Syntheses and anti-inflammatory activity of azamollugin derivatives.

Oxomollugin (2) is a degradation product of mollugin (1) and a potent inhibitor of NO-production including nuclear factor kappa B signals. In our endeavor to develop a potent anti-inflammatory compound, we synthesized several aza-derivatives of oxomollugin (2) and evaluated their NO-production inhibitory activity. Azamollugin (3) showed a potent inhibitory activity, and its activity (IC50 0.34µM) was proved to be more potent than that of oxomollugin (2, IC50 1.3µM).

Improvement in a post-stroke pediatric patient with hemiplegia: Use of a hand-arm bimanual intensive therapy with hybrid assistive limb.

BACKGROUND: Pediatric stroke is a rare medical condition that often leads to long-lasting motor and cognitive impairments. Although therapies for adults after a stroke are well described, treatments for motor deficits following a pediatric stroke are yet to be investigated. We report a case of pediatric stroke in the chronic phase, in which a combination of novel treatments resulted in a significant improvement in physical function. CASE REPORT: A seven-year-old girl with a left hemispheric cerebral infarction lost almost all right upper extremity motor function. Following onabotulinumtoxinA treatment, she underwent hand-arm bimanual intensive therapy augmented with a hybrid assistive limb for 90 h over 15 days. Evaluation after the training revealed significant improvements in physical function, daily activities, and occupational performance. CONCLUSIONS: This report highlights the importance of innovative combinations of techniques in the treatment of pediatric stroke.

Oxomollugin, an oxidized substance in mollugin, inhibited LPS-induced NF-κB activation via the suppressive effects on essential activation factors of TLR4 signaling.

Oxomollugin is a degraded product of mollugin and was found to be an active compound that inhibits LPS-induced NF-κB activation. In this study, we investigated the inhibitory activity of oxomollugin, focusing on TLR4 signaling pathway, resulting in NF-κB activation. Oxomollugin inhibited the LPS-induced association of essential factors for initial activation of TLR4 signaling, MyD88, IRAK4 and TRAF6. Furthermore, oxomollugin showed suppressive effects on LPS-induced modification of IRAK1, IRAK2 and TRAF6, LPS-induced association of TRAF6-TAK1/TAB2, and followed by IKKα/ß phosphorylation, which critical in signal transduction leading to LPS-induced NF-κB activation. The consistent results suggested that oxomollugin inhibits LPS-induced NF-κB activation via the suppression against signal transduction in TLR4 signaling pathway.The activities of oxomollugin reported in this study provides a deeper understanding on biological activity of mollugin derivatives as anti-inflammatory compounds.

[Reliability of the measurement of stature in subjects with severe motor and intellectual disabilities].

OBJECTIVE: The objective of this study was to examine the reliability of the measurement of stature in individuals with severe motor and intellectual disabilities. METHODS: Using a stratified sampling method 12 subjects (mean age of 28.7 +/- 11.5), were selected from 73 subjects with severe motor and intellectual disabilities (mean age of 37.1 +/- 15.0). Each subjects'stature was measured using two measurement methods. One measured the tibia length (TL-method) and, the other measured the whole body by measuring three sections. (Division-method). 3 examiners measured all subjects using the TL-method and Division-method two times repeatedly. In addition, one examiner measured all subjects within two weeks following the initial measurement. Intra-rater reliability was calculated from single and two times measurements using each method. The Inter-rater reliability was calculated using measurement results from 2 or 3 examiners in both single and two times measurements. The correlation between values measured by the TL-method and the Division-method was calculated using Spearman rank correlation coefficient. RESULTS: The TL-method and Division-method had good intra-rater reliability (ICC>0.90, 95%CI>0.80) and good inter-rater reliability (ICC>0.90, 95%CI>0.70) in both measurement methods. For both measurement methods, inter-rater reliability was more preferable when 3 examiners measured two times repeatedly (ICC>0.90, 95%CI>0.90). There was good correlation between values measured by TL-method and the Division-method (r=0.83). CONCLUSIONS: Both TL-method and Division-method had good reliability. However, the TL-method could be considered a more useful measurement method as it can be completed more easily and in a short period of time.

[The effects of lamotrigine on refractory epilepsies in persons with severe motor and intellectual disabilities].

