Search

The inner ear is a structurally complex vertebrate organ built to encode sound, motion, and orientation in space. Given its complexity, it is not surprising that inner ear dysfunction is a relatively common consequence of human genetic mutation. Studies in model organisms suggest that many genes currently known to be associated with human hearing impairment are active during embryogenesis. Hence, the study of inner ear development provides a rich context for understanding the functions of genes implicated in hearing loss. This chapter focuses on molecular mechanisms of inner ear development derived from studies of model organisms.

Novel autoregulatory metabolites, arthrosporols A-C (1-3), involved in regulating the morphological switch in fungi, were purified and characterized from the carnivorous fungus Arthrobotrys oligospora. These compounds possess a novel hybrid carbon skeleton consisting of an epoxy-cyclohexenol combined with a rare monocyclic sesquiterpenol substructure. This is the first report of a monocyclic sesquiterpenol of this type of fungal origin. Compounds 1-3 displayed significant inhibitory activities toward the formation of conidiophores, while compounds 1 and 3 showed the opposite effects on the formation of a two-dimensional network with increasing rates of 40-90% and inhibiting rates of 30-90%, respectively.

Development of the mammalian inner ear requires coordination of cell proliferation, cell fate determination and morphogenetic movements. While significant progress has been made in identifying developmental signals required for inner ear formation, less is known about how distinct signals are coordinated by their downstream mediators. Members of the Rac family of small GTPases are known regulators of cytoskeletal remodeling and numerous other cellular processes. However, the function of Rac GTPases in otic development is largely unexplored. Here, we show that Rac1 and Rac3 redundantly regulate many aspects of inner ear morphogenesis. While no morphological defects were observed in Rac3(-/-) mice, Rac1(CKO); Rac3(-/-) double mutants displayed enhanced vestibular and cochlear malformations compared to Rac1(CKO) single mutants. Moreover, in Rac1(CKO); Rac3(-/-) mutants, we observed compromised E-cadherin-mediated cell adhesion, reduced cell proliferation and increased cell death in the early developing otocyst, leading to a decreased size and malformation of the membranous labyrinth. Finally, cochlear extension was severely disrupted in Rac1(CKO); Rac3(-/-) mutants, accompanied by a loss of epithelial cohesion and formation of ectopic sensory patches underneath the cochlear duct. The compartmentalized expression of otic patterning genes within the Rac1(CKO); Rac3(-/-) mutant otocyst was largely normal, however, indicating that Rac proteins regulate inner ear morphogenesis without affecting cell fate specification. Taken together, our results reveal an essential role for Rac GTPases in coordinating cell adhesion, cell proliferation, cell death and cell movements during otic development., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Gbx2 encodes a DNA-binding transcription factor that plays pivotal roles during embryogenesis. Gain-and loss-of-function studies in several vertebrate species have demonstrated a requirement for Gbx2 in development of the anterior hindbrain, spinal cord, inner ear, heart, and neural crest cells. However, the target genes through which GBX2 exerts its effects remain obscure. Using chromatin immunoprecipitation coupled with direct sequencing (ChIP-Seq) analysis in a human prostate cancer cell line, we identified cis-regulatory elements bound by GBX2 to provide insight into its direct downstream targets. The analysis revealed more than 286 highly significant candidate target genes, falling into various functional groups, of which 51% are expressed in the nervous system. Several of the top candidate genes include EEF1A1, ROBO1, PLXNA4, SLIT3, NRP1, and NOTCH2, as well as genes associated with the Usher syndrome, PCDH15 and USH2A, and are plausible candidates contributing to the developmental defects in Gbx2(-/-) mice. We show through gel shift analyses that sequences within the promoter or introns of EEF1A1, ROBO1, PCDH15, USH2A and NOTCH2, are directly bound by GBX2. Consistent with these in vitro results, analyses of Gbx2(-/-) embryos indicate that Gbx2 function is required for migration of Robo1-expressing neural crest cells out of the hindbrain. Furthermore, we show that GBX2 activates transcriptional activity through the promoter of EEF1A1, suggesting that GBX2 could also regulate gene expression indirectly via EEF1A. Taken together, our studies show that GBX2 plays a dynamic role in development and diseases.

Inner ear hair cells convert the mechanical stimuli of sound, gravity, and head movement into electrical signals. This mechanotransduction process is initiated by opening of cation channels near the tips of hair cell stereocilia. Since the identity of these ion channels is unknown, and mutations in the gene encoding transmembrane channel-like 1 (TMC1) cause hearing loss without vestibular dysfunction in both mice and humans, we investigated the contribution of Tmc1 and the closely related Tmc2 to mechanotransduction in mice. We found that Tmc1 and Tmc2 were expressed in mouse vestibular and cochlear hair cells and that GFP-tagged TMC proteins localized near stereocilia tips. Tmc2 expression was transient in early postnatal mouse cochlear hair cells but persisted in vestibular hair cells. While mice with a targeted deletion of Tmc1 (Tmc1(Δ) mice) were deaf and those with a deletion of Tmc2 (Tmc2(Δ) mice) were phenotypically normal, Tmc1(Δ)Tmc2(Δ) mice had profound vestibular dysfunction, deafness, and structurally normal hair cells that lacked all mechanotransduction activity. Expression of either exogenous TMC1 or TMC2 rescued mechanotransduction in Tmc1(Δ)Tmc2(Δ) mutant hair cells. Our results indicate that TMC1 and TMC2 are necessary for hair cell mechanotransduction and may be integral components of the mechanotransduction complex. Our data also suggest that persistent TMC2 expression in vestibular hair cells may preserve vestibular function in humans with hearing loss caused by TMC1 mutations.

