Multifaceted array-based keloidal gene expression profiling reveals specific MDFI upregulation in keloid lesions.

BACKGROUND: Keloid lesions are characterized by mesenchymal cell proliferation and excessive extracellular matrix deposition. Previous microarray analyses have been performed to investigate the mechanism of keloid development. However, the molecular pathology that contributes to keloid development remains obscure. AIM: To explore the underlying essential molecules of keloids using microarrays. METHODS: We performed microarray analyses of keloid and nonlesional skin tissues both in vivo and in vitro. Gene expression levels were compared between tissues and cells. Quantitative reverse transcription (qRT)-PCR and immunohistochemical staining were used to determine the expression levels of molecules of interest in keloid tissues. RESULTS: Several common molecules were upregulated in both keloid tissues and keloid-lesional fibroblasts. PTPRD and NTM were upregulated both in vivo and in vitro. The genes MDFI and ITGA4 were located at the centre of the gene coexpression network analysis using keloid tissues. qRT-PCR revealed significant expression levels of PTPRD and MDFI in keloid tissues. Immunopathological staining revealed that MDFI-positive cells, which have fibroblast characteristics, were located in the keloid-associated lymphoid tissue (KALT) portion of the keloid tissue. CONCLUSION: Our gene expression profiles of keloids could distinguish the difference between lesional tissue and cultured lesional fibroblasts, and MDFI was found to be commonly expressed in both tissues and cells. Thus, MDFI-positive cells, which were located in the KALT, may play an important role in keloid pathogenesis and thus might be useful for in vitro keloid studies.

Utility of a diffusion kurtosis model in the differential diagnosis of orofacial tumours.

AIM: To compare the goodness of fit and correlations between diffusion kurtosis imaging (DKI) and a mono-exponential (ME) model, to compare the corrected apparent diffusion coefficient (Dapp) and apparent kurtosis (Kapp) of the DKI model, and the apparent diffusion coefficient (ADC) of the ME model among the various orofacial lesions, and to evaluate the diagnostic performances between the two models. MATERIALS AND METHODS: A total of 100 orofacial lesions underwent echo-planar diffusion magnetic resonance imaging (MRI) with four b-values. The goodness of fit was evaluated using Akaike information criterion. The correlations of the diffusion-derived parameters were evaluated. The diagnostic performance was analysed by receiver operating characteristics (ROC). RESULTS: The DKI model showed a significantly better goodness of fit than the ME model (p<0.0001). The Kapp had a strongly negative correlation with the Dapp (ρ=-0.749) and ADC (ρ=-0.938). A strongly positive correlation existed between the Dapp and ADC (ρ=0.906). All parameters differed significantly between benign tumours and malignant tumours (p<0.05). In differentiating benign tumours from the malignant tumours, the AUC of Dapp (0.871) was larger than that of ADC (0.805); however, a significant difference was not found (p=0.102). CONCLUSION: The DKI model had better goodness of fit than the ME model. Furthermore, the Dapp and Kapp were also characteristic for each pathological category; however, the DKI model did not yield a significantly higher diagnostic performance than the ME model, which might be related to the high correlation among the diffusion-derived parameters and wide variation among categories.

Dynamic contrast-enhanced MRI of oral squamous cell carcinoma: a preliminary study of the correlations between quantitative parameters and the clinical stage.

OBJECTIVE: To probe the utility of dynamic contrast-enhanced MRI (DCE-MRI) parameters in assessing the clinical characteristics of oral squamous cell carcinoma. METHODS: A total of 85 tumours were included. We applied the Tofts and Kermode model for the DCE-MRI data and obtained three dependent parameters: the influx forward volume transfer constant into the extravascular extracellular space (EES) from the plasma (K(trans)), the fractional volume of EES per unit volume of tissue (ve) and the fractional volume of plasma (vp). We evaluated the correlations between these parameters and the clinical stages. RESULTS: The T stage showed a negative correlation with the K(trans) (r = -0.2272; p = 0.0365), but it did not show a significant correlation with the other parameters. The N stage showed a negative correlation with K(trans) (r = -0.1948; p = 0.0404), and there were significant differences between N1 and N2+3 (0.119 ± 0.027 vs 0.096 ± 0.023 min(-1); p = 0.0198) and between N0 and N2+3 (0.114 ± 0.29 vs 0.096 ± 0.023 min(-1); p = 0.0288). CONCLUSION: A decrease in the K(trans) at the primary site was found in advanced N stage cases, which might indicate that the hypoxic status cause a high possibility of the metastasis. ADVANCES IN KNOWLEDGE: A decrease in the K(trans) at the primary site suggested the high possibility of an advanced N stage.

