Grazing impact of filter-feeding zoobenthos in a Danish fjord

The grazing impact has been evaluated of filter-feeding animals in Odense Fjord (Denmark) that is characterized by large biomass of filter feeding polychaetes (Nereis diversicolor), clams (Mya arenaria) and cockles (Cerastoderma glaucum). The filtration rate of each population (F_tot) was related to the total water volume (V_tot) in the different areas of the fjord and expressed as ‘the grazing impact’ Q=V_tot/F_tot. This quantity is related to the half-time for phytoplankton by t_1/2=Q×ln 2, representing the time to reduce the concentration by a factor of 2 in the water column above the filter-feeding populations, assuming efficient vertical mixing. Based on recorded population densities it was found that the potential grazing impact exerted by the three dominating species could be expressed as Q=0.17 d and t_1/2=0.12 d. This indicates a strong filter-feeder control of the phytoplankton although it should be emphasised that the potential grazing impact may only be realised when the filter-feeding zoobenthos is exposed to the whole water column by effective vertical mixing of the water on windy days.     

Negative interstitial pressure in the peritendinous region during exercise

In the presentstudy, tissue pressure in the peritendinous area ventral to the humanAchilles tendon was determined. The pressure was measured during restand intermittent isometric calf muscle exercise at three torques (56, 112, and 168 Nm) 20, 40 and 50 mm proximal to the insertion of thetendon in 11 healthy, young individuals. In allexperiments a linear significant decrease in pressure was obtained withincreasing torque [e.g., at 40 mm: 0.4 ± 0.3 mmHg(rest) to 135 ± 12 mmHg (168 Nm)]. No significant differences were obtained among the three areas measured. On the basisof these observations, microdialysis was performed in the peritendinousregion with a colloid osmotic active substance (Dextran 70, 0.1 g/ml)added to the perfusate with the aim of counteracting the negativetissue pressure. Dialysate volume was found to be fully restored (100 ± 4%) during exercise. It is concluded that a marked negativetissue pressure is generated in the peritendinous space around theAchilles tendon during exercise in humans. Negative tissue pressurecould lead to fluid shift and could be involved in the increase inblood flow previously noted in the peritendinous tissue during exercise(H. Langberg, J. Bülow, and M. Kjær. Acta Physiol. Scand. 163: 149-153, 1998; H. Langberg,J. Bülow, and M. Kjær. Clin.Physiol. 19: 89-93, 1999).

     

Bovine embryos contain a higher proportion of polyploid cells in the trophectoderm than in the embryonic disc

The frequency of polyploid cells in the embryonic disc (ED) and in the trophectoderm (TE) was assessed in 50 in vitro produced bovine embryos fixed at days 7-8 post insemination (pi) and in 20 in vitro produced embryos that were transferred to uteri of recipients at day 7 and then recovered and fixed at day 12 pi. Separation of TE and ED cells was obtained by microdissection and the frequency of polyploid cells was determined by interphase cytogenetic analysis using fluorescence in situ hybridization (FISH) with chromosome 6- and chromosome 7-specific probes. The results show that 96% of day 7 embryos contain polyploid cells in the TE, whereas only 58% contain polyploid cells in the ED. In day 12 embryos 85% of TE and 40% of ED preparations contain polyploid cells. Statistical analysis revealed that the frequency of polyploid cells was significantly higher in the TE than in the ED in embryos containing less than 25% polyploid cells (n = 65). The few embryos (n = 5), which contained more than 25% polyploid cells, did not show this difference. Further, it was revealed that the level of polyploidy on day 7-8 was significantly higher than on day 12, both in the TE (two-fold) and in the ED (seven-fold).     