The purpose of this study was to report our experience of percutaneous retrieval of dislodged port catheters with concurrent use of pigtail and loop snare catheters. During a 5-year period at our institute (June 2005 to July 2010), a total of 23 dislodged port catheters were retrieved. The interval between port catheter implantation and dislodged catheter retrieval ranged from 43 days to 1,414 days (mean 586.7 days). The time of delayed retrieval ranged from 1 day to 45 days (mean 4.6 days). All dislodged catheters were retrieved with the concurrent use of pigtail and loop snare catheters via femoral venous route. The prevalence of port catheter dislodgement at our institute was 3.4%. All dislodged port catheters were removed successfully with pigtail and loop snare catheters together. No procedure-related complications were encountered, except for transient arrhythmia in two patients, which required no medication. In conclusion, the concurrent use of pigtail and loop snare catheters is a feasible and easy way for percutaneous retrieval of a dislodged central venous port catheter., (Copyright © 2011. Published by Elsevier B.V.)

Objective: The present study was performed to investigate competitive interaction between arenobufagin and verapamil hydrochloride with serum albumin., Methods: Equilibrium dialysis and high-performance liquid chromatography were used to analyze the binding rates of the two medicines to serum protein. The interactions based on bovine serum albumin (BSA) and human serum albumin (HSA) were investigated by using spectrofluorimetry. The interaction mode of arenobufagin and verapamil hydrochloride binding to serum proteins was simulated by molecular docking., Results: The rate of arenobufagin (0.1μg/mL) binding to bovine serum was (61.1±0.2)%. Verapamil hydrochloride (0.025 to 0.1μg/mL) significantly reduced the bovine serum binding rate of arenobufagin, from (60.2±3.7)% to (36.9±3.4)%. However, arenobufagin at the tested doses had no marked effects on the binding rate of verapamil hydrochloride. Furthermore, the verapamil hydrochloride had an active effect on the arenobufagin-induced fluorescence quenching of BSA and HSA. The molecular docking results showed that verapamil hydrochloride and arenobufagin binded to HSA at site I. Molecular interaction energy of verapamil hydrochloride binding to site I was stronger than that of arenobufagin., Conclusion: Verapamil hydrochloride reduces the binding of arenobufagin to bovine serum. The mechanism may be a competitive interaction of arenobufagin and verapamil hydrochloride at site I on HSA.

Thirteen different magnetitum samples were analyzed by FTIR, and the FTIR fingerprint was set up with them. Processed samples and the crudes were compared with the fingerprint. It was found that all the similarities of the samples are more than 0.97. The similarities and the correlation coefficients of the processed samples are decreased. The FTIR fingerprint could be used for evaluating the quality of magnetitum for commodities.

Vertebrate hearing and balance are based in complex asymmetries of inner ear structure. Here, we identify retinoic acid (RA) as an extrinsic signal that acts directly on the ear rudiment to affect its compartmentalization along the anterior-posterior axis. A rostrocaudal wave of RA activity, generated by tissues surrounding the nascent ear, induces distinct responses from anterior and posterior halves of the inner ear rudiment. Prolonged response to RA by posterior otic tissue correlates with Tbx1 transcription and formation of mostly nonsensory inner ear structures. By contrast, anterior otic tissue displays only a brief response to RA and forms neuronal elements and most sensory structures of the inner ear.

Two new cerebrosides, 1-O-(beta-d-glucopyranosyloxy)-(2S,3S,4R,8Z)-2-[(2'R)-2'-hydroxytricosanoylamino]-8-nonadecene-3,4-diol (1) and 1-O-(beta-D-glucopyranosyloxy)-(2S,3R,4E,8Z)-2-[(2'R)-2'-hydroxynonadecanoylamino]-4,13-nonadecene-3-diol (2), were isolated from the pollen of Typha angustifolia. Their structures were elucidated by chemical and spectral means. This is the first report on the occurrence of cerebroside in Typha (Typhaceae). Compounds 1 and 2 exhibited effect on the proliferation of cultured vascular smooth muscle cell (VSMCs) induced by fatal bovine serum (FBS)., (Copyright (c) 2009 Elsevier B.V. All rights reserved.)

Kaposiform hemangioendothelioma is a very infrequent, locally aggressive vascular neoplasm, characterized by fascicular spindle cell proliferation. It occurs almost exclusively in infants and adolescents, and is often associated with Kasabach-Merritt phenomenon. The tumor is predominantly located subcutaneously or in the deep soft tissue of the extremities and trunk, peritoneum, or retroperitoneum. However, this tumor can sometimes be located on the head and neck. We report a case of kaposiform hemangioendothelioma of the maxillary sinus in a 4-month-old female infant presenting with cheek swelling, thrombocytopenia, anemia, and disseminated intravascular coagulation. Sinus computed tomography presented an enhancing, bone-destructing tumor. Magnetic resonance images showed an uncommon appearance as mostly low signal intensity on T2-weighted images., (Copyright 2010 Elsevier. Published by Elsevier B.V. All rights reserved.)

The mammalian organ of Corti of the inner ear is a highly sophisticated sensory end organ responsible for detecting sound. Noggin is a secreted glycoprotein, which antagonizes bone morphogenetic proteins 2 and 4 (Bmp2 and Bmp4). The lack of this antagonist causes increased rows of inner and outer hair cells in the organ of Corti. In mice, Bmp2 is expressed transiently in nascent cochlear hair cells. To investigate whether Noggin normally modulates the levels of Bmp2 for hair cell formation, we deleted Bmp2 in the cochlear hair cells using two cre strains, Foxg1(cre/+) and Gfi1(cre/+). Bmp2 conditional knockout cochleae generated using these two cre strains show normal hair cells. Furthermore, Gfi1(cre/+);Bmp2(lox/-) mice are viable and have largely normal hearing. The combined results of Noggin and Bmp2 mutants suggest that Noggin is likely to regulate other Bmps in the cochlea such as Bmp4.

TiO2 nanoceramic films were deposited on glasses by rf magnetron sputtering and corresponded to nanocrystalline anatase. The porosity and surface roughness decreased with the oxygen pressure. The optical transmission of TiO2 nanoceramic films obviously increased with the decrease of film thickness or the increase of oxygen pressure, especially in the visible region. Moiré deflectometry was used to measure the nonlinear refractive indices of TiO2 films deposited in a mixed Ar-O2 atmosphere. The nonlinear refractive index was measured to be of the order of 10(-8) cm2 W(-1) and the change in refractive index was of the order of 10(-5). As the oxygen pressure increased, the transparent TiO2 film exhibited a high linear refractive index, a low stress and a low stress-optical coefficient.

