Short communication: Reliability of single-step genomic BLUP breeding values by multi-trait test-day model analysis.

The purpose of our study was to develop an approximation procedure to estimate reliabilities of single-step genomic BLUP breeding values in a test-day model for routine evaluation of milk yield in a dairy cattle population. Input data consisted of 20,220,047 first-, second-, and third-lactation test-day milk yield records of 1,126,102 Czech Holstein cows (each lactation being considered a separate trait), with 1,844,679 animals in the pedigree file and with genomic data from 2,236 bulls. Evaluation was according to a multi-lactation model. The procedure was based on the effective number of records per animal from milk recording as well as from genomic and pedigree relationships. Traits were analyzed individually, and genetic covariances among traits were subsequently taken into account. The use of genomic information increased average reliability in young bulls from 0.276 to 0.505, but increased reliability in proven bulls only from 0.828 to 0.855. The reliabilities of genomic breeding values in multi-trait evaluation for first, second and third lactations, respectively, averaged 0.652, 0.673, and 0.633 for young bulls and 0.907, 0.894, and 0.852 for proven bulls. For an index combining all 3 lactations, the average reliability of a single-step genomic BLUP prediction was 0.712 and 0.925 for younger and proven bulls, respectively. Increased reliability due to genotyping in the population of all genotyped and nongenotyped animals was very small (<0.01) because of the small proportion of genotyped animals in the population.

Contribution of domestic production records, Interbull estimated breeding values, and single nucleotide polymorphism genetic markers to the single-step genomic evaluation of milk production.

Estimated breeding values (EBV) for first-lactation milk production of Holstein cattle in the Czech Republic were calculated using a conventional animal model and by single-step prediction of the genomic enhanced breeding value. Two overlapping data sets of milk production data were evaluated: (1) calving years 1991 to 2006, with 861,429 lactations and 1,918,901 animals in the pedigree and (2) calving years 1991 to 2010, with 1,097,319 lactations and 1,906,576 animals in the pedigree. Global Interbull (Uppsala, Sweden) deregressed proofs of 114,189 bulls were used in the analyses. Reliabilities of Interbull values were equivalent to an average of 8.53 effective records, which were used in a weighted analysis. A total of 1,341 bulls were genotyped using the Illumina BovineSNP50 BeadChip V2 (Illumina Inc., San Diego, CA). Among the genotyped bulls were 332 young bulls with no daughters in the first data set but more than 50 daughters (88.41, on average) with performance records in the second data set. For young bulls, correlations of EBV and genomic enhanced breeding value before and after progeny testing, corresponding average expected reliabilities, and effective daughter contributions (EDC) were calculated. The reliability of prediction pedigree EBV of young bulls was 0.41, corresponding to EDC=10.6. Including Interbull deregressed proofs improved the reliability of prediction by EDC=13.4 and including genotyping improved prediction reliability by EDC=6.2. Total average expected reliability of prediction reached 0.67, corresponding to EDC=30.2. The combination of domestic and Interbull sources for both genotyped and nongenotyped animals is valuable for improving the accuracy of genetic prediction in small populations of dairy cattle.

Estimation of genetic parameters for hip dysplasia in Czech Labrador Retrievers.

The objective of this study was to estimate the genetic parameters, genetic trends and breeding values using linear model (LM) and threshold model (TM) for the development of hip dysplasia (HD) in Labrador Retrievers in the Czech Republic (n = 3151). The right and left hip joints were evaluated separately using the Fédération Cynologique Internationale scoring system. Four linear and four TMs were tested for the correct estimation of genetic parameters. All the tested models utilized fixed effects of sex, assessor, year of birth, regression of age at evaluation, random direct genetic effects and the effect of the animals' permanent environments. The models differed in the inclusion of the following effects: fixed effects of regression of inbreeding coefficient, random maternal effect and random effect of the maternal permanent environment. Compared to the TM, the LM provided lower coefficients of direct (0.25-0.29 versus 0.26-0.35) and maternal heritability (0.01-0.02 versus 0.03-0.05), repeatability (0.76-0.77 versus 0.78-0.83) and of the correlation between direct and maternal effects (-0.55 to -0.21 versus -0.80 to -0.27). In the tested models, no statistical significance was found for fixed regression of inbreeding coefficients or for the random effect of the permanent maternal environment. In spite of the similarity of the LM and TM results, the TM is recommended as the more suitable model for estimating genetic parameters and subsequent breeding values for HD in Labrador Retrievers in the Czech Republic.

