Evaluation of serologic and viral detection methods for diagnosing feline herpesvirus-1 infection in cats with acute respiratory tract or chronic ocular disease

OBJECTIVE: To determine the value of virus isolation (VI), immunofluorescent antibody (IFA) assay, serum neutralization (SN), and ELISA for the diagnosis of clinical feline herpesvirus-1 (FHV-1) infection in cats. ANIMALS: 46 clinically normal cats, 17 cats with signs of acute respiratory tract disease, and 38 cats with signs of chronic ocular disease. PROCEDURE: Conjunctival swabs for VI, conjunctival scrapings for IFA testing, and venous blood samples for SN or ELISA testing were obtained from all cats. RESULTS: FHV-1 was detected in 10.9 and 28.3% of clinically normal cats and in 18.2 and 33.3% of cats with FHV-1-associated disease by VI and the IFA assay, respectively. There were no significant differences in the viral detection rate between cats with acute respiratory tract disease and cats with chronic ocular disease or between diseased cats and clinically normal cats; however, FHV-1 was never detected by both methods in clinically normal cats. Overall FHV-1 seroprevalence was 97% when tested by ELISA and 66% when tested by SN. Seroprevalence did not vary significantly among the 3 groups for either serologic test. Magnitude of SN and ELISA titers varied greatly but independently of presence or absence of clinical signs of FHV-1-associated disease. Sensitivity, specificity, and positive and negative predictive values were assessed for VI and the IFA assay--jointly and individually--and for each SN and ELISA titer magnitude. Values never all exceeded 50%. CLINICAL IMPLICATIONS: Because FHV-1 can be detected commonly in clinically normal cats by the IFA assay or VI, neither test appears to aid in the clinical diagnosis of FHV-1 infection. Seroprevalence does not appear to vary between affected and clinically normal cats. SN, ELISA, VI, and the IFA assay appear to be of limited value in the diagnosis of FHV-1-associated disease in cats. Concurrent assessment of the IFA assay and VI results may permit exclusion of FHV-1 as an etiologic agent if results of both tests are negative.     

miRNA and piRNA expression profiles of breeder cock testes detected by next‐generation sequencing

miRNAs are small non‐coding regulatory RNAs that play key roles in diverse biological processes. In this study, we used the Solexa sequencing technique to profile miRNAs in breeder cock testes to illustrate their functions. A total of 663 co‐expressed miRNAs and 3,180 co‐expressed piRNAs were detected in three libraries. Based on Mir‐X^? miRNA qRT‐PCR, three miRNAs representing low, medium and high expression levels according to the sequencing results were selected randomly to validate the miRNAs' expression profiles. Results suggested that the miRNA expression profiles data could represent actual miRNA expression levels. Moreover, target genes prediction of the co‐expressed miRNAs and further Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses were performed, which revealed that some candidate miRNAs were involved in the regulation of the spermatogenesis process, spermatozoa function and testicular metabolism. In conclusion, we provided a useful resource for further elucidation of the miRNAs' regulatory role in spermatogenesis, contributing to a preliminary database for functional and molecular mechanistic studies in testicular metabolism, spermatogenesis and other testes functions.     

Fullerene pipes

Single-wall fullerene nanotubes were converted from nearly endless, highly tangled ropes into short, open-ended pipes that behave as individual macromolecules. Raw nanotube material was purified in large batches, and the ropes were cut into 100- to 300-nanometer lengths. The resulting pieces formed a stable colloidal suspension in water with the help of surfactants. These suspensions permit a variety of manipulations, such as sorting by length, derivatization, and tethering to gold surfaces.     

Circular polarization in star- formation regions: implications for biomolecular homochirality

Strong infrared circular polarization resulting from dust scattering in reflection nebulae in the Orion OMC-1 star-formation region has been observed. Circular polarization at shorter wavelengths might have been important in inducing chiral asymmetry in interstellar organic molecules that could be subsequently delivered to the early Earth by comets, interplanetary dust particles, or meteors. This could account for the excess of L-amino acids found in the Murchison meteorite and could explain the origin of the homochirality of biological molecules.     

