tIgE] (P = .006). There was no difference in AOAH expression levels by AOAH genotype for any of the markers. Comparing genotypic distributions at both the AOAH marker rs2727831 and CD14(-260)C >T raises the possibility of gene-gene interaction (P = .006-.036). CONCLUSION: Our results indicate that polymorphisms in markers within the AOAH gene are associated with risk of asthma and associated quantitative traits (IgE and cytokine levels) among asthmatic subjects and their families in Barbados, and there is an interactive effect on tIgE and asthma concentrations between an AOAH marker and the functional CD14(-260)C >T polymorphism. CLINICAL IMPLICATIONS: AOAH is a novel innate immunity candidate gene associated with asthma and related phenotypes in an African ancestry population.

Polymorphisms in the novel gene acyloxyacyl hydoxylase (AOAH) are associated with asthma and associated phenotypes

BACKGROUND: The gene encoding acyloxyacyl hydrolase (AOAH), an enzyme that hydrolyzes secondary fatty acyl chains of LPS, is localised on chromosome 7p14-p12, where evidence for linkage to total IgE (tIgE) concentrations and asthma has been previously reported. OBJECTIVE: We hypothesized that variants in AOAH are associated with asthma and related phenotypes. Because both AOAH and soluble CD14 respond to LPS, we tested for gene-gene interaction. METHODS: We investigated the association between 28 single nucleotide polymorphisms throughout the AOAH gene and asthma, concentrations of tIgE, the ratio of IL-13/IFN-y, and soluble CD14 levels among 125 African Caribbean, multiplex asthmatic pedigrees (n=834). Real-time PCR was used to assess whether AOAH cDNA expression differed with AOAH genotype. RESULTS: Significant effects were observed for all 4 phenotypes and AOAH markers in 3 distinct regions (promoter, introns 1-6, and the intron 12/exon 13 boundary/intron 13 region) by means of single-marker and haplotype analyses, with the strongest evidence for a 2-single-nucleotide-polymorphism haplotype and log [tIgE] (P=.006). There was no difference in AOAH expression levels by AOAH genotype for any of the markers. Comparing genotypic distributions at both the AOAH marker rs2727831 and CD14(-260)C>T raises the possibility of gene-gene interaction (P=.006-.036). CONCLUSION: Our results indicate that polymorphisms in markers within the AOAH gene are associated with risk of asthma and associated quantitative traits (IgE and cytokine levels) among asthmatic subjects and their families in Barbados, and there is an interactive effect on tIgE and asthma concentrations between an AOAH marker and the functional CD14(-260)C>T polymorphism. CLINICAL IMPLICATIONS: AOAH is a novel innate immunity candidate gene associated with asthma and related phenotypes in an African ancestry population.

Evaluation of the CD14/-260 polymorphism and house dust endotoxin exposure in the Barbados Asthma Genetics Study.

BACKGROUND: Both a functional promoter polymorphism in the gene encoding CD14 (C-260T) and exposure to endotoxin are believed to play key roles in modulating the immune response and expression of atopic disease. OBJECTIVE: We aimed to evaluate the role of the CD14 C-260T polymorphism in a population of African descent and to test for interaction between this genotype and house dust endotoxin (HDE) exposure on atopic phenotypes. METHODS: Asthmatic probands and their families were recruited as part of the Barbados Asthma Genetics Study. The C-260T polymorphism and two additional CD14 promoter markers (G-1461T, C-1721T) were genotyped. Endotoxin was measured in house dust samples. RESULTS: Using a Family-Based Association Test, the C-260T allele appeared to be protective against asthma ( z = -2.444; P = .015) and asthma severity ( z = -2.615; P = .009) under a recessive model. No significant associations were observed for the G-1461T and C-1721T markers both individually and in haplotypes. In a case-control analysis, the CD14 TT genotype was found to reduce risk of asthma compared with the CD14 CC/CT genotypes (odds ratio [OR], 0.26; 95% CI, 0.14-0.49) and was associated with lower asthma severity scores ( P < .002). The TT genotype might protect against asthma for individuals with low HDE (OR, 0.09; 95% CI, 0.03-0.24), but may be a risk factor for individuals with high HDE (OR, 11.66; 95% CI, 1.03-131.7), suggesting a gene-environment interaction. CONCLUSION: These data suggest that the CD14-260 polymorphism may play a role in controlling risk to atopic disease and underscore the importance of incorporating key environmental exposures into studies of genetic risk factors.