Cell type specific adhesion to surfaces functionalised by amine plasma polymers.

Our previously-obtained impressive results of highly increased C2C12 mouse myoblast adhesion to amine plasma polymers (PPs) motivated current detailed studies of cell resistance to trypsinization, cell proliferation, motility, and the rate of attachment carried out for fibroblasts (LF), keratinocytes (HaCaT), rat vascular smooth muscle cells (VSMC), and endothelial cells (HUVEC, HSVEC, and CPAE) on three different amine PPs. We demonstrated the striking difference in the resistance to trypsin treatment between endothelial and non-endothelial cells. The increased resistance observed for the non-endothelial cell types was accompanied by an increased rate of cellular attachment, even though spontaneous migration was comparable to the control, i.e., to the standard cultivation surface. As demonstrated on LF fibroblasts, the resistance to trypsin was similar in serum-supplemented and serum-free media, i.e., medium without cell adhesion-mediating proteins. The increased cell adhesion was also confirmed for LF cells by an independent technique, single-cell force spectroscopy. This method, as well as the cell attachment rate, proved the difference among the plasma polymers with different amounts of amine groups, but other investigated techniques could not reveal the differences in the cell behaviour on different amine PPs. Based on all the results, the increased resistance to trypsinization of C2C12, LF, HaCaT, and VSMC cells on amine PPs can be explained most probably by a non-specific cell adhesion such as electrostatic interaction between the cells and amine groups on the material surface, rather than by the receptor-mediated adhesion through serum-derived proteins adsorbed on the PPs.

Comparative studies of different stromal cell microenvironments in support of human B-cell development.

This study compared human murine stromal cells for their capacity to support human hematopoietic stem cell (HSC) development into the B lineage. FACS sorted human fetal bone marrow (BM) HSC (CD34+CD19- or CD34+/CD10-/CD19-/CD45RA) were cultured on human fetal BM stromal cells, human skin fibroblasts, or murine S17 stromal cells and analyzed by flow cytometry or reverse transcriptase polymerase chain reaction. CD34+CD19- HSC on human BM stromal cells or fibroblasts differentiated into B-lineage cells with a continuum in density of surface CD19 expression, and some cells expressing micro/kappa or micro/lambda B-cell receptors. In contrast, CD19+ cells from S17 cultures had two- to fourfold higher levels of CD19, but no cells expressing B-cell receptors. The number and percentage of CD19+ cells was high, intermediate, or low in the human BM, human fibroblast, or murine S17 stromal cell cultures, respectively. Reverse transcriptase polymerase chain reaction analysis showed that TdT, CD19, and DHQ52-J(H) rearrangements were expressed at comparable levels when CD34+/CD19- HSC were plated on human or murine stromal cells. In contrast, CD34+/CD10-/CD19-/CD45RA HSC plated on human or murine stromal cells expressed CD19 in both cultures, but TdT was only expressed in human stromal cell cultures. We conclude that human BM stromal cell, human skin fibroblasts, and murine S17 stromal cell cultures can provide complementary and comparative tools for identification of stromal cell ligands with potentially unique functions in regulating human B-cell development.

Domestic estimated breeding values and genomic enhanced breeding values of bulls in comparison with their foreign genomic enhanced breeding values.

