Dystrophin protein and rflp analyis for fetal diagnosis and carrier confirmation of duchenne muscular dystrophy

A pregnant woman with indeterminate Duchenne muscular dystrophy (DMD) carrier status, but with DMD diagnosed in her deceased brother (unavailable for study), presented for prenatal diagnosis, intending to continue the pregnancy only if proven unaffected with DMD with near absolute certainty. Creatine kinase (CK) assays to clarify carrier status were inconclusive. Male sex in the fetus was identified, but DNA restriction fragment length polymorphism (RFLP) analysis was not yet available to this centre to investigate the possible transmission of the DMD gene, and the pregnancy was terminated. Tissue histology and dystrophin protein analysis demonstrated the absence of DMD. In a situation with proven maternal carrier status, future fetal inheritance of the opposite maternal X chromosome would indicate the presence of DMD. However, maternal carrier status remained in doubt through a second pregnancy, even with RFLP studies, and was finally established when dystrophin analysis confirmed the presence of DMD in the second fetus. Histologic findings are presented, contrasting features in the two fetuses. The value of dystrophin analysis for establishing the diagnosis of fetal DMD, in this case proving maternal carrier status in a difficult situation, and for demonstrating DMD gene:RFLP haplotype relationships is illustrated.     

The effect of physico-chemical parameters and chemical compounds on the activity of beta-d-galactosidase (B-GAL), a marker enzyme for indicator microorganisms in water

The presence of coliforms in polluted water was determined enzymatically (in situ) by directly monitoring the activity of beta-d-galactosidase (B-GAL) through the hydrolysis of the yellow chromogenic subtrate, chlorophenol red beta-d-galactopyranoside (CPRG), which produced a red chlorophenol red (CPR) product. The objectives of this study were to monitor the effect of compounds commonly found in the environment and used in water treatment on a B-GAL CPRG assay and to investigate the differences between the environmental B-GAL enzyme and the pure commercial enzyme. Environmental B-GAL was optimally active at pH 7.8. Two temperature optima were observed at 35 and 55 degrees C, respectively. B-GAL activity was strongly inhibited by silver and copper ions. While calcium and ferrous ions at lower concentrations (50-100mgl(-1)) increased the enzyme activity, a reduction was observed at higher concentrations (200mgl(-1)). Sodium hypochlorite, normally used in rural areas to disinfect water gradually decreased B-GAL activity at concentrations between 0 and 5600ppm for both the commercial and environmental enzymes. B-GAL from the environment behaved differently from its commercially available counterpart.     

Generation of cyanide ion by the reaction of phenol with nitrous acid in wastewater

It was found that phenol reacts with nitrous acid to produce cyanide ions. Cyanide ion generation is attributed to the conversion of phenol to nitrosophenol through the well-known nitrosation reaction, and decomposition of benzoquinonoxim to form cyanide and aliphatic compound.     

Use of lees materials as an adsorbent for removal of organochlorine compounds or benzene from wastewater

Lees materials such as wheat bran, rapeseed, linseed, okara (lees of bean curd), and sakekasu (sake lees) were found to effectively adsorb organochlorine compounds. The amounts of these compounds such as chloroform, dichloromethane, and benzene adsorbed were plotted against the equilibrium concentration of substances in solution on a logarithmic scale. A linear relationship was obtained, indicating that the adsorption reactions were of the Freundlich type. When the lees materials were applied to wastewater (pH: 10) containing 0.1 g/l of dichloromethane, dichloromethane was removed from the wastewater in the range of 70-90% efficiency after 90 min. There was a high correlation between the removal efficiency and the number of spherosomes, which are intracellular particles attributed to the uptake of organochlorine compounds.     

Contamination and bioaccumulation of benzotriazole ultraviolet stabilizers in fish from Manila Bay, the Philippines using an ultra-fast liquid chromatography-tandem mass spectrometry

Methyl methacrylate (MMA) is grafted on multiwalled carbon nanotubes (MWCNTs) by using N₂ plasma technique. The MMA grafted MWCNTs (MWCNT-g-pMMA) are characterized by using Raman spectroscopy, powder X-ray diffraction, X-ray photoelectron spectroscopy, thermo gravimetric analysis-differential thermal analysis (TGA-DTA), and potentiometric acid-base titration method. The application of MWCNT-g-pMMA in the removal of 4,4′-dichlorinated biphenyl (4,4′-DCB) from large volumes of aqueous solutions is investigated under ambient conditions. The results indicate that the adsorption of 4,4′-DCB on MWCNT-g-pMMA is much higher than that of 4,4′-DCB on MWCNTs, and the adsorbed 4,4′-DCB is difficult to be thermally decomposed from MWCNT-g-pMMA according to the TGA-DTA analysis. MWCNT-g-pMMA are suitable materials in the preconcentration and immobilization of polychlorinated biphenyls (PCBs) from large volumes of aqueous solutions in environmental pollution cleanup.     

