Levels of cadmium and mercury in the hair of Atlantic walruses (Odobenus rosmarus rosmarus) from Svalbard,Norway

Hair samples of 15 adult male Atlantic walruses (Odobenus rosmarus rosmarus) collected from anaesthetized individuals at Svalbard, Norway, were analysed for cadmium and total mercury. The mean level of cadmium was 0.860 ± 0.321 μg/g dry weight (median = 0.811, range = 0.349–1.51 μg/g dry weight) and the mean level of mercury was 0.235 ± 0.100 μg/g dry weight (median = 0.251, range = 0.121–0.424 μg/g dry weight). Levels of cadmium and mercury in hair of walruses from other areas are not known. Both cadmium and mercury levels in hair of walruses from Svalbard are relatively low compared to the levels found in the hair of other marine mammal species. It has been documented from a number of marine species, including marine mammals such as ringed seals and polar bears, that both cadmium and mercury levels at Svalbard are lower than in other areas. It is uncertain as to what degree levels in hair reflect levels in internal organs in walruses. In rare and highly endangered species or populations tissue samples can be difficult to collect. In walruses, it is possible to collect hair from anaesthetized individuals or at the haul-out sites during moult, to monitor heavy metal levels of the population. Accepted: 6 December 1998     

Patterns of genetic variation in isolated Danish populations of the endangered butterfly Euphydryas aurinia

In the present study, we report an investigation on molecular variation in the endangered univoltine butterfly Euphydryas aurinia (Rottemburg, 1775), a species heavily affected by habitat degradation and fragmentation in Denmark. Levels of genetic variation in extant populations were estimated using six variable number tandem repeat loci and were found to be low compared to other butterfly species with low migration rates. An analysis of genetic structure, based on both allele frequencies and genotype distributions, divided the entire sample into four distinct clusters. This was partially concordant with the a priori subdivision based on collection areas. An overall F_ST value of 0.16 (pairwise values ranging from 0.087–0.276) indicated restrictions of gene flow. Especially two populations had higher F_ST values than the others, suggesting their isolation, and showed signs of bottlenecks/founder events. One population deviated significantly from Hardy–Weinberg equilibrium, suggesting a possible Wahlund effect or the presence of null alleles. The results suggest habitat fragmentation, resulting in genetic drift and possibly inbreeding. Future management is therefore recommended to increase gene flow between the remaining populations while habitats are restored in order to increase carrying capacity. © 2008 The Linnean Society of London, Biological Journal of the Linnean Society, 2008, 95 , 677–687.     

Heterologous expression and purification of the infectious salmon anemia virus hemagglutinin esterase

This study presents the heterologous production and purification of a soluble and functional form of the hemagglutinin esterase (HE) of the infectious salmon anemia virus (ISAV) isolate 4 (Glesvaer/2/90). The HE possesses receptor binding and receptor destroying enzyme (RDE) activity and is probably involved in the infection process.The recombinant HE protein (recHE 4) was expressed in insect cells (Sf9) using the baculovirus expression vector system. Both the transmembrane region and the cytoplasmic tail were deleted, and a C-terminal His₆-tag was attached to facilitate identification and purification of the recHE 4 protein. As determined by Western analysis the recHE 4 was secreted at 20 °C and not at 28 °C. By testing three HE constructs differing in their promoter and secretion signal sequences it was clear that the HE’s own secretion signal sequence is more important than the promoter with respect to the amount of secreted recHE 4 obtained under the conditions used. A one-step purification by nickel-affinity chromatography resulted in a highly purified recHE 4, identified by matrix assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) analysis. Also, the recHE 4 is glycosylated and contains disulfide bridges within the molecule. Functional studies including the verification of the receptor destroying enzyme (RDE) activity as well as the binding to Atlantic salmon erythrocytes (hemagglutination) indicate that the recHE 4 has similar functions as its native counterpart.In conclusion, insect cells secrete a functional form of the ISAV 4 HE. This is suitable for further analyses on its function and immunogenicity.     

Expa: a program for calculating extreme pathways in biochemical reaction networks

SUMMARY: The set of extreme pathways, a generating set for all possible steady-state flux maps in a biochemical reaction network, can be computed from the stoichiometric matrix, an incidence-like matrix reflecting the network topology. Here, we describe the implementation of a well-known algorithm to compute these pathways and give a summary of the features of the available software. AVAILABILITY: The C-code, along with a Windows executable and sample network reaction files, are available at http://systemsbiology.ucsd.edu CONTACT: palsson@ucsd.edu.     