Autosomal recessive cystinuria caused by genome-wide paternal uniparental isodisomy in a patient with Beckwith-Wiedemann syndrome.

Approximately 20% of Beckwith-Wiedemann syndrome (BWS) cases are caused by mosaic paternal uniparental disomy of chromosome 11 (pUPD11). Although pUPD11 is usually limited to the short arm of chromosome 11, a small minority of BWS cases show genome-wide mosaic pUPD (GWpUPD). These patients show variable clinical features depending on mosaic ratio, imprinting status of other chromosomes, and paternally inherited recessive mutations. To date, there have been no reports of a mosaic GWpUPD patient with an autosomal recessive disease caused by a paternally inherited recessive mutation. Here, we describe a patient concurrently showing the clinical features of BWS and autosomal recessive cystinuria. Genetic analyses revealed that the patient has mosaic GWpUPD and an inherited paternal homozygous mutation in SLC7A9. This is the first report indicating that a paternally inherited recessive mutation can cause an autosomal recessive disease in cases of GWpUPD mosaicism. Investigation into recessive mutations and the dysregulation of imprinting domains is critical in understanding precise clinical conditions of patients with mosaic GWpUPD.

Clinical applications of analysis of plasma circulating complete hydatidiform mole pregnancy-associated miRNAs in gestational trophoblastic neoplasia: a preliminary investigation.

The aim of this study was to investigate the clinical application of plasma complete hydatidiform mole pregnancy-associated microRNAs (CHM-miRNAs: hsa-miR-520b, hsa-miR-520f and hsa-miR-520c-3p). We measured plasma CHM-miRNA concentration by real-time quantitative reverse transcriptase polymerase chain reaction in two cases of CHM resulting in gestational trophoblastic neoplasia later. As progress of treatments in both cases, the plasma concentrations of CHM-miRNAs showed a decreasing tendency similar to the pattern for serum hCG concentration, but exhibited a transient increasing tendency after each course of chemotherapy, suggesting that the plasma CHM-miRNAs could be an additional follow-up marker for malignant changes of CHM.

Circulating levels of maternal plasma cell-free pregnancy-associated placenta-specific microRNAs are associated with placental weight.

The aim of this study was to investigate the relationship between plasma concentration of cell-free pregnancy-associated placenta-specific microRNAs and clinical variables (placental weight, maternal body mass index, and neonatal birth weight). Circulating levels of cell-free pregnancy-associated placenta-specific microRNAs (miR-515-3p, miR-517a, miR-517c and miR-518b) in maternal plasma were measured by quantitative real-time RT-PCR in sixty-two pregnant women. The levels of cell-free pregnancy-associated placenta-specific microRNAs were significantly associated with placental weight, but not associated with body mass index or birth weight. Therefore, the measurement of cell-free pregnancy-associated placenta-specific miRNAs levels in maternal plasma may reflect the pregnancy status related to placenta volume.

A new method for displaying the lingual artery using high-resolution three-dimensional phase-contrast magnetic resonance angiography.

