Migiwa Asano

Rapid determination of glyphosate, glufosinate, bialaphos, and their major metabolites in serum by liquid chromatography―tandem mass spectrometry using hydrophilic interaction chromatography

We developed a simple and rapid method for the simultaneous determination of phosphorus-containing amino acid herbicides (glyphosate, glufosinate, bialaphos) and their major metabolites, aminomethylphosphonic acid (AMPA) and 3-methylphosphinicopropionic acid (MPPA), in human serum. Serum samples were filtrated through an ultrafiltration membrane to remove proteins. The filtrate was then washed with chloroform, and injected into a liquid chromatography-tandem mass spectrometry (LC-MS/MS) system. Chromatographic separation was achieved on a hydrophilic interaction chromatography (HILIC) column. Determination of the target herbicides and metabolites was successfully carried out without derivatization or solid phase extraction (SPE) cartridge clean-up. The recoveries of these compounds, added to human serum at 0.2μg/mL, ranged from 94% to 108%, and the relative standard deviations (RSDs) were within 5.9%. The limits of detection (LODs) were 0.01μg/mL for MPPA, 0.02μg/mL for AMPA, 0.03μg/mL for both glyphosate and glufosinate, and 0.07μg/mL for bialaphos, respectively.

Chemiluminescent Determination of Cholesterol Hydroperoxides in Human Erythrocyte Membrane

A method for separating, detecting, and quantifying cholesterol hydroperoxide (Ch-OOH) based on extraction, purification by solid-phase extraction cartridge, high-performance liquid chromatography with chemiluminescent detection (HPIC-CI), and liquid chromatography mass spectrometry has been developed for human erythrocyte membrane. We prepared standard compounds of the cholesterol 5α-, 7α-, and 7β-hydroperoxides (Ch 5α-OOH, Ch 7α-OOH, and Ch 7β-OOH). An octyl silica column with methanol/water/acetonitrile 89∶9∶2 (by vol) as eluent was used to determine Ch-OOH. HPLC-CL that incorporated cytochrome c and luminol as the post-column luminescent reagent was used. We also investigated the optimal assay conditions and how to prevent formation of artifact Ch-OOH. Analysis of erythrocyte membranes from seven healthy volunteers identified Ch 7α-OOH and Ch 7β-OOH, but not Ch 5α-OOH, as commonly occurring components. The respective mean concentrations of Ch 7α-OOH and Ch 7β-OOH were 2,5±1.6 and 5A±3.5 pmol/mL blood.

Association of FKBP5 gene haplotypes with completed suicide in the Japanese population

BACKGROUND The hypothalamus-pituitary-adrenal (HPA) axis is known to have a role in suicidal behaviors in patients with affective disorders. However, the incomplete overlapping of the genetic factors of suicidal behaviors and the genetic factors of affective disorders suggest that the genes associated with predisposition to suicidal behaviors and affective disorders are different. There is increasing evidence that genes regulating the HPA axis have effects on suicidal behaviors. To test this idea, we examined the association of three HPA axis-related genes (glucocorticoid receptor (NR3C1), mineralocorticoid receptors (NR3C2), and FK506 binding protein 5 (FKBP5)) with suicide. METHODS We selected 3 SNPs of the FKBP5 (rs3800373, rs1360780, and rs2395635), 2 SNPs of the NR3C1 (rs6196 and rs10052957), and 3 SNPs of the NR3C2 genes (rs5525, rs5522, and rs2070951) based on their frequency in the Japanese population. Using TaqMan probe assays, we determined these SNPs in 219 completed suicide victims and 228 age- and gender-matched healthy control subjects. RESULTS No significant differences in genotypic distribution or allelic frequency of any single SNPs between the completed suicide and control groups were observed. The distributions of TT, TC, and GT haplotypes of the FKBP5 gene (comprised of rs3800373 and rs1360780) between the completed suicide and control groups were significantly different (p<0.05 for each haplotype). The TC haplotype withstood correction for multiple comparisons (corrected p = 0.034). CONCLUSION Our results suggest that haplotypes in FKBP5 gene are associated with completed suicide. This finding needs to be confirmed using rigorous SNPs selection in a larger sample.

