The retrosigmoid approach using auditory monitoring for a small AN can accomplish total tumour removal with an excellent hearing preservation rate. CNAP provides reliable auditory monitoring more frequently than ABR, reflects the intraoperative auditory function almost in real-time, predicts postoperative hearing with excellent sensitivity and specificity, and is more useful for monitoring in the removal of small AN with hearing preservation.

Our findings suggest that TRPV1 is involved in bladder overactivity via afferent nerve activation but it is not associated with normal voiding function. A TRPV1 antagonist would be a useful drug for bladder overactivity with a different pharmacological profile than antimuscarinic agents.

We previously reported emergence and disappearance of circadian molecular oscillations during differentiation of mouse embryonic stem (ES) cells and reprogramming of differentiated cells, respectively. Here we present a robust and stringent in vitro circadian clock formation assay that recapitulates in vivo circadian phenotypes. This assay system first confirmed that a mutant ES cell line lacking Casein Kinase I delta (CKIδ) induced ∼3 hours longer period-length of circadian rhythm than the wild type, which was compatible with recently reported results using CKIδ null mice. In addition, this assay system also revealed that a Casein Kinase 2 alpha subunit (CK2α) homozygous mutant ES cell line developed significantly longer (about 2.5 hours) periods of circadian clock oscillations after in vitro or in vivo differentiation. Moreover, revertant ES cell lines in which mutagenic vector sequences were deleted showed nearly wild type periods after differentiation, indicating that the abnormal circadian period of the mutant ES cell line originated from the mutation in the CK2α gene. Since CK2α deficient mice are embryonic lethal, this in vitro assay system represents the genetic evidence showing an essential role of CK2α in the mammalian circadian clock. This assay was successfully applied for the phenotype analysis of homozygous mutant ES cells, demonstrating that an ES cell-based in vitro assay is available for circadian genetic screening.

Retinoylation (acylation of proteins by retinoic acid) is considered as one mechanism of retinoic acid (RA) action occurring in cells in vitro and in vivo. Previously, our studies showed that in rat tissues the formation of retinoyl-CoA from RA, the first step of retinoylation, required ATP, CoA and MgCl(2). In the current study, we examined whether the transfer of retinoyl-CoA into proteins, the second step of retinoylation, occurs in rat tissues. [(3)H]-Labeled-retinoyl-CoA bound covalently to proteins in rat liver, kidney, testis, and brain. The levels of incorporation of retinoyl-CoA into proteins were higher in vitamin A-deficient rats than in normal ones. The formation of retinoylated proteins depended on the incubation time, and the concentrations of retinoyl-CoA and homogenate. The reaction was suppressed by fatty acyl-CoAs and palmitic acid, but not by arachidonic acid. The Vmax and Km values for retinoyl-CoA in the formation of retinoylated proteins using a crude liver extract were estimated to be 2,597.3 pmol/min/mg protein and 9.5 x 10(-5) M, respectively. Retinoylated proteins formed from retinoyl-CoA, including a 17 kDa protein exhibiting high radioactivity, disappeared in the presence of 2-mercaptoethanol, indicating that RA was linked to the proteins through a thioester bond. These results demonstrate that retinoylation in rat tissues occurs via retinoyl-CoA formed from RA. This process may play a significant physiological role in cells.

Retinoylation (retinoic acid acylation) is a posttranslational modification of proteins occurring in a variety of cell types in vitro and in tissues in vivo. The widespread occurrence of retinoylation suggests that it may play a role in many effects of retinoic acid (RA) on cells. One metabolic pathway for retinoylation involves the intermediate formation of retinoyl-CoA and subsequent transfer and covalent binding of the retinoyl moiety to protein. However, such reactions are not well known. To gain further insight into retinoylation, we studied the synthesis of retinoyl-CoA, the first step in this multi-stage process. The formation of [(3)H]-retinoyl-CoA was determined in incubation mixtures containing rat liver extract, [(3)H]-RA, ATP, CoA, and MgCl(2). No retinoyl-CoA was formed in the presence of boiled extract, or in the absence of ATP, CoA, or MgCl(2) (a divalent cation). A greater amount of retinoyl-CoA was obtained from microsomal fractions of rat liver than from other subfractions. The presence of retinoyl-CoA was also detected in extracts prepared from rat testis, kidney, brain, spleen, and pancreas. The level of retinoylation in various tissue extracts was related directly to the amount of retinoyl-CoA formed. V(max) and K(m) values for RA in the formation of liver retinoyl-CoA were estimated to be 1.0 x 10(-4) micromol/min/mg protein and 24 nM, respectively. Synthesis of retinoyl-CoA was suppressed by fatty acids and fatty acyl-CoAs. These results indicate that ATP-dependent generation of retinoyl-CoA occurs in rat tissues and may play a significant physiological role in RA actions mediated by retinoylation.