Field identification of ‘types’ A and B of the ascidian Ciona intestinalis in a region of sympatry

The ascidian species Ciona intestinalis is a major model chordate in developmental and evolutionary biology, and an important fouling organism and invasive species. However, genomic investigation has recently revealed the existence of two cryptic species, genetically distinct yet without obvious morphological differences, currently referred to as types A and B. Here, we show that they are externally distinctive in a zone of sympatry in the western English Channel. Examining genotyped specimens, we found that types A and B of C. intestinalis can generally be distinguished by body colour, pigmentation at the distal end of the siphons and the presence or absence of tubercles on the sides of the siphons. Detecting specimens of hybrid descent still requires detailed molecular analysis, but these visual characters in combination will identify living specimens of types A and B with high probability. These differences are shown to be inherited.     

Validation of diffusive mini-samplers for aldehyde and VOC and its feasibility for measuring the exposure levels of elementary school children

Exposure to various chemicals can cause adverse effects to health, such as asthma and allergies, especially in children. Data on personal exposure levels in children are scarce, thus small lightweight diffusive mini-samplers for aldehydes and volatile organic compounds (VOCs) were designed to measure the exposure level of children to these chemicals. The aim of the study was to validate and examine the applicability of these mini-samplers for measuring daily chemical exposure. The diffusive mini-samplers are 20 mm in length, 11 mm in diameter, and 1.67 g in weight. The devices are cylindrically shaped with polytetrafluoroethylene membrane filters placed at each end. To measure aldehydes and acetone, 20 mg of 2,4-dinitrophenylhydrazine was used as an absorbent. To measure VOCs, a carbon molecular sieve was used. The sampling rate for each chemical was determined by parallel sampling with active samplers in a closed exposure bag. The blank levels of the chemicals and the storage stability of the device were tested. The mini-samplers were compared to commercially available diffusive samplers. To examine the applicability of the samplers, 65 elementary school children carried them for 24 h. The sampling rates for formaldehyde, acetaldehyde, and acetone were 20.9, 22.9, and 19.7 mL min(-1), respectively. The limits of quantification (LOQ) for the 24-hour sampling by high-performance liquid chromatography/ultraviolet (HPLC/UV) analysis were 8.3, 7.6, and 8.8 μg m(-3) for formaldehyde, acetaldehyde, and acetone, respectively. The sampling rates for the 11 VOCs were determined and ranged from 3.3 mL min(-1) for styrene and 2-ethyl-1-hexanol to 11.7 mL min(-1) for benzene. The LOQ for the 24-hour sampling by gas chromatography-mass spectrometry (GC-MS) analysis ranged from 5.9-105.2 μg m(-3), 1.1-24.7 parts per billion. The storage stability after 5 days ranged from 94.8 to 118.2%. Formaldehyde, acetone, benzene, and toluene were detected above the LOQ in more than 90% of the children, and the median concentrations were 21.7, 20.9, 10.1, and 21.5 μg m(-3), respectively. This study shows that the diffusive samplers developed were suitable for children to carry and were capable of measuring the children's daily chemical exposure.     

Biomagnification profiles of tributyltin (TBT) and triphenyltin (TPT) in Japanese coastal food webs elucidated by stable nitrogen isotope ratios

The measurement of organotins in the various biotas of coastal food webs with stable nitrogen isotope ratios (δ¹⁵N), which increase 3.4‰ per trophic level, can provide a biomagnification profile of organotins through food web. In this study, various biological samples were collected from three localities in Western Japan between 2002 and 2003 for analyses. Tributyltin (TBT) and triphenyltin (TPT) were still detected with a maximum of 99.5 and 8.7 ng wet weight g⁻¹, respectively. Unlike TBT, significant biomagnification of TPT through the food web (expressed by δ¹⁵N) was found in all three localities. The log transformed octanol–water partition coefficient (log K_ow) of TPT of 2.11–3.43 was overlapped by, but was slightly lower than, that of TBT of 3.70–4.70. Thus, this study demonstrates that although these chemicals have a log K_ow lower than 5, at least TPT undergoes significant biomagnification through the food web.