The viability of cryopreserved PBPC depends on the DMSO concentration and the concentration of nucleated cells in the graft

BACKGROUND: DMSO is widely used as a cryoprotectant for PBPC. It is desirable to reduce the amount of DMSO without jeopardizing the quality of the stem cell product. The present study was undertaken to investigate whether recovery and survival of CD34+ cells would be significantly altered when PBPC used for autologous transplantations were cryopreserved with four different DMSO concentrations. METHODS: Apheresis samples of PBPC from 20 consecutive patients were mixed in parallel with 2%, 4%, 5% and 10% DMSO, frozen with identical cell concentrations at a controlled rate, and stored in liquid nitrogen for 6-8 weeks. PBPC samples from 11 consecutive patients were also cryopreserved with two different cell concentrations (150 and 300 x 10(6) nucleated cells/mL) to investigate the effect of increasing the cell concentrations while decreasing the DMSO concentration. The flow cytometric absolute count method, based on ISHAGE guidelines, was used to measure the absolute count of total and viable CD34+ cells in the post-thaw samples. RESULTS: PBPC cryopreserved at 150 x 10(6) cells/mL with 2% DMSO yielded significantly inferior CD34+ cell recovery (P < 0.001) and survival (P < 0.001) compared with cryopreservation with 4% and 5% DMSO. This was also observed when comparing higher cell concentrations. However, a reduced cell survival (P = 0.02) was observed when the nucleated cell concentration was increased from 150 to 300 x 10(6) cells/mL in samples cryopreserved with 5% DMSO. DISCUSSION: We conclude that 5% DMSO may be the optimal dose for cryopreserving PBPC as long as the cells have not been concentrated at much more than 200 x 10(6) nucleated cells/mL.     

Measurement of the uncomplexed fraction of tissue inhibitor of metalloproteinases-1 in the prognostic evaluation of primary breast cancer patients

Several studies have demonstrated an association between high tumor tissue levels of total tissue inhibitor of metalloproteinases-1 (TIMP-1) and a poor prognosis of primary breast cancer patients. In the present study we investigated whether measurements of the uncomplexed fraction of TIMP-1 added prognostic information to that already obtained from total TIMP-1. We measured the uncomplexed fraction of TIMP-1, using a thoroughly validated ELISA specific for this fraction, in 341 tumor tissue extracts obtained from patients with primary breast cancer. These measurements were related to previously performed measurements of total TIMP-1 as well as to patient outcome. The observation time was 8.3 years (range, 7.3-11.3 years). During this period 136 patients died, and 153 patients experienced recurrence of disease. Cox regression analysis of recurrence-free survival (RFS) suggested that a score based on both uncomplexed and total TIMP-1, reflecting the tumor level of TIMP-1/MMP complexes, would be a more precise estimate of prognosis than total TIMP-1 alone. Univariate survival analysis showed a highly significant relationship between high values of the score and poor outcomes for RFS (p = 0.0002; hazard ratio = 2.7; 95% confidence interval, 1.5-4.8). Similar results were found for overall survival (p = 0.0001; hazard ratio = 3.3; 95% confidence interval, 1.8-6.3). Multivariate analysis of RFS and overall survival demonstrated that the score was significant including the classical prognostic factors used in breast cancer (p < 0.0001). The present study raises the hypothesis that it is the tumor level of TIMP-1/MMP complexes (i.e. activated matrix metalloproteinases) rather than TIMP-1 itself that determines prognosis, supporting the use of the combined score and not only total TIMP-1 in stratification of breast cancer patients.     

Freshwater bryozoans (Bryozoa) of Norway IV: Distribution and ecology of four species of Plumatella with notes on Hyalinella punctata

Bryozoans were investigated during field studies of 601 lakes and other surface water bodies throughout Norway from 1960 to 1978. The frequency of occurrence of Plumatella repens was evaluated in relation to 12 environmental variables. Statistically significant deviations from the frequencies expected on the basis of random distribution were described using the categories preference, avoidance and absence. P. repens is one of the most common bryozoan species in Norway. It is represented at 227 localities in our material and by 29 additional records. The species occurred frequently all over the country, north to 71°09N (the northernmost record on the European mainland). Maximum elevation above sea level was 1052 m (maximum for Northern Europe). P. repens preferred lakes, avoided rivers, and was absent from the smallest water bodies. The species preferred (1) low elevation (2–200 m above sea level), (2) medium water temperature, (3) rich aquatic vegetation (eutrophic conditions), (4) small wave action, (5) high content of calcium and magnesium, (6) pH 6.4–9.6, and (7) rather high water colour. Otherwise P. repens avoided (1) habitats above 500 m but occurred up to 1052 m, (2) the lowest water temperature intervals, (3) sites with poor aquatic vegetation and stony shores, (4) medium wave action, (5) sites with a low calcium and magnesium content, (6) lakes of low water colour, and (7) lakes with pH below 6.6: it was absent from lakes with pH below 5.2. For many environmental variables, the species had a wider tolerance range than reported from elsewhere. P. fungosa, P. casmiana, and P. emarginata are all rare in Norway. The total numbers of known localities, with our records in (), are 18 (11), 7 (7), and 7 (5), respectively. These sites are all from lowland areas up to 133 m above sea level. Environmental variables in the habitats are described. P. fungosa was represented by a single floatoblast in each of the four northernmost lakes where it was recorded. One of these lakes was in North Norway some 1000 km away from the nearest known sites with living colonies. The presence of large colonies of P. fungosa in some parts of southern Norway and only `single floatoblast lakes' further north is discussed in relation to the equilibrium theory of island biogeography: birds facilitate immigration by carrying floatoblasts and environmental conditions prevent their germination and the development of living colonies. All records of P. casmiana and P. emarginata were from South Norway. Results for P. fruticosa have been published previously. Notes are given on an old and now missing sample from ca. 1900, identified by Lacourt (1968) as Hyalinella punctata – the only record of this species from Norway.