Gegen Qinlian dispensing granule, a favorite composite formula, is a combination of Radix Puerariae Lobatae, Radix Scutellariae, Rhizoma Coptidis and Radix et Rhizoma Glycyrrhizae Praeparata cum Melle. To develop a method to overall evaluate correlation between the formula and its four raw herbs, LC fingerprints of the formula and its raw herbs were developed and LC-DAD-MS was employed to identify the components in the formula fingerprint. According to the characteristic fragmentation behavior of known flavonoids, alkaloids and saponins isolated from the four raw herbs as well as retention time, UV and MS data of detected compounds, a total of 23 constituents in the formula fingerprint were structurally characterized. Chemical correlation between the formula and the four crude herbs was evaluated qualitatively and quantitatively by the developed LC fingerprints. The results showed that 8 components in the formula fingerprint were addressed to Radix Puerariae Lobatae, 11 to Radix Scutellariae, 7 to Rhizoma Coptidis, 3 to Radix et Rhizoma Glycyrrhizae Praeparata cum Melle, and that the relative area ratios of the common peaks in the formula vary slightly in comparison with corresponding ratios in its crude herbs, demonstrating the chemical constituents in the formula have patterns similar to those in its crude herbs., (Copyright 2009 Elsevier GmbH. All rights reserved.)

A mature inner ear is a complex structure consisting of vestibular and auditory components. Microsurgical ablations, rotations, and translocations were performed in ovo to identify the tissues that control inner ear morphogenesis. We show that mesenchyme/ectoderm adjacent to the developing ear specifically governs the shape of vestibular components - the semicircular canals and ampullae - by conferring anteroposterior axial information to these structures. In contrast, removal of individual hindbrain rhombomeres adjacent to the developing ear preferentially affects the growth and morphogenesis of the auditory subdivision, the cochlear duct, or basilar papilla. Removal of rhombomere 5 affects cochlear duct growth, while rhombomere 6 removal affects cochlear growth and morphogenesis. Rotating rhombomeres 5 and 6 along the anteroposterior axis also impacts cochlear duct morphogenesis but has little effect on the vestibular components. Our studies indicate that discrete tissues, acting at a distance, control the morphogenesis of distinct elements of the inner ear. These results provide a basis for identifying factors that are essential to vestibular and auditory development in vertebrates., (Copyright 2009. Published by Elsevier Inc.)

Accurate and consistent visualization of the entire coronary system with high-grade imaging quality is crucial for routine applications of multi-detector-computed tomography (MDCT) coronary angiography. To determine the imaging quality of 64-slice-MDCT coronary angiography, we respectively explored the quantitative parameters of imaging quality in 105 consecutive subjects (71 men, 34 women; aged 58.66 +/- 10.62 years) who underwent 64-slice-MDCT coronary angiography to screen for coronary disease. The interobserver agreement for semi-quantitative image quality, visible length, signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) of the coronary arteries was good. The SNR and CNR of the proximal segments of the coronary arteries were superior to that of the distal segments of coronary arteries (p < 0.001). The visible length of the stenosed right coronary artery was significantly shorter than that of the non-stenosed right coronary artery (p = 0.03). The SNR and CNR of the stenosed and non-stenosed coronary arteries revealed no significant difference (p > 0.05). Body weight and body mass index were inversely related to the SNR and CNR of the aorta (p < 0.001). In conclusion, 64-slice-MDCT coronary angiography can provide excellent imaging quality of coronary arteries in subjects undergoing screening for coronary disease, although the SNR and CNR were relatively low at the distal segments of coronary arteries.

San-ao decoction (SAD), comprising Herba Ephedrae, Radix et Rhizoma Glycyrrhizae and Seneb Armeniacae Amarum, is one of the most popular traditional Chinese medicine (TCM) formulae for asthma. Peroxisome proliferator-activated receptors (PPARs) areey regulators of lipid and glucose metabolism and have become important therapeutic targets for various deseases, PPARgamma activation might exhibit anti-inflammatory properties in different chronic inflammatory processes. The EtOAc fraction of SAD showed a significant effect on PPARgamma activation. A simple and rapid method has been established for separation and characterization of the main compounds in the PPARgamma-activating fraction of SAD by ultra-fast HPLC coupled with quadropole time-of-flight mass pectrometry (UPLC-Q-TOF/MS). A total of 10 compounds were identified in the activating fraction of SAD, including amygdalin (1), liquiritin (2), 6'-acetyliquiritin (3), liquiritigenin (4), isoliquiritigenin (5), formononetin (6), licoisoflavanone (7), glycycoumarin (8), glycyrol (9) and uercetin (10). The results also characterized formononetin as a predominant component in this fraction. The dose-effect relationship comparison study of formononetin and the EtOAc fraction of SAD by adding formononetin was performed, the results suggested that formononetin was the major component of the EtOAc fraction of SAD responsible for activating PPARgamma, and the method will possibly be applied to study the complex biological active constituents of other TCMs.

Background: The International Panel on the Diagnosis of Multiple Sclerosis first incorporated abnormalities demonstrated by brain and spinal cord MRI into the diagnostic criteria (McDonald criteria) for multiple sclerosis (MS), which were later revised in 2005. In 2006, Swanton and colleagues modified the MRI criteria to simplify and speed the diagnosis., Objective: The purpose of this study was to compare the ability of two sets of criteria (the revised McDonald MRI criteria and Swanton's modified criteria) to predict conversion from clinically isolated syndrome (CIS) to clinically definite multiple sclerosis (CDMS) from baseline MRI findings., Methods: Sixty-four patients presenting with CIS suggestive of multiple sclerosis were recruited from 2001 to 2006 and followed up for at least 2 years. Their baseline brain and spinal cord MRI studies were retrospectively evaluated. The patients who developed CDMS during follow-up were treated as positive cases. The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and accuracy of the two MRI dissemination-in-space criteria were calculated., Results: Thirty patients (46.9%) converted to CDMS. The sensitivity specificity, PPV, NPV and accuracy (%) of the revised McDonald criteria were 53, 100, 100, 71 and 78, respectively, while those for Swanton's modified criteria were 60, 100, 100, 74 and 81., Conclusions: In conclusion, Swanton's modified criteria are more sensitive and accurate (but not significantly so). However, Swanton's criteria are simpler to use and have equally high specificity and PPV.