Estimated breeding values (EBVs) and genomic enhanced breeding values (GEBVs) for milk production of young genotyped Holstein bulls were predicted using a conventional BLUP - Animal Model, a method fitting regression coefficients for loci (RRBLUP), a method utilizing the realized genomic relationship matrix (GBLUP), by a single-step procedure (ssGBLUP) and by a one-step blending procedure. Information sources for prediction were the nation-wide database of domestic Czech production records in the first lactation combined with deregressed proofs (DRP) from Interbull files (August 2013) and domestic test-day (TD) records for the first three lactations. Data from 2627 genotyped bulls were used, of which 2189 were already proven under domestic conditions. Analyses were run that used Interbull values for genotyped bulls only or that used Interbull values for all available sires. Resultant predictions were compared with GEBV of 96 young foreign bulls evaluated abroad and whose proofs were from Interbull method GMACE (August 2013) on the Czech scale. Correlations of predictions with GMACE values of foreign bulls ranged from 0.33 to 0.75. Combining domestic data with Interbull EBVs improved prediction of both EBV and GEBV. Predictions by Animal Model (traditional EBV) using only domestic first lactation records and GMACE values were correlated by only 0.33. Combining the nation-wide domestic database with all available DRP for genotyped and un-genotyped sires from Interbull resulted in an EBV correlation of 0.60, compared with 0.47 when only Interbull data were used. In all cases, GEBVs had higher correlations than traditional EBVs, and the highest correlations were for predictions from the ssGBLUP procedure using combined data (0.75), or with all available DRP from Interbull records only (one-step blending approach, 0.69). The ssGBLUP predictions using the first three domestic lactation records in the TD model were correlated with GMACE predictions by 0.69, 0.64 and 0.61 for milk yield, protein yield and fat yield, respectively.

The effect of oils on PAH, PCDD, PCDF, and PCB emissions from a spark engine fueled with leaded gasoline.

The effect of synthetic and mineral oils on the formation of polyaromatic hydrocarbons (PAHs), polychlorinated dibenzo-p-dioxins (PCDDs), dibenzofurans (PCDFs), and biphenyls (PCBs) in emissions from a spark ignition engine was studied on a Skoda Favorit engine fueled with leaded gasoline. The test cycle simulated urban traffic conditions on a chassis dynamometer, in accordance with the ECC 83.00 test. The data for selected PAHs as well as PCDDs, PCDFs, and PCBs congener profiles are presented. PCDD/Fs emissions for an unused oil and the oil after 10000-km operation varied from 300 to 2000 fmol/m3, PCBs emissions from 75 to 178 pmol/m3, and PAHs emissions from 150 to 420 microg/m3. The content of PCBs in oils varied from 2 to 920 mg/kg.

Impact of milk pricing system on the economic response to selection on milk components.

The impact of various milk pricing systems on the economic weights and the resulting selection responses for milk yield and fat and protein contents in Holstein and Czech Fleckvieh were investigated. The calculations were done for the situations with and without quotas on milk yield and fat content. There was a substantial influence of the milk pricing system on the selection response. The use of index weights based on a 'wrong' pricing system reduced the total economic selection response by up to 51% of the selection response expected for an index based on the 'correct' pricing system. Financial losses in the selection response were greater with quotas than without quotas. Higher losses were also calculated for Holstein compared with Czech Fleckvieh. Incorrect assumptions on the output limitation (absence or presence of milk quotas) led only to minor reductions in the total selection response (mostly <2%) if the other parameters were correct. The results indicate that customized subindices for milk production traits would increase farmers' profit from sire selection when selling milk on the basis of different pricing systems.

Multichannel SPR biosensor for detection of endocrine-disrupting compounds.

A surface plasmon resonance (SPR) biosensor for simultaneous detection of multiple organic pollutants exhibiting endocrine-disrupting activity, namely atrazine, benzo[a]pyrene, 2,4-dichlorophenoxyacetic acid (2,4-D) and 4-nonylphenol, is reported. The biosensor utilizes a multichannel SPR sensor based on wavelength modulation of SPR and wavelength division multiplexing (WDM) of sensing channels, antibodies as biorecognition element and a competitive immunoassay detection format. An analysis time of 45 min (including 30-min incubation of the sample with antibodies) and limits of detection as low as 0.05, 0.07, 0.16 and 0.26 ng mL(-1) are demonstrated for benzo[a]pyrene, atrazine, 2,4-D and 4-nonylphenol, respectively. The biosensor is also shown to be regenerable and suitable for repeated use.

Effect of subsidy regimes on economic values of functional traits in beef cattle breeding.