The aim of this study was to display the lingual artery superimposed on the anatomical image and to confirm its course and relation to the adjacent structures, noninvasively. Nineteen volunteers participated in the magnetic resonance imaging (MRI) study and one was excluded for excessive movement during scanning. A three-dimensional phase-contrast sequence (3D-PC) of magnetic resonance angiography (MRA) was used for vessel images, and a 3D-T1 high-resolution volume examination (THRIVE) was used for anatomical images. Colour-coded vessel images from 3D-PC MRA were superimposed on the 3D volume anatomical images, and the arterial course and relation to the adjacent structures were confirmed with multiplanar reconstructed cross-sectional (MPR) images. 3D-PC MRA images visualized the lingual artery in all 18 subjects and the sublingual artery in 14 subjects. In seven of 18 cases the bilateral sublingual arteries were shown to run side by side but had no contact with the sublingual veins. They ran together with the sublingual veins in four cases. Three cases showed irregular patterns. The bilateral sublingual arteries could not be identified in four cases. 3D-PC MRA images of the lingual artery superimposed on the anatomical images may be clinically useful to confirm its course and relationship to the adjacent structures before surgery, in order to prevent haemorrhage.

Imaging findings of neurogenic tumours in the head and neck region.

OBJECTIVE: The aim of this study was to describe the CT, MRI and ultrasonography findings of five cases of neurogenic tumours in the head and neck region. METHODS: Five neurogenic tumours were analysed with respect to their CT value, the presence of cystic change, target sign, lobulation, connection to the nerve and vascularity. RESULTS: The contrast-enhanced CT (ECT) of the schwannomas demonstrated either a mass with low enhancement (two out of three cases), which reflected the predominant Antoni B components, or a mass with cystic changes, which was an Antoni A-based schwannoma displaying cystic changes (one out of three cases). On MRI, all tumours showed homogeneous and isointense signals for muscle on T1 weighted images (T1 WIs). T2 weighted images (T2 WIs) and gadolinium (Gd)-enhanced T1 WIs demonstrated target sign in both schwannomas. Ultrasound examination showed a well-defined, ovoid or round hypoechoic mass. The direct connection to the nerve was demonstrated in two of the five cases. Lobulation was observed in only one of the five cases and cystic changes were observed in one of the five cases. In all of the cases, no vascularity was seen in power Doppler images (PDIs) obtained percutaneously. CONCLUSIONS: Low-enhanced areas on ECTs can be specific for schwannomas, which suggests the predominance of Antoni B components. The target sign on T2 WIs and Gd-enhanced T1 WIs can be specific, which can be used to differentiate the two different components (Antoni A and Antoni B). The direct connection to the nerve can be a specific finding for neurogenic tumours; however, at present the sensitivity is 40%.

Surfactant protein C G100S mutation causes familial pulmonary fibrosis in Japanese kindred.

Several mutations in the surfactant protein C (SP-C) gene (SFTPC) have been reported as causing familial pulmonary fibrosis (FPF). However, the genetic background and clinical features of FPF are still not fully understood. We identified one Japanese kindred, in which at least six individuals over three generations were diagnosed with pulmonary fibrosis. We examined the patients radiologically and histopathologically and sequenced their SFTPC and ABCA3 genes. We also established a cell line stably expressing the mutant gene. All the patients had similar radiological and histopathological characteristics. Their histopathological pattern was that of usual interstitial pneumonia, showing numerous fibroblastic foci even in areas without abnormal radiological findings on chest high-resolution computed tomography. No child had respiratory symptoms in the kindred. Sequencing of SFTPC showed a novel heterozygous mutation, c.298G>A (G100S), in the BRICHOS domain of proSP-C, which co-segregated with the disease. However, in the ABCA3 gene, no mutation was found. In vitro expression of the mutant gene revealed that several endoplasmic reticulum stress-related proteins were strongly expressed. The mutation increases endoplasmic reticulum stress and induces apoptotic cell death compared with wild-type SP-C in alveolar type II cells, supporting the significance of this mutation in the pathogenesis of pulmonary fibrosis.

Evaluation of T2 values and apparent diffusion coefficient of the masseter muscle by clenching.

