Serum level of soluble (pro)renin receptor is modulated in chronic kidney disease.

BACKGROUND: Prorenin, the precursor of renin, binds to the (pro)renin receptor [(P)RR] and triggers intracellular signaling. The ligand binding sites of (P)RR are disconnected and are present in the soluble form of the receptor in serum. Given that the clinical significance of serum prorenin and soluble (P)RR in chronic kidney disease (CKD) is unclear, we investigated the relationship between serum prorenin, soluble (P)RR, and various clinical parameters in patients with CKD. METHODS: A total of 374 patients with CKD were enrolled. Serum samples were collected, and the levels of soluble (P)RR and prorenin were measured using ELISA kits. Serum creatinine (Cr), blood urea nitrogen (BUN), uric acid (UA), hemoglobin (Hb), soluble secreted α-Klotho, and the urine protein/Cr ratio were also measured. Similarly, clinical parameters were also evaluated using serum and urine sample collected after 1 year (n = 204). RESULTS: Soluble (P)RR levels were positively associated with serum Cr (P < 0.0001, r = 0.263), BUN (P < 0.0001, r = 0.267), UA (P < 0.005, r = 0.168) levels, CKD stage (P < 0.0001, r = 0.311) and urine protein/Cr ratio (P < 0.01, r = 0.157), and inversely with estimated glomerular infiltration rate (eGFR) (P < 0.0001, r = -0.275) and Hb (P < 0.005, r = -0.156). Soluble (P)RR levels were inversely associated with α-Klotho levels (P < 0.001, r = -0.174) but did not correlate with prorenin levels. With respect to antihypertensive drugs, soluble (P)RR levels were significantly lower in patients treated with an angiotensin II receptor blocker (ARB) than in those without ARB therapy (P < 0.005). Soluble (P)RR levels were significantly lower in CKD patients with diabetes mellitus or primary hypertension than in those without these conditions (P < 0.05). In contrast, serum levels of prorenin did not correlate with parameters related to renal function. Serum prorenin levels were significantly higher in CKD patients with diabetes mellitus than in nondiabetic patients (P < 0.05), but not in CKD patients with hypertension (P = 0.09). Finally, with respect to the relationship between basal soluble (P)RR levels and the progression rates of renal function, soluble (P)RR levels were positively associated with ΔCr (P < 0.05, r = 0.159) and inversely associated with ΔeGFR (P < 0.05, r = -0.148). CONCLUSION: Serum levels of soluble (P)RR correlated with the stage of CKD. Our findings suggest that soluble (P)RR may be involved in renal injury and influence the progression of CKD.

Serum levels of soluble secreted α-Klotho are decreased in the early stages of chronic kidney disease, making it a probable novel biomarker for early diagnosis.

BACKGROUND: α-Klotho was first identified as an aging gene and was later shown to be a regulator of phosphate metabolism. Fibroblast growth factor 23 (FGF23) is the key regulator of phosphate metabolism. Serum levels of soluble α-Klotho in chronic kidney disease (CKD) patients have not previously been determined, especially in relation with FGF23 and creatinine levels. This study was designed to investigate whether serum soluble α-Klotho levels are modulated by renal function, age, and FGF23 level in CKD patients. This study is the first report on the utility of measuring soluble α-Klotho levels in human CKD. METHODS: A total of 292 CKD patients were enrolled. Serum samples were collected, and FGF23 and soluble α-Klotho levels were measured using enzyme-linked immunosorbent assay kits. In addition, serum creatinine, hemoglobin, albumin, calcium, and phosphate levels were measured. RESULTS: Serum soluble α-Klotho levels were associated positively with estimated glomerular filtration rate (eGFR) (P < 0.0001) and inversely with serum creatinine level (P < 0.01). Interestingly, α-Klotho levels were significantly decreased in stage 2 CKD compared with stage 1 (P = 0.0001). Serum FGF23 levels were associated positively with serum creatinine and negatively with eGFR. FGF23 levels were significantly increased in stage 5 compared with stage 1 CKD. Soluble α-Klotho was associated inversely with log-transformed FGF23 level (P < 0.01). CONCLUSION: Our data indicate that soluble α-Klotho levels are significantly decreased in stage 2 CKD compared to stage 1, and not only in the advanced stages of the disease. Soluble α-Klotho may thus represent a new biomarker for the diagnosis of CKD, especially in the early stage.

