Prenatal interphase FISH diagnosis of PLP1 duplication associated with Pelizaeus–Merzbacher disease

A submicroscopic genomic duplication in Xq22.2 that contains the entire proteolipid protein 1 gene (PLP1) is responsible for the majority of Pelizaeus–Merzbacher disease (PMD) patients. We previously developed an interphase FISH assay to screen for PLP1 duplications in PMD patients using peripheral blood and lymphoblastoid cell lines. This assay has been utilized as a clinical diagnostic test in our cytogenetics laboratory. To expand usage of the interphase FISH assay to prenatal diagnosis of PLP1 duplications, we examined three PMD families with PLP1 duplications utilizing aminiotic fluid samples. In two families the FISH assay revealed fetuses with PLP1 duplications, whereas the other fetus showed a normal copy number of PLP1. Haplotype analyses, as well as an additional FISH analysis using postnatal blood samples, confirmed the results of the prenatal analyses. Our study demonstrates utility of the interphase FISH assay in the prenatal diagnosis of PLP1 duplications in PMD. Copyright © 2001 John Wiley & Sons, Ltd.     

18S rRNA phylogeny of sea spiders with emphasis on the position of Rhynchothoracidae

The phylogenetic relationships among all living families of sea spiders (Arthropoda: Pycnogonida) are investigated using nearly complete 18S rRNA sequences from 57 ingroup species and five chelicerates under the Bayesian and maximum likelihood methods. Monophyly of Colossendeidae, Pycnogonidae, Phoxichilidiidae, Endeidae and Pallenopsidae is consistently supported. However, the genera formerly classified in the family Ammotheidae are split up into two distantly related groups. The genera Ascorhynchus and Eurycyde (here recognized as Ascorhynchidae) are possibly an early offshoot of sea spiders, whereas other ammotheids constitute a robust terminal clade with Pallenopsidae, Phoxichilidiidae and Endeidae. This topology also opposes the prevalent assumption of successive losses and simplification of three kinds of cephalic appendages like in a previous cladistic analysis. At least three independent losses are suggested for palps by the inferred topology, and both chelifores and female ovigers may have been lost twice. Our knowledge of early ontogeny and internal anatomy is more congruent with the present 18S rRNA data. The families Callipallenidae and Nymphonidae with unique “attaching larvae” are grouped together in present molecular trees, suggesting that extended paternal care of offspring evolved only once in Pycnogonida. Confident clustering of Pycnogonidae and Rhynchothoracidae indicates that the number of female genital pores is an evolutionary conservative character.     

Influence of americium-241 on the microbial population and biodegradation of organic waste

The present study investigated the influence of ²⁴¹Am on microbial growth and the degradation of organic waste. Leachate samples collected in a lysimeter were periodically analyzed for bacterial growth, under both aerobic and anaerobic conditions. ²⁴¹Am inhibited bacterial growth, and the degradation of organic matter was delayed in comparison with the control. Minimal inhibitory concentration assays and survival curves revealed that it inhibits the growth of Pseudomonas putida F1. The assay also revealed that ²⁴¹Am is more toxic than ²³⁸U, Zn²⁺ and Cd²⁺. This study further led to the finding of four new radionuclide-tolerant bacterial strains: Flavobacterium spp., Pseudomonas gladioli, Chryseobacterium indologenes and Ochrobactrum anthropi. The survival curves of P. gladioli, C. indologenes revealed that these bacteria are resistant to metal as consortia.     

Life cycle of meats: an opportunity to abate the greenhouse gas emission from meat industry in Japan

The food industry is one of the world's largest industrial sectors, hence a large contributor of greenhouse gases (GHG) which cause global warming. This study evaluates the life cycle of various types of meat to determine if the GHG emission from the meat industry in Japan could be reduced if the population makes different dietary choices. It was confirmed that the GHG emission of beef was greater than that of pork or chicken. The GHG emission from meat in general also depends on the per capita caloric intake (if meat supplies the recommended animal protein or contributes to it at the present rate). In a healthy and balanced diet (9.2 MJ i.e., 2200 kcal in total, where either mixed meat or chicken or pork or beef contributes 2.2%), the GHG emission is estimated to be 0.28 or 0.17 or 0.15 or 0.77 kg CO? eq/person/day, respectively. A change in consumption patterns (from beef to chicken or pork) and the adoption of a healthy and balanced diet would help to abate about 2.5-54.0 million tons (CO? eq) produced by the meat industry each year in Japan.     