OBJECTIVE: The aim of this study was to evaluate the changes in T2 values and apparent diffusion coefficient (ADC) in the masseter muscle by clenching in healthy volunteers. METHODS: 37 volunteers were enrolled in the study. We measured bite force using pressure-sensitive paper and a T2 map. The ADC map was obtained at rest, during clenching, immediately after and 5 min after clenching. The spin-echo sequence was used to calculate T2, and single-shot spin-echo echo planar imaging was used to calculate the ADC. The motion-probing gradients (MPGs) were applied separately along the posterior-to-anterior (PA), right-to-left (RL) and superior-to-inferior (SI) directions, with b values of 0, 300 and 600 s mm(-2) in each direction. ADC-PA, ADC-RL, and ADC-SI values were obtained, and we calculated the ADC-iso for the mean diffusivity. RESULTS: There were no significant differences between the stronger and weaker sides of bite force before, during or 5 min after clenching for T2 and ADC. The bite force had little effect on these parameters; thus, we used the average of the two sides for the following analyses. Time course analysis of ADC-iso, ADC-PA, ADC-RL and ADC-SI demonstrated a marked increase after clenching and a rapid decrease immediately after clenching, although they did not completely return to the initial values; however, the change in ADC-RL was significantly greater than those in ADC-PA or ADC-SI (P<0.001 each). The changes in T2 were similar to those of ADC, although not as marked. CONCLUSIONS: ADC (especially ADC-RL) was altered by contraction of the masseter muscle.

Maternal uniparental isodisomy and heterodisomy on chromosome 6 encompassing a CUL7 gene mutation causing 3M syndrome.

We report a case of segmental uniparental maternal hetero- and isodisomy involving the whole of chromosome 6 (mat-hUPD6 and mat-iUPD6) and a cullin 7 (CUL7) gene mutation in a Japanese patient with 3M syndrome. 3M syndrome is a rare autosomal recessive disorder characterized by severe pre- and postnatal growth retardation that was recently reported to involve mutations in the CUL7 or obscurin-like 1 (OBSL1) genes. We encountered a patient with severe growth retardation, an inverted triangular gloomy face, an inverted triangle-shaped head, slender long bones, inguinal hernia, hydrocele testis, mild ventricular enlargement, and mild mental retardation. Sequence analysis of the CUL7 gene of the patient revealed a homozygous missense mutation, c.2975G>C. Genotype analysis using a single nucleotide polymorphism array revealed two mat-hUPD and two mat-iUPD regions involving the whole of chromosome 6 and encompassing CUL7. 3M syndrome caused by complete paternal iUPD of chromosome 6 involving a CUL7 mutation has been reported, but there have been no reports describing 3M syndrome with maternal UPD of chromosome 6. Our results represent a combination of iUPDs and hUPDs from maternal chromosome 6 involving a CUL7 mutation causing 3M syndrome.

Occlusal pressure, contact area, force and the correlation with the morphology of the jaw-closing muscles in patients with skeletal mandibular asymmetry.

The purpose of this study was to investigate the difference in the occlusal force between deviated and non-deviated sides of the mandible in adult patients with skeletal mandibular asymmetry, and then also compare the findings to those obtained from controls. The absolute and balance data of the occlusal pressure, occlusal contact area and occlusal force of 23 patients and the controls were examined. Correlations between the occlusal force and the morphology of the jaw-closing muscles were also analysed. The occlusal pressure of patients was not smaller than controls, however, the occlusal contact area and occlusal force in patients were significantly lower than those in the controls. There was no significant difference in the balance of the occlusal contact area and the occlusal force between the right and left sides in the controls, while the balance was shifted to the deviated side in the patients. Interestingly, the balance of the occlusal pressure was very similar between the patients and the controls. Most parameters of the morphology of the jaw-closing muscles did not show a linear correlation with either the occlusal pressure or force. In conclusion, the occlusal contact area and occlusal force in patients were significantly lower than those in the controls, and also the balance was shifted to the deviated side in patients with skeletal mandibular asymmetry. It is assumed that the morphology and orientation of jaw-closing muscles may have not linear but complex correlation to the weaker and unbalanced occlusal force in patients.