OBJECTIVE: We studied the effects and tolerability of lamotrigine (LTG) on refractory epilepsies in persons with severe motor and intellectual disabilities (SMID). METHODS: We introduced LTG to 40 subjects all who were severely retarded and suffered from symptomatic epilepsies consisting of six cases of generalized epilepsy (GE), 33 of partial epilepsy (PE), and one of unclassified epilepsy (according to the 1989 ILAE classification standards). We evaluated the efficacy of LTG in seizure reduction and its tolerability for persons with SMID. RESULTS: Following treatment, 83% of GE and 42% of PE cases showed a 50% reduction in seizure frequency. From the seizures observed in our study cohort, a response rate (RR) with an over 50% reduction was found in 54% of secondarily generalized tonic-clonic (n = 24), 30% of complex partial (n = 18), and 25% of myoclonic seizures (n = 16). As for adverse effects, nine subjects (22.5%) experienced sleep problems, six subjects (15%) experienced excitement mainly during night-time, and four cases each (10%) experienced involuntary movements and sleepiness. As for additional effects on patients' quality of life (QOL), even in poor responder (< 50% RR) patients, LTG improved "alertness" in six cases, improved "activity of daily life" in four cases, and helped to diminish "appetite loss" in three cases. CONCLUSIONS: LTG therapy for epileptic conditions in persons with SMID has potential for not only reducing seizure frequency, but also for improving QOL.

Review of Cognitive Characteristics of Autism Spectrum Disorder Using Performance on Six Subtests on Four Versions of the Wechsler Intelligence Scale for Children.

This study was a systematic review of research using the Wechsler Intelligence Scale for Children (WISC) with Autism Spectrum Disorder (ASD) to examine cognitive characteristics of children with ASD beyond the impact of revisions based on WISC and diagnostic criteria changes. The classic "islets of ability" was found in individuals with full-scale IQs < 100. The "right-descending profiles" were observed among high IQ score individuals. High levels on the Block Design and low Coding levels were consistently found regardless of the variation in intellectual functioning or diagnosis. This review identified patterns of cognitive characteristics in ASD individuals using empirical data that researchers may have previously been aware of, based on their experiences, owing to the increased prevalence of ASD.

Genetic Analysis of UGT1A1 Polymorphisms Using Preserved Dried Umbilical Cord for Assessing the Potential of Neonatal Jaundice as a Risk Factor for Autism Spectrum Disorder in Children.

Neonatal jaundice has been suggested as a perinatal risk factor for autism spectrum disorder (ASD). We examined UGT1A1 polymorphisms to assess the potential of neonatal jaundice as a risk factor for ASD in children by using DNA extracted from preserved umbilical cord. In total, 79 children with ASD were genotyped for UGT1A1*28 (c.-41-40dup), UGT1A1*6 (c.211 G > A), and UGT1A1*27 (c.686 C > A). The allele frequency of UGT1A1*6 (OR = 1.34, p = 0.26) and UGT1A1*28 (OR = 0.80, p = 0.54) and the prevalence of UGT1A1*28/*6 diplotypes did not differ significantly from those in the control population. No UGT1A1*27 allele was detected in the subjects. ASD symptom assessment scores were not associated with UGT1A1*28/*6/*27 genotypes or UGT1A1*28/*6 diplotypes. These results suggest that neonatal jaundice is not significantly associated with ASD.

Effect of CPS14217C>A genotype on valproic-acid-induced hyperammonemia.

BACKGROUND: In order to clarify the factors causing hyperammonemia and to predict occurrences during treatment with valproic acid (VPA), we investigated the effect of the genetic polymorphism of carbamoyl-phosphate synthase 1 (CPS14217C>A) on susceptibility of hyperammonemia, together with the effect of coadministration of other anticonvulsants. METHODS: Seventy-nine patients with epilepsy were enrolled, and five of them had hyperammonemia. Univariate and multivariate logistic regression analyses were performed. RESULTS: The aspartate aminotransferase level in the patients with hyperammonemia was significantly higher than that in those without hyperammonemia. The risk of hyperammonemia was significantly influenced by the number of anticonvulsants concomitantly administered with VPA. Also, the distribution of the CPS14217C>A genotype differed depending on whether the patients had hyperammonemia or not. No significant effects of CPS14217 genotypes and the number of anticonvulsants coadministered with VPA on the serum concentrations of VPA were observed. The multivariate logistic regression analysis showed that the concomitant administration of two or more anticonvulsants with VPA and the heterozygous or homozygous carrier state of the A allele of the CPS14217C>A polymorphism were independent risk factors for developing hyperammonemia. CONCLUSIONS: These findings suggested that in epileptic patients undergoing VPA therapy, CPS14217A polymorphism and the number of coadministered anticonvulsants would be considered as risk factors for hyperammonemia, even if the serum VPA concentrations were controlled.