Determination of dioxin concentrations in fish and seafood samples using a highly sensitive reporter cell line, DR-EcoScreen cells

There is a strong need for the development of relatively rapid and low-cost bioassays for the determination of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs), and dioxin-like polychlorinated biphenyls (dl-PCBs) in environmental and food samples. In this study, we applied a reporter gene assay using DR-EcoScreen cells (DR-cell assay), which is highly sensitive to dioxins, to the determination of PCDD/Fs and dl-PCBs in fish and seafood samples. The PCDD/Fs and dl-PCBs were extracted from homogenated samples (10 g) of 30 fish and shellfish, purified by clean-up procedure using a multilayered silica gel column and an alumina column, and applied to DR-cell assay. Interestingly, the bioanalytical equivalent (BEQ) values obtained from the DR-cell assay [<0.1 ∼ 5.4 pg BEQ g⁻¹ wet weight (ww)] were closely correlated with the toxicity equivalent (TEQ) values from conventional high-resolution gas chromatography/high-resolution mass spectrometry (HRGC-HRMS) analysis (r² = 0.912), and the slope of regression line was 0.913. Therefore, we multiplied the BEQ values from the DR-cell assay by a conversion coefficient (1.095, the reciprocal of 0.913) to approximate the TEQ values from the HRGC-HRMS analysis. Furthermore, we used this DR-cell assay to perform a prescreening test of PCDD/Fs and dl-PCBs in 16 fish and seafood samples purchased from a supermarket, revealing that a sample from the fatty flesh of a bluefin tuna exceeded 8 pg TEQ g⁻¹ ww (the European Union-tolerance limit). Taken together, these results suggest that the DR-cell assay might be applicable as a rapid and low-cost prescreening method to determine dioxin levels in fish and seafood samples.     

Effects of dietary habits and CYP1A1 polymorphisms on blood dioxin concentrations in Japanese men

The major source of dioxin impurities in Japan in the past was agrochemical formulations; more recently, it has been exhaust from waste incinerators. To examine the environmental and genetic factors that influence blood dioxin concentration, we investigated the relationship among dioxin concentrations, dietary habits and cytochorome P450 1A1 (CYP1A1) polymorphisms (MspI type and Ile-Val type) in Japanese fishermen and farmers, including also a group of office workers as controls. The mean dioxin concentrations in the fishermen, the farmers and the controls were 161369, 79079 and 100500 pg/g fat, respectively. The elevated dioxin concentration with polychlorinated dibenzo-p-dioxins, polychlorinated dibenzofurans and coplanar-PCBs found in the fishermen may be due to the frequent consumption of fish; no such relationship was found both in the farmers and the controls. We found that the concentrations of congeners found as impurities in certain chemicals such as those previously used in agriculture showed no significant differences among the three groups; we consider it unlikely that the farmers would be directly exposed to dioxins from such chemicals. Thus, it is probable that the primary route of dioxin exposure in the Japanese population is through the food chain via fish consumption, regardless of occupation. No meaningful relationship between blood dioxin concentration and CYP1A1 polymorphisms was found in this study, although there was a significant difference between the concentration of total non-ortho-PCBs in genotypes A and B. Further studies on more subjects, including those of genotype C, are needed to confirm the relationship between blood dioxin concentrations and MspI polymorphisms.     

Recycling, landfill consumption, and CO2 emission: analysis by waste input–output model

The emission of waste in an economy, including landfill, is to a large extent determined by its patterns of technology, institutions, and lifestyle. A mathematical model (the waste input–output model) is presented that gives a simple analytical representation of this interdependence. The model was used to evaluate the effects of alternative waste disposal and recycling options on the levels of industrial production, landfill consumption, and the emission of carbon dioxide, and also to analyze the overall dependence on landfill of individual industries. It was found that a systematic combination of the options could be effective in reducing the overall landfill consumption and carbon dioxide emission. Received: November 4, 2000 / Accepted: August 2, 2001     

Evaluation of the impact of matrix effects in LC/MS measurement on the accurate quantification of neonicotinoid pesticides in food by isotope-dilution mass spectrometry

The use of isotope-labeled internal standards is the most widely accepted approach to overcome the matrix effects on quantification of pesticides in food by LC/MS. We evaluated the impact of the matrix effects on quantification of six neonicotinoid pesticides, acetamiprid, clothianidin, dinotefuran, imidacloprid, thiacloprid, and thiamethoxam, in food by using deuterated internal standards. The calibration curves for each pesticide were obtained by using matrix-free and matrix-matched calibration solutions with blank brown rice, carrot, and green onion extracts. For brown rice and carrot, the matrix effects were not observed. In contrast, the slopes of calibration curves for each pesticide were influenced by presence of green onion extracts in calibration solutions (variability of the slopes was 4–9%), because the ratios of peak area for native pesticide to those for internal standards were influenced by matrix. The spike-and-recovery test with green onion was also performed. The analytical values obtained by using matrix-free calibration solution were biased from the spiked concentration, whereas those obtained by using matrix-matched calibration solution were comparable to the spiked concentration. These results indicate that matrix-matched calibration solution should be used for accurate quantification of neonicotinoid pesticides in food by LC/MS using deuterated internal standards.