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Acetyl-CoA Carboxylase α (ACACA), Fatty Acid Synthase (FASN) and Stearoyl-CoA Desaturase (SCD) genes encode key enzymes in fatty acid synthesis. The aim of this research was to verify if ACACA, FASN and SCD expression changes with lactation stages, and to analyze their expression levels and their relationship with the milk fatty acid profile. Milk samples were collected three times from each cow (n=11): at 50–100 DIM, 170–200 DIM and >250 DIM. RNA for gene expression analysis was extracted from milk somatic cells. ACACA expression was significantly greater at the 3rd compared to the 1st lactation stage. FASN and SCD expression significantly increased in the 3rd compared to the 1st and 2nd stages. The strongest positive, highly significant correlations were found in the 3rd lactation stage between ACACA and FASN expression, and between FASN and SCD expression. Few significant correlations were found between gene expression and fatty acid content in milk. Interestingly, no significant correlation was found between the ACACA or FASN genes and fatty acids synthesized de novo. No significant correlation was observed between the SCD gene and monounsaturated fatty acids. It was demonstrated that ACACA, FASN and SCD mRNA expression changes during lactation and is strongly coordinated. Poor correlation was found between fatty acid contents and gene expression. [ABSTRACT FROM AUTHOR]

Abstract Background Solid tumours comprise various cells, including cancer cells, resident stromal cells, migratory haemopoietic cells and other. These cells regulate tumour growth and metastasis. Macrophages constitute probably the most important element of all interactions within the tumour microenvironment. However, the molecular mechanism, that guides tumour environment, still remains unknown. Exploring the underlying molecular mechanisms that orchestrate these phenomena has been the aim of our study. A co-culture of canine mammary cancer cells and macrophages was established and maintained for 72 hrs. Having sorted the cells, gene expression in cancer cells and macrophages, using DNA microarrays, was examined. The results were confirmed using real-time qPCR and confocal microscopy. Moreover, their ability for migration and invasion has been assessed. Results Microarray analysis showed that the up-regulated genes in the cancer cell lines are involved in 15 highly over-manifested pathways. The pathways that drew our diligent attention included: the inflammation pathway mediated by chemokine and cytokine, the Toll receptor signalling pathway and the B cell activation. The up-regulated genes in the macrophages were involved in only 18 significantly over-manifested pathways: the angiogenesis, the p53 pathway feedback loops2 and the Wnt signalling pathway. The microarray analysis revealed that co-culturing of cancer cells with macrophages initiated the myeloid-specific antigen expression in cancer cells, as well as cytokine/chemokine genes expression. This finding was confirmed at mRNA and protein level. Moreover, we showed that macrophages increase cancer migration and invasion. Conclusions The presence of macrophages in the cancer environment induces acquisition of the macrophage phenotype (specific antigens and chemokines/cytokines expression) in cancer cells. We presumed that cancer cells also acquire other myeloid features, such as: capabilities of cell rolling, spreading, migration and matrix invasion (what has also been confirmed by our results). It may, perhaps, be the result of myeloid-cancer cell hybrid formation, or cancer cells mimicking macrophages phenotype, owing to various proteins secreted by macrophages.

Majority of the Polish dairy goat population consists of two breeds: Polish White Improved (some 50% of the whole population) and Polish Fawn Improved (some 25% of the whole population). In a previous study no differences in basic performance parameters (milk yield, fat, protein, and lactose content) were found between these two breeds, though a difference in milk somatic cell score was noted. To examine the genomic differences, gene expression profiles of three types of biological material were investigated. Somatic cells isolated from milk, milk fat globules, and peripheral blood nuclear cells were subjected to bovine DNA microarrays. Hierarchical clustering of selected tissues' transcriptomic profiles revealed the presence of three distinct sub-clusters, representing each type of sample, but not the breed sampled. The only gene that was significantly different between the two breeds in terms of regulation was Bos taurus agouti signaling protein gene. In summary, no significant differences in the expression of 44,000 genes, including those involved in milk production and the immune system, were identified between the two studied Polish goat breeds, while, as expected, the differences in gene expression in the studied types of biological material were significant. These findings suggest that productively and immunologically these two main Polish dairy goat breeds consist one population. [ABSTRACT FROM AUTHOR]

Laboratory and veterinary research often produces very specific data, which are difficult to collect and standardize, hence to interpret. The multi-stage character and complexity of our project, the variety of laboratory equipment used and the involvement of external service providers made it even more challenging to collect all data in a unified electronic form and store them in one place. A research project conducted on a living organism requires a holistic approach to the results of different health indicators, to the efficiency of each animal in a study group and to data analysis - so as to fully and correctly evaluate the state of health, or the stage of illness. In order to facilitate the work of experts who evaluate the parameters and the research data obtained in a series of experiments, a database was designed and made accessible on-line to logged-in users. While designing this database, interesting solutions and ready-made functionalities were applied, so as to efficiently collect and display data. Database design tools such as MSSQL Express and MS Access were used. The MSSQL database was managed by the SQL Server Management Studio. To create and display data on the website, MS Excel, Visual Studio 2008 Express and IIS Express were used. This new data management system provides for an efficient storage of and insight into all data records, and can be taken advantage of in a variety of comprehensive scientific and research projects in the area of animal science especially veterinary researches. [ABSTRACT FROM AUTHOR]

Due to the presence and activity of mammary stem cells (MaSC), growth and remodelling of mammary gland during puberty and lactation cycles is possible. In this study the number of putative mammary stem/progenitor cells was examined in 20 months old non-pregnant Holstein-Friesian heifers. Cells were double-stained with fluorescent dye-conjugated antibodies against stem cell antigen-1 (Sca-1) and fibronectin type III domain containing 3B (FNDC3B), and were analysed using scanning cytometry and flow cytometry. Nuclei were counterstained with Hoechst 33342. Scanning and flow cytometry revealed 2.43±0.32% and 1±0.37% of MaSC in total cell number, respectively. Sca-1posFNDC3Bpos cells did not express estrogen receptor (ERα), confirming their undifferentiated phenotype. In conclusion, scanning cytometry is a preferable method for evaluation of the number and localization of MaSC in situ, whereas, flow cytometry with cell sorting enables further genomic and biochemical analyses of isolated cells. [ABSTRACT FROM AUTHOR]

Microarray-based transcriptomic profiling enables simultaneous measurement of expression of multiple genes from one biological sample. Here we describe a detailed protocol, which serves to examine global gene expression using whole genome oligonucleotide microarrays. We also provide examples of bioinformatics tools, which are helpful in analyses and interpretation of microarray data, and propose further biological assays, to warrant conclusions drawn from transcriptomic signature.