Biochemical examination of fetal skin biopsy specimens obtained by fetoscopy: Use of the method for analysis of keratins and filaggrin

This paper describes a method for biochemical analysis of proteins from fetal skin biopsy samples. The method has wide potential application for diagnosis of disorders with a known protein abnormality detectable by protein staining or a specific antibody. Analysis requires a single 1 mm biopsy, is rapid (2 days) and extremely sensitive. In the present study, fetal skin biopsies from normal fetuses and a fetus at risk for lamellar ichthyosis were obtained. The epidermis or hairs with attached follicular cells were dissected from the remaining skin. Proteins were extracted and separated by SDS-polyacrylamide gel electrophoresis. Proteins from duplicate gels were transferred to nitrocellulose and immunostained for the acidic and basic keratins and for the keratin filament associated protein, filaggrin, using monoclonal antibodies. All samples contained keratins typical of fetal epidermis at 20 weeks gestation. Presence of filaggrin is variable at this age and depends on the presence of keratinized cells of hair canals. No keratin abnormalities in the fetus at risk for lamellar ichthyosis were detected, however, in one presumably normal biopsy, an abnormally low proportion of the 67 kd keratin and the presence of follicular keratins were evident. These results demonstrate that biochemical analysis of fetal biopsies is possible, thus increasing the diagnostic potential of the fetal biopsy procedure for disorders in which a known protein or antigen is altered in utero.     

Genetic study of the extent and consequences of sexual and asexual reproduction in the deep-sea epizoic anemones Amphianthus inornata and Kadosactis commensalis (Cnidaria: Anthozoa)

The anemone Amphianthus inornata is found at bathyal depths living on colonies of the gorgonian Acanella arbuscula. Previous studies of the morphology and reproductive stage of this anemone, during different times of the year, have indicated that it reproduces sexually on a seasonal basis. A small proportion of the study population were also reported to be undergoing asexual reproduction by fission. The anemone Kadosactis commensalis is also bathyal, but lives mainly on the holothurian Paroriza prouhoi. Previous morphological studies have indicated that K. commensalis is a protandrous hermaphrodite that exhibits non-seasonal sexual reproduction only. In the present study, allozyme electrophoresis was used to examine the prevalence and genetic consequences of asexual reproduction in a population of Amphianthus inornata from 2 200 m in the Rockall Trough, North Atlantic Ocean. Genetic evidence, from five randomly selected polymorphic enzyme loci, for asexual reproduction in this species was weak. Exact tests indicated that genotype frequencies did not differ significantly from those expected under Hardy–Weinberg equilibrium. F _IS (correlation of homologous alleles with reference to local population, assuming random mating) values also did not differ significantly from zero, and observed heterozygosity (H _o =0.446) and genotypic diversity (G _o =17.0387) were very similar to Hardy–Weinberg expected frequencies (H _e =0.446; G ^* _e =17.0010). Evidence suggests that the contribution of asexual reproduction to recruitment in the study population of A. inornata is low. For a single population of K. commensalis from 4 850 m on the Porcupine Abyssal Plain, North Atlantic Ocean, the hypothesis that inbreeding due to reduced occurrence of outcrossing between anemones on a single holothurian host was examined by electrophoresis of ten randomly selected enzyme loci. Single-locus genotypic frequencies were significantly different from expected frequencies for one locus P≤ 0.05, hexokinase-1 (Hex-1)]. F _IS values were significantly different from zero for two enzyme loci (Hex-1 and Hex-2, P≤ 0.01 and P≤ 0.05, respectively), and the overall observed heterozygosity was lower than the expected heterozygosity (H _o =0.125, H _e =0.140). The hypothesis of inbreeding could not be rejected by the present study, although sample size was small (N=55), leading to possible bias in tests of significance. Genetic variation in A. inornata was higher than that recorded for most eukaryotes, although interlocus sampling error for only five loci is high. High genetic variability has been found in other sea anemones, and has been related to high longevity and mixed reproductive modes. Genetic variation in K. commensalis was in the higher range of that found in other eukaryotes, and is not unusual for anemones. Received: 5 August 1996 / Accepted: 11 December 1996     

Extent and nature of extracellular organic production by the marine coccolithophorid Hymenomonas carterae

