Maximum-likelihood estimation of sensitivity and specificity of ELISAs and faecal culture for diagnosis of paratuberculosis

The accuracy of three diagnostic tests for paratuberculosis was evaluated using maximum-likelihood estimation of sensitivity and specificity. We also explored the variety of estimates that can be obtained if the tests are to be used in populations of different composition with regard to infection and disease states. Two enzyme-linked immunosorbent assays (ELISAs) were evaluated separately with the faecal culture (FC). The study was carried out as a cross-sectional field study to cover all likely states of infection with Mycobacterium avium subsp. paratuberculosis.The three basic assumptions for the maximum-likelihood technique were evaluated to validate the results. Our accuracy estimates for the ELISAs were not very different from those previously published, but those for faecal culture differed if a different cut-off value was chosen for the ELISA. If faecal culture was used for screening in a Danish dairy region where the median ELISA reading was a measure of the general disease situation, the sensitivity of the faecal culture was 20-25%. If faecal culture was used as a confirmatory test on cows with a high ELISA reading (and thus high level of antibodies), the sensitivity of the faecal culture would be in the range 60-70%. These results emphasise the importance of the composition of a target population before selecting a specific diagnostic test for a given purpose. We concluded that faecal culture is useful for confirmation but not for screening purposes.     

Wood modification with alkoxysilanes

Wood was treated with three different alkoxysilanes which are able to undergo a sol–gel process: tetraethoxysilane (TEOS), methyl triethoxysilane (MTES) and propyl triethoxysilane (PTEO). Two types of treatments were compared: impregnation of fibre saturated wood with monomeric silane solutions, and impregnation with pre-hydrolysed partly oligomeric silanes. Wood properties such as cell wall bulking, anti-swelling efficiency (ASE), moisture uptake and durability were more significantly improved in samples treated with monomeric silanes than in samples treated with oligomeric silanes. SEM-EDX mapping showed that this treatment resulted in a higher degree of silicon incorporation into the cell wall, although the weight percent gain (WPG) was lower compared to the treatment with pre-hydrolysed partly oligomeric silanes. The resistance against soil micro-organisms was enhanced in the initial phase of incubation especially in those samples treated with organo-functional-alkoxysilanes. In miniaturised block tests with the white-rot basidiomycete Trametes versicolor, an improved durability was observed within the test period. During an air conditioning step, a weight loss of the treated samples occurred which was accompanied by a reduction in bulking and ASE. The initial reduction of moisture uptake observed after treatment diminished almost completely. This effect was explained by an ageing of the gels in the wood cell wall which is a consequence of uncompleted hydrolysis and condensation of the silanes during the treatment.     

体重决定猪回肠内的蛋白吸收率

<正>近期伊利诺伊大学的J.A.Almeida博士、 Hans H.Stein博士以及Carsten Pedersen博士联手展开体重对于标准化回肠蛋白质和氨基酸吸收率的影响试验,结论证明,20 kg以下的仔猪回肠对于粗蛋白和氨基酸的吸收率比20~50 kg的育成猪和50~110 kg的育肥猪低。而且同一猪群的不同猪只也表现出吸收率的差异性。例如,在测定的300只仔猪中粗蛋白和氨基酸的吸收率     

Ash and biochar amendment of coarse sandy soil for growing crops under drought conditions

Appropriate soil amendments may increase plant available water and crop yields on coarse sandy soils under drought conditions. In this study, we applied straw ash or straw biochar from gasification to a Danish coarse sandy subsoil to assess the effects on soil water retention, evapotranspiration and crop yields. Spring barley (2016, 2017) and winter wheat (2018) were grown over three years in columns containing 25cm of organic matter-rich topsoil, 80 cm of amended coarse sandy soil (1.5%, 3%, 6% wt. ash or 1% wt. biochar or control soil) and 45 cm of un-amended subsoil. Precipitation, evaporative demands and soil moisture were recorded across the growth seasons, with 2018 having severe drought conditions. This year evapotranspiration levels increased with increasing ash and biochar content (by 54% and 33% for the 6% ash- and 1% biochar-amended soils, respectively), and plant dry matter increased by 18% in both the 1% biochar- and 6% ash-treated soils compared to the untreated control. A linear relationship was established between in situ field capacity and ash dosage (R² = .96), showing an increase of 2.2% per percentage (wt.) of ash added, while the 1% biochar treatment increased the capacity by 3.5%, indicating a higher efficiency than for ash. However, we did not find significant positive effects on grain yields. The results show that ash and biochar have the potential to significantly increase soil water retention, evapotranspiration and total dry matter yield in drought conditions, but that this may not correspond to an increase in grain yield.     