Lmx1a is a LIM homeodomain-containing transcription factor, which is required for the formation of multiple organs. Lmx1a is broadly expressed in early stages of the developing inner ear, but its expression is soon restricted to the non-sensory regions of the developing ear. In an Lmx1a functional null mutant, dreher (dr(J)/dr(J)), the inner ears lack a non-sensory structure, the endolymphatic duct, and the membranous labyrinth is poorly developed. These phenotypes are consistent with Lmx1a's role as a selector gene. More importantly, while all three primary fates of the inner ear - neural, sensory, and non-sensory - are specified in dr(J)/dr(J), normal boundaries among these tissues are often violated. For example, the neurogenic domain of the ear epithelium, from which cells delaminate to form the cochleovestibular ganglion, is expanded. Within the neurogenic domain, the demarcation between the vestibular and auditory neurogenic domains is most likely disrupted as well, based on the increased numbers of vestibular neuroblasts and ectopic expression of Fgf3, which normally is associated specifically with the vestibular neurogenic region. Furthermore, aberrant and ectopic sensory organs are observed; most striking among these is vestibular-like hair cells located in the cochlear duct.

We introduce a microfluidic refractive index sensor based on a directional coupler architecture using solid-core photonic crystal fibers. The sensor achieves very high sensitivity by coupling the core mode to a mode in the adjacent fluid-filled waveguide that is beyond modal cutoff, and with strong field overlap. We demonstrate the device through the selective infiltration of a single hole with fluid along a microstructured optical fiber. A detection limit of 4.6x10(-7) refractive index units has been derived from measurements with a sensitivity of 30,100 nm per refractive index unit, which is the highest for a fiber device to date.

A long vertical vestibule with a diverticulum-like morphology of cavotricuspid isthmus (CTI) was demonstrated by right atrial angiogram in a 47-year-old male patient in whom his typical atrial flutter was difficult to be ablated. Compatible with the right atrial angiography, the diverticulum-like CTI with a long vertical vestibule, pouch-like recess, and a long transverse posterior component was also demonstrated on volume-rendering reconstruction computed tomography and the virtual endoscopy showed prominent trabeculation inside the diverticulum-like part of CTI. The bidirectional CTI conduction block was successfully achieved after ablating along the CTI guided by the right atrial angiography.

We demonstrate the spectral and spatial reconfigurability of photonic crystal double-heterostructure cavities in silicon by microfluidic infiltration of selected air holes. The lengths of the microfluidic cavities are changed by adjusting the region of infiltrated holes in steps of several microns. We systematically investigate the spectral signature of these cavities, showing high Q-factor resonances for a broad range of cavity lengths. The fluid can be removed by immersing the device in toluene, offering complete reconfigurability. Our cavity writing technique allows for tolerances in the infiltration process and provides flexibility as it can be employed at any time after photonic crystal fabrication.

Background: The main pulmonary arterial (PA) distensibility in patients with pulmonary hypertension (PH) and pulmonary embolism (PE) is uncertain., Methods and Results: We enrolled 45 patients with echocardiographic signs of PH and without imaging evidence of PE, and another 45 who were found by multidetector-row computed tomography to have PE. Fifty normal patients served as a control group. The PA distensibility was calculated from the change in main PA diameter between diastole and systole, as the maximal systolic diameter minus the minimal diastolic diameter divided by the minimal diastolic diameter. The PA distensibility is lowest in PH (6.0+/-2.7%), followed by PE (12.9+/-3.4%) and then the normal controls (25.9+/-5.7%). Statistical analysis of data obtained from patients with PE or PH reveals that a PA distensibility of >8.3% could be used to identify PE with a sensitivity of 83% and a specificity of 82%. After a 3-month anticoagulation, 22 of 45 PE patients had complete resolution of thrombus; 23 had residual thrombus. The PE patients, regardless of residual thrombus presence, had significant improvement of PA distensibility after a 3-month anticoagulation, although the diameters of main PA did not shrink., Conclusion: PA distensibility is a method to distinguish acute PE from chronic PH.

Objective: To investigate the inhibitory effect of the small interfering RNA targeting mdm2 gene on the growth of osteosarcoma cells., Methods: PGCsilencerTM-mdm2 siRNA was constructed and transfected into the osteosarcoma cell line U2OS cells. The inhibitory effects on mdm2 were determined by RT-PCR and Western blot analysis. The cell growth activity was determined by MTT assay, and the cell apoptosis was examined by flow cytometry. The therapeutic effects of simdm2 was assessed on the nude mouse model of transplanted tumor., Results: The simdm2 plasmid was successfully constructed. After simdm2 being transfected into the U2OS cells, the expressions of mdm2 gene and protein were significantly inhibited. The ability of cell growth activity decreased greatly and cell apoptosis occurred apparently. There was no significant difference between the negative control group and non-transfected group. The growth of xenograft tumor in simdm2 transfected nude mice was inhibited and the expressions of mdm2 gene and protein were down-regulated remarkably., Conclusion: siRNA targeting mdm2 gene inhibits the mdm2 expression in osteosarcoma U2OS cells and the growth of osteosarcoma in nude mice.