Alcoholic muscle disease and biomembrane perturbations (review)

Excessive alcohol ingestion is damaging and gives rise to a number of pathologies that influence nutritional status. Most organs of the body are affected such as the liver and gastrointestinal tract. However, skeletal muscle appears to be particularly susceptible, giving rise to the disease entity alcoholic myopathy. Alcoholic myopathy is far more common than overt liver disease such as cirrhosis or gastrointestinal tract pathologies. Alcohol myopathy is characterised by selective atrophy of Type II (anaerobic, white glycolic) muscle fibres: Type I (aerobic, red oxidative) muscle fibres are relatively protected. Affected patients have marked reductions in muscle mass and impaired muscle strength with subjective symptoms of cramps, myalgia and difficulty in gait. This affects 40-60% of chronic alcoholics (in contrast to cirrhosis, which only affects 15-20% of chronic alcohol misuers).Many, if not all, of these features of alcoholic myopathy can be reproduced in experimental animals, which are used to elucidate the pathological mechanisms responsible for the disease. However, membrane changes within these muscles are difficult to discern even under the normal light and electron microscope. Instead attention has focused on biochemical and other functional studies. In this review, we provide evidence from these models to show that alcohol-induced defects in the membrane occur, including the formation of acetaldehyde protein adducts and increases in sarcoplasmic-endoplasmic reticulum Ca(2+)-ATPase (protein and enzyme activity). Concomitant increases in cholesterol hydroperoxides and oxysterol also arise, possibly reflecting free radical-mediated damage to the membrane. Overall, changes within muscle membranes may reflect, contribute to, or initiate the disturbances in muscle function or reductions in muscle mass seen in alcoholic myopathy. Present evidence suggest that the changes in alcoholic muscle disease are not due to dietary deficiencies but rather the direct effect of ethanol or its ensuing metabolites.

Improvement of the accuracy of references in the Canadian Journal of Anaesthesia.

A previous study indicated that there were many citation errors in the Canadian Journal of Anaesthesia After this report, editors of the Journal requested any contributors, whose papers were accepted for publication, to verify the accuracy of reference citation by including a photocopy of the first page of each reference. The present study examined if the accuracy of the reference list had improved. We compared citation errors between volumes of 1990 and 1994. One hundred references from each year’s publication were randomly selected. After citations of nonjournal articles were excluded, the remaining 190 citations were carefully scrutinized. Authors’ names, article title, journal title, volume number, page numbers, and year were examined in each selected reference. A reference was deemed correct if each element of the citation was identical to its source. Of the examined references, 48% and 22% contained one or more errors in 1990 and 1994, respectively. Errors in the title and author field of citation were most common in the either of the two years, occurring in about 70% of the references which contained some errors. Citation errors in Canadian Journal of Anaesthesia were considerably improved after the request to verify citation accuracy. Although this check-system probably contributes to the improvement of accuracy of reference citation, the rate of citation errors remains high. We believe that contributors’efforts will enhance the value of the journal.RésuméUne étude antérieure a décelé, dans le Journal Canadien d’Anesthésie, un grand nombre d’erreurs bibliographiques. Par la suite, les éditeurs ont demandé aux collaborateurs de la revue dont les articles étaient acceptés pour publication, de vérifier l’exactitude des références et de fournir une photocopie de la première page de chacune des celles-ci. L’étude actuelle avait pour but de vérifier s’il en avait résulté une plus grande exactitude pour les références. Nous avons comparé les erreurs bibliographiques entre les volumes 1990 et 1994. Cent références pour chacune des années de publication ont été sélectionnées au hasard. Après l’élimination de celles qui ne provenaient pas de revues, les 190 références résiduelles ont été examinées soigneusement. Le nom des auteurs, le titre de l’article, le litre du journal, le numéro du volume, les numéros de pages, et l’année ont été vérifiés dans cet ordre. Une référence était jugée correcte si chacun des ses éléments était identique à sa source. Des références étudiées, 48% et 22% contenaient respectivement une erreur ou plus en 1990 et en 1994. Les erreurs de litres et d’auteurs étaient les plus fréquentes pour chacune des deux années et survenaient dans 70% des cos de références erronées. Les erreurs bibliographiques ont considérablement diminué dans le Journal Canadien d’Anesthésie après cette demande de vérification. Bien que ce système aide à améliorer la fidélité des références, le taux d’erreurs demeure élevé. Nous croyons que les efforts de la part des collaborateurs du Journal continueront d’améliorer son calibre.