Evidence for association between a Toll-like receptor 4 gene polymorphism and moderate/severe periodontitis in the Japanese population.

BACKGROUND AND OBJECTIVE: Chronic periodontitis is an inflammatory disease caused by bacteria in subgingival pockets. Because Toll-like receptor 2 and Toll-like receptor 4 have been shown to play an important role in the recognition of periodontal pathogens, we investigated the relevance of genetic variations in TLR2 and TLR4 to susceptibility to periodontitis. MATERIAL AND METHODS: A total of 97 patients with chronic periodontitis and 100 control subjects were examined for mutations in TLR2 and TLR4. Case-control analysis was performed using individual single nucleotide polymorphisms detected during the mutation search. RESULTS: The missense mutations reported previously in TLR2 (677 Arg>Trp and 753 Arg>Gln) and in TLR4 (299 Asp>Gly and 399 Thr>Ile) were not detected in 97 of the Japanese patients with chronic periodontitis or in 100 of the Japanese control subjects. Nine single nucleotide polymorphisms were identified in exons of TLR2 and TLR4. The case-control analysis revealed that the frequency of the C/C genotype at base-pair position +3725 in TLR4 was significantly higher in both the moderate and the severe periodontitis patient group than in the control group. CONCLUSION: A genetic variation of TLR4 might be associated with moderate and severe periodontitis in the Japanese population.

A novel missense mutation in Van der Woude syndrome: usefulness of fingernail DNA for genetic analysis.

Van der Woude syndrome (VWS) is an autosomal-dominant oral facial disorder. To find a gene mutation in a Japanese family using fingernail DNA samples, we performed this study. We hypothesized that a gene mutation in IRF6 might be involved in VWS, and that fingernail DNA samples may be valuable for detecting such mutations. Linkage and haplotype analyses of the family mapped the disease locus to the 1q32-q41 region. Mutation analysis with an improved extraction method for fingernail DNA detected a novel missense mutation (1046A>T, E349V) in exon 7 of IRF6 in all the affected members of the family. Since the E349V change may disturb the hydrophobic core and affect regulatory activity of IRF6, it is most likely that the mutation is causative for VWS in this family. Fingernail DNA is thus useful for linkage and mutation analyses, since the fingernail can be easily obtained non-invasively, sent through the mail, and stored for a long period. We emphasize here the usefulness of fingernail DNA for the genetic analysis of a disease.

Patched homologue 1 mutations in four Japanese families with basal cell nevus syndrome.

AIM: To search for patched homologue 1 (PTCH1) mutations in four families with basal cell nevus syndrome (BCNS). METHODS: Mutation analysis of PTCH1 in unrelated Japanese families affected with BCNS was carried out by direct sequencing. RESULTS: Six novel PTCH1 mutations, 833G-->A in exon 6, 1415C-->A and 1451G-->T in exon 10, 2798delC in exon 17, 2918-2925dupAGTTCCCT in exon 18 and 3956C-->A in exon 23, were identified. CONCLUSIONS: Among the six PTCH1 mutations, two frameshift mutations (2798delC and 2918-2925dupAGTTCCCT) and one nonsense mutation (833G-->A) are predicted to lead to premature termination of PTCH1 protein translation. Three simultaneous mutations, 1415C-->A (A472D) and 1451G-->T (G484V) in exon 10, and 3956G-->A (R1319H) in exon 23, were found on one allele in only affected members in one family and none of them were found among 90 unrelated healthy Japanese. The three mutations on one chromosome may have resulted from errors in the recombinational repair process and this is the first report on the PTCH1 mutations due to such a mechanism.

Size and orientation of masticatory muscles in patients with mandibular laterognathism.