Angular head movements in vertebrates are detected by the three semicircular canals of the inner ear and their associated sensory tissues, the cristae. Bone morphogenetic protein 4 (Bmp4), a member of the Transforming growth factor family (TGF-beta), is conservatively expressed in the developing cristae in several species, including zebrafish, frog, chicken, and mouse. Using mouse models in which Bmp4 is conditionally deleted within the inner ear, as well as chicken models in which Bmp signaling is knocked down specifically in the cristae, we show that Bmp4 is essential for the formation of all three cristae and their associated canals. Our results indicate that Bmp4 does not mediate the formation of sensory hair and supporting cells within the cristae by directly regulating genes required for prosensory development in the inner ear such as Serrate1 (Jagged1 in mouse), Fgf10, and Sox2. Instead, Bmp4 most likely mediates crista formation by regulating Lmo4 and Msx1 in the sensory region and Gata3, p75Ngfr, and Lmo4 in the non-sensory region of the crista, the septum cruciatum. In the canals, Bmp2 and Dlx5 are regulated by Bmp4, either directly or indirectly. Mechanisms involved in the formation of sensory organs of the vertebrate inner ear are thought to be analogous to those regulating sensory bristle formation in Drosophila. Our results suggest that, in comparison to sensory bristles, crista formation within the inner ear requires an additional step of sensory and non-sensory fate specification., Competing Interests: The authors have declared that no competing interests exist.

Conductive hearing loss occurs when sound waves are not relayed efficiently to the inner ear. Mutations of the NOGGIN (NOG) gene in humans are associated with several autosomal dominant disorders such as proximal symphalangism and multiple synostoses. These syndromes are characterized by skeletal defects and synostoses, which include conductive hearing loss. Noggin is an antagonist of bone morphogenetic proteins (BMPs), and balanced levels of BMPs and Noggin are required for proper skeletal formation. Depending on the genetic background, some of the Nog(+/-) mice display mild hearing loss, that is, conductive in nature. Since Noggin is a single exon gene, this data strongly suggest that the autosomal dominant disorders associated with NOG mutations are due to haploinsufficiency of NOGGIN. The conductive hearing loss in Nog(+/-) mice is caused by an ectopic bone bridge located between the stapes and the posterior wall of the tympanum, which affects the normal mobility of the ossicle. Our analyses suggest that the ectopic bone formation is caused by a failure of the stapes and styloid process to separate completely during development. This failure of bone separation in the Nog(+/-) mice reveals another consequence of chondrocyte hyperplasia due to unopposed Bmp activities in these mutants such as Bmp4 and Bmp14 (Gdf5). More importantly, these results establish Nog(+/-) mice as the first animal model for the study of conductive rather than neurosensory hearing loss that has direct relevance to human genetic disorders.

Signaling from rhombomeres 5 and 6 of the hindbrain is thought to be important for inner ear patterning. In Noggin -/- embryos, the gross anatomy of the inner ear is distorted and malformed, with cochlear duct outgrowth and coiling most affected. We attributed these defects to a caudal shift of the rhombomeres caused by the shortened body axis and the kink in the neural tube. To test the hypothesis that a caudal shift of the rhombomeres affects inner ear development, we surgically generated chicken embryos in which rhombomeres 5 and 6 were similarly shifted relative to the position of the inner ears, as in Noggin mutants. All chicken embryos with shifted rhombomeres showed defects in cochlear duct formation indicating that signaling from rhombomeres 5 and 6 is important for cochlear duct patterning in both chicken and mice. In addition, the size of the otic capsule is increased in Noggin -/- mutants, which most likely is due to unopposed BMP signaling for chondrogenesis in the peri-otic mesenchyme.

Organization of the vertebrate inner ear is mainly dependent on localized signals from surrounding tissues. Previous studies demonstrated that sonic hedgehog (Shh) secreted from the floor plate and notochord is required for specification of ventral (auditory) and dorsal (vestibular) inner ear structures, yet it was not clear how this signaling activity is propagated. To elucidate the molecular mechanisms by which Shh regulates inner ear development, we examined embryos with various combinations of mutant alleles for Shh, Gli2 and Gli3. Our study shows that Gli3 repressor (R) is required for patterning dorsal inner ear structures, whereas Gli activator (A) proteins are essential for ventral inner ear structures. A proper balance of Gli3R and Gli2/3A is required along the length of the dorsoventral axis of the inner ear to mediate graded levels of Shh signaling, emanating from ventral midline tissues. Formation of the ventral-most otic region, the distal cochlear duct, requires robust Gli2/3A function. By contrast, the formation of the proximal cochlear duct and saccule, which requires less Shh signaling, is achieved by antagonizing Gli3R. The dorsal vestibular region requires the least amount of Shh signaling in order to generate the correct dose of Gli3R required for the development of this otic region. Taken together, our data suggest that reciprocal gradients of GliA and GliR mediate the responses to Shh signaling along the dorsoventral axis of the inner ear.

The positional cues for formation of individual inner ear components are dependent on pre-established axial information conferred by inductive signals from tissues surrounding the developing inner ear. This review summarizes some of the known molecular pathways involved in establishing the three axes of the inner ear, anterior-posterior (AP), dorsal-ventral (DV) and medial-lateral (ML). Signals required to establish the AP axis of the inner ear are not known, but they do not appear to be derived from the hindbrain. In contrast, the hindbrain is essential for establishing the DV axis of the inner ear by providing inductive signals such as Wnts and Sonic hedgehog. Signaling from the hindbrain is also required for the formation of the ML axis, whereas formation of the lateral wall of the otocyst may be a result of first establishing both the AP and DV axes. In addition, this review addresses how genes induced within the otic epithelium as a result of axial specification continue to mediate inner ear morphogenesis.