Evaluation of diffusion parameters and T2 values of the masseter muscle during jaw opening, clenching, and rest.

BACKGROUND: Diffusion-weighted imaging (DWI) and diffusion-tensor imaging (DTI) can be used to evaluate changes that accompany skeletal muscle contraction. PURPOSE: To investigate whether jaw opening or closure affect the diffusion parameters of the masseter muscles (MMs). MATERIAL AND METHODS: Eleven healthy volunteers were evaluated. Diffusion-tensor images were acquired to obtain the primary (λ(1)), secondary (λ(2)), and tertiary eigenvalues (λ(3)). We estimated these parameters at three different locations: at the level of the mandibular notch for the superior site, the level of the mandibular foramen for the middle site, and the root apex of the mandibular molars for the inferior site. RESULTS: Both λ(2) and λ(3) during jaw opening were significantly lower than that at rest at the superior (P = 0.006, P < 0.0001, respectively) and middle site (P = 0.004, P = 0.0001, respectively); however, the change in λ(1) was not significant. At the lower site, no parameter was significantly different at rest and during jaw opening. There was no significant difference in T2 between at rest (40.3 ± 4.4 ms) and during jaw opening (39.2 ± 2.7 ms; P = 0.12). The changes induced by jaw closure were marked at the inferior site. In the middle and inferior sites, the three eigenvalues were increased by jaw closure, and the changes in λ(1) (P = 0.0145, P = 0.0107, respectively) and λ(2) (P = 0.0003, P = 0.0001) were significant (especially λ(2)). CONCLUSION: The eigenvalues for diffusion of the MM were sensitive to jaw position. The recruitment of muscle fibers, specific to jaw position, reflects the differences in changes in muscle diffusion parameters.

Multiple congenitally missing teeth treated with autotransplantation and orthodontics.

Treatment of children with several congenitally missing teeth is challenging, because growth and development of the oral structures must be considered. The treatment options include retaining the deciduous teeth and postponing treatment until later or extracting the deciduous teeth and doing one of the following: allowing the space to close spontaneously, closing the space orthodontically, or in patients whose growth is finished, using a prosthetic or implant replacement. One other viable option, if donor teeth are available, is autotransplantation. The treatment plan for patients with missing teeth should be based on a comprehensive evaluation of the patient's age, occlusion, and space requirements as well as on the size and shape of the adjacent teeth. This case report presents the management of a patient in the early mixed dentition with multiple missing teeth. The treatment consisted of a combination of autotransplantation of the maxillary right first premolar to the mandibular right first premolar region and orthodontic treatment with a 5-year follow-up after autotransplantation.

Molecular imaging in masseter muscle observed by muscle function magnetic resonance imaging and 31 P-magnetic resonance spectroscopy in patients with a jaw deformity.

BACKGROUND: Skeletal mandibular protrusion would influence to the muscle fatigue of the masticatory muscles. Establishing a diagnostic procedures combining physiological and biochemical information is necessary for quantitative evaluation of masticatory muscle fatigue. OBJECTIVE: The transverse relaxation time (T2 time) of muscle functional magnetic resonance imaging (mfMRI), and 31 P-magnetic resonance spectroscopy (MRS) were used to investigate the reliability as parameters for measuring the masseter muscle in patients with skeletal mandibular prognathism. METHOD: The subjects were 19 patients diagnosed as skeletal mandibular protrusions and 19 healthy subjects as a control group. Transverse relaxation time (T2 value) determined by mfMRI along with creatine phosphate (PCr) and inorganic phosphorus (Pi) determined by 31 P-MRS before, during, and after clenching were used for molecular imaging of muscle fatigue. RESULTS: The average T2 value of the patient group was significantly higher than that of the healthy control group at rest. Furthermore, the average T2 value transiently increased in both groups during experimental clenching. The PCr and Pi showed a tendency toward a transient decrease and increases, respectively. The pH in the masseter muscle showed a transient decrease in both groups prior to and following experimental clenching. The pH in the masseter muscle of the patient group was significantly lower than that in the healthy control group at rest and recovery. CONCLUSION: We showed mfMRI and 31 P-MRS are useful for evaluating masseter fatigue during clenching, and the masseter muscle in the prognathic patients showed more severe fatigue than the healthy controls.