Attenuation of the catecholamine response to tracheal intubation with oral clonidine in children

We conducted a prospective, randomized, double-blind, controlled clinical trial to examine (1) whether plasma catecholamine (CA) concentrations increased in response to tracheal intubation in children, and (2) the effects of clonidine on the CA responses. Sixty children (ASA physical status I) aged 7– 13 yr were allocated to one of three groups (n = 20 for each group): diazepam 0.4 mg · kg− 1 (active control), clonidine 2 μg · kg− 1, or clonidine 4 μg · kg− 1 po. These agents were administered 105 min before induction of anaesthesia followed by oral atropine 0.03 mg · kg− 1 given 60 min before anaesthesia which was induced with thiamylal 5 mg · kg− 1 and tracheal intubation was facilitated with vecuronium 0.2 mg · kg− 1. Laryngoscopy, lasting 30 sec, was attempted two minutes after administration of the induction agents. Serial values for blood pressure, heart rate, and venous plasma CA concentrations were compared among the three groups and with the respective preinduction measurements. Children receiving diazepam or clonidine 2 μg · kg− 1 showed remarkable increases in systolic and diastolic blood pressures, heart rate, and plasma CA concentrations in response to tracheal intubation (P < 0.05). The increases were similar for the two regimens. These haemodynamic and CA changes were smaller in children receiving clonidine 4 μg · kg− 1 (P < 0.05). The haemodynamic responses were positively correlated with the CA responses. These findings indicate that tracheal intubation following rapid sequence induction of anaesthesia in children provokes a reflex increase in sympathetic activity characterized by increased plasma CA concentrations, and that attenuation of the cardiovascular changes with a high oral dose of clonidine may be due to suppression of the increase in sympathetic activity evoked by the intubation.RésuméAu cours d’une étude contrôlée prospective randomisée et à double insu, les auteurs examinent 1) si la concentration plasmatique des catécholamine (CA) augmente chez les enfants en réponse à l’intubation de la trachée et 2) quels sont les effets de la clonidine sur la réponse des CA. Soixante enfants (ASA 1) âgés de 7 à 13 ans sont répartis en trois groupes (n = 20 pour chacun) pour recevoir: diazepam 0,4 mg · kg− 1 (contrôle actif), clonidine 2 μg · kg− 1, ou clonidine 4 μg · kg− 1 po. Ces agents sont administrés 105 min avant l’induction de l’anesthésie et sont suivis d’atropine 0,03 mg · kg− 1 po 60 min avant celleci. L’anesthésie est induite avec du thiamylal 5 mg · kg− 1 et la trachée est intubée après l’administration de vécuronium 0,2 mg · kg− 1. La laryngoscopie d’une durée de 30 sec est tentée deux min après l’administration des agents d’induction. Les mesures en série de la pression artérielle, de la fréquence cardiaque et de la concentration plasmatique des CA sont comparées entre les trois groupes et avec les valeurs respectives précédant l’induction. Les enfants qui reçoivent diazépam ou clonidine 2 μg· kg− 1 manifestent des augmentations importantes des pressions systolique et diastolique, de la fréquence cardiaque et de la concentration des CA en réponse à l’intubation de la trachée (P < 0,05). Les augmentations constatées ne diffèrent pas entre ces deux traitements. Les changements hémodynamiques et de CA sont moindres chez les enfants qui reçoivent de la clonidine 4 μg · kg− 1 (P < 0,05). La réponse hémodynamique a une corrélation positive avec celle des CA. Ces données montrent que l’intubation de la trachée après une induction à séquence rapide provoque chez l’enfant une augmentation réflexe de l’activité sympathique caractérisée par une augmentation des CA plasmatiques, et que l’atténuation de la réponse cardiovasculaire avec une dose orale élevée de clonidine peut être due à la suppression de l’augmentation de l’activité sympathique évoquée par l’intubation.