The ability to transport cations and anions across epithelia is critical for the regulation of pH, ionic homeostasis, and volume of extracellular fluids. Although the transporters and channels that facilitate ion and water movement across cell membranes are well known, the molecular mechanisms and signal transduction events that regulate these activities remain poorly understood. The Eph family of receptor tyrosine kinases and their membrane-anchored ephrin ligands are well known to transduce bidirectional signals that control axon guidance and other cell migration/adhesion events during development. However, these molecules are also expressed in non-motile epithelial cells, including EphB2 in K(+)-secreting vestibular dark cells and ephrin-B2 in the adjacent transitional cells of the inner ear. Consistent with these expression patterns, mice with cytoplasmic domain mutations that interfere with EphB2 forward signaling or ephrin-B2 reverse signaling exhibit a hyperactive circling (waltzing) locomotion associated with a decreased amount of endolymph fluid that normally fills the vestibular labyrinth. Endolymph is unusual as an extracellular fluid in that it is normally high in K(+) and low in Na(+). Direct measurement of this fluid in live animals revealed significant decreases in K(+) concentration and endolymphatic potential in both EphB2 and ephrin-B2 mutant mice. Our findings provide evidence that bidirectional signaling mediated by B-subclass Ephs and ephrins controls the production and ionic homeostasis of endolymph fluid and thereby provide the first evidence that these molecules can control the activities of mature epithelial cells.

Purpose: To prospectively investigate the apparent diffusion coefficient (ADC) and choline levels measured at hydrogen 1 ((1)H) magnetic resonance (MR) spectroscopy, to monitor therapeutic responses of hepatocellular carcinoma (HCC) to transcatheter arterial chemoembolization (TACE)., Materials and Methods: Institutional review board approval was obtained, and all patients and control subjects provided informed consent. Histologically proved large HCCs (>3 cm in diameter) were evaluated in 20 patients (16 men and four women; mean age, 59 years; range, 34-80 years) before TACE and 2-3 days after TACE. A control group of eight adults (five men and three women; mean age, 43 years; range, 24-76 years) with normal livers was examined by using the same protocol. Hepatic choline levels were measured by means of an external phantom replacement method, quantifying the peak at 3.2 ppm at (1)H MR spectroscopy. ADCs were measured for all lesions. A Wilcoxon rank sum test was used to compare absolute choline concentrations and ADCs at baseline between HCCs and normal liver parenchyma. Changes in choline levels and ADCs in the tumors before and after TACE were analyzed by using the Wilcoxon signed rank test., Results: The median preoperative choline level in patients with HCC (measured in 18 of the 20 patients) was 4.0 mmol/L (range, 0.0-17.2 mmol/L), which was significantly higher than that in patients with normal livers (n = 8) (median, 1.6 mmol/L; range, 0.0-2.1 mmol/L; P < .01). Among 18 patients with HCC, choline levels decreased significantly from before TACE to after TACE (P < .01). A significant increase in ADC from before TACE to after TACE in the 20 patients with HCC was also found (P < .01)., Conclusion: Hepatic choline levels and ADCs may allow monitoring of therapeutic responses of HCC to TACE although larger, more definitive and quantitative studies with clinical end points are needed., ((c) RSNA, 2006.)

Technical success rates for percutaneous restoration of thrombosed arteriovenous grafts are high. However, in thrombosed grafts without restorable original outflow veins, percutaneous salvage is usually not possible. In this situation, patients are referred for bypass grafting or recreation of their vascular access sites. This report describes a patient in whom the original outflow vein at the venous anastomosis was completely obliterated and in whom vascular access was successfully salvaged by percutaneously reanastomosing the venous stump of the thrombosed graft with an adjacent patent vein. This procedure is technically feasible for the salvage of a thrombosed graft.

Gbx2 is a homeobox-containing transcription factor that is related to unplugged in Drosophila. In mice, Gbx2 and Otx2 negatively regulate each other to establish the mid-hindbrain boundary in the neural tube. Here, we show that Gbx2 is required for the development of the mouse inner ear. Absence of the endolymphatic duct and swelling of the membranous labyrinth are common features in Gbx2-/- inner ears. More severe mutant phenotypes include absence of the anterior and posterior semicircular canals, and a malformed saccule and cochlear duct. However, formation of the lateral semicircular canal and its ampulla is usually unaffected. These inner ear phenotypes are remarkably similar to those reported in kreisler mice, which have inner ear defects attributed to defects in the hindbrain. Based on gene expression analyses, we propose that activation of Gbx2 expression within the inner ear is an important pathway whereby signals from the hindbrain regulate inner ear development. In addition, our results suggest that Gbx2 normally promotes dorsal fates such as the endolymphatic duct and semicircular canals by positively regulating genes such as Wnt2b and Dlx5. However, Gbx2 promotes ventral fates such as the saccule and cochlear duct, possibly by restricting Otx2 expression.

An early and crucial event in vertebrate inner ear development is the acquisition of axial identities that in turn dictate the positions of all subsequent inner ear components. Here, we focus on the role of the hindbrain in establishment of inner ear axes and show that axial specification occurs well after otic placode formation in chicken. Anteroposterior (AP) rotation of the hindbrain prior to specification of this axis does not affect the normal AP orientation and morphogenesis of the inner ear. By contrast, reversing the dorsoventral (DV) axis of the hindbrain results in changing the DV axial identity of the inner ear. Expression patterns of several ventrally expressed otic genes such as NeuroD, Lunatic fringe (Lfng) and Six1 are shifted dorsally, whereas the expression pattern of a normally dorsal-specific gene, Gbx2, is abolished. Removing the source of Sonic Hedgehog (SHH) by ablating the floor plate and/or notochord, or inhibiting SHH function using an antibody that blocks SHH bioactivity results in loss of ventral inner ear structures. Our results indicate that SHH, together with other signals from the hindbrain, are important for patterning the ventral axis of the inner ear. Taken together, our studies suggest that tissue(s) other than the hindbrain confer AP axial information whereas signals from the hindbrain are necessary and sufficient for the DV axial patterning of the inner ear.