Assessment of the sequential change of the masseter muscle by clenching: a quantitative analysis of T1, T2, and the signal intensity of the balanced steady-state free precession.

BACKGROUND: The persistent muscle contractions during clenching are thought to cause some temporomandibular disorders. However, no report has so far evaluated the effect of clenching on the masticatory muscles by magnetic resonance imaging (MRI). PURPOSE: To investigate the effect of clenching with maximum voluntary contraction on the T(1), T(2), and signal intensity (SI) of the balanced fast field-echo (b FFE) of the masseter muscle. MATERIAL AND METHODS: A total of 11 volunteers participated. Multi-echo spin-echo echo-planar imaging was used for T(2) measurements, and multi-shot Look-Locker sequence for T(1) measurements. The Look-Locker sequence has been used for fast T(1) mapping and this method has been applied for the imaging of various tissues. In addition, the b FFE was used due to the high temporal resolution. These three sequences lasted for 10 min and the participants were instructed to clench from 60 s to 80 s after the start of the data acquisition. T(2), T(1), and SI were normalized compared to pre-clenching values. RESULTS: T(2) decreased by clenching, which reflected a decrease of tissue perfusion due to the mechanical pressure. It increased rapidly after the clenching (peak value, 1.11+/-0.03; peak time, 16.8+/-7.6 s after the clenching), which corresponded to the reactive hyperemia and later, it gradually returned to the initial values (half period, 2.22+/-0.84 min). The change in the SI of the b FFE was triphasic and similar to that of T(2) clenching. T(1) increased after the cessation of the clenching and later gradually decreased during the recovery periods. However, the change of T(1) was quite different from that of T(2), with a lower peak value (1.04+/-0.02), a later peak time (36.0+/-28.0 s), and a longer half period (4.76+/-3.40 min) (P<0.0001, 0.0066, 0.02, respectively). CONCLUSION: The change in T(2) was triphasic and we considered that it predominantly reflected the tissue perfusion.

Effect of clenching on T2 and diffusion parameters of the masseter muscle.

BACKGROUND: Persistent muscle contractions during clenching are considered to be one reason for temporomandibular disorders. However, no report has evaluated the effect of clenching on the masticatory muscles, as measured by magnetic resonance imaging (MRI). PURPOSE: To investigate whether clenching has an effect on either T(2) or the coefficients for diffusion of the masseter muscles (MM), and to evaluate the effect of the distribution of bite force on such indices. MATERIAL AND METHODS: Twenty-three subjects were examined. Bite force was measured by a pressure-sensitive sheet, and the force of the right and the left sides was calculated. MRI was used to evaluate T(2), the apparent diffusion coefficient (ADC), and the primary (lambda(1)), secondary (lambda(2)), and tertiary eigenvalues (lambda(3)). These indices on the stronger side of the bite force were compared to those on the weaker side. Thereafter, the indices were compared between at rest and during clenching. RESULTS: There was no significant difference in any of the indices (T(2), ADC, lambda(1), lambda(2), and lambda(3)) between the side of stronger bite force and the side with weaker. T(2) increased by clenching, and the difference was significant in the side with stronger bite force (P = 0.006). ADC, lambda(1), lambda(2), and lambda(3) increased significantly by clenching (P <0.01, P <0.01, P <0.01, and P <0.01, respectively) on both sides. The percentage of change of lambda(2) by clenching was 26.2+/-15.7% on the stronger side and 26.9+/-18.6% on the weaker side, which was significantly greater than either that of lambda(1) or lambda(3). CONCLUSION: The coefficients for diffusion of the MM were sensitive to change by clenching, and lambda(2) was the most sensitive. Moreover, the relative distribution of the bite forces had no effect on any of the indices.

Effect of exposure parameters and gutta-percha cone size on fracture-like artifacts in endodontically treated teeth on cone-beam computed tomography images.