[Determination of sodium chlorite in processed herring roe by ion chromatography with a conductivity detector].

An analytical method for residual sodium chlorite in several kinds of processed herring roe treated with sodium chlorite was studied. Sodium chlorite was extracted with 9 mmol/L sodium carbonate. After centrifugation, the supernatant was filtered through a 0.2 microm nylon filter. The filtrate was deproteinized by ultrafiltration and chloride ion was removed with an On-Guard Ag cartridge column. The eluate was subjected to conductivity detector-ion chromatography. Recoveries of sodium chlorite from herring roe spiked at the level of 5 mg/kg were 88 +/- 3.7% (n = 5, CV 4.2%). The method had a quantitation limit of 5 mg/kg for processed herring roes.

Relationship between stature and tibial length for children with moderate-to-severe cerebral palsy.

OBJECTIVE: To derive the equation for estimating stature, based on tibial length, for children with moderate-to-severe cerebral palsy (CP) and lower limb joint contracture or scoliosis. METHODS: The participants (3-12-years-old) included 50 children with moderate-to-severe CP (mean age, 8.3±2.4 years) and 38 typically developed (TD) children (mean age, 7.5±2.6 years). Thirty-four (68%) of the children with CP had a gross motor function classification system level of V. Furthermore, 40 (80%) had definite lower limb joint contracture or scoliosis. The stature and the tibial length measurements of all participants were determined. Regression equations to estimate stature, based on tibial lengths, were determined for both TD children and children with CP. Moreover, regression equations defining the relationship between tibial length and age were compared between the two groups of children, using multiple regression analysis. RESULTS: The regression equations for estimating stature, based on tibial length, were stature=tibial length×3.25+34.45 [cm], R(2)=0.91 (TD children), and stature=tibial length×3.42+31.82 [cm], R(2)=0.81 (CP children). In children with CP, tibial lengths were significantly shorter than those in similarly aged TD children. CONCLUSION: The stature of children with moderate-to-severe CP can be estimated from their tibial lengths, regardless of the presence of joint contracture or scoliosis. The tibial length may be a proxy for estimating stature during the growth assessment of children with moderate-to-severe CP.

Oxomollugin, a potential inhibitor of lipopolysaccharide-induced nitric oxide production including nuclear factor kappa B signals.

Mollugin, a naphthoquinone derivative, was reported to possess various biological activities such as anti-inflammatory and anti-tumor activity. Mollugin isolated from Rubia tinctorum roots inhibited lipopolysaccharide-induced nitric oxide (NO) production in RAW264.7 macrophages. However, mollugin synthesized for further investigation of its anti-inflammatory mechanism showed weak activity in addition to unstable assay results. From the result of analysis on a degradation product of mollugin, oxomollugin was found to be the main active substance of mollugin degradation, showing a potent inhibitory activity on NO-production including nuclear factor kappa B signals.

[Improved sensitive determination method for bromate in bread].

An effective clean-up procedure was developed to determine trace levels of bromate in bread by high-performance liquid chromatography with post-column flow reactor detection. Bromate was extracted from bread with deionized pure water. After centrifugation, the supernatant was filtered through a paper filter. The filtrate was filtered through a 0.2 micron nylon filter and chloride ion was removed by an IC-SP M Ag cartridge column or On-Guard Ag cartridge column. The eluate was applied to an Oasis MAX anion exchange cartridge column. The column was washed with 20% acetic acid and water. Bromate was then eluted with 0.5% sodium nitrate solution. The eluate was determined by HPLC with post column flow reactor detection. The method had a quantitation limit of 2 ng/g in bread products. Recoveries of bromate from bread ranged from 68 to 72% at a spiked bromate level of 2-10 ng/g.

Structure of acid-stable carmine.

Acid-stable carmine has recently been distributed in the U.S. market because of its good acid stability, but it is not permitted in Japan. We analyzed and determined the structure of the major pigment in acid-stable carmine, in order to establish an analytical method for it. Carminic acid was transformed into a different type of pigment, named acid-stable carmine, through amination when heated in ammonia solution. The features of the structure were clarified using a model compound, purpurin, in which the orientation of hydroxyl groups on the A ring of the anthraquinone skeleton is the same as that of carminic acid. By spectroscopic means and the synthesis of acid-stable carmine and purpurin derivatives, the structure of the major pigment in acid-stable carmine was established as 4-aminocarminic acid, a novel compound.