In the vertebrate inner ear, the ability to detect angular head movements lies in the three semicircular canals and their sensory tissues, the cristae. The molecular mechanisms underlying the formation of the three canals are largely unknown. Malformations of this vestibular apparatus found in zebrafish and mice usually involve both canals and cristae. Although there are examples of mutants with only defective canals, few mutants have normal canals without some prior sensory tissue specification, suggesting that the sensory tissues, cristae, might induce the formation of their non-sensory components, the semicircular canals. We fate-mapped the vertical canal pouch in chicken that gives rise to the anterior and posterior canals, using a fluorescent, lipophilic dye (DiI), and identified a canal genesis zone adjacent to each prospective crista that corresponds to the Bone morphogenetic protein 2 (Bmp2)-positive domain in the canal pouch. Using retroviruses or beads to increase Fibroblast Growth Factors (FGFs) for gain-of-function and beads soaked with the FGF inhibitor SU5402 for loss-of-function experiments, we show that FGFs in the crista promote canal development by upregulating Bmp2. We postulate that FGFs in the cristae induce a canal genesis zone by inducing/upregulating Bmp2 expression. Ectopic FGF treatments convert some of the cells in the canal pouch from the prospective common crus to a canal-like fate. Thus, we provide the first molecular evidence whereby sensory organs direct the development of the associated non-sensory components, the semicircular canals, in vertebrate inner ears.

Objective: To optimize the conditions of supercritical fluid extraction (SFE) for curcumin in Curcuma longa., Method: Optimum extraction conditions were studied by orthogonal tests. The extracts were analyzed by HPLC., Result: The optimal extraction conditions were pressure 25 MPa, temperature 55 degrees C, static time 4 h, dynamic time 5 h, flow rate of CO2 3.5 L x min(-1), co-solvent ethanol 30% (mL x g(-1))., Conclusion: It is feasible to extract curcumin by SFE.

The paired box transcription factor, Pax2, is important for cochlear development in the mouse inner ear. Two mutant alleles of Pax2, a knockout and a frameshift mutation (Pax21Neu), show either agenesis or severe malformation of the cochlea, respectively. In humans, mutations in the PAX2 gene cause renal coloboma syndrome that is characterized by kidney abnormalities, optic nerve colobomas and mild sensorineural deafness. To better understand the role of Pax2 in inner ear development, we examined the inner ear phenotype in the Pax2 knockout mice using paint-fill and gene expression analyses. We show that Pax2-/- ears often lack a distinct saccule, and the endolymphatic duct and common crus are invariably fused. However, a rudimentary cochlea is always present in all Pax2 knockout inner ears. Cochlear outgrowth in the mutants is arrested at an early stage due to apoptosis of cells that normally express Pax2 in the cochlear anlage. Lack of Pax2 affects tissue specification within the cochlear duct, particularly regions between the sensory tissue and the stria vascularis. Because the cochlear phenotypes observed in Pax2 mutants are more severe than those observed in mice lacking Otx1 and Otx2, we postulate that Pax2 plays a key role in regulating the differential growth within the cochlear duct and thus, its proper outgrowth and coiling.

Purpose: To investigate the value of in vivo proton magnetic resonance spectroscopy (MRS) in the assessment of large focal hepatic lesions and to measure the metabolite change of hepatocellular carcinoma (HCC) after transcatheter arterial chemoembolization (TACE) using 3.0-T scanner., Materials and Methods: In this prospective study, 43 consecutive patients with large (not less than 3 cm in diameter) hepatic tumors and eight normal volunteer were included. MRS of the lesions in addition to uninvolved liver parenchyma was carried out using a whole-body 3.0-T scanner. Among the patients with proven HCC, eight lesions were evaluated before and two to five days after TACE. The choline-to-lipid (cho/lipid) ratio was measured by dividing the peak area of choline at 3.2 ppm by the peak area of lipid at 1.3 ppm. The sensitivity and specificity profiles of MRS in the diagnosis of malignant hepatic tumors were determined by plotting empirical receiver operating characteristic (ROC) curve. The mean cho/lipid ratios in different groups before and after TACE were also measured., Results: The technical success rate for MRS was 90% (53/59). The ROC curve showed proton MRS has moderate discriminating ability in diagnosing malignant hepatic tumors, although the sensitivity was less than 50% while 1-specificity was less than 20%. The area under the curve was 0.71 (P < 0.05). The mean +/- 1 standard error (SE) of cho/lipid ratios for uninvolved liver (N = 8), benign tumor (N = 8), and malignant tumor (N = 21; 19 HCC, one angiosarcoma, and one lymphoma) were 0.06 +/- 0.02, 0.02 +/- 0.02, and 0.17 +/- 0.05, respectively. A significantly statistical difference (ANOVA planned contrast test, P = 0.01 and Games-Howell procedure, P = 0.03) was achieved in the mean cho/lipid ratio between malignant and benign tumors. The mean cho/lipid ratios were significantly decreased from 0.23 +/- 0.11 before TACE to 0.01 +/- 0.00 after the treatment (t = 2.01, P < 0.05, one-tail paired t-test; z = -2.37, P < 0.05, Wilcoxon Signed Ranks Test)., Conclusion: In vivo proton MRS is technically feasible for the evaluation of focal hepatic lesions. The technique has potential in the detection of early metabolite change in malignant liver tumors after TACE but limitation still exists in clear differentiation between normal liver and benign and malignant tumor., (Copyright 2004 Wiley-Liss, Inc.)

We report the case of a 63-year-old female who had chronic hepatitis C and who was diagnosed with hepatocellular carcinoma. Hepatic angiography showed one visible and tortuous falciform artery arising as the terminal branch of the left hepatic artery. Transcatheter arterial chemoembolization (TACE) was performed via the left hepatic artery. The patient developed supraumbilical skin rash with local tenderness on the following day. After supportive treatment by a dermatologist, the skin rash subsided gradually with sequelae of irregular skin surface and one small subcutaneous nodule. Skin biopsy of the lesion 1 year later showed fat necrosis with foreign body reaction and fibrosis. We discuss this rare complication of TACE and review the literature.