Free radicals in alcoholic myopathy: indices of damage and preventive studies

Chronic alcoholic myopathy affects up to two-thirds of all alcohol misusers and is characterized by selective atrophy of Type II (glycolytic, fast-twitch, anaerobic) fibers. In contrast, the Type I fibers (oxidative, slow-twitch, aerobic) are relatively protected. Alcohol increases the concentration of cholesterol hydroperoxides and malondialdehyde-protein adducts, though protein-carbonyl concentration levels do not appear to be overtly increased and may actually decrease in some studies. In alcoholics, plasma concentrations of α-tocopherol may be reduced in myopathic patients. However, α-tocopherol supplementation has failed to prevent either the loss of skeletal muscle protein or the reductions in protein synthesis in alcohol-dosed animals. The evidence for increased oxidative stress in alcohol-exposed skeletal muscle is thus inconsistent. Further work into the role of ROS in alcoholic myopathy is clearly warranted.Chronic alcoholic myopathy affects up to two-thirds of all alcohol misusers and is characterized by selective atrophy of Type II (glycolytic, fast-twitch, anaerobic) fibers. In contrast, the Type I fibers (oxidative, slow-twitch, aerobic) are relatively protected. Alcohol increases the concentration of cholesterol hydroperoxides and malondialdehyde-protein adducts, though protein-carbonyl concentration levels do not appear to be overtly increased and may actually decrease in some studies. In alcoholics, plasma concentrations of alpha-tocopherol may be reduced in myopathic patients. However, alpha-tocopherol supplementation has failed to prevent either the loss of skeletal muscle protein or the reductions in protein synthesis in alcohol-dosed animals. The evidence for increased oxidative stress in alcohol-exposed skeletal muscle is thus inconsistent. Further work into the role of ROS in alcoholic myopathy is clearly warranted.

Lansoprazole reduces preoperative gastric fluid acidity and volume in children.

The purpose of this study was to explore the efficacy of lansoprazole, a proton pump inhibitor, in reducing the acidity and volume of gastric aspirate in children immediately following the induction of anaesthesia. One hundred healthy in-patients aged 3–11 yr undergoing elective surgery were randomly allocated to four groups (n = 25 each): lansoprazole-lansoprazole, placebo-placebo, placebo-lansoprazole, and lansoprazole-placebo. For each treatment regimen, the first medication was administered at 9:00 pm on the night before surgery and the second at 5:30 am on the morning of the day of surgery (three hours preoperatively). The dose of lansoprazole was 30 mg (approximately 1.4 mg · kg−1 mean). Children were offered 10 ml · kg−1 apple juice three hours before induction of anaesthesia. After induction of anaesthesia and tracheal intubation, gastric fluid was aspirated through a large-bore, multiorifice orogastric tube and analyzed for pH and total fluid volume. Lansoprazole increased gastric fluid pH and decreased gastric fluid volume regardless of whether it was administered before or after placebo. Two consecutive doses of lansoprazole was the most effective means of increasing the pH and reducing the volume of gastric aspirate; in this group, there were no subjects with gastric aspirate volume >0.4 ml · kg−1 and pH <2.5. Oral lansoprazole, at least 30 mg, given on the night before surgery or on the morning of surgery will improve the gastric environment at the time of induction of paediatric anaesthesia. The most effective regimen was two doses (at bedtime and on the morning) of lansoprazole.RésuméCette étude a pour objectif l’évaluation de l’efficacité du lansoprazole, un inhibiteur de la pompe à proton, sur la réduction de l’acidité et du volume du contenu gastriques chez l’enfant mesurés immédiatement après l’induction de l’anesthésie. Cent sujets bien portants âgés de 3 à 11 ans hospitalisés pour une chirurgie non urgente sont répartis au hasard en quatre groupes (n = 25 par groupe) de la façon suivante: lansaprazole-lansaprazole, placebo-placebo, placebo-lansaprazole, et lansaprazole-placebo. Dans tous les cas, la première médication est administrée à 21:00 la veille de la chirurgie et la deuxième à 5:30 le matin de la chirurgie (trois heures avant l’intervention). La dose de lansaprazole est de 30 mg (environ 1,4 mg · kg−1 en moyenne). On offre aux enfant 10 ml · kg−1 de jus de pomme trois heures avant l’induction de l’anesthésie. Après l’induction et l’intubation de la trachée, le liquide gastrique est aspiré avec une sonde gastrique de gros calibre à plusieurs orifices et on analyse son pH et son volume. La lansaprazole augmente le pH et diminue le volume du contenu gastrique qu’il soit administré avant ou après le placebo. Le moyen le plus efficace pour augmenter le pH et diminuer le volume est d’administrer deux doses de lansaprazole successives: dans ce groupe, le volume du contenu gastrique est toujours inférieur à 0,4 ml · kg−1 et le pH supérieur à 2,5. Le lansaprazole, à la dose de 30 mg, administré par la bouche la veille ou le matin de la chirurgie améliore les paramètres gastrique au moment de l’induction de l’anesthésie. La méthode la plus sûre est constituée par l’administration de deux doses de lansaprazole, au coucher et le matin.