OBJECTIVES: To ascertain the effects of exposure parameters (tube current and tube voltage) and the gutta-percha cone (GPC) size on root fracture-like artifacts obtained with cone-beam computed tomography (CBCT). METHODS: Fracture-like artifacts appearing on CBCT images of nine extracted human mandibular premolars filled with GPCs of size #50 or #80 were analyzed using six exposure factors: two tube voltages (80 kV and 110 kV); and three tube currents (4 mA, 7 mA, and 10 mA). On axial images, the gray value (GV) was recorded at three points: the mesiobuccal portion (MBP) as the sound dentin, the mesial portion (MP) as the artifact line, and the water area (WA). The rate of decrease in the GV (RDGV) of the artifact line was calculated using the formula: RDGV (%) = (GV of MBP - GV of MP) × 100/(GV of MBP - GV of WA). RESULTS: Comparison of the #80 group and the #50 group with equal tube voltages and tube currents shows that artifact lines in the #80 group were more obvious than those in the #50 group. The artifact lines with 80 kV were markedly more visible than those with 110 kV for each tube current and GPC size. Tube current changes did not affect the artifact line for any tube voltage or GPC size. CONCLUSIONS: For the reduction of artifacts, we recommend selection of higher tube voltages and lower tube currents when taking CBCT images of teeth with each GPC size.

Assessment of the relationship between impacted mandibular third molars and inferior alveolar nerve with dental 3-dimensional computed tomography.

PURPOSE: The purpose of this study was to assess the capacity of dental 3-dimensional computed tomography (3D-CT; limited cone-beam CT) to predict the exposure and injury of the inferior alveolar nerve (IAN) after mandibular third molar extractions. MATERIALS AND METHODS: This study was a retrospective case series of patients who presented for extraction of mandibular third molars. Subjects eligible for study enrollment were those who underwent preoperative dental 3D-CT because the mandibular third molars were determined to be extremely close to the IAN on panoramic radiogram. The predictive variable was the anatomic relation of the IAN and third molar apices and was a binary variable, contact or noncontact. The primary outcome variable was IAN exposure, and the secondary outcome variable was IAN injury. RESULTS: From January 2006 to August 2007, 1,853 mandibular third molars in 1,539 patients were extracted. Among them, dental 3D-CT was performed on 53 third molars in 47 patients. The mandibular third molars were judged to make contact with the mandibular canal on dental 3D-CT images in 35 cases (66%). Intraoperative IAN exposure was observed in 17 (49%) contact cases and 2 (11%) noncontact cases on dental 3D-CT images. Of 53 cases extracted after dental 3D-CT examinations, IAN injury occurred in 8 cases (15%). IAN exposure led to IAN injury in 36.8% of cases, whereas IAN injury occurred in only 2.9% of cases without IAN exposure. Although the incidence of IAN injury in the molar-canal contact cases was 23%, all 8 cases with IAN injury (100%) were included in these contact cases. CONCLUSION: When viewing the anatomic relation between the IAN and mandibular third molar root apices using dental 3D-CT, contact of the 2 anatomic structures results in an increased risk for IAN exposure or injury.

Amelogenin is a negative regulator of osteoclastogenesis via downregulation of RANKL, M-CSF and fibronectin expression in osteoblasts.

Amelogenin is a novel enamel matrix protein. Knockout mice showed enhanced osteoclast formation and resorption of tooth cementum. This study investigated the effects of amelogenin on osteoclastogenesis. In co-cultures with calvaria osteoblasts and purified bone marrow cells, amelogenin inhibited osteoclastogenesis dramatically. Furthermore, amelogenin inhibited the expression of receptor activator of nuclear factor kappaB ligand (RANKL), macrophage-colony stimulating factor (M-CSF) and fibronectin in osteoblasts, while RANKL expression was induced by fibronectin and inhibited by treatment with fibronectin small interfering RNA. These results suggest that the inhibitory effects of amelogenin on osteoclastogenesis lead to downregulation of RANKL, M-CSF and fibronectin production in osteoblasts.

Identification of a Novel Alternatively Spliced Form of Inflammatory Regulator SWAP-70-Like Adapter of T Cells.