The vertebrate inner ear consists of a complex labyrinth of epithelial cells that is surrounded by a bony capsule. The molecular mechanisms coordinating the development of the membranous and bony labyrinths are largely unknown. Previously, using avian retrovirus encoding Noggin (RCAS-Noggin) or beads soaked with Noggin protein, we have shown that bone morphogenetic proteins (BMPs) are important for the development of the otic epithelium in the chicken inner ear. Here, using two additional recombinant avian retroviruses, dominant negative and constitutively active forms of BMP receptors IB (BMPRIB), we show that BMPs, possibly acting through BMPRIB, are important for otic capsule formation. We also show that Bmp2 is strongly expressed in the prospective semicircular canals starting from the canal outpouch stage, suggesting that BMP2 plays an important role in canal formation. In addition, by correlating expression patterns of Bmps, their receptors, and localization of phosphorylated R-Smad (phospho R-Smad) immunoreactivity, an indicator of BMP activation, we show that BMPs emanating from the otic epithelium influence chondrogenesis of the otic capsule including the cartilage surrounding the semicircular canals.

Organization of the inner ear into auditory and vestibular components is dependent on localized patterns of gene expression within the otic vesicle. Surrounding tissues are known to influence compartmentalization of the otic vesicle, yet the participating signals remain unclear. This study identifies Sonic hedgehog (Shh) secreted by the notochord and/or floor plate as a primary regulator of auditory cell fates within the mouse inner ear. Whereas otic induction proceeds normally in Shh(-/-) embryos, morphogenesis of the inner ear is greatly perturbed by midgestation. Ventral otic derivatives including the cochlear duct and cochleovestibular ganglia failed to develop in the absence of Shh. The origin of the inner ear defects in Shh(-/-) embryos could be traced back to alterations in the expression of a number of genes involved in cell fate specification including Pax2, Otx1, Otx2, Tbx1, and Ngn1. We further show that several of these genes are targets of Shh signaling given their ectopic activation in transgenic mice that misexpress Shh in the inner ear. Taken together, our data support a model whereby auditory cell fates in the otic vesicle are established by the direct action of Shh.

Transcatheter occlusion of a small or moderate patent ductus arteriosus (PDA) using Gianturco coils is safe and effective. Several investigators have reported using different techniques with variable results. To eliminate the serious potential complications of inadvertent coil embolization to undesirable sites and to improve control of the coil throughout the procedure, a modified snare-assisted method with approach from the main pulmonary artery was developed. However, articles discussing this technique are still limited. Herein, we report our successful experience of the snare-assisted technique to occlude small or moderate PDAs in three children aged from 1 to 8 years old. We achieved immediate total occlusion in all three cases with no complications. All three patients were discharged the next day and there was no coil dislodgement, left pulmonary stenosis, or residual shunting during the follow-up period of up to 3 years. This technique had the advantages of safety, optimal positioning, and improved stability of coil implantation.

A total of 110 central venous access procedures using Port-A catheters were performed in our center for pediatric patients with oncologic or hematologic diseases over a period of 6 years from November 1994 to June 2000. Spontaneous dislodgement of the outlet catheters was noted in two cases. Both dislodged catheters were located in the left pulmonary artery and were successfully retrieved via the right femoral vein by nonsurgical transvenous snares. Awareness of the possibility of spontaneous catheter dislodgement and embolization of Port-A catheters is important as prompt retrieval will prevent further serious complications. When the procedure is performed by an open surgical method, to reduce the possibility of spontaneous catheter dislodgement, the use of a guidewire is recommended while a Port-A catheter is being inserted into the subclavian vein.

Nor-1 belongs to the nur subfamily of nuclear receptor transcription factors. The precise role of Nor-1 in mammalian development has not been established. However, recent studies indicate a function for this transcription factor in oncogenesis and apoptosis. To examine the spatiotemporal expression pattern of Nor-1 and the developmental and physiological consequences of Nor-1 ablation, Nor-1-null mice were generated by insertion of the lacZ gene into the Nor-1 genomic locus. Disruption of the Nor-1 gene results in inner ear defects and partial bidirectional circling behavior. During early otic development, Nor-1 is expressed exclusively in the semicircular canal forming fusion plates. After formation of the membranous labyrinth, Nor-1 expression in the vestibule is limited to nonsensory epithelial cells localized at the inner edge of the semicircular canals and to the ampullary and utricular walls. In the absence of Nor-1, the vestibular walls fuse together as normal; however, the endolymphatic fluid space in the semicircular canals is diminished and the roof of the ampulla appears flattened due to defective continual proliferative growth of the semicircular canals.

Generating the diversity of cell types in the inner ear may require an interplay between regional compartmentalization and local cellular interactions. Recent evidence has come from gene targeting, lineage analysis, fate mapping and gene expression studies. Notch signaling and neurogenic gene regulation are involved in patterning or specification of sensory organs, ganglion cells and hair cell mechanoreceptors.

Portal vein thrombosis (PVT) in patients with hepatocellular carcinoma (HCC) has a poor impact on prognosis. Many of these tumors may cause intrahepatic and extrahepatic metastases. From January 1991 to December 1996, 41 unresectable HCC patients with PVT underwent transcatheter arterial chemoembolization (TACE) and external beam radiation therapy (EBRT) to the portion of PVT. The irradiated field, with a mean equivalent field size of 6.6 x 7.1 cm2, was localized and simulated by abdominal sonography, angiography and computed tomography. Radiation dose ranged from 36 to 66 Gy (mean dose: 51.4 Gy), in a daily fraction of 1.8 to 2 Gy. The response of EBRT was evaluated by abdominal sonography within 3 months of completion of EBRT. The response rates of the PVT after treatment were 39% for complete response (CR), 41% for partial response (PR), and 19% for no response (NR), respectively. The median overall survival time from start of radiotherapy was 10 months for all patients, 17 months for CR patients, 8 months for PR patients and 4 months for NR patients. By multivariate analysis, response of PVT resulted in a significant improvement in survival. (P = 0.001) There was no occurrence of severe complication of radiation-induced liver disease. The results obtained with combined treatment modality of EBRT and TACE in the treatment of HCC patients with PVT are encouraging.