Association of CRHR1 and CRHR2 with major depressive disorder and panic disorder in a Japanese population.

Major depressive disorder (MDD) and panic disorder (PD) are common and disabling medical disorders with stress and genetic components. Dysregulation of the stress response of the hypothalamic-pituitary-adrenal axis, including the corticotrophin-releasing hormone (CRH) signaling via primary receptors (CRHR1 and CRHR2), is considered to play a major role for onset and recurrence in MDD and PD. To confirm the association of CRHR1 and CRHR2 with MDD and PD, we investigated 12 single nucleotide polymorphisms (SNPs) (rs4076452, rs7209436, rs110402, rs242924, rs242940, and rs173365 for CRHR1 and rs4722999, rs3779250, rs2267710, rs1076292, rs2284217, and rs226771 for CRHR2) in MDD patients (n = 173), PD patients (n = 180), and healthy controls (n = 285). The SNP rs110402 and rs242924 in the CRHR1 gene and the rs3779250 in the CRHR2 gene were associated with MDD. The SNP rs242924 in the CRHR1 gene was also associated with PD. The T-A-T-G-G haplotype consisting of rs7209436 and rs173365 in CRHR1 was positively associated with MDD. The T-A haplotype consisting of rs7209436 and rs110402 in CRHR1 was positively associated with MDD. The C-C haplotype consisting of rs4722999 and rs37790 in CRHR1 was associated with PD. These results provide support for an association of CRHR1 and CRHR2 with MDD and PD.

[Measurement of the transport activities of bile salt export pump using chemiluminescence detection method].

Monovalent bile acids, such as taurine- and glycine-conjugated bile acids, are excreted into bile by bile salt export pumps (BSEP, ABCB11). Human BSEP (hBSEP) is physiologically important because it was identified as the gene responsible for the genetic disease: progressive familial intrahepatic cholestasis type 2 (PFIC-2). The evaluation of the inhibitory effect of hBSEP transport activity provides significant information for predicting toxic potential in the early phase of drug development. The role and function of hBSEP have been investigated by the examination of the ATP-dependent transport of radioactive isotopically (RI)-labeled bile acid such as a tritium labeled taurocholic acid, in membrane vesicles obtained from hBSEP-expressing cells. The chemiluminescence detection method using 3alpha-hydroxysteroid dehydrogenase (3alpha-HSD) had been developed for a simple analysis of bile acids in human biological fluids. This method is extremely sensitive and it may be applicable for the measurements of bile acid transport activities by hBSEP vesicles without using RI-labeled bile acid. The present paper deals with an application of the chemiluminescence detection method using 3alpha-HSD with enzyme cycling method to the measurement of ATP-dependent transport activities of taurocholic acid (T-CA) in membrane vesicles obtained from hBSEP-expressing Sf9 cells. Calibration curves for T-CA was linear over the range from 10 to 400 pmol/ml. The values of the kinetic parameters for hBSEP vesicles obtained by the chemiluminescence detection method were comparable with the values of that obtained by liquid chromatography-mass spectrometry method. This assay method was highly useful for the measurements of bile acid transport activities.

Incidence of anesthesia-related medication errors over a 15-year period in a university hospital.

To clarify the incidence of anesthesia-related medication errors in Kyushu University Hospital, a retrospective analysis of anesthesia-related incidents from 1993 to 2007 was conducted based on the "Investigation of anesthesia-related medication incidents" by the Japanese Society of Anesthesiologists. Out of a total of 64,285 anesthesia cases, drug errors occurred in 50 cases (0.078%), but none of the incidents led to serious sequelae. Wrong medication was the most common type of drug error (48%), followed by overdose (38%), underdose (4%), omission (2%), and incorrect administration route (8%). The most commonly involved drugs were opioids, cardiac stimulants, and vasopressors. Syringe swap was the leading cause of wrong medication, accounting for 42%, drug ampoule swap occurred in 33%, and the wrong choice of drug was made in 17%. The first, second, and third most frequent causes of overdose involved a misunderstanding or preconception of the dose (53%), pump misuse (21%), and dilution error (5%). The error frequency did not decrease over the 15-year period. The responsible anesthesiologists were most likely to be doctors with a little experience. To reduce anesthesia-related medication errors, improvements of protocols for handling medication and instruction, and an improved education system for the anesthesia trainees are essential.

Measurement of transport activities of bile acids in human multidrug resistance-associated protein 3 using liquid chromatography-tandem mass spectrometry.

A method has been developed for the measurement of transport activities in membrane vesicles obtained from human multidrug resistance-associated protein 3-expressing Sf9 cells for 1beta-hydroxy-, 6alpha-hydroxy- and unsaturated bile acids by high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry. Calibration curves for the bile acids were linear over the range of 10 to 2000 pmol/mL, and the detection limit was less than 2 pmol/mL for all bile acids using selected reaction monitoring analysis. The method was applied to measurements of adenosine triphosphate-dependent transport activities of the membrane vesicles for the above-mentioned hydroxylated and unsaturated bile acids. The present study demonstrated that the human multidrug resistance-associated protein 3 vesicles accepted 1beta-, 6alpha-hydroxylated and unsaturated bile acids along with common bile acids, such as glycocholic acid and taurolithocholic acid 3-sulfate. The developed method is useful for measurements of bile acid transport activities.

Measurement of bile salt export pump transport activities using a fluorescent bile acid derivative.

A novel fluorescent bile acid derivative, 4-N,N-dimethylaminosulfonyl-2,1,3-benzoxadiazole- conjugated bile acid was synthesized as a probe to develop a rapid screening method for function analysis of bile salt export pump (BSEP, ABCB 11). The transport properties of the synthetic fluorescent bile acid derivative in membrane vesicles obtained from hBSEP-expressing Sf9 cells were examined using the liquid chromatography-electrospray ionization-mass spectrometry method. The Michaelis-Menten constant and maximum uptake rate for the synthetic fluorescent bile acid derivative by hBSEP were 23.1+/-1.6 microM and 623.2+/-22.4 pmol/min/mg protein, respectively. These kinetic parameters of the synthetic fluorescent bile acid derivative were comparable with those of an unlabeled bile acid, taurocholic acid. Moreover, we examined inhibitory effects of various drugs on hBSEP-mediated uptake of the fluorescent bile acid derivative using a fluorescence detection method. The relative uptake activities (percent of control) for the fluorescent bile acid derivative in the presence of an inhibitor were in accordance with previous findings using (3)H-labeled taurocholic acid. Our results suggest that the synthetic fluorescent bile acid derivative may be useful for evaluation of the inhibitory effects of various drugs on hBSEP-mediated uptake.

Measurement of the transport activities of bile salt export pump using LC-MS.

The high performance liquid chromatography-electrospray ionization-mass spectrometry method has been applied to the measurement of bile acid transport activities in membrane vesicles obtained from a human bile salt export pump expressing Sf9 cells. The amounts of bile acids transported using the human bile salt export pump expressing Sf9 cells were determined using liquid chromatography-electrospray ionization-mass spectrometry method and the values of the kinetic parameters were determined to be comparable with those obtained using radioisotope-labeled substrates. The developed method was highly useful for the measurements of bile acid transport activities.