Size measurements of jaw muscles reflect their force capabilities and correlate with facial morphology. Using MRI, we examined the size and orientation of jaw muscles in patients with mandibular laterognathism in comparison with a control group. We hypothesized that the muscles of the deviated side would be smaller than those of the non-deviated side, and that the muscles of both sides would be smaller than in controls. In patients, a comparison of deviated and non-deviated sides showed, in orientation, differences for masseter and medial pterygoid muscles, but, in size, differences only for the masseter muscle. Nevertheless, muscle sizes in patients were much smaller than in controls. Lateral displacement of the mandible can explain the orientation differences, but not the smaller muscle size, in patients. It is possible that the laterodeviation initiates an adaptive process in the entire jaw system, resulting in extensive atrophy of the jaw muscles.

CT analysis of the Stafne's bone defects of the mandible.

OBJECTIVE: To investigate the aetiology of Stafne's bone defects by analysing the CT findings of two types of defects, which appeared differently on panoramic radiographs. METHODS: 32 lesions with suspicion of the Stafne's bone defect on a panoramic radiograph were categorised into two groups: typical "Stafne type", which showed the connection to the base of the mandible, and non-typical "Cyst type", which showed no connection. Age, sex, the existence of hypertension and the following CT findings: location, size and inner content of the defect, existence of expansion of the buccal cortical bone, and location of submandibular glands, were analysed. RESULTS: There were 14 "Stafne type" and 17 "Cyst type" on the panoramic radiographs. One lesion in the "Cyst type" was a true cystic lesion and was excluded from further review. The "Stafne type" was seen primarily in the posterior and inferior locations, while the "Cyst type" was seen in the anterior and superior locations on the CT. The size of the defect was significantly larger in the "Stafne type". Buccal cortical expansion was observed only in three lesions in the "Stafne type". While the "Cyst type" contained mainly fatty tissue, the "Stafne type" contained other soft tissues. Though the submandibular glands on the defect side were located anteriorly in both types compared with those on the contralateral side and on the control patients, they spread mainly outwards in the "Stafne type". CONCLUSION: The Stafne's bone defects are thought to be caused by the dislocated submandibular gland. The differences between the two types might occur as a result of a different location of the submandibular gland.

Correlation between diagnostic accuracy and perceptibility.

OBJECTIVES: To correlate diagnostic accuracy for proximal caries with perceptibility of low contrast image details using regression analysis. The other purpose was to determine the attenuation range required for proximal caries diagnosis. METHODS: The results of the two types of observer performance tests described above were retrieved from previous studies. Recording media included in those studies were the Compuray and the Dixel, direct digital radiographic systems, and Ektaspeed Plus film. The average numbers of perceptibility of image details from five observers were calculated for each step and for every combination of contiguous steps of the aluminium test phantom from the perceptibility test. The average diagnostic accuracy for proximal caries from the same five observers was correlated with the total number of perceptible details from the phantom using regression analysis. Finally, attenuation range required for proximal caries diagnosis was calculated from the attenuation range of the phantom where the maximum correlation coefficient was obtained. RESULTS: Maximum correlation (r=0.68) was obtained at the combination of five contiguous steps of the aluminium test phantom. Attenuation range required for proximal caries diagnosis corresponded to the 2 mm to 6 mm thickness of aluminium with acrylic block of 12 mm thickness. CONCLUSIONS: There is a correlation between perceptibility of low contrast image details and diagnostic accuracy for proximal caries. There may be a possibility to simplify observer performance tests for proximal caries diagnosis by using the standardized phantom simulating its attenuation range.

In vivo cross-sectional area of human jaw muscles varies with section location and jaw position.

Muscle cross-sectional area (CSA) is used as a measure for maximum muscle force. This CSA is commonly determined at one location within the muscle and for one jaw position. The purpose of this study was to establish a method to standardize the analysis of the CSA of the masticatory muscles in vivo, and to compare the CSAs along their entire length for two different jaw positions (opened and closed). The CSAs in the planes perpendicular to the long axes of the masseter, medial, and lateral pterygoid muscles were measured in ten normal young adult subjects by magnetic resonance imaging. Our results showed large differences among the muscles and a non-uniform change in CSA after jaw-opening. The method enables the CSA measurement to be standardized in vivo, and allows for a correct comparison of CSAs in different skull morphologies.