Circulating microRNAs as Biomarkers for Detection of Autologous Blood Transfusion

MicroRNAs (miRNAs) are small non-coding RNAs that regulate various biological processes. Cell-free miRNAs measured in blood plasma have emerged as specific and sensitive markers of physiological processes and disease. In this study, we investigated whether circulating miRNAs can serve as biomarkers for the detection of autologous blood transfusion, a major doping technique that is still undetectable. Plasma miRNA levels were analyzed using high-throughput quantitative real-time PCR. Plasma samples were obtained before and at several time points after autologous blood transfusion (blood bag storage time 42 days) in 10 healthy subjects and 10 controls without transfusion. Other serum markers of erythropoiesis were determined in the same samples. Our results revealed a distinct change in the pattern of circulating miRNAs. Ten miRNAs were upregulated in transfusion samples compared with control samples. Among these, miR-30b, miR-30c, and miR-26b increased significantly and showed a 3.9-, 4.0-, and 3.0-fold change, respectively. The origin of these miRNAs was related to pulmonary and liver tissues. Erythropoietin (EPO) concentration decreased after blood reinfusion. A combination of miRNAs and EPO measurement in a mathematical model enhanced the efficiency of autologous transfusion detection through miRNA analysis. Therefore, our results lay the foundation for the development of miRNAs as novel blood-based biomarkers to detect autologous transfusion.     

Fat metabolism is regulated by altered gene expression of lipogenic enzymes and regulatory factors in liver and adipose tissue but not in semimembranosus muscle of pigs during the fattening period

It has been shown previously that lipid metabolism is regulated by fatty acids (FA) and that thyroid hormones are important regulators of energy metabolism. The effects of weight, dietary fat level and dietary FA profile on thyroid hormone levels and expression of lipogenic genes and tissue FA composition were studied. Sixty-one crossbred gilts weighing 62 ± 5.2 kg BW average were either slaughtered at the beginning of the trial (n = 5) or fed one of seven diets (n = 8 pigs per diet): a semi-synthetic diet formulated to contain a very low level of fat (NF) and six diets based on barley-soybean meal supplemented with approximately 10% fat of different origin and slaughtered at 100 kg BW. The supplemental fats were tallow, high-oleic sunflower oil, sunflower oil (SFO), linseed oil, fat blend (55% tallow, 35% sunflower oil, 10% linseed oil) and fish oil blend (40% fish oil, 60% linseed oil). In general, the dietary FA profiles altered the FA composition of liver, semimembranosus muscle and adipose tissues. Pigs fed the NF diet had the highest free and total triiodothyronine (T3) values followed by pigs fed SFO. Total T3 levels were higher in pigs at 60 kg than in pigs at 100 kg. Correlations between thyroid hormones and genes encoding enzymes of fat synthesis in adipose tissue (acetyl CoA carboxylase (ACACA), fatty acid synthase and stearoyl CoA desaturase (SCD)) and the large differences in expression of lipogenic genes at different weights (60 and 100 kg BW), suggest a role for thyroid hormones and for T3, in particular, in regulating whole animal fat metabolism, with effects brought about by altered expression of lipogenic genes. Liver sterol receptor element binding protein-1 (SREBP1) mRNA content was affected by dietary treatment (P < 0.001) and was correlated with ACACA and SCD, whereas adipose tissue SREBP1 was not correlated with the mRNA abundance of any lipogenic enzyme. Weight and tissue factors showed greater influence on mRNA abundance of genes related with lipid metabolism than diet and tissue FA composition. In the pig, FA synthesis appear to be of greater magnitude in adipose tissue than in the liver as suggested by the higher expression of lipogenic genes in adipose tissue.     

Studies of the association of Arg72Pro of tumor suppressor protein p53 with type 2 diabetes in a combined analysis of 55,521 Europeans

Aims

A study of 222 candidate genes in type 2 diabetes reported association of variants in RAPGEF1, ENPP1, TP53, NRF1, SLC2A2, SLC2A4 and FOXC2 with type 2 diabetes in 4,805 Finnish individuals. We aimed to replicate these associations in a Danish case-control study and to substantiate any replicated associations in meta-analyses. Furthermore, we evaluated the impact on diabetes-related intermediate traits in a population-based sample of middle-aged Danes.

Methods

We genotyped nine lead variants in the seven genes in 4,973 glucose-tolerant and 3,612 type 2 diabetes Danish individuals. In meta-analyses we combined case-control data from the DIAGRAM+ Consortium (n = 47,117) and the present genotyping results. The quantitative trait studies involved 5,882 treatment-naive individuals from the Danish Inter99 study.

