The degree of microRNA-34b/c methylation in serum-circulating DNA is associated with malignant pleural mesothelioma

Objectives

Malignant pleural mesothelioma (MPM) is an aggressive tumor with a poor prognosis. microRNA-34b/c (miR-34b/c), which plays an important role in the pathogenesis of MPM, is frequently downregulated by DNA methylation in approximately 90% of MPM cases. In this study, we estimated the degree of miR-34b/c methylation in serum-circulating DNA using a digital methylation specific PCR assay (MSP).

Materials and methods

A real-time MSP assay was performed using the SYBR Green method. The melting temperature (Tm) of each PCR product was examined using a melting curve analysis. For a digital MSP assay, 40 wells were analyzed per sample. A total of 110 serum samples from 48 MPM cases, 21 benign asbestos pleurisy (BAP) cases, and 41 healthy volunteers (HVs) were examined.

Results

Positive range of Tm value for miR-34b/c methylation was defined as 77.71–78.79 °C which was the mean ± 3 standard deviations of 40 wells of a positive control. The number of miR-34b/c methylated wells was counted per sample according to this criterion. The number of miR-34b/c methylated wells in MPM cases was significantly higher than that in BAP cases (P = 0.03) or HVs (P < 0.001). Advanced MPM cases tended to have higher number of miR-34b/c methylated wells than early MPM cases. Receiver–operating characteristic (ROC) curve analysis revealed that three number of miR-34b/c methylated wells per sample was the best cut-off of positivity of MPM with a 67% of sensitivity and a 77% specificity for prediction. The area under the ROC curve was 0.77.

Conclusions

Our digital MSP assay can quantify miR-34b/c methylation in serum-circulating DNA. The degree of miR-34b/c methylation in serum-circulating DNA is associated with MPM, suggesting that this approach might be useful for the establishment of a new detection system for MPM.     

Computer-assisted simulations in orthodontic diagnosis and the application of a new cone beam X-ray computed tomography

Computational simulations which include three-dimensional (3-D) image processing and biomechanical calculations should provide useful information to our research and orthodontic clinic as a clinical tool defined as 'thinking'. In this review, 1) biomechanical simulations applied to predict the mandibular growth; 2) mathematical models of virtual bone cells and 3) 3-D images and solid model simulations for surgical planning are introduced. In biomechanical simulation, biting force, electromyographic (EMG) activity and cephalograms of 32 subjects were applied. Computational results of mathematical model were compared with actual bone growth in a rat. Three-dimensional image and solid model of 14 patients were utilized for their treatment planning. From the results, several concepts of our simulations were confirmed: 1) reaction forces generated by masticatory muscles at the condyle control the direction of mandibular growth; 2) some mathematical models have the possibility to describe the process of bone growth; 3) 3-D image processing software and solid models are necessary for diagnosis and planning of orthognathic surgery. We also believe that the orthodontists can more accurately predict the affects of surgical procedures and orthodontic tooth movement using the new cone beam X-ray computed tomography (CT) (CB MercuRay; Hitachi Medico Technology, Tokyo, Japan) and its advanced application software.     

Gene expression profiling of mouse condylar cartilage during mastication by means of laser microdissection and cDNA array

Little is known about the mechanisms of mandibular condylar growth. In this study, gene expression in the mandibular condylar cartilage of young post-natal mice was monitored by means of a cDNA microarray, real-time PCR, and laser microdissection before and after the initiation of mastication (newborn, 7 days, 21 days, initiation of mastication, and 35 days). Insulin-like growth factor-1 (IGF-I), transforming-growth-factor-beta-2 (TGFbeta2), and aggrecan mRNAs were clearly expressed at 21 days, while the expression of osteopontin mRNAs was most clear at 35 days. Parathyroid-hormone-related protein (PTHrP), Indian-hedgehog (Ihh), and insulin-like growth factor-2 (IGF-2) mRNAs were clearly expressed during lactation (newborn and 7 days). Heat-shock-protein 84 (HSP-84) and heat-shock-protein 86 (HSP-86) were clearly expressed at 35 days. These results revealed that gene expression changed during mandibular condylar cartilage growth, and that, interestingly, these changes coincided with the initiation of mastication.     

Three-dimensional changes in the condyle during development of an asymmetrical mandible in a rat: a microcomputed tomography study.

