Mutation analysis of the ADAR1 gene in dyschromatosis symmetrica hereditaria and genetic differentiation from both dyschromatosis universalis hereditaria and acropigmentatio reticularis

Dyschromatosis symmetrica hereditaria (DSH) (also called "reticulate acropigmentation of Dohi") is a pigmentary genodermatosis of autosomal dominant inheritance. We have clarified for the first time four pathological mutations of the double-stranded RNA-specific adenosine deaminase gene (ADAR1 or DSRAD) in four DSH pedigrees. In this paper, we report 16 novel mutations containing six missense substitutions (p.V906F, p.K1003R, p.G1007R, p.C1036S, p.S1064F, p.R1078C), two splice site mutations (IVS2+2T>G, IVS8+2T>A), six frameshift mutations (p.H216fs, p.K433fs, p.G507fs, p.P727fs, p.V955fs, p.K1201fs), and two nonsense mutations (p.R426X, p.Q600X) found in Japanese patients with DSH. We did not establish any clear correlation between the clinical phenotypes and the genotypes of ADAR1 gene mutations in our examination of 16 cases plus four pedigrees. None of the different mutations identified in our studies of 20 cases suggested any founder effect. Furthermore, we did not identify any mutations in the ADAR1 gene of three patients with dyschromatosis universalis hereditaria or three patients with acropigmentatio reticularis, indicating that the two diseases are completely different from DSH, although they have sometimes been suggested to be phenotypical variations of DSH.     

Development of a high-resolution YSO gamma camera system that employs 0.8-mm pixels

Objective

YSO (Ce-doped Y₂SiO₅) is a promising scintillator for a single-photon imaging system since it has relatively high light output and does not contain any natural radioactivity. Since YSO is not hygroscopic, it may be possible to fabricate a block with small pixels for a high-resolution system. For this purpose, we developed a high-resolution gamma camera system that employs smaller than 1-mm YSO pixels.

Methods

The gamma camera’s detector used 0.8 × 0.8 × 7-mm YSO pixels. All the surfaces of these YSO pixels were mechanically polished, combined with a 0.1-mm-thick BaSO₄ reflector to form a 48 × 48 matrix, and optically coupled to a high quantum efficiency, 2-inch square position sensitive photomultiplier tube (Hamamatsu Photonics H10966 A-100). The YSO block was 43.2 × 43.2 mm. The YSO gamma camera was encased in a 5-mm-thick tungsten container, and a parallel collimator was mounted on its front. The parallel hole collimator was made of a 3-layer (each layer was 5-mm thick) tungsten plate, and each plate had 48 × 48, 0.6-mm holes that were positioned by one-to-one coupling with the YSO pixels.

Results

Even with the 0.8-mm YSO pixels, we clearly resolved most of the pixels in a 2-dimensional histogram with a peak-to-valley ratio of 2.9 for the 122-keV gamma photons. The energy resolution was 20.4 % FWHM. The spatial resolutions with a parallel hole collimator 2 mm from the collimator surface were 0.7- and 1.3-mm FWHM for the 122- and ~35-keV gamma photons, respectively. We successfully obtained phantoms and small animal images with our YSO gamma camera system.

Conclusion

Our high-resolution system has a potential to be useful for molecular imaging research.     

MR imaging of primary leiomyosarcoma of the thyroid gland

Primary leiomyosarcoma of the thyroid gland is extremely rare and radiological information on this tumor is scant. We presented radiological findings on primary thyroid leiomyosarcoma in a 66-year-old woman in which anaplastic carcinoma was suspected based on clinical and cytological features and discussed the radiological clues to distinguish between the two diseases. Ultrasonography showed an ill-defined hypoechoic mass without halo in the left lobe and the isthmus of the thyroid gland. Computed tomography depicted a low-density mass with calcification and necrosis, which invaded the thyroid cartilage. No lymphadenopathy was seen. The tumor was demonstrated as an isointense mass on T1-weighted MR images and a mass of intermediate signal on T2-weighted images. The tumor showed a fair enhancement on gadolinium-enhanced T1-weighted images. Although the radiological picture was nonspecific, primary thyroid leiomyosarcoma appeared less invasive and far less frequent in developing nodal metastasis than anaplastic carcinoma in light of the literature.     

Disruption of the anterior commissure in Olig2 deficient mice

In the present study, we examined neural circuit formation in the forebrain of the Olig2 knockout (Olig2-KO) mouse model and found disruption of the anterior commissure at the late foetal stage. Axon bundles of the anterior commissure encountered the wall of the third ventricle and ceased axonal extension. L1-CAM immunohistochemistry showed that Olig2-KO mice lose decussation formation in the basal forebrain. DiI tracing revealed that the thin bundles of the anterior commissure axons crossed the midline but ceased further extension into the deep part of the contralateral side. Furthermore, some fractions of DiI-labelled axons were oriented dorsolaterally, which was not observed in the control mouse forebrain. The rostral part of the third ventricle was much wider in the Olig2-KO mice than in wild-type mice, which likely resulted in the delay of midline fusion and subsequent delay and malformation of the anterior commissure. We analysed gene expression alterations in the Olig2-KO mice using a public database and found multiple genes, which are related to axon guidance and epithelial-mesenchymal transition, showing subtle expression changes. These results suggest that Olig2 is essential for anterior commissure formation, likely by regulating multiple biological processes.     