Activation of naive CD4+ T cells results in the development of several distinct subsets of effector Th cells, including Th2 cells that play a pivotal role in allergic inflammation and helminthic infections. SWAP-70-like adapter of T cells (SLAT), also known as Def6 or IBP, is a guanine nucleotide exchange factor for small GTPases, which regulates CD4+ T cell inflammatory responses by controlling Ca2+/NFAT signaling. In this study, we have identified a novel alternatively spliced isoform of SLAT, named SLAT2, which lacks the region encoded by exons 2-7 of the Def6 gene. SLAT2 was selectively expressed in differentiated Th2 cells after the second round of in vitro stimulation, but not in differentiated Th1, Th17, or regulatory T (Treg) cells. Functional assays revealed that SLAT2 shared with SLAT the ability to enhance T cell receptor- (TCR-) mediated activation of NFAT and production of IL-4 but was unable to enhance TCR-induced adhesion to ICAM-1. Ectopic expression of SLAT2 or SLAT in Jurkat T cells resulted in the expression of distinct forms of filopodia, namely, short versus long ones, respectively. These results demonstrate that modulating either SLAT2 or SLAT protein expression could play critical roles in cytokine production and actin reorganization during inflammatory immune responses.

Vascularity as assessed by Doppler intraoral ultrasound around the invasion front of tongue cancer is a predictor of pathological grade of malignancy and cervical lymph node metastasis.

OBJECTIVES: To quantitatively evaluate the relationship of vascularity of tongue cancer as demonstrated on intraoral ultrasonography images and tumour thickness with pathological grade of malignancy and the presence of cervical lymph node metastases. METHODS: 18 patients with tongue cancer were enrolled in this retrospective study. Using Doppler ultrasonography images of the invasion front of the cancers along the length of their tumour boundaries, three vascular indexes were analysed quantitatively, namely ratio of blood flow signal area within the cancer to whole tumour area (BAR), blood flow signal number ratio (BNR) and blood flow signal width ratio (BWR). The associations between these three indexes and occurrence of cervical lymph node metastasis and pathological grade of malignancy [Yamamoto-Kohama (YK) classification] were assessed. Furthermore, the relationship between tumour thickness and occurrence of cervical lymph node metastasis was evaluated on B-mode intraoral ultrasonography images. RESULTS: There was no significant association between BAR and tumour thickness or occurrence of cervical lymph node metastasis. The BNRs and BWRs of patients with cervical lymph node metastasis were significantly higher than those of patients without nodal involvement. The BWRs of patients with high-grade malignancy (YK-4C) were significantly higher than those of patients with low-grade malignancy (YK-2 or 3). CONCLUSIONS: BNR and BWR on the invasion front of the tongue cancer are predictors of pathological grade of malignancy and cervical lymph node metastasis.

Interaction between fibronectin and ß1 integrin is essential for tooth development.

The dental epithelium and extracellular matrix interact to ensure that cell growth and differentiation lead to the formation of teeth of appropriate size and quality. To determine the role of fibronectin in differentiation of the dental epithelium and tooth formation, we analyzed its expression in developing incisors. Fibronectin mRNA was expressed during the presecretory stage in developing dental epithelium, decreased in the secretory and early maturation stages, and then reappeared during the late maturation stage. The binding of dental epithelial cells derived from postnatal day-1 molars to a fibronectin-coated dish was inhibited by the RGD but not RAD peptide, and by a ß1 integrin-neutralizing antibody, suggesting that fibronectin-ß1 integrin interactions contribute to dental epithelial-cell binding. Because fibronectin and ß1 integrin are highly expressed in the dental mesenchyme, it is difficult to determine precisely how their interactions influence dental epithelial differentiation in vivo. Therefore, we analyzed ß1 integrin conditional knockout mice (Intß1lox-/lox-/K14-Cre) and found that they exhibited partial enamel hypoplasia, and delayed eruption of molars and differentiation of ameloblasts, but not of odontoblasts. Furthermore, a cyst-like structure was observed during late ameloblast maturation. Dental epithelial cells from knockout mice did not bind to fibronectin, and induction of ameloblastin expression in these cells by neurotrophic factor-4 was inhibited by treatment with RGD peptide or a fibronectin siRNA, suggesting that the epithelial interaction between fibronectin and ß1 integrin is important for ameloblast differentiation and enamel formation.