A rapidly growing postnatal animal model was used to study changes in the calcified tissue of the mandibular condyle during altered muscle function. A maxillary occlusal splint was designed to shift the mandible laterally (left) during closure. Groups of 5 Wistar rats were killed at 5, 9, 15, 21, 30, and 40 weeks (n = 30), with an equal number of controls. The experimental animals developed shorter, asymmetrical mandibles compared with the control animals. The left condyle became larger and thicker than the right condyle. Microcomputed tomography assessment of the left and right condylar trabecular bone indicated that both had less bone volume than the control condyle. The right masseter muscle significantly lost fiber size and type IIA oxidative fibers, suggesting that the right masseter muscle was used with less tension development. In contrast, the left masseter maintained its fiber size and was similar to the control masseter fiber diameters. Comparison in the sequence of changes indicated that the morphologic changes occurred first in the ramus (age, 5 weeks), before the corpus (age, 15 weeks), and before changes in masseter fiber size and composition (age, 9 weeks). This study showed that both the mandible and the condyle modified their shape and size, as well as the trabecular bone of the condyle, during shifting of the mandible to one side as it closed.     

Effects of plaque disclosing agents on esthetic restorative materials used in pediatric dentistry

The aim of study was to evaluate the color stability of tooth-colored restorative materials usually used in pediatric dentistry after the application of two plaque disclosing agents. Twenty specimens of each material: a resin-modified glass ionomer, a composite resin and an ion-releasing composite resin, were prepared. Baseline color evaluation was performed, samples were exposed to the plaque disclosing agents: a basic fuchsin solution and a fluorescent dye, and new color evaluations were made. The resin-modified glass ionomer stained with basic fuchsin presented the greatest color change in the present study, and the fluorescent dye did not show statistically significant changes among the restorative materials. In conclusion, basic fuchsin dyes should be carefully used in children with a great number of tooth-colored restorations.     

Genome-wide linkage analysis of mandibular prognathism in Korean and Japanese patients

The existence of familial aggregation of mandibular prognathism (MP) suggests that genetic components play an important role in its etiology. In this study, a genome-wide linkage analysis to identify loci susceptible to MP was conducted with 90 affected sibling-pairs in 42 families, comprised of 40 Korean sibling-pairs and 50 Japanese sibling-pairs. Two non-parametric linkage analyses, GENEHUNTER-PLUS and SIBPAL, were applied and detected nominal statistical significance of linkage to MP at chromosomes 1p36, 6q25, and 19p13.2. The best evidence of linkage was detected near D1S234 (maximum Z(lr) = 2.51, P = 0.0012). In addition, evidence of linkage was observed near D6S305 (maximum Z(lr) = 2.23, P = 0.025) and D19S884 (maximum Z(lr) = 1.93, P = 0.0089). Identification of the susceptible genes in the linkage regions will pave the way for insights into the molecular pathways that cause MP, especially overgrowth of the mandible, and may lead to the development of novel therapeutic tools.     

Root fracture in immature anterior teeth followed for 15 years

Abstract –  We report a case of injury to an immature tooth, observed over a period of 15 years. In 1987, a 9-year-old boy fell down in a schoolyard. The right central incisor demonstrated palato-version and radiographic observations revealed that the roots of both central incisors were incomplete. Further, a root fracture in the apical region of the central incisors was observed. During the first treatment visit, the right central incisor was repositioned and both teeth splinted. After confirming that the line of fracture was aligned, the fixation was continued for 2 months. The teeth were examined periodically for the next 15 years. Both teeth had favorable outcomes with continued root development of both the apical and coronal segments with good apposition of the fracture lines.     

Effects of dietary consistency on the mandible of rats at the growth stage: computed X-ray densitometric and cephalometric analysis

Thirty 3-week-old male Wistar rats were grouped into a hard diet control group, a kneaded-diet group, and a powdered-diet group. After 6 weeks of growth, all rats were killed under deep anesthesia. The effects of dietary physical consistency on the mandible were investigated with respect to morphology and bone mineral content using lateral cephalometric analysis and computed X-ray densitometry. Significant differences between the experimental and control groups were determined by measuring the means and standard deviation of the coordinates and differences between 15 points selected on the mandibles. Gonion (Go), which is a measurement of depth to the X-axis, was significantly less in the powdered-diet group, while the kneaded-diet group showed no significant differences. The most posterior point of the coronoid process (Cr) and the most posterior point of condylar process (Cd), which are measures of height to the Y-axis, were less in the kneaded-diet group than in the control group. In the powdered-diet group, Cr, Cd, Go and Infradentale were significantly less than in the control group. On the other hand, the bone mineral content was significantly lower in the coronoid process and angle of mandible in only the powdered-diet group.