TPH2 is not a susceptibility gene for suicide in Japanese population

BACKGROUND Serotonergic systems mediate a control of aggression and/or impulsivity in human and are suggested to be involved in suicidal behavior. The newly identified neuronal tryptophan hydroxylase isoform 2 (TPH2), the rate-limiting enzyme in serotonin synthesis, represents a prime candidate in numerous genetic association analyses of suicidal behavior; however, the results are still inconclusive. The discrepancy may result from the heterogeneity of pathogenesis of suicidal behavior and/or methodological mismatches. We, therefore, attempted to replicate the association of TPH2 gene with suicide using a case-control study of 234 completed suicides and 260 control subjects in Japanese population. METHODS We genotyped 15 tagging-single nucleotide polymorphisms (SNPs) including 4 SNPs, which were previously reported to be associated with suicidal behavior, using the TaqMan probe assays and the polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) method. RESULTS We found no significant differences in genotypic distributions (uncorrected p=0.06-0.98) or allelic frequencies (uncorrected p=0.09-0.95) of the fifteen SNPs between the completed suicides and control groups. Haplotypes constructed with these SNPs were also not associated with suicide (uncorrected p=0.03-0.96 and corrected p=0.20-1.00). Even when we took sex and suicidal methods (violent or non-violent) into account for the analyses, no significant differences in genotypic distributions, allelic/haplotypic frequencies were found in the two groups. CONCLUSION Our results suggest that the common SNPs and haplotypes of the TPH2 gene are unlikely to contribute to the genetic susceptibility to suicidal behavior in Japanese population.

Serial review: alcohol, oxidative stress, and cell injuryFree radicals in alcoholic myopathy: indices of damage and preventive studies1

Chronic alcoholic myopathy affects up to two-thirds of all alcohol misusers and is characterized by selective atrophy of Type II (glycolytic, fast-twitch, anaerobic) fibers. In contrast, the Type I fibers (oxidative, slow-twitch, aerobic) are relatively protected. Alcohol increases the concentration of cholesterol hydroperoxides and malondialdehyde-protein adducts, though protein-carbonyl concentration levels do not appear to be overtly increased and may actually decrease in some studies. In alcoholics, plasma concentrations of α-tocopherol may be reduced in myopathic patients. However, α-tocopherol supplementation has failed to prevent either the loss of skeletal muscle protein or the reductions in protein synthesis in alcohol-dosed animals. The evidence for increased oxidative stress in alcohol-exposed skeletal muscle is thus inconsistent. Further work into the role of ROS in alcoholic myopathy is clearly warranted.Chronic alcoholic myopathy affects up to two-thirds of all alcohol misusers and is characterized by selective atrophy of Type II (glycolytic, fast-twitch, anaerobic) fibers. In contrast, the Type I fibers (oxidative, slow-twitch, aerobic) are relatively protected. Alcohol increases the concentration of cholesterol hydroperoxides and malondialdehyde-protein adducts, though protein-carbonyl concentration levels do not appear to be overtly increased and may actually decrease in some studies. In alcoholics, plasma concentrations of alpha-tocopherol may be reduced in myopathic patients. However, alpha-tocopherol supplementation has failed to prevent either the loss of skeletal muscle protein or the reductions in protein synthesis in alcohol-dosed animals. The evidence for increased oxidative stress in alcohol-exposed skeletal muscle is thus inconsistent. Further work into the role of ROS in alcoholic myopathy is clearly warranted.