Effect of Transgene Introduction and Recloning on Efficiency of Porcine Transgenic Cloned Embryo Production In Vitro

Retrovirus-mediated exogenous gene transfection of somatic cells is an efficient method to produce transgenic embryos by somatic cell nuclear transfer (SCNT). This study evaluated whether efficiency of transgenic embryos production, by SCNT using fibroblast cells transfected by retrovirus vector, is influenced by the introduced transgene and whether recloning could further improve its efficiency. Transgenic cloned embryos were produced by SCNT of porcine foetal fibroblast cells transfected by either LNβ-Z or LNβ-enhanced green fluorescent protein (EGFP) retrovirus vector and evaluated for their developmental ability in vitro . Blastomeres from four-cell stage porcine embryos, produced by SCNT of foetal fibroblast cells transfected with LNβ-EGFP retroviral vector, were subsequently recloned into enucleated metaphase II oocytes and evaluated for changes in chromatin configuration, in vitro embryo development and gene expression. Analysis of results showed that cleavage and blastocyst rates of porcine SCNT embryos, using LacZ (53.6 ± 6.4%; 12.0 ± 5.7%) or EGFP (57.5 ± 6.3%; 10.1 ± 4.1%) transfected fibroblasts, did not differ (p > 0.05) from those of non-transfected controls (60.9 ± 8.2%; 12.3 ± 4.0%). Recloning of blastomeres did not further improve the in vitro development rate. Interestingly, the nuclei of blastomere underwent slower remodelling process than somatic cell nuclei. Both cloned and recloned embryos showed 100% transgene expression and there were no evidence of mosaicism. In conclusion, our data shows that the efficiency of transgenic cloned embryos production by SCNT of somatic cells transfected with replication-defective retrovirus vector is not influenced by the transgene introduction into donor cells and recloning of four-cell stage blastomere could not further improve its efficiency.     

Genomic prediction of sunflower hybrid performance

Predicting single‐cross performance is of high importance to improve the efficiency of sunflower (Helianthus annuus L.) hybrid breeding programmes. We used experimental data from inter‐ and intragroup sunflower hybrids and their parental lines adapted to Central Europe to (i) study the genetic diversity and combining ability and (ii) examine the accuracy to predict hybrid performance based on phenotypic and genomic data. We evaluated 133 intragroup and 104 intergroup hybrids with their parental lines in replicated trials at four environments for grain yield, oil yield and oil content. Furthermore, the parental lines were fingerprinted with 572 AFLP markers. Variance due to specific combining ability was comparable for intergroup and intragroup crosses. This suggested a lack of clearly defined heterotic groups for the sample of studied sunflower lines. Prediction accuracy of hybrid performance based on general combining ability effects was high and could not be increased using genomic selection approaches. For situations where no information on GCA effects of parental lines was available, hybrid prediction based on genomic selection methods was accurate for groups of related lines. For groups of unrelated lines, however, we observed a strong decrease in the prediction accuracy. This suggests that prediction of hybrid performance for crosses based on genetically distant parents remains challenging.     

Best linear unbiased prediction of triticale hybrid performance

Predicting single-cross performance is of special interest in hybrid breeding of triticale. We used molecular and phenotypic data of factorial triticale crosses and compared several approaches to predict their single-cross performance. Twenty-three inbred lines and their 76 incomplete factorial crosses were field evaluated for grain yield, plant height, and heading time at five locations in Central Europe. In addition, the parental lines were genotyped with 52 SSR markers. Plant height and heading time were predicted with high accuracy based on mid-parent performance. In contrast, prediction of hybrid performance based on mid-parent value was not accurate for grain yield. Using general combining ability effects led to an enhanced prediction accuracy of hybrid grain yield performance. This accuracy could be slightly improved using best linear unbiased prediction approaches. The prediction accuracy was considerably high even if the number of tested hybrids was small. Consequently, best linear unbiased prediction of hybrid performance is a promising tool for hybrid triticale breeding programs.