Estimation of concentrations of antifungal agents allowed as food additives in foods and their daily intake based on official inspection results in Japan in fiscal year 1998.

The mean concentrations and daily intake of four antifungal agents were estimated based on the results of an analysis of 7,005 samples of food obtained in official inspections by Japanese local governments in fiscal year 1998. The mean concentration of diphenyl was 0.0004% of the allowable limit, and those of imazalil, o-phenylphenol, and thiabendazole were 14.0%, 3.5%, and 5.7%, respectively. The daily intakes of these antifungal agents per person, estimated from their concentrations and the daily consumption of the foods, were 0.000326, 1.89, 11.5, and 23.3 micrograms, respectively, and assuming a body weight of 50 kg, the amounts of these antifungal agents consumed were 0.000013%, 0.15%, 0.12%, and 0.47% of the acceptable daily intake, respectively. These values are similar to the values obtained on the basis of the results of the official inspection in fiscal years 1994 and 1996, except that the amount of diphenyl is much lower (1/100).

[Identification of acid-stable carmine in imported apple syrup product].

An unknown red pigment was purified from an apple syrup product imported from Canada, using a DIAION HP-20 column with methanol as the eluent. By spectroscopic means and chemical synthesis, the isolated pigment was identified as 4-aminocarminic acid, which is the major pigment of acid-stable carmine (a red colorant illegal in Japan). In addition, HPLC and TLC methods were proposed to detect this illegal colorant. While the color of carminic acid changed from yellow to red in the pH range of McIlvaine buffer (3.0-7.0), the color of 4-aminocarminic acid was always red, and also the ultraviolet/visible (UV/Vis) spectra did not change. These characteristics are useful to distinguish 4-aminocarminic acid from carminic acid.

Specific determination of bromate in bread by ion chromatography with ICP-MS.

A sensitive method for detecting bromate in bread by ion chromatography with inductively-coupled plasma mass spectrometry (IC/ICP-MS) was developed. Bromate was extracted from bread with water. The clean-up procedure included a 0.2 micron filter, a C18 cartridge for defatting, a silver cartridge to remove halogen anions, a centrifugal ultrafiltration unit to remove proteins, and a cation-exchange cartridge to remove silver ions. A 500 microL sample solution was applied to IC/ICP-MS. The detection limit and the quantitation limit of bromate in the solution were 0.3 ng/mL and 1.0 ng/mL, expressed as HBrO3, respectively, which corresponded to 2 ng/g and 5 ng/g, respectively, in bread. Recovery of bromate was about 90%, and the CV was about 2%. Based on the detection limit in solution and recovery from bread, the detection limit of bromate in bread was estimated to be 2 ng/g.

[Determination of six p-hydroxybenzoic acid esters in laver (nori) by HPLC].

A rapid and simple method was developed for simultaneous determination of methyl, ethyl, isopropyl, npropyl, isobuthyl and n-buthyl p-hydroxybenzoic acid esters (PHBA-Es) in laver by HPLC. Six PHBA-Es were extracted from laver with n-hexane-ethyl acetate (1:1) by shaking. The extract was evaporated. The residue was dissolved in methyl alcohol and determined by HPLC. Recoveries of six PHBA-Es spiked in laver were 93.6-101.2% at the level of 2 micrograms/g.

[Improvement of carbon monoxide analysis in fish meat].

Carbon monoxide (CO) treatment of fish meat of tuna, yellowtail, tilapia etc. is not allowed in Japan, since it can maintain the red color for a longer period than the microbiological shelf life of fish meat. The official method for quantification of CO has a problem, in that a part of the CO is lost during the preparation of the fish sample. To solve this problem, we modified the official method in this study. We also applied this modified method to survey the contents of CO in tuna, yellowtail, young yellowtail, and tilapia. As a result, the modified method was found to be more suitable for CO quantification than the official method. An inter-laboratory study by 4 laboratories confirmed that the CO content of many samples of tilapia exceeded the regulation value, apparently due to the higher recovery of CO, compared to the official method. Therefore, it was suggested that the regulation value in the case of tilapia should be changed if this method is introduced as an official method.