Intraoral sonographic features of tongue cancer after radical radiotherapy.

PURPOSE: The purpose was to illustrate the sonographic changes of tongue cancer after radical radiotherapy. MATERIALS AND METHODS: In 24 patients with tongue cancer treated by interstitial brachytherapy (BRT) (uneventful recovery n = 16 , recurrence n = 5, severe soft tissue complication n = 3), follow-up examination were performed and both the margin and the vascular pattern were retrospectively assessed. We basically performed US examination at least once every 3 months after BRT within 1 year during follow-up period. RESULTS: In the healing process, intraoral sonography shows an unclear margin immediately after brachytherapy and a transient increase of the vascularity lasted within 6 months after BRT, followed by a decrease in the vascularity. The large difference in echogenicity between the primary site and the surrounding tissue and the increased difference suggested the possibility of either radiation ulceration or recurrence. CONCLUSION: Intraoral sonography could depict the sequence changes of the tongue after BRT, and it was thus useful to confirm the clinical findings of either radiation ulcers or recurrence.

Change of angiostructure and hemodynamics in lymph node metastases in rabbits.

OBJECTIVES: Experimental models of lymph node metastasis were developed for Doppler sonography. METHODS: Cervical lymph node (LN) metastasis was induced by the implantation of a VX-2 tumor on the oral floor of 10 rabbits. Twenty metastatic LNs were observed weekly by power Doppler sonography, and the presence of an avascular area and the peripheral vessels were evaluated. The time-averaged maximum velocity (TAMx) at the hilum was also measured. RESULTS: The percentage of the metastatic LNs presenting with an avascular area increased over time. The peripheral vessels were detected in 14 of 20 LNs. The peak time of the TAMx significantly correlated to that of the initial detection of the avascular area. CONCLUSIONS: In the beginning, metastatic LNs were depicted as hypervascular structures while the TAMx at the hilum increased. Later, an avascular area was detected and the TAMx at the hilum decreased, which resulted in a blood supply to the node from the peripheral vessels.

Value of computed tomography findings in differentiating between intraosseous malignant tumors and osteomyelitis of the mandible affecting the masticator space.

OBJECTIVES: We sought to evaluate the diagnostic efficacy of computed tomography (CT) images in the differentiation between intraosseous malignant tumors and osteomyelitis spreading into the masticator space. STUDY DESIGN: A retrospective evaluation was carried out by using CT images from 12 patients with intraosseous malignant tumors and 9 patients with osteomyelitis involving the masticator space and accompanying mandibular bone destruction. The following CT observations are discussed: (1) bone destruction pattern subdivided into spotty, gross, or permeative; (2) cortical bone expansion; (3) diffuse osteosclerotic changes; (4) periosteal reaction; (5) masticator muscle involvement; (6) enlargement of the facial muscle; and (7) attenuation in the subcutaneous adipose tissue. RESULTS: The pattern of permeative bone destruction, cortical bone expansion, and the enlargement of both the masseter and medial pterygoid muscles were all observed in patients with malignant tumors. In contrast, diffuse sclerotic change and a periosteal reaction were significant observations in patients with osteomyelitis. CONCLUSION: The efficacy of CT in establishing a differential diagnosis of malignant tumors or osteomyelitis is supported by this study.

Cervical lymph nodes with or without metastases from oral squamous carcinoma: a correlation of MRI findings and histopathologic architecture.