Isolation and characterization of bone marrow-derived equine mesenchymal stem cells

OBJECTIVE: To isolate and characterize bone marrow-derived equine mesenchymal stem cells (MSCs) for possible future therapeutic applications in horses. SAMPLE POPULATION: Equine MSCs were isolated from bone marrow aspirates obtained from the sternum of 30 donor horses. PROCEDURES: Cells were cultured in medium (alpha-minimum essential medium) with a fetal calf serum content of 20%. Equine MSC features were analyzed to determine selfrenewing and differentiation capacity. For potential therapeutic applications, the migratory potential of equine MSCs was determined. An adenoviral vector was used to determine the transduction rate of equine MSCs. RESULTS: Equine MSCs can be culture-expanded. Equine MSCs undergo cryopreservation in liquid nitrogen without altering morphologic characteristics. Furthermore, equine MSCs maintain their ability to proliferate and differentiate after thawing. Immunocytochemically, the expression of the stem cell marker CD90 can be detected on equine MSCs. The multilineage differentiation potential of equine MSCs was revealed by their ability to undergo adipogenic, osteogenic, and chondrogenic differentiation. CONCLUSIONS AND CLINICAL RELEVANCE: Our data indicate that bone marrow-derived stromal cells of horses can be characterized as MSCs. Equine MSCs have a high transduction rate and migratory potential and adapt to scaffold material in culture. As an autologous cell population, equine MSCs can be regarded as a promising cell population for tissue engineering in lesions of the musculoskeletal system in horses.     

The effect of lupins as compared with peas and oats on the yield of the subsequent winter barley crop

New high yielding early maturing cultivars of lupins have been introduced in north-west Europe as grain protein crops in crop rotations. This paper reports on a comparative study of lupins with peas and oats, and of their effect on yield of subsequent winter barley crops. These crops were given five levels of N under irrigated and non-irrigated conditions on sand and loam. Under rain fed conditions the grain yield of pea, oat and lupin varied between 24–36, 34–53 and 18–37 hkg DM ha⁻¹, respectively. Supplemental irrigation raised grain yield of oat to 50–60 hkg DM ha⁻¹, while grain yield in pea was not affected and grain yield in lupin in most cases decreased due to gray mould attack and excessive vegetative growth in the indeterminate lupin variety. Under rain fed conditions, the grain nitrogen content of pea, oat and lupin varied between 137–172, 61–80 and 189–226 kg N ha⁻¹, respectively, and was significantly higher in lupin as compared with pea. On sandy soil, similar low-root densities were found for pea, oat and lupin below 30 cm depth. On sand, at final harvest the residual soil-N of lupin and pea, as measured in a subsequent winter barley crop not supplied with N fertilizer, was 15 and 8–10 kg N ha⁻¹ higher than in winter barley following oat, respectively. The nature of the probably more N-root residues of lupin is discussed. On loam, the residual N of lupin and pea was similar, 18–27 kg N ha⁻¹. On sand, under rain fed conditions preceding lupin and pea as compared with oat, increased the barley grain yield at zero N-application 77 and 49%, respectively; the effect of lupin was significantly higher than that of pea until the highest N-level 120 kg N-application ha⁻¹. On loam under rain fed conditions preceding lupin and pea increased the barley grain yield at zero N-application by 36 and 62%, respectively, as compared with oat; at N-application>60 kg N ha⁻¹ the grain yield was similar after all three crops. For both soil types the same level of effect was found under irrigated conditions. Conclusions: Supplemental irrigation might result in lower grain yield in lupin due to gray mould attack and excessive growth if indeterminate lupin varieties are used. Grain nitrogen yield of lupin is significantly higher than that of pea. On sand, the effect of lupin on the subsequent winter barley grain yield is significantly higher than that of pea, probably due to greater N-root nitrogen residues. On loam, lupin and pea have similar effects on the subsequent winter barley crop.     

Diverse plant and animal genetic records from Holocene and Pleistocene sediments

Genetic analyses of permafrost and temperate sediments reveal that plant and animal DNA may be preserved for long periods, even in the absence of obvious macrofossils. In Siberia, five permafrost cores ranging from 400,000 to 10,000 years old contained at least 19 different plant taxa, including the oldest authenticated ancient DNA sequences known, and megafaunal sequences including mammoth, bison, and horse. The genetic data record a number of dramatic changes in the taxonomic diversity and composition of Beringian vegetation and fauna. Temperate cave sediments in New Zealand also yielded DNA sequences of extinct biota, including two species of ratite moa, and 29 plant taxa characteristic of the prehuman environment. Therefore, many sedimentary deposits may contain unique, and widespread, genetic records of paleoenvironments.     

Bypass of DNA lesions generated during anticancer treatment with cisplatin by DNA polymerase eta

DNA polymerase eta (Pol eta) is a eukaryotic lesion bypass polymerase that helps organisms to survive exposure to ultraviolet (UV) radiation, and tumor cells to gain resistance against cisplatin-based chemotherapy. It allows cells to replicate across cross-link lesions such as 1,2-d(GpG) cisplatin adducts (Pt-GG) and UV-induced cis-syn thymine dimers. We present structural and biochemical analysis of how Pol eta copies Pt-GG-containing DNA. The damaged DNA is bound in an open DNA binding rim. Nucleotidyl transfer requires the DNA to rotate into an active conformation, driven by hydrogen bonding of the templating base to the dNTP. For the 3'dG of the Pt-GG, this step is accomplished by a Watson-Crick base pair to dCTP and is biochemically efficient and accurate. In contrast, bypass of the 5'dG of the Pt-GG is less efficient and promiscuous for dCTP and dATP as a result of the presence of the rigid Pt cross-link. Our analysis reveals the set of structural features that enable Pol eta to replicate across strongly distorting DNA lesions.