Pre- and post-transcriptional modifications of gene expression are emerging as foci of disease studies, with some studies revealing the importance of non-coding transcripts, like long non-coding RNAs (lncRNAs) and microRNAs (miRNAs). We hypothesize that transcription factors (TFs), lncRNAs and miRNAs modulate immune response in bovine mastitis and could potentially serve as disease biomarkers and/or drug targets. With computational analyses, we identified candidate genes potentially regulated by miRNAs and lncRNAs base pair complementation and thermodynamic stability of binding regions. Remarkably, we found six miRNAs, two being bta-miR-223 and bta-miR-24-3p, to bind to several targets. LncRNAs NONBTAT027932.1 and XR_003029725.1, were identified to target several genes. Functional and pathway analyses revealed lipopolysaccharide-mediated signaling pathway, regulation of chemokine (C-X-C motif) ligand 2 production and regulation of IL-23 production among others. The overarching interactome deserves further in vitro/in vivo explication for specific molecular regulatory mechanisms during bovine mastitis immune response and could lay the foundation for development of disease markers and therapeutic intervention.

Entamoeba histolytica possesses a 24.5 kilobase plasmid-like molecule which encodes for the organism's ribosomal RNAs. Sequence analysis of this extrachromosomal element revealed the presence of AT rich sequences which show homology to the origin of replication of other lower eucaryotes. An 802 bp fragment containing these sequences was cloned into a yeast shuttle vector lacking the origin of replication and the construct tested for its ability to replicate autonomously in yeast. Mitotic stability tests as well as evidence for plasmid maintenance indicate that the transformed cells contained self-replicating episomes and not stably integrated molecules. The nucleotide sequence of this ARS-containing fragment is presented.

Bovine mastitis is mainly caused by Escherichia coli and Streptococcus uberis. It is associated with complex multifactorial phenotypes, requiring an integrative approach to elucidate the molecular networks underlying variable disease outcome. The association of lncRNAs, miRNAs and their target as key players in immunity regulation has not been studied in bovine mastitis. We hypothesize that non-coding genes could mediate immune response in bovine mastitis and have potential as disease markers or drug targets. Through bioinformatic analyses, we investigated the networks of lncRNAs, miRNAs and mRNAs, and identified key regulatory elements driving immune response in bovine mastitis. Our analyses reveal 16 highly significant immune response genes including MYD88, IL-10, IL-4, ICAM1, CXCL8, IL-18 and CSF2. Notably, of the 2904 miRNAs reported, six - bta-miR-24-3p, bta-miR-149-5p, bta-miR-223, bta-miR-185, bta-miR-328, and bta-miR-874, were predicted to bind multiple regions of target genes. Likewise, eight out of 22 lncRNAs including NONBTAT001181.2, XR_003030515.1 and XR_003030515.1 bind 13 mRNA targets. Of interest, some of these are conserved in 15 different species, including Homo sapiens. Our functional analyses show that these lncRNAs and miRNAs may regulate pathogenesis of bovine mastitis through lipopolysaccharide-mediated signaling pathway, regulation of chemokine (C-X-C motif) ligand 2 production, regulation of IL-23 production, positive regulation of chemokine production and pattern recognition receptor activity. This interactome lays a foundation for molecular interconnectivity of regulatory elements in bovine mastitis, deserving further elucidation for potential vaccine and therapeutics.

Background: Bovine mastitis is a mammary gland infectious disease caused by a variety of pathogens with a devasting economic impact worldwide. Pre- and post-transcriptional modifications, including transcription factors (TFs), altering gene expression are emerging foci of disease studies, with minimal studies revealing the importance of non-coding transcripts, like long non-coding RNAs (lncRNAs) and microRNAs (miRNAs). We hypothesize TFs, lncRNAs and miRNAs can modulate the immune response in bovine mastitis and can potentially serve as disease biomarkers and/or drug targets.Methods: With computational analyses, we aimed to identify candidate bovine mastitis genes and construct the networks of miRNA, lncRNA and TFs regulating the gene’s mRNA, affecting disease pathogenesis. Experimentally validated genes associated with bovine mastitis were obtained through an extensive search for significantly mentioned genes, utilizing several databases. Prediction of miRNA and lncRNA binding bovine mastitis candidate genes were performed through several algorithms and software that relied on base pair complementation, evolutionary conservation, and thermodynamic stability of binding regions. Combined interactome network of lncRNAs, miRNAs, TFs and immune gene targets were constructed.Results: Sixteen of 923 genes were found to be highly significant in bovine mastitis disease pathway including, CD4, IL-10, IFNγ, IL-4, TLR4, TNFα, and CD14. Remarkably, we found six miRNAs, two being bta-miR-223 and bta-miR-24-3p, to bind to several targets. Eight out of 22 lncRNA, such as NONBTAT027932.1 and XR_003029725.1, were identified as regulatory elements that target the genes based on the normalized binding free energies ranging from -0.1774 to -2.875. Similarly identified were 17 TFs, including JUN and CREB. Our functional and pathway analyses revealed several pathways like lipopolysaccharide-mediated signaling pathway, regulation of chemokine (C-X-C motif) ligand 2 production and regulation of IL-23 production among others. Conclusions: The overarching interactome in this study is the first of its kind regarding bovine mastitis, deserving further in vitro/in vivo explication for specific molecular regulatory mechanisms during bovine mastitis immune response, which could lay the foundation for development of disease markers and therapeutic intervention.

Pre- and post-transcriptional modifications of gene expression are emerging as foci of disease studies, with some studies revealing the importance of non-coding transcripts, like long non-coding RNAs (lncRNAs) and microRNAs (miRNAs). We hypothesize that TFs, lncRNAs and miRNAs can modulate the immune response in bovine mastitis and potentially serve as disease biomarkers and/or drug targets. With computational analyses, we identified candidate genes potentially regulated by miRNAs and lncRNAs base pair complementation and thermodynamic stability of binding regions. Remarkably, we found six miRNAs, two being bta-miR-223 and bta-miR-24-3p, to bind to several targets. NONBTAT027932.1 and XR_003029725.1, were identified to target several genes. Functional and pathway analyses revealed lipopolysaccharide-mediated signaling pathway, regulation of chemokine (C-X-C motif) ligand 2 production and regulation of IL-23 production among others. The overarching interactome deserves further in vitro/in vivo explication for specific molecular regulatory mechanisms during bovine mastitis immune response and could lay the foundation for development of disease markers and therapeutic intervention.