Results

None of the nine investigated variants were significantly associated with type 2 diabetes in the Danish samples. However, for all nine variants the estimate of increase in type 2 diabetes risk was observed for the same allele as previously reported. In a meta-analysis of published and online data including 55,521 Europeans the G-allele of rs1042522 in TP53 showed significant association with type 2 diabetes (OR = 1.06 95% CI 1.02–1.11, p = 0.0032). No substantial associations with diabetes-related intermediary phenotypes were found.

Conclusion

The G-allele of TP53 rs1042522 is associated with an increased prevalence of type 2 diabetes in a combined analysis of 55,521 Europeans.     

Methane emissions from wetlands and their relationship with vascular plants: an Arctic example

This paper investigates the relationship between vascular plant production and CH₄ emissions from an arctic wet tundra ecosystem in north‐east Greenland. Light intensity was manipulated by shading during three consecutive growing seasons (1998–2000). The shading treatment resulted in lower carbon cycling in the ecosystem as mean seasonal net ecosystem exchange (NEE) decreased from ?336 to ?196 mg CO₂ m^?2 h^?1 and from ?476 to ?212 mg CO₂ m^?2 h^?1 in 1999 and 2000, respectively, and total ecosystem respiration decreased from 125 to 94 mg CO₂ m^?2 h^?1 in 1999 and from 409 to 306 mg CO₂ m^?2 h^?1 in 2000. Seasonal mean CH₄ emissions in controls and shaded plots were, respectively, 6.5 and 4.5 mg CH₄ m^?2 h^?1 in 1999 and 8.3 and 6.2 mg CH₄ m^?2 h^?1 in 2000. We found that CH₄ emission was sensitive to NEE and carbon turnover, and it is reasonable to assume that the correlation was due to a combined effect of vegetative CH₄ transport and substrate quality coupled to vascular plant production. Total above‐ground biomass was correlated to mean seasonal CH₄ emission, but separation into species showed that plant‐mediated CH₄ transport was highly species dependent. Potential CH₄ production peaked at the same depth as maximum root density (5–15 cm) and treatment differences further suggest that substrate quality was negatively affected by decreased NEE in the shaded plots. The concentration of dissolved CH₄ decreased in the control plots as the growing season progressed while it was relatively stable in the shaded plots. This suggests that a progressively better developed root system in the controls increased the capacity to transport CH₄ from the soil to the atmosphere. In conclusion, vascular plant photosynthetic rate and subsequent allocation of recently fixed carbon to below‐ground structures seemed to influence both vegetative CH₄ transport and substrate quality.     

A 2-Megabase Physical Contig Incorporating 43 DNA Markers on the Human X Chromosome at p11.23–p11.22 from ZNF21 to DXS255

A comprehensive physical contig of yeast artificial chromosomes (YACs) and cosmid clones between ZNF21 and DXS255 has been constructed, spanning 2 Mb within the region Xp11.23–p11.22. As a portion of the region was found to be particularly unstable in yeast, the integrity of the contig is dependent on additional information provided by the sequence-tagged site (STS) content of cosmid clones and DNA marker retention in conventional and radiation hybrids. The contig was formatted with 43 DNA markers, including 19 new STSs from YAC insert ends and an internalAlu-PCR product. The density of STSs across the contig ranges from one marker every 20 kb to one every 60 kb, with an average density of one marker every 50 kb. The relative order of previously known genes and expressed sequence tags in this region is predicted to be Xpter–ZNF21–DXS7465E (MG66)–DXS7927E (MG81)–WASP, DXS1011E, DXS7467E (MG21)–DXS- 7466E (MG44)–GATA1–DXS7469E (Xp664)–TFE3–SYP (DXS1007E)–Xcen. This contig extends the coverage in Xp11 and provides a framework for the future identification and mapping of new genes, as well as the resources for developing DNA sequencing templates.     

A Three Years' Study of Life Cycle,Population Dynamics and Production of Asellus aquaticus L. in a Macrophyte Rich Stream

Populations of A. aquaticus were sampled quantitatively in 1979–82 at two localities in the river Suså which has a slope of <1 m km⁻¹ and large variations in macrophyte biomass, discharge and stream velocity. The latter two differed significantly between years. A. aquaticus had (1)−2 life cycles per year and summer and winter cohort production intervals of 85 and 290 days. Populations of A. aquaticus varied between 0 and 28,000 ind. m⁻² with an exponential increase in the spring and an exponential decrease in the autumn-winter. The A. aquaticus rate of decrease varied between 0.62 and 5.61 % d⁻¹ and increased with increasing rate of elimination of the macrophyte biomass Differences between the two localities were due to differences in physical heterogeneity. Production varied between 2.8 and 9.1 g DW m⁻² yr⁻¹ and between 12.9 and 55 g DW m⁻² yr⁻¹ at the two localities and P/B ratios were 7.4–9.9 yr⁻¹. Physical limitations are thought to be most important for the populations of A. aquaticus in the Suså, and the macrophyte biomass played an important role in modifying the physical environment. Differences between streams and lake populations are discussed.     