OBJECTIVES: The aim of this study was to describe the magnetic resonance images (MRIs) of the architecture of metastatic lymph nodes as well as healthy lymph nodes and to correlate the images with the histopathologic architecture of the lymph nodes from oral squamous cell carcinoma. STUDY DESIGN: The signal intensities of 98 cervical lymph nodes from 20 patients with oral squamous cell carcinoma were evaluated on T2-weighted and diffusion-weighted whole-body imaging with background body signal suppression (DWIBS). The MRI findings were compared with the histopathologic architecture. RESULTS: On T2-weighted images, very hyperintense and/or isointense areas were specific findings for lymph nodes containing metastases from oral squamous cell carcinoma. Histopathology indicated that these areas corresponded to cystic degeneration or keratinization. Using DWIBS images with inverted black-and-white image contrast, cystic degeneration, keratinization, fibrous tissue, tumor tissue, and lymphoid tissue were either hypointense or intermediate in intensity. CONCLUSIONS: On T2-weighted images, very hyperintense and/or isointense areas were characteristic findings for lymph nodes containing metastases from oral squamous cell carcinoma. This suggests cystic degeneration or keratinization within the affected lymph nodes. Using DWIBS images, it was difficult to differentiate metastatic from benign lymph nodes.

Influences of interferon-gamma on cell proliferation and interleukin-6 production in Down syndrome derived fibroblasts.

OBJECTIVE: Down syndrome, a frequently encountered genetic disorder, is usually associated with medical problems related to infectious disease, such as periodontal diseases and prolonged wound healing. Although affected individuals are considered to have clinical problems related to high interferon (IFN) sensitivity, the molecular mechanisms of IFN activities are not completely understood. DESIGN: Down syndrome derived fibroblasts, Detroit 539 (D1) and Hs 52.Sk (D2) cells, were used. To analyse the expressions of interferon (IFN) receptors and downstream of IFN-gamma, western blotting was performed. Cell proliferation was determined by counting cells following trypan blue staining. Media levels of IL-1beta, TNF-alpha, and IL-6 were quantified using ELISA. RESULTS: IFN-gamma receptor 2 and IFN-alpha receptor 1, but not IFN-gamma receptor 1, were highly expressed in D1 and D2 cells, as compared to the control fibroblast cells. Cell proliferation by D1 and D2 cells was lower than that by the control fibroblasts, further, IFN-gamma had a greater effect to inhibit cell proliferation by D1 and D2 cells. In addition, IFN-gamma treatment increased the phosphorylation of STAT1 and MAPK in D1 cells as compared to normal fibroblasts. Also, the presence of exogenous IFN-gamma in the growth medium significantly induced IL-6, but not IL-1beta or TNF-alpha, in D1 and D2 cells. CONCLUSION: Taken together, our results are consistent with hypersensitive reactions to IFN-gamma seen in patients with Down syndrome and may provide useful information to elucidate the mechanisms of IFN-gamma activities in those individuals.

Neurotrophic factor neurotrophin-4 regulates ameloblastin expression via full-length TrkB.

Neurotrophic factors play an important role in the development and maintenance of not only neural but also nonneural tissues. Several neurotrophic factors are expressed in dental tissues, but their role in tooth development is not clear. Here, we report that neurotrophic factor neurotrophin (NT)-4 promotes differentiation of dental epithelial cells and enhances the expression of enamel matrix genes. Dental epithelial cells from 3-day-old mice expressed NT-4 and three variants of TrkB receptors for neurotrophins (full-length TrkB-FL and truncated TrkB-T1 and -T2). Dental epithelial cell line HAT-7 expressed these genes, similar to those in dental epithelial cells. We found that NT-4 reduced HAT-7 cell proliferation and induced the expression of enamel matrix genes, such as ameloblastin (Ambn). Transfection of HAT-7 cells with the TrkB-FL expression construct enhanced the NT-4-mediated induction of Ambn expression. This enhancement was blocked by K252a, an inhibitor for Trk tyrosine kinases. Phosphorylation of ERK1/2, a downstream molecule of TrkB, was induced in HAT-7 cells upon NT-4 treatment. TrkB-FL but not TrkB-T1 transfection increased the phosphorylation level of ERK1/2 in NT-4-treated HAT-7 cells. These results suggest that NT-4 induced Ambn expression via the TrkB-MAPK pathway. The p75 inhibitor TAT-pep5 decreased NT-4-mediated induction of the expression of Ambn, TrkB-FL, and TrkB-T1, suggesting that both high affinity and low affinity neurotrophin receptors were required for NT-4 activity. We found that NT-4-null mice developed a thin enamel layer and had a decrease in Ambn expression. Our results suggest that NT-4 regulates proliferation and differentiation of the dental epithelium and promotes production of the enamel matrix.