Programs of the IAEA utilizing nuclear analytical techniques in the life sciences

Biological Trace Element Research - Nuclear analytical techniques have an important role in the IAEA in life sciences programs relating to human health. A major effort has recently been devoted to...     

The identification and partial characterization of a plasmid containing the gene for the membrane-associated hydrogenase from E. coli

Escherichia coli DNA was digested with restriction endonuclease PstI and ligated into the PstI site of plasmid pBR322. Recombinant plasmids that were constructed in this manner were used to transform E. coli H61, a mutant with a decreased level of hydrogenase activity. Complementation of this hydrogenase mutation identified a bacterial clone carrying the gene for the membrane-associated E. coli hydrogenase in plasmid pBL101. In E. coli minicells, the pBL101 DNA directed the synthesis of a protein of a size corresponding to that of the precursor of the E. coli membrane-associated hydrogenase, which appears to contain an uncleaved leader peptide. A restriction map of the cloned DNA was determined for 14 endonucleases.     

Introduction

Biological Trace Element Research -     

Application of polyacrylamide gel electrophoresis/neutron activation analysis for protein quantification

Biological Trace Element Research - A combination of two methods, polyacrylamide gel electrophoresis (PAGE) and neutron activation analysis (NAA), has been applied to solutions containing...     

Alaskan marine mammal tissue archival project

A project to establish an archive of Alaskan marine mammal tissues was conceived in 1987 to be a part of the National Biomonitoring Specimen Bank (NBSB). Protocols and field collection of marine mammals, long-term storage, and analysis are summarized in this paper. Instrumental neutron activation analysis has been used for an initial evaluation of trace element content in samples of northern fur seal (Callorhinus ursinus) from the Pribilof Islands. The findings agree with previously observed trace element levels in northern fur seals. The archived specimens can be used in future studies when comparisons of past and present pollution levels are needed.

     

Molecular prey identification in Central European piscivores

Diet analysis is an important aspect when investigating the ecology of fish‐eating animals and essential for assessing their functional role in food webs across aquatic and terrestrial ecosystems. The identification of fish remains in dietary samples, however, can be time‐consuming and unsatisfying using conventional morphological analysis of prey remains. Here, we present a two‐step multiplex PCR system, comprised of six assays, allowing for rapid, sensitive and specific detection of fish DNA in dietary samples. This approach encompasses 78 fish and lamprey species native to Central European freshwaters and enables the identification of 31 species, six genera, two families, two orders and two fish family clusters. All targeted taxa were successfully amplified from 25 template molecules, and each assay was specific when tested against a wide range of invertebrates and vertebrates inhabiting aquatic environments. The applicability of the multiplex PCR system was evaluated in a feeding trial, wherein it outperformed morphological prey analysis regarding species‐specific prey identification in faeces of Eurasian otters. Additionally, a wide spectrum of fish species was detected in field‐collected faecal samples and regurgitated pellets of Common Kingfishers and Great Cormorants, demonstrating the broad applicability of the approach. In conclusion, this multiplex PCR system provides an efficient, easy to use and cost‐effective tool for assessing the trophic ecology of piscivores in Central Europe. Furthermore, the multiplex PCRs and the primers described therein will be applicable wherever DNA of the targeted fish species needs to be detected at high sensitivity and specificity.     

Kinetic analysis and comparison of uptake, distribution, and excretion of 48V-labeled compounds in rats

Setyawati, I. A., K. H. Thompson, V. G. Yuen, Y. Sun, M. Battell, D. M. Lyster, C. Vo, T. J. Ruth, S. Zeisler, J. H. McNeill, and C. Orvig. Kinetic analysis and comparison of uptake,distribution, and excretion of⁴⁸V-labeled compounds in rats.J. Appl. Physiol. 84(2): 569-575, 1998.Vanadium has been found to be orally active in lowering plasmaglucose levels; thus it provides a potential treatment for diabetesmellitus. Bis(maltolato)oxovanadium(IV) (BMOV) is a well-characterizedorganovanadium compound that has been shown in preliminarystudies to have a potentially useful absorption profile. Tissuedistributions of BMOV compared with those of vanadyl sulfate (VS) werestudied in Wistar rats by using⁴⁸V as a tracer. In this study,the compounds were administered in carrier-added forms by either oralgavage or intraperitoneal injection. Data analyzed by a compartmentalmodel, by using simulation, analysis, and modeling (i.e., SAAM II)software, showed a pattern of increased tissue uptake with use of⁴⁸V-BMOV compared with⁴⁸VS. The highest⁴⁸V concentrations at 24 h aftergavage were in bone, followed by kidney and liver. Most ingested⁴⁸V was eliminated unabsorbed byfecal excretion. On average, ⁴⁸Vconcentrations in bone, kidney, and liver 24 h after oraladministration of ⁴⁸V-BMOV weretwo to three times higher than those of⁴⁸VS, which is consistent with theincreased glucose-lowering potency of BMOV in acute glucose loweringcompared with VS.

     

Geographical distribution of haplotypes in Swedish families with Huntington's disease

This study was planned to determine the number of origins of the mutation underlying Huntington's disease (HD) in Sweden. Haplotypes were constructed for 23 different HD families, using six different polymorphisms [(CCG) _n , GT70, 674, BS1, E2 and 4.2], including two within the gene. In addition, extensive genealogical investigations were performed, and the geographical origin of the haplotypes was studied. Ten different haplotypes were observed suggesting multiple origins for the HD mutation in Sweden. Analysis of the two polymorphic markers within the HD gene (the CCG repeat and GT70) indicates that there are at least three origins for the HD mutation in Sweden. One of these haplotypes (7/A) accounts for 89% of the families, suggesting that the majority of the Swedish HD families are related through a single HD mutation of ancient origin. Furthermore, three of the families that were previously considered to be unrelated could be traced to a common ancestor in the 15th century, a finding that is consistent with this hypothesis.     

Enhanced sensitivity for the determination of selenium by INAA

A working analytical procedure for selenium determinations by INAA, using γ spectrometry with a germanium well-type detector, is presented. The spectral line at 400.7 keV is employed, enhanced with the effect of energy summing in true coincidence of cascade photons. The main advantage of the method is high efficiency and reliable, interference-free results; potential drawbacks are increases in input count rate and pile-up losses, decrease in resolution, and sensitivity to errors in sample-to-detector geometry. The procedure is applied to certification analysis of reference materials, large-area biomonitoring by oak leave samples, and determination of separated proteins.     

Indole-3-acetic acid in Douglas fir seedlings: A reappraisal

The use of ring-labelled, pentadeutero IAA as an internal standard in selected ion monitoring analysis of Douglas fir seedlings revealed an estimate of IAA which was nearly an order of magnitude smaller than that reported earlier.