Simultaneous determination of lofepramine and desipramine by a high-performance liquid chromatographic method used for therapeutic drug monitoring

A simple reversed-phase HPLC method with ultraviolet detection for the simultaneous measurement of lofepramine and desipramine is described. Only a single alkaline extraction was used, with clomipramine as internal standard. The column used was to Supelco PCN column, and the mobile phase was acetonitrile-methanol-0.015 M phosphate buffer (120:35:100, v/v). The average recoveries were 78.8% for desipramine and 103.8% for lofepramine, and limits of quantitation were 25 and 5 nmol/1, respectively. The inter-assay C.V.s for lofepramine and desipramine were 6.0 and 7.6%, respectively. The method is specific and has excellent accuracy, and has been used for therapeutic drug monitoring of patients with depressions treated with lofepramine. Mean steady-state plasma concentrations found for lofepramine and desipramine were 8.5 ± 6.1 and 123.6 ± 120.6 nmol/l, respectively. It is concluded that lofepramine in itself has an antidepressive effect.     

Exposure affects the risk of an owl being mobbed – experimental evidence

Mobbing is a widespread anti‐predator strategy in birds, and predators are generally expected to avoid mobbing. For example, observational studies suggest that the cryptic roosting behaviour of nocturnal predators, such as many owls, may be a strategy to limit mobbing. In this paper, we present the results of the first experimental study investigating to what degree roost exposure influences the risk of being mobbed, and the intensity of a mobbing incidence once initiated. To determine these factors, we used an experimental setup with taxidermic mounts of tawny owls Strix aluco in Grib Skov forest, Denmark. The risk of an owl being mobbed during a 50 min morning survey period increased with the exposure of its roosting position, from 24% when hidden to 85% when openly exposed. The corresponding increase in the afternoon was from 6% to 36%. This suggests that an owl may minimize the mobbing rate by reducing the encounter rate with potential mobbers through its choice of roost. Once initiated, the duration of the mobbing (a proxy for the presumed cost of being mobbed) was independent of the roosting position of the mounted owl, but was positively correlated with the number of birds in the mob.     

Denitrification in stream epilithon: Seasonal variation in Gelbæk and Rabis Bæk, Denmark

Abstract Seasonal variations of denitrification activity were determined and compared with various environmental parameters in undisrupted epilithon communities (biofilm) from two Danish lowland streams. In the nutrient-rich Gelbæk, denitrification activity followed extensive changes in biofilm thickness, dry weight and chlorophyll a content during the season. The absolute maximum of denitrification (1.4 mmol N m⁻² d⁻¹, dark incubation) was recorded when the biofilm was best developed in the spring (April and May). Activity decreased dramatically after most of the biofilm suddenly disintegrated and peeled off in early summer. Photosynthetic O₂ production was an important controlling factor of denitrification on a diurnal scale, since the activities were always 2–3 fold lower in the light (50 μE m⁻² s⁻¹) than in the dark. In the more nutrient-poor Rabis Bæk, the biofilm was much less developed and denitrification activities were relatively small (maximum of 0.2 mmol N m⁻² d⁻¹ under dark incubation). The results indicate that productivity of the micro-algae regulates both the seasonal and diurnal patterns of denitrification in the biofilms.     

X-linked retinitis pigmentosa: New map studies of XLRP2, and a possible human centromere effect

Summary A new large Danish family with X-linked retinitis pigmentosa was studied for linkage analysis. Carrier diagnosis was performed using full-field electro-retinogram combined with a careful fundus examination. Multipoint linkage analysis, employing DNA markers from the proximal short arm of the X chromosome and the cytogenetic centromere marker, revealed the highest location score distally to DXS255 and proximal to the ornithine carbamoyl transferase locus. In comparison with the first Danish family that we studied, the pericentromeric recombination fraction was increased; it is speculated that the observed difference in genetic distances from the centromere in the 2 Danish families is correlated with a difference in the size and location of the centromeric heterochromatin.