Bilateral cholesteatoma and habitual sniffing

OBJECTIVE: To examine the clinical findings of acquired bilateral cholesteatoma with special reference to incidence of habitual sniffing and sniff-related negative middle ear pressure. METHODS: Eighty-eight fresh cases of unilateral cholesteatoma and 33 fresh cases of bilateral cholesteatoma, which were operated on at Department of Otolaryngology, Hyogo College of Medicine, were examined in this study. Responses to a detailed questionnaire were obtained from the patients concerning about the habit of habitual sniffing to relieve aural symptoms such as aural fullness, autophonia or hyperacusis. The same questionnaire was obtained from unilateral cholesteatoma patients to compare the incidence of habitual sniffing with that of patients with bilateral cholesteatoma. We measured the negative middle ear pressure at the time of sniffing by using TTAG (tubo-tympano aerodynamic graphy, Nagashima Co. Ltd, Tokyo). We also compared the positive percentage of the sniff test in bilateral cholesteatoma with in unilateral cholesteatoma and normal controls. Sniff test was performed in 30 patients with bilateral cholesteatoma, 20 patients with unilateral cholesteatoma and 20 normal controls. RESULTS: In 33 patients with bilateral cholesteatoma (66 ears), 57 ears had the pars flaccida type (86.4%) and 9 ears had the pars tensa type (13.6%). Cholesteatoma of pars flaccida type were predominant in bilateral cholesteatoma. The rate of habitual sniffing of bilateral cholesteatoma (23/33, 69.7%) was significantly higher than that of unilateral cholesteatoma (21/88, 23.9%). The incidence of positive sniff test in bilateral cholesteatoma (19/30, 63.3%) was significantly higher than in unilateral one (6/20, 30%) and normal control (3/20, 15%). CONCLUSIONS: Habitual sniffing was closely related to the pathogenesis of bilateral cholesteatoma, especially in cases with bilateral pars flaccida type.     

By inhibiting Src,verapamil and dasatinib overcome multidrug resistance via increased expression of Bim and decreased expressions of MDR1 and survivin in human multidrug-resistant myeloma cells

The calcium channel blocker verapamil inhibits the transport function of multidrug resistance protein 1 (MDR1). Although verapamil acts to reverse MDR in cancer cells, the underlying mechanism remains unclear. In the present study, we investigated the mechanism of reversing MDR by verapamil in anti-cancer drug-resistant multiple myeloma (MM) cell lines. We found that verapamil suppresses MDR1 and survivin expressions and increases Bim expression via suppression of Src activation. Furthermore, dasatinib reversed the drug-resistance of the drug-resistant cell lines. These findings suggest that Src inhibitors are potentially useful as an anti-MDR agent for the treatment of malignant tumor cells.     

Expression and regulation of tumor suppressor gene maspin in human bladder cancer

Maspin is a member of serine protease inhibitor family with tumor suppressing activity for breast and prostate cancers, acting at the level of tumor invasion and metastasis. However, there have been no published data regarding the role of maspin in human bladder cancer. We evaluated maspin expression in 65 series of bladder cancer samples (22 transurethral resection (TUR) and 43 radical cystectomy) and studied the regulatory mechanism of maspin gene activation in bladder cancer cells. Maspin expression was immunohistochemically detected in four (18.2%) patients with TUR and 22 (51.2%) patients with radical cystectomy whereas no expression was observed in normal transitional cells located at tumor-free area in bladder. The maspin expression was significantly correlated with the development of muscle invasive bladder cancer (P=0.00008). Using a luciferase reporter system, maspin promoter activity was induced in the maspin-positive bladder cancer cell lines as well as maspin-negative RT4 cells. Furthermore, treatment with the DNA methyltransferase inhibitor, 5-aza-2' deoxycytidine, and histone deacetylase inhibitor, trichostatin A, led to re-expression of maspin in RT4 cells. Our results indicate that maspin may contribute to bladder cancer development and that DNA methylation and histone deacetylation may be important for regulating maspin gene activation in bladder cancer cells.     

Utility of spinal MRI in children with anorectal malformation

Background  The association between spinal cord anomalies and imperforate anus is well recognized. Until now, the incidence of tethered cord has been assumed to be higher in patients with high-type imperforate anus. However, recent reports suggest that tethered cord is as common in patients with a low lesion as in those with a high lesion. Objective  To review the incidence of spinal cord anomalies in those with a low lesion and those with a high (including intermediate) anorectal malformation (ARM), and to determine the best diagnostic imaging strategy. Materials and methods  A group of 50 consecutive patients with postoperative ARM and in whom spinal MRI had been performed were identified retrospectively. We reviewed and compared the following factors between those with a high lesion and those with a low lesion: (1) clinical symptoms, (2) spinal cord anomalies, and (3) vertebral anomalies. Results  The incidence of spinal cord anomalies was no different between those with a high lesion and those with a low lesion, and spinal cord anomalies were present regardless of the presence of vertebral anomalies or symptoms. Conclusion  Owing to the high incidence of spinal cord anomalies in patients with imperforate anus, MRI is the best imaging tool for detecting such anomalies regardless of the level of the lesion.     

Image evaluation of endolymphatic space in fluctuating hearing loss without vertigo

The objective of the present study was to investigate endolymphatic space images in patients with fluctuating hearing loss without vertigo, and to elucidate its underlying pathophysiology. Eight patients with fluctuating hearing loss without vertigo were included in this study. 3T MRI was taken, 24 h after intratympanic injection of gadolinium-diethylenetriamine pentaacetic acid (Gd-DTPA). Electrocochleography and VEMP tests were performed to evaluate cochlear and vestibular functions. Endolymphatic hydrops were observed both in the cochlea and in the vestibule of all eight patients. Three patients out of six whose summating potential/action potential (SP/AP) ratio was recordable showed an elevation of SP/AP ratio. In the two patients with remarkable endolymphatic hydrops in the vestibule, VEMP was absent from the affected ear. In conclusion, 3T MRI after intratympanic injection of Gd-DTPA revealed endolymphatic hydrops both in the cochlea and in the vestibule in the patients with fluctuating hearing loss without vertigo.