Magnetic resonance urography by virtual reality modelling

PURPOSE: The purpose of this study was to create a 3D visualization of the urinary tract by a novel virtual reality approach, and to evaluate the usefulness of this method for papillary classification as compared with 2D urogram obtained by maximum intensity projection (MIP). MATERIALS AND METHODS: In one healthy pig, magnetic resonance urography was performed using a T1-weighted 3D gradient echo pulse sequence. Post-processing was performed by means of an MIP algorithm and by using 3D virtual reality modelling, followed by manual classification of papillae as being either simple or compound. RESULTS: The 2D MIP urogram demonstrated 6 simple and 6 compound papillae, whereas the 3D urogram demonstrated 5 simple and 7 compound papillae. In both urograms, some papillae were unsuccessfully classified. CONCLUSION: The possibility of using virtual reality devices allowed 3D rotation and offered additional diagnostic information. However, further studies should reveal its feasibility in diseased kidneys.     

Improvements for comparative analysis of changes in diversity of microbial communities using internal standards in PCR-DGGE

The use of internal standards both during DNA extraction and PCR-DGGE procedure gives the opportunity to analyse the relative abundance of individual species back to the original sample, thereby facilitating relative comparative analysis of diversity. Internal standards were used throughout the DNA extraction and PCR-DGGE to compensate for experimental variability. Such variability causes decreased reproducibility among replicate samples as well as compromise comparisons between samples, since experimental errors cannot be differentiated from actual changes in the community abundance and structure. The use of internal standards during DNA extraction and PCR-DGGE is suitable for ecological and ecotoxicological experiments with microbial communities, where relative changes in the community abundance and structure are studied. We have developed a protocol Internal Standards in Molecular Analysis of Diversity (ISMAD) that is simple to use, inexpensive, rapid to perform and it does not require additional samples to be processed. The internal standard for DNA extraction (ExtrIS) is a fluorescent 510-basepair PCR product which is added to the samples prior to DNA extraction, recovered together with the extracted DNA from the samples and analysed with fluorescence spectrophotometry. The use of ExtrIS during isolation of sample DNA significantly reduced variation among replicate samples. The PCR internal standard (PCR(IS)) originates from the Drosophila melanogaster genome and is a 140-basepair long PCR product, which is amplified by non-competitive primers in the same PCR reaction tubes as the target DNA and analysed together with the target PCR product on the same DGGE gel. The use of PCR(IS) during PCR significantly reduced variation among replicate samples both when assessing total PCR product and when comparing bands representing species on a DGGE gel. The entire ISMAD protocol was shown to accurately describe changes in relative abundance in an environmental sample using PCR-DGGE. It should, however, be mentioned that despite the use of ISMAD some inherent biases still exist in DNA extraction and PCR-DGGE and these should be taken into consideration when interpreting the diversity in a sample based on a DGGE gel.     

Unfolding, aggregation, and seeded amyloid formation of lysine-58-cleaved beta 2-microglobulin

Beta(2)-microglobulin (beta(2)m) is the amyloidogenic protein in dialysis-related amyloidosis, but the mechanisms underlying beta(2)m fibrillogenesis in vivo are largely unknown. We study a structural variant of beta(2)m that has been linked to cancer and inflammation and may be present in the circulation of dialysis patients. This beta(2)m variant, DeltaK58-beta(2)m, is a disulfide-linked two-chain molecule consisting of amino acid residues 1-57 and 59-99 of intact beta(2)m, and we here demonstrate and characterize its decreased conformational stability as compared to wild-type (wt) beta(2)m. Using amide hydrogen/deuterium exchange monitored by mass spectrometry, we show that DeltaK58-beta(2)m has increased unfolding rates compared to wt-beta(2)m and that unfolding is highly temperature dependent. The unfolding rate is 1 order of magnitude faster in DeltaK58-beta(2)m than in wt-beta(2)m, and at 37 degrees C the half-time for unfolding is more than 170-fold faster than at 15 degrees C. Conformational changes are also reflected by a very prominent Congo red binding of DeltaK58-beta(2)m at 37 degrees C, by the evolution of thioflavin T fluorescence, and by changes in intrinsic fluorescence. After a few days at 37 degrees C, in contrast to wt-beta(2)m, DeltaK58-beta(2)m forms well-defined high molecular weight aggregates that are detected by size-exclusion chromatography. Atomic force microscopy after seeding with amyloid-beta(2)m fibrils under conditions that induce minimal fibrillation in wt-beta(2)m shows extensive amyloid fibrillation in DeltaK58-beta(2)m samples. The results highlight the instability and amyloidogenicity under near physiological conditions of a slightly modified beta(2)m variant generated by limited proteolysis and illustrate stages of amyloid formation from early conformational variants to overt fibrillation.     