Production of extracellular dissolved organic substances by cultures of the marine coccolithophorid Hymenomonas carterae SMBA 254 was investigated by the ¹⁴C-tracer technique. In 4 h incubations, extracellular ¹⁴C production represented 20 to 64% of ¹⁴C incorporation into cell material for cultures nearing, or during stationary growth; for rapidly growing cultures with relatively low cell densities the ¹⁴C-production was only 4 to 10% of incorporation. Intra-and extra-cellular material was fractionated by ion-exchange membrane electrodialysis. Ionic compounds accounted for 40 to 60% of extracellular material; in contrast only 16 to 27% of intracelular metabolites were in this category. Of a range of methods which were investigated, separation of compounds by gel-filtration in conjunction with two-dimensional thin-layer chromatography proved most effective. Up to 80% of extracellular material was of less than 1 800 mol. wt and included carbohydrates and amino acids, with glutamic acid predominating. The intracellular material was more heterogeneous, with a substantially greater proportion of high molecular weight material. The quite distinct compositions show clearly that extracellular production observed in these experiments arose through excretion and was not attributable to cell lysis.     

Prenatal diagnosis of mosaicism 46, XX/46, XX, −21, +t(21q21q)

Mosaicism for a structural chromosome abnormality in amniotic cell cultures indicative of true fetal mosaicism is a rare event. In addition to the laboratory findings the clinical interpretation for counselling in such cases is based on observation of the same abnormality in liveborns as well as previous experience with prenatal diagnosis of the same or similar abnormalities. We report here the prenatal diagnos is of 46,XX/46,XX,−21,+t(21q21q) which was confirmed in fetal skin cell and amnion cell cultures.     

Structural marsh management effects on coastal fishes and crustaceans

Structural marsh management, using levees and water-control structures, is used in the coastal zone for many objectives, for example, to reduce marsh loss, to enhance waterfowl habitat, to revegetate open-water areas, and to reduce saltwater intrusion. The literature was evaluated to categorize structural marsh management and to determine some of its effects on fishes and crustaceans. Structural marsh management had positive effects on standing stock of most resident organisms and negative effects on marine-transient organisms. Emigration was negatively affected for both resident and marine-transient organisms. Techniques such as opening structures at critical migration times, designing structures that offer the greatest management flexibility, and using structures to create a flow-through system could reduce these impacts. More effort should be put into monitoring managed areas to determine if the objectives are being met and to evaluate the effects on fishes and crustaceans. Although frequent manipulation could reduce these impacts, the costs and problems incurred may outweigh the anticipated benefits.     

PARAMETER DETERMINATION FOR THE MUSKINGUM-CUNGE FLOOD ROUTING METHOD1

ABSTRACT: The proportionality coefficient, K, and the weighing parameter, X, required for the Muskingum-Cunge Flood Routing Method are dependent on the hydraulic characteristics of the channel and the dynamic characteristic of the flood wave. This work focuses on the determination of the Muskingum-Cunge Flood Routing Method parameters for streams where measured hydrographs are not available (i.e., ungaged streams) with floods that stay within the channel banks. In the present work, a gaged creek was used and a dynamic wave was routed to test the reliability of the parameters determined through the Schaefer and Stevens technique (Schaefer and Stevens, 1978). The predicted outflow hydrographs are compared to the hydrographs obtained for the same stream determined with the Muskingum Routing option of the HEC-1 program. Cypress Creek in Harris County, Texas, was the model for this work; and the corresponding data were extracted from the Grant Road and Westfield, Texas, USGS gaging stations.     

Sources and levels of potentially toxic elements in the biodegradable fraction of autoclaved non-segregated household waste and its compost/digestate

Waste that reflected the average UK composition of household waste was treated by autoclaving at the three set pressure/temperature levels of 2.7 bar/130 degrees C, 6.2 bar/160 degrees C and 15.5 bar/200 degrees C. The biodegradable fraction of the autoclaved household waste (;floc') was manually separated by screening and underwent characterization for its Cd, Cr, Cu, Pb, Hg, Ni, and Zn content. Autoclaving did not guarantee the production of compost/digestate that met the UK specification for compost, BSi PAS100, without restrictions being made on the composition of the waste feedstock. Results indicate that the levels of Zn and Cd associated with floc materials alone could lead to compost limit values being exceeded. For all other potentially toxic elements (PTEs), the estimated excessive (i.e. above levels of compliance) PTEs levels for compost/digestate were mainly due to external (i.e. non-floc) materials, primarily electronic/electrical waste. Batteries may have also contributed to the high levels of Zn and Hg. In this study, for all PTEs examined, with the exception of Cd and Zn, autoclaving had a performance comparable to that of the most effective mechanical biological treatment systems.