We investigated the impact of five monetary subsidy regimes on economic values of traits in a cow-calf pasture production system with surplus calves fed for slaughter. The following regimes were analysed: (1) maximum prices for slaughter animals actually received in the Czech Republic during 2004, with no subsidies of any kind; (2) prices as in (1), with subsidies awarded per hectare of permanent grassland and per calf born; (3) prices as in (1), with subsidies awarded per hectare of agricultural land, per hectare of pasture and meadow, per beef cow in a forage system and per livestock unit; (4) prices as in (1), with subsidies awarded per hectare of agricultural land; (5) no subsidies, but prices received for slaughter animals that covered production costs and resulted in 1% profitability. The modelled farm showed negative profit under real price conditions with no subsidies (regime 1), which led to an underestimation of economic values for functional traits. The same results were obtained in regimes in which subsidies did not depend on the number of animals (3) or on meat production from the enterprise (4). Economic values of production traits (growth and carcass traits) did not vary among subsidy regimes. To determine optimum economic values for functional traits in beef cattle, we advocate using the method applied in subsidy regime 5, no subsidies but prices for slaughter animals that cover production cost and a small profit.

[Liquid chromatographic parameters of groups of herbicidal active substances. I. Urea herbicides]. / Flüssigchromatographische parameter herbizider wirkstoffgruppen

Because of the polarity of herbicidal urea derivatives, liquid chromatography is the most suitable method for their determination. The separation of a number of active substances is described. A versatile applicable ternary solvent mixture is used as the mobile phase. Several columns are tested for their separation performances and their capability to retain active substances and some known degradation products. To fully utilize the sensitivity of the photometric detector the UV spectra of the tested compounds are determined.

Correlates of successful outcome in crisis intervention therapy.

The present study examines the correlates of successful outcome (subsequent inpatient or outpatient treatment) of 305 outpatient cases treated by crisis intervention therapy. Although the two measures of outcome were correlated with each, the respective relationships with the predictor variables were sometimes different. Demographic variables and therapist's characteristics were not correlated with either outcome measures. Past responses to stress (prior state hospitalization and suicidal behavior) were predictors of subsequent inpatient treatment, but not subsequent outpatient treatment. On the other hand, current precipitating events did not predict inpatient treatment, but did predict subsequent outpatient treatment. Several statistically nonsignificant relationships which either replicate or disagree with previous findings are also discussed.

Hydraulic conductivity of the endothelial and outer layers of the rabbit aorta.

Pressure-driven fluid flow across the arterial wall was measured to determine wall hydraulic conductivity (Lp) before and after removal of the endothelium. The thoracic aortas of rabbits, anesthetized with Nembutal, were cannulated, perfused with oxygenated Ringer solution, and removed. With one cannula connected to a capillary manometer and the other closed, the manometer meniscus shift could be used as an indication of fluid loss through the wall plus vessel volume increase (creep). The latter effect, when measured, accounted for about one-fourth of the total volume displacement. The Lp given in cm/(s.cmH2O) +/- SD, was 3.30 +/- 0.96 x 10(-8). Another method employed continuous weighing of a closed aortic segment to obtain fluid loss, and yielded an Lp of 4.07 +/- 1.3 x 10(-8), and after mechanically removing the endothelium, the Lp became 7.73 +/- 2.8 x 10(-8). Using the above data, an Lp could be calculated for aortic endothelium of 8.6 x 10(-8). This suggests that about half the total transmural pressure drop occurs across the endothelium. Scanning electronmicrographs were used to check the condition of the endothelium.

Microenvironmental influences on human B-cell development.

Mammalian B-cell development can be viewed as a developmental performance with several acts. The acts are represented by checkpoints centered around commitment to the B-lineage and functional Ig gene rearrangement--culminating in expression of the pre-B-cell receptor (pre-BCR) and the BCR. Progression of cells through these checkpoints is profoundly influenced by the fetal liver and adult bone marrow (BM) stromal cell microenvironments. Our laboratory has developed a model of human B-cell development that utilizes freshly isolated/non-transformed human BM stromal cells as an in vitro microenvironment. Human CD34+ hematopoietic stem cells plated in this human BM stromal cell microenvironment commit to the B lineage and progress through the pre-BCR and BCR checkpoints. This human BM stromal cell microenvironment also provides survival signals that prevent apoptosis in human B-lineage cells. Human B-lineage cells exhibit differential expression of Notch receptors and human BM stromal cells express the Notch ligand Jagged-1. These results suggest a potential role for Notch in regulating B-lineage commitment and/or progression through the pre-BCR and BCR checkpoints.