γ-Glutamyltransferase and Breast Cancer Risk Beyond Alcohol Consumption and Other Life Style Factors – A Pooled Cohort Analysis

Objective

Elevated γ-Glutamyltransferase serum levels are associated with increased risk of overall cancer incidence and several site-specific malignancies. In the present prospective study we report on the associations of serum γ-Glutamyltransferase with the risk of breast cancer in a pooled population-based cohort considering established life style risk factors.

Methods

Two cohorts were included in the present study, i.e. the Vorarlberg (n = 97,268) and the Malmoe cohort (n = 9,790). Cox proportional hazards regression models were fitted to estimate HRs for risk of breast cancer.

Results

In multivariate analysis adjusted for age, body mass index and smoking status, women with γ-Glutamyltransferase levels in the top quartile were at significantly higher risk for breast cancer compared to women in the lowest quartile (HR 1.21, 95% CI 1.09 to 1.35; p = 0.005). In the subgroup analysis of the Malmoe cohort, γ-Glutamyltransferase remained an independent risk factor for breast cancer when additionally considering alcohol intake. A statistically significant increase in risk was seen in women with γ-Glutamyltransferase-levels in the top versus lowest quartile in a multivariate model adjusted for age, body mass index, smoking status, physical activity, parity, oral contraceptive-use and alcohol consumption (HR 1.37, 95% CI 1.11–1.69, p = 0.006).

Conclusion

Our findings identified γ-Glutamyltransferase as an independent risk factor for breast cancer beyond the consumption of alcohol and other life style risk factors.     

Characterisation of integrons and antibiotic resistance genes in Danish multiresistant Salmonella enterica Typhimurium DT104

The presence and genetic content of integrons was investigated in eight Salmonella enteritica Typhimurium DT104 isolates from different pig herds in Denmark. Two different integrons were identified using PCR and sequencing. Each of the integrons carried a single resistance cassette in addition to the sul1 and qacEΔ1 genes characteristic of integrons. The first integron encoded the ant (3″)-Ia gene that specified resistance to spectinomycin and streptomycin. The second contained the pse-1 β-lactamase gene. All the multiresistant strains contained both integrons. The presence of these two integrons did not account for the total phenotypic resistance of all the isolates and does not exclude the presence of other mobile DNA elements.     

Small-angle neutron scattering study of the ultrastructure of chloroplast thylakoid membranes — Periodicity and structural flexibility of the stroma lamellae

The multilamellar organization of freshly isolated spinach and pea chloroplast thylakoid membranes was studied using small-angle neutron scattering. A broad peak at ~ 0.02 Å^? 1 is ascribed to diffraction from domains of ordered, unappressed stroma lamellae, revealing a repeat distance of 294 Å ± 7 Å in spinach and 345 Å ± 11 Å in pea. The peak position and hence the repeat distance of stroma lamellae is strongly dependent on the osmolarity and the ionic strength of the suspension medium, as demonstrated by varying the sorbitol and the Mg⁺⁺-concentration in the sample. For pea thylakoid membranes, we show that the repeat distance decreases when illuminating the sample with white light, in accordance with our earlier results on spinach, also regarding the observation that addition of an uncoupler prohibits the light-induced structural changes, a strong indication that these changes are driven by the transmembrane proton gradient. We show that the magnitude of the shrinkage is strongly dependent on light intensity and that the repeat distance characteristic of the dark state after illumination is different from the initial dark state. Prolonged strong illumination leads to irreversible changes and swelling as reflected in increased repeat distances. The observed reorganizations are discussed within the frames of the current structural models of the granum-stroma thylakoid membrane assembly and the regulatory mechanisms in response to variations in the environmental conditions in vivo. This article is part of a Special Issue entitled: Photosynthesis Research for Sustainability: from Natural to Artificial.