Evaluation of the CD14/-260 polymorphism and house dust entotoxin exposure in the Barbados Asthma Genetics Study

BACKGROUND: Both a functional promoter polymorphism in the gene encoding CD14 (C-260T) and exposure to endotoxin are believed to play key roles in modulating the immune response and expression of atopic disease. OBJECTIVE: We aimed to evaluate the role of the CD14 C-260T polymorphism in a population of African descent and to test for interaction between this genotype and house dust endotoxin (HDE) exposure on atopic phenotypes. METHODS: Asthmatic probands and their families were recruited as part of the Barbados Asthma Genetics Study. The C-260T polymorphism and two additional CD14 promoter markers (G-1461T, C-1721T) were genotyped. Endotoxin was measured in house dust samples. RESULTS: Using a Family-Based Association Test, the C-260T allele appeared to be protective against asthma (z=−2.444; P=.015) and asthma severity (z=−2.615; P=.009) under a recessive model. No significant associations were observed for the G-1461T and C-1721T markers both individually and in haplotypes. In a case-control analysis, the CD14 TT genotype was found to reduce risk of asthma compared with the CD14 CC/CT genotypes (odds ratio [OR], 0.26; 95% CI, 0.14-0.49) and was associated with lower asthma severity scores (P < .002). The TT genotype might protect against asthma for individuals with low HDE (OR, 0.09; 95% CI, 0.03-0.24), but may be a risk factor for individuals with high HDE (OR, 11.66; 95% CI, 1.03-131.7), suggesting a gene-environment interaction. CONCLUSION: These data suggest that the CD14-260 polymorphism may play a role in controlling risk to atopic disease and underscore the importance of incorporating key environmental exposures into studies of genetic risk factors.

Evidence for asthma susceptibility genes on chromosome 11 in an African-American population.

Initial genome-wide scan data provided suggestive evidence for linkage of the asthma phenotype in African-American (AA), but not Caucasian, families to chromosome 11q markers (peak at D11S1985; LOD=2). To refine this region, mapping analysis of 91 AA families (51 multiplex families and 40 asthmatic case-parent trios) was performed with an additional 17 markers flanking the initial peak linkage marker. Multipoint analyses of the 51 multiplex families yielded significant evidence of linkage with a peak non-parametric linkage score of 4.38 at marker D11S1337 (map position 68.6 cM). Furthermore, family-based association and transmission disequilibrium tests conducted on all 91 families showed significant evidence of linkage in the presence of disequilibrium for several individual markers in this region. A putative susceptibility locus was estimated to be at map position 70.8 cM with a confidence interval spanning the linkage peak. Evidence from both linkage and association analyses suggest that this region of chromosome 11 contains one or more susceptibility genes for asthma in these AA families.

Dense mapping of chromosome 12q13.12-q23.3 and linkage to asthma and atopy

BACKGROUND: Asthma is a complex disease characterized by a high prevalence of allergic diathesis and the almost ubiquitous presence of upper airway disease (eg rhinitis). Previously, we observed linkage of asthma among Afro-Caribbean families to markers in chromosome 12q, which contains a number of genes encoding for products closely related to allergic airway inflammation and disease. OBJECTIVE: To identify susceptibility loci in chromosome 12q contributing to the genetics of upper and lower airway diseases and to expand the region to include genes encoding IFN- ã(IFNG) and one of the signal transducers and activators of transcription (STAT6), we conducted further linkage studies among 33 multiplex families. METHODS: We characterized 528 subjects from Barbados for asthma; 82 percent were characterized for allergic rhinitis. Two-point and multipoint linkage analysis of 22 microsatellite markers (spanning ~79 centimorgan) was performed. RESULTS: Affected sib-pair analysis revealed significant evidence for linkage to asthma over approximately 30 cM (P < .05 to .002), with the best evidence for linkage at a CA repeat polymorphism in the first intron of IFNG in 12q21.1 (P = .002). Evidence of linkage to allergic rhinitis was observed in the same region (D12S313, P = .006, and IFNGCA, P = .01. respectively). Multipoint linkage analysis also provided evidence for linkage to asthma, with the best nonparametric linkage analysis score at D12S326 (nonparametric linkage score = 3.8, P = .0008). Modest evidence for linkage to allergic rhinitis was observed next to D12S326 at D12S1052 (p = .036) CONCLUSIONS: Our findings suggest that (1) one or more loci in the chromosome 12q13.12-q23.3 region are contributing to the expresstion of the clinical phenotype asthma and the strongest evidence for linkage is in a region near the gene encoding IFNG and (2) a susceptibility locus for both asthma